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Sample records for 13c-metabolic flux analysis

  1. 13C metabolic flux analysis.

    PubMed

    Wiechert, W

    2001-07-01

    Metabolic flux analysis using 13C-labeled substrates has become an important tool in metabolic engineering. It allows the detailed quantification of all intracellular fluxes in the central metabolism of a microorganism. The method has strongly evolved in recent years by the introduction of new experimental procedures, measurement techniques, and mathematical data evaluation methods. Many of these improvements require advanced skills in the application of nuclear magnetic resonance and mass spectrometry techniques on the one hand and computational and statistical experience on the other hand. This minireview summarizes these recent developments and sketches the major practical problems. An outlook to possible future developments concludes the text.

  2. (13)C metabolic flux analysis of recombinant expression hosts.

    PubMed

    Young, Jamey D

    2014-12-01

    Identifying host cell metabolic phenotypes that promote high recombinant protein titer is a major goal of the biotech industry. (13)C metabolic flux analysis (MFA) provides a rigorous approach to quantify these metabolic phenotypes by applying isotope tracers to map the flow of carbon through intracellular metabolic pathways. Recent advances in tracer theory and measurements are enabling more information to be extracted from (13)C labeling experiments. Sustained development of publicly available software tools and standardization of experimental workflows is simultaneously encouraging increased adoption of (13)C MFA within the biotech research community. A number of recent (13)C MFA studies have identified increased citric acid cycle and pentose phosphate pathway fluxes as consistent markers of high recombinant protein expression, both in mammalian and microbial hosts. Further work is needed to determine whether redirecting flux into these pathways can effectively enhance protein titers while maintaining acceptable glycan profiles.

  3. 13C metabolic flux analysis in complex systems.

    PubMed

    Zamboni, Nicola

    2011-02-01

    Experimental determination of in vivo metabolic rates by methods of (13)C metabolic flux analysis is a pivotal approach to unravel structure and regulation of metabolic networks, in particular with microorganisms grown in minimal media. However, the study of real-life and eukaryotic systems calls for the quantification of fluxes also in cellular compartments, rich media, cell-wide metabolic networks, dynamic systems or single cells. These scenarios drastically increase the complexity of the task, which is only partly dealt by existing approaches that rely on rigorous simulations of label propagation through metabolic networks and require multiple labeling experiments or a priori information on pathway inactivity to simplify the problem. Albeit qualitative and largely driven by human interpretation, statistical analysis of measured (13)C-patterns remains the exclusive alternative to comprehensively handle such complex systems. In the future, this practice will be complemented by novel modeling frameworks to assay particular fluxes within a network by stable isotopic tracer for targeted validation of well-defined hypotheses.

  4. 13C metabolic flux analysis at a genome-scale.

    PubMed

    Gopalakrishnan, Saratram; Maranas, Costas D

    2015-11-01

    Metabolic models used in 13C metabolic flux analysis generally include a limited number of reactions primarily from central metabolism. They typically omit degradation pathways, complete cofactor balances, and atom transition contributions for reactions outside central metabolism. This study addresses the impact on prediction fidelity of scaling-up mapping models to a genome-scale. The core mapping model employed in this study accounts for (75 reactions and 65 metabolites) primarily from central metabolism. The genome-scale metabolic mapping model (GSMM) (697 reaction and 595 metabolites) is constructed using as a basis the iAF1260 model upon eliminating reactions guaranteed not to carry flux based on growth and fermentation data for a minimal glucose growth medium. Labeling data for 17 amino acid fragments obtained from cells fed with glucose labeled at the second carbon was used to obtain fluxes and ranges. Metabolic fluxes and confidence intervals are estimated, for both core and genome-scale mapping models, by minimizing the sum of square of differences between predicted and experimentally measured labeling patterns using the EMU decomposition algorithm. Overall, we find that both topology and estimated values of the metabolic fluxes remain largely consistent between core and GSM model. Stepping up to a genome-scale mapping model leads to wider flux inference ranges for 20 key reactions present in the core model. The glycolysis flux range doubles due to the possibility of active gluconeogenesis, the TCA flux range expanded by 80% due to the availability of a bypass through arginine consistent with labeling data, and the transhydrogenase reaction flux was essentially unresolved due to the presence of as many as five routes for the inter-conversion of NADPH to NADH afforded by the genome-scale model. By globally accounting for ATP demands in the GSMM model the unused ATP decreased drastically with the lower bound matching the maintenance ATP requirement. A non

  5. A scientific workflow framework for (13)C metabolic flux analysis.

    PubMed

    Dalman, Tolga; Wiechert, Wolfgang; Nöh, Katharina

    2016-08-20

    Metabolic flux analysis (MFA) with (13)C labeling data is a high-precision technique to quantify intracellular reaction rates (fluxes). One of the major challenges of (13)C MFA is the interactivity of the computational workflow according to which the fluxes are determined from the input data (metabolic network model, labeling data, and physiological rates). Here, the workflow assembly is inevitably determined by the scientist who has to consider interacting biological, experimental, and computational aspects. Decision-making is context dependent and requires expertise, rendering an automated evaluation process hardly possible. Here, we present a scientific workflow framework (SWF) for creating, executing, and controlling on demand (13)C MFA workflows. (13)C MFA-specific tools and libraries, such as the high-performance simulation toolbox 13CFLUX2, are wrapped as web services and thereby integrated into a service-oriented architecture. Besides workflow steering, the SWF features transparent provenance collection and enables full flexibility for ad hoc scripting solutions. To handle compute-intensive tasks, cloud computing is supported. We demonstrate how the challenges posed by (13)C MFA workflows can be solved with our approach on the basis of two proof-of-concept use cases.

  6. Parallel labeling experiments for pathway elucidation and (13)C metabolic flux analysis.

    PubMed

    Antoniewicz, Maciek R

    2015-12-01

    Metabolic pathway models provide the foundation for quantitative studies of cellular physiology through the measurement of intracellular metabolic fluxes. For model organisms metabolic models are well established, with many manually curated genome-scale model reconstructions, gene knockout studies and stable-isotope tracing studies. However, for non-model organisms a similar level of knowledge is often lacking. Compartmentation of cellular metabolism in eukaryotic systems also presents significant challenges for quantitative (13)C-metabolic flux analysis ((13)C-MFA). Recently, innovative (13)C-MFA approaches have been developed based on parallel labeling experiments, the use of multiple isotopic tracers and integrated data analysis, that allow more rigorous validation of pathway models and improved quantification of metabolic fluxes. Applications of these approaches open new research directions in metabolic engineering, biotechnology and medicine.

  7. Elucidating the role of copper in CHO cell energy metabolism using (13)C metabolic flux analysis.

    PubMed

    Nargund, Shilpa; Qiu, Jinshu; Goudar, Chetan T

    2015-01-01

    (13)C-metabolic flux analysis was used to understand copper deficiency-related restructuring of energy metabolism, which leads to excessive lactate production in recombinant protein-producing CHO cells. Stationary-phase labeling experiments with U-(13)C glucose were conducted on CHO cells grown under high and limiting copper in 3 L fed-batch bioreactors. The resultant labeling patterns of soluble metabolites were measured by GC-MS and used to estimate metabolic fluxes in the central carbon metabolism pathways using OpenFlux. Fluxes were evaluated 300 times from stoichiometrically feasible random guess values and their confidence intervals calculated by Monte Carlo simulations. Results from metabolic flux analysis exhibited significant carbon redistribution throughout the metabolic network in cells under Cu deficiency. Specifically, glycolytic fluxes increased (25%-79% relative to glucose uptake) whereas fluxes through the TCA and pentose phosphate pathway (PPP) were lower (15%-23% and 74%, respectively) compared with the Cu-containing condition. Furthermore, under Cu deficiency, 33% of the flux entering TCA via the pyruvate node was redirected to lactate and malate production. Based on these results, we hypothesize that Cu deficiency disrupts the electron transport chain causing ATP deficiency, redox imbalance, and oxidative stress, which in turn drive copper-deficient CHO cells to produce energy via aerobic glycolysis, which is associated with excessive lactate production, rather than the more efficient route of oxidative phosphorylation.

  8. SUMOFLUX: A Generalized Method for Targeted 13C Metabolic Flux Ratio Analysis.

    PubMed

    Kogadeeva, Maria; Zamboni, Nicola

    2016-09-01

    Metabolic fluxes are a cornerstone of cellular physiology that emerge from a complex interplay of enzymes, carriers, and nutrients. The experimental assessment of in vivo intracellular fluxes using stable isotopic tracers is essential if we are to understand metabolic function and regulation. Flux estimation based on 13C or 2H labeling relies on complex simulation and iterative fitting; processes that necessitate a level of expertise that ordinarily preclude the non-expert user. To overcome this, we have developed SUMOFLUX, a methodology that is broadly applicable to the targeted analysis of 13C-metabolic fluxes. By combining surrogate modeling and machine learning, we trained a predictor to specialize in estimating flux ratios from measurable 13C-data. SUMOFLUX targets specific flux features individually, which makes it fast, user-friendly, applicable to experimental design and robust in terms of experimental noise and exchange flux magnitude. Collectively, we predict that SUMOFLUX's properties realistically pave the way to high-throughput flux analyses.

  9. SUMOFLUX: A Generalized Method for Targeted 13C Metabolic Flux Ratio Analysis

    PubMed Central

    Kogadeeva, Maria

    2016-01-01

    Metabolic fluxes are a cornerstone of cellular physiology that emerge from a complex interplay of enzymes, carriers, and nutrients. The experimental assessment of in vivo intracellular fluxes using stable isotopic tracers is essential if we are to understand metabolic function and regulation. Flux estimation based on 13C or 2H labeling relies on complex simulation and iterative fitting; processes that necessitate a level of expertise that ordinarily preclude the non-expert user. To overcome this, we have developed SUMOFLUX, a methodology that is broadly applicable to the targeted analysis of 13C-metabolic fluxes. By combining surrogate modeling and machine learning, we trained a predictor to specialize in estimating flux ratios from measurable 13C-data. SUMOFLUX targets specific flux features individually, which makes it fast, user-friendly, applicable to experimental design and robust in terms of experimental noise and exchange flux magnitude. Collectively, we predict that SUMOFLUX's properties realistically pave the way to high-throughput flux analyses. PMID:27626798

  10. A Peptide-Based Method for 13C Metabolic Flux Analysis in Microbial Communities

    PubMed Central

    Ghosh, Amit; Nilmeier, Jerome; Weaver, Daniel; Adams, Paul D.; Keasling, Jay D.; Mukhopadhyay, Aindrila; Petzold, Christopher J.; Martín, Héctor García

    2014-01-01

    The study of intracellular metabolic fluxes and inter-species metabolite exchange for microbial communities is of crucial importance to understand and predict their behaviour. The most authoritative method of measuring intracellular fluxes, 13C Metabolic Flux Analysis (13C MFA), uses the labeling pattern obtained from metabolites (typically amino acids) during 13C labeling experiments to derive intracellular fluxes. However, these metabolite labeling patterns cannot easily be obtained for each of the members of the community. Here we propose a new type of 13C MFA that infers fluxes based on peptide labeling, instead of amino acid labeling. The advantage of this method resides in the fact that the peptide sequence can be used to identify the microbial species it originates from and, simultaneously, the peptide labeling can be used to infer intracellular metabolic fluxes. Peptide identity and labeling patterns can be obtained in a high-throughput manner from modern proteomics techniques. We show that, using this method, it is theoretically possible to recover intracellular metabolic fluxes in the same way as through the standard amino acid based 13C MFA, and quantify the amount of information lost as a consequence of using peptides instead of amino acids. We show that by using a relatively small number of peptides we can counter this information loss. We computationally tested this method with a well-characterized simple microbial community consisting of two species. PMID:25188426

  11. Hybrid optimization for 13C metabolic flux analysis using systems parametrized by compactification

    PubMed Central

    Yang, Tae Hoon; Frick, Oliver; Heinzle, Elmar

    2008-01-01

    Background The importance and power of isotope-based metabolic flux analysis and its contribution to understanding the metabolic network is increasingly recognized. Its application is, however, still limited partly due to computational inefficiency. 13C metabolic flux analysis aims to compute in vivo metabolic fluxes in terms of metabolite balancing extended by carbon isotopomer balances and involves a nonlinear least-squares problem. To solve the problem more efficiently, improved numerical optimization techniques are necessary. Results For flux computation, we developed a gradient-based hybrid optimization algorithm. Here, independent flux variables were compactified into [0, 1)-ranged variables using a single transformation rule. The compactified parameters could be discriminated between non-identifiable and identifiable variables after model linearization. The developed hybrid algorithm was applied to the central metabolism of Bacillus subtilis with only succinate and glutamate as carbon sources. This creates difficulties caused by symmetry of succinate leading to limited introduction of 13C labeling information into the system. The algorithm was found to be superior to its parent algorithms and to global optimization methods both in accuracy and speed. The hybrid optimization with tolerance adjustment quickly converged to the minimum with close to zero deviation and exactly re-estimated flux variables. In the metabolic network studied, some fluxes were found to be either non-identifiable or nonlinearly correlated. The non-identifiable fluxes could correctly be predicted a priori using the model identification method applied, whereas the nonlinear flux correlation was revealed only by identification runs using different starting values a posteriori. Conclusion This fast, robust and accurate optimization method is useful for high-throughput metabolic flux analysis, a posteriori identification of possible parameter correlations, and also for Monte Carlo

  12. (13)C-metabolic flux analysis for mevalonate-producing strain of Escherichia coli.

    PubMed

    Wada, Keisuke; Toya, Yoshihiro; Banno, Satomi; Yoshikawa, Katsunori; Matsuda, Fumio; Shimizu, Hiroshi

    2017-02-01

    Mevalonate (MVA) is used to produce various useful products such as drugs, cosmetics and food additives. An MVA-producing strain of Escherichia coli (engineered) was constructed by introducing mvaES genes from Enterococcus faecalis. The engineered strain produced 1.84 mmol/gDCW/h yielding 22% (C-mol/C-mol) of MVA from glucose in the aerobic exponential growth phase. The mass balance analysis revealed that the MVA yield of the engineered strain was close to the upper limit at the biomass yield. Since MVA is synthesized from acetyl-CoA using NADPH as a cofactor, the production of MVA affects central metabolism in terms of carbon utilization and NADPH requirements. The reason for this highly efficient MVA production was investigated based on (13)C-metabolic flux analysis. The estimated flux distributions revealed that the fluxes of acetate formation and the TCA cycle in the engineered strain were lower than those in the control strain. Although the oxidative pentose phosphate pathway is considered as the NADPH generating pathway in E. coli, no difference of the flux was observed between the control and engineered strains. The production/consumption balance of NADPH suggested that additional requirement of NADPH for MVA synthesis was obtained from the transhydrogenase reaction in the engineered strain. Comparison between the measured flux distribution and the ideal values for MVA production proposes a strategy for further engineering to improve the MVA production in E. coli.

  13. (13)C-metabolic flux analysis in S-adenosyl-L-methionine production by Saccharomyces cerevisiae.

    PubMed

    Hayakawa, Kenshi; Kajihata, Shuichi; Matsuda, Fumio; Shimizu, Hiroshi

    2015-11-01

    S-Adenosyl-L-methionine (SAM) is a major biological methyl group donor, and is used as a nutritional supplement and prescription drug. Yeast is used for the industrial production of SAM owing to its high intracellular SAM concentrations. To determine the regulation mechanisms responsible for such high SAM production, (13)C-metabolic flux analysis ((13)C-MFA) was conducted to compare the flux distributions in the central metabolism between Kyokai no. 6 (high SAM-producing) and S288C (control) strains. (13)C-MFA showed that the levels of tricarboxylic acid (TCA) cycle flux in SAM-overproducing strain were considerably increased compared to those in the S228C strain. Analysis of ATP balance also showed that a larger amount of excess ATP was produced in the Kyokai 6 strain because of increased oxidative phosphorylation. These results suggest that high SAM production in Kyokai 6 strains could be attributed to enhanced ATP regeneration with high TCA cycle fluxes and respiration activity. Thus, maintaining high respiration efficiency during cultivation is important for improving SAM production.

  14. (13)C-metabolic flux analysis of co-cultures: A novel approach.

    PubMed

    Gebreselassie, Nikodimos A; Antoniewicz, Maciek R

    2015-09-01

    In this work, we present a novel approach for performing (13)C metabolic flux analysis ((13)C-MFA) of co-culture systems. We demonstrate for the first time that it is possible to determine metabolic flux distributions in multiple species simultaneously without the need for physical separation of cells or proteins, or overexpression of species-specific products. Instead, metabolic fluxes for each species in a co-culture are estimated directly from isotopic labeling of total biomass obtained using conventional mass spectrometry approaches such as GC-MS. In addition to determining metabolic fluxes, this approach estimates the relative population size of each species in a mixed culture and inter-species metabolite exchange. As such, it enables detailed studies of microbial communities including species dynamics and interactions between community members. The methodology is experimentally validated here using a co-culture of two E. coli knockout strains. Taken together, this work greatly extends the scope of (13)C-MFA to a large number of multi-cellular systems that are of significant importance in biotechnology and medicine.

  15. (13) C-metabolic flux analysis of human adenovirus infection: Implications for viral vector production.

    PubMed

    Carinhas, Nuno; Koshkin, Alexey; Pais, Daniel A M; Alves, Paula M; Teixeira, Ana P

    2017-01-01

    Adenoviruses are human pathogens increasingly used as gene therapy and vaccination vectors. However, their impact on cell metabolism is poorly characterized. We performed carbon labeling experiments with [1,2-(13) C]glucose or [U-(13) C]glutamine to evaluate metabolic alterations in the amniocyte-derived, E1-transformed 1G3 cell line during production of a human adenovirus type 5 vector (AdV5). Nonstationary (13) C-metabolic flux analysis revealed increased fluxes of glycolysis (17%) and markedly PPP (over fourfold) and cytosolic AcCoA formation (nearly twofold) following infection of growing cells. Interestingly, infection of growth-arrested cells increased overall carbon flow even more, including glutamine anaplerosis and TCA cycle activity (both over 1.5-fold), but was unable to stimulate the PPP and was associated with a steep drop in AdV5 replication (almost 80%). Our results underscore the importance of nucleic and fatty acid biosynthesis for adenovirus replication. Overall, we portray a metabolic blueprint of human adenovirus infection, highlighting similarities with other viruses and cancer, and suggest strategies to improve AdV5 production. Biotechnol. Bioeng. 2017;114: 195-207. © 2016 Wiley Periodicals, Inc.

  16. Parallel labeling experiments validate Clostridium acetobutylicum metabolic network model for (13)C metabolic flux analysis.

    PubMed

    Au, Jennifer; Choi, Jungik; Jones, Shawn W; Venkataramanan, Keerthi P; Antoniewicz, Maciek R

    2014-11-01

    In this work, we provide new insights into the metabolism of Clostridium acetobutylicum ATCC 824 obtained using a systematic approach for quantifying fluxes based on parallel labeling experiments and (13)C-metabolic flux analysis ((13)C-MFA). Here, cells were grown in parallel cultures with [1-(13)C]glucose and [U-(13)C]glucose as tracers and (13)C-MFA was used to quantify intracellular metabolic fluxes. Several metabolic network models were compared: an initial model based on current knowledge, and extended network models that included additional reactions that improved the fits of experimental data. While the initial network model did not produce a statistically acceptable fit of (13)C-labeling data, an extended network model with five additional reactions was able to fit all data with 292 redundant measurements. The model was subsequently trimmed to produce a minimal network model of C. acetobutylicum for (13)C-MFA, which could still reproduce all of the experimental data. The flux results provided valuable new insights into the metabolism of C. acetobutylicum. First, we found that TCA cycle was effectively incomplete, as there was no measurable flux between α-ketoglutarate and succinyl-CoA, succinate and fumarate, and malate and oxaloacetate. Second, an active pathway was identified from pyruvate to fumarate via aspartate. Third, we found that isoleucine was produced exclusively through the citramalate synthase pathway in C. acetobutylicum and that CAC3174 was likely responsible for citramalate synthase activity. These model predictions were confirmed in several follow-up tracer experiments. The validated metabolic network model established in this study can be used in future investigations for unbiased (13)C-flux measurements in C. acetobutylicum.

  17. Integrated 13C-metabolic flux analysis of 14 parallel labeling experiments in Escherichia coli.

    PubMed

    Crown, Scott B; Long, Christopher P; Antoniewicz, Maciek R

    2015-03-01

    The use of parallel labeling experiments for (13)C metabolic flux analysis ((13)C-MFA) has emerged in recent years as the new gold standard in fluxomics. The methodology has been termed COMPLETE-MFA, short for complementary parallel labeling experiments technique for metabolic flux analysis. In this contribution, we have tested the limits of COMPLETE-MFA by demonstrating integrated analysis of 14 parallel labeling experiments with Escherichia coli. An effort on such a massive scale has never been attempted before. In addition to several widely used isotopic tracers such as [1,2-(13)C]glucose and mixtures of [1-(13)C]glucose and [U-(13)C]glucose, four novel tracers were applied in this study: [2,3-(13)C]glucose, [4,5,6-(13)C]glucose, [2,3,4,5,6-(13)C]glucose and a mixture of [1-(13)C]glucose and [4,5,6-(13)C]glucose. This allowed us for the first time to compare the performance of a large number of isotopic tracers. Overall, there was no single best tracer for the entire E. coli metabolic network model. Tracers that produced well-resolved fluxes in the upper part of metabolism (glycolysis and pentose phosphate pathways) showed poor performance for fluxes in the lower part of metabolism (TCA cycle and anaplerotic reactions), and vice versa. The best tracer for upper metabolism was 80% [1-(13)C]glucose+20% [U-(13)C]glucose, while [4,5,6-(13)C]glucose and [5-(13)C]glucose both produced optimal flux resolution in the lower part of metabolism. COMPLETE-MFA improved both flux precision and flux observability, i.e. more independent fluxes were resolved with smaller confidence intervals, especially exchange fluxes. Overall, this study demonstrates that COMPLETE-MFA is a powerful approach for improving flux measurements and that this methodology should be considered in future studies that require very high flux resolution.

  18. (13)C Metabolic Flux Analysis of acetate conversion to lipids by Yarrowia lipolytica.

    PubMed

    Liu, Nian; Qiao, Kangjian; Stephanopoulos, Gregory

    2016-11-01

    Volatile fatty acids (VFAs) are an inexpensive and renewable carbon source that can be generated from gas fermentation and anaerobic digestion of fermentable wastes. The oleaginous yeast Yarrowia lipolytica is a promising biocatalyst that can utilize VFAs and convert them into triacylglycerides (TAGs). However, currently there is limited knowledge on the metabolism of Y. lipolytica when cultured on VFAs. To develop a better understanding, we used acetate as the sole carbon source to culture two strains, a control strain and a previously engineered strain for lipid overaccumulation. For both strains, metabolism during the growth phase and lipid production phase were investigated by metabolic flux analysis using two parallel sodium acetate tracers. The resolved flux distributions demonstrate that the glyoxylate shunt pathway is constantly active and the flux through gluconeogenesis varies depending on strain and phase. In particular, by regulating the activities of malate transport and pyruvate kinase, the cells divert only a portion of the glyoxylate shunt flux required to satisfy the needs for anaplerotic reactions and NADPH production through gluconeogenesis and the oxidative pentose phosphate pathway (PPP). Excess flux flows back to the tricarboxylic acid (TCA) cycle for energy production. As with the case of glucose as the substrate, the primary source for lipogenic NADPH is derived from the oxidative PPP.

  19. 13C metabolic flux analysis in Clostridium acetobutylicum during growth on L-arabinose

    NASA Astrophysics Data System (ADS)

    Hurley, Margaret; Sund, Christian; Liu, Sanchao; Germane, Katherine; Servinsky, Matthew; Gerlach, Elliot

    2015-03-01

    Clostridium acetobutylicum's metabolic pathways have been studied for decades due to its metabolic diversity and industrial value, yet many details of its metabolism are continuing to emerge. To elucidate the role of xylulose-5-P/fructose-6-P phosphoketolase (XFP), and the recently discovered Pentose Phosphate Pathway (PKP) in C. acetobutylicum, experimental and computational metabolic isotope analysis was performed under growth on glucose, xylose, and arabinose. Results indicate that PKP utilization increased with increasing xylose concentration and this trend was further pronounced during growth on arabinose. This was confirmed by mutation of the gene encoding XFP, which almost completely abolished flux through the PKP during growth on arabinose and resulted in decreased acetate:butyrate ratios. We discuss these experimental and computational results here, and the implications for our understanding of sugar metabolism in C. acetobutylicum.

  20. 13 C-metabolic flux analysis in heterologous cellulase production by Bacillus subtilis genome-reduced strain.

    PubMed

    Toya, Yoshihiro; Hirasawa, Takashi; Morimoto, Takuya; Masuda, Kenta; Kageyama, Yasushi; Ozaki, Katsuya; Ogasawara, Naotake; Shimizu, Hiroshi

    2014-06-10

    The great potential of Bacillus subtilis to produce biomaterials would be further enhanced by the development of strains with deletions of non-essential genomic regions. Here, using stationary (13)C-metabolic flux analysis ((13)C-MFA), we investigated the metabolism during cellulase production by the genome-reduced B. subtilis strain MGB874. We transformed MGB874 and wild-type strains with the heterologous cellulase gene, and cultured these on a synthetic medium containing glucose as carbon source. The addition of glutamate and the genome reduction enhanced cellulase production, which led us to use (13)C-MFA to assess the effects of glutamate addition and gene deletions on metabolism. We found that there was a significant increase in the flux in the pentose phosphate (PP) pathway, whereas the fluxes of reactions from acetyl-CoA to α-ketoglutarate were repressed in the presence of glutamate. We hypothesize that the increase in the PP pathway flux was caused by the decrease of citrate synthase flux through the accumulation of glycolytic intermediates. Excess NADPH produced by the PP pathway may affect the increase in cellulase production. Furthermore, the fluxes on glycolysis and the acetate formation of the cellulase-producing wild-type strain were significantly larger than that of the cellulase-producing MGB874 strain when the strains were cultured with glucose and glutamate.

  1. Cutting the Gordian Knot: Identifiability of anaplerotic reactions in Corynebacterium glutamicum by means of (13) C-metabolic flux analysis.

    PubMed

    Kappelmann, Jannick; Wiechert, Wolfgang; Noack, Stephan

    2016-03-01

    Corynebacterium glutamicum is the major workhorse for the microbial production of several amino and organic acids. As long as these derive from tricarboxylic acid cycle intermediates, the activity of anaplerotic reactions is pivotal for a high biosynthetic yield. To determine single anaplerotic activities (13) C-Metabolic Flux Analysis ((13) C-MFA) has been extensively used for C. glutamicum, however with different network topologies, inconsistent or poorly determined anaplerotic reaction rates. Therefore, in this study we set out to investigate whether a focused isotopomer model of the anaplerotic node can at all admit a unique solution for all fluxes. By analyzing different scenarios of active anaplerotic reactions, we show in full generality that for C. glutamicum only certain anaplerotic deletion mutants allow to uniquely determine the anaplerotic fluxes from (13) C-isotopomer data. We stress that the result of this analysis for different assumptions on active enzymes is directly transferable to other compartment-free organisms. Our results demonstrate that there exist biologically relevant metabolic network topologies for which the flux distribution cannot be inferred by classical (13) C-MFA.

  2. (13)C-metabolic flux analysis of lipid accumulation in the oleaginous fungus Mucor circinelloides.

    PubMed

    Zhao, Lina; Zhang, Huaiyuan; Wang, Liping; Chen, Haiqin; Chen, Yong Q; Chen, Wei; Song, Yuanda

    2015-12-01

    The oleaginous fungus Mucor circinelloides is of industrial interest because it can produce high levels of polyunsaturated fatty acid γ-linolenic acid. M. circinelloides CBS 277.49 is able to accumulate less than 15% of cell dry weight as lipids, while M. circinelloides WJ11 can accumulate lipid up to 36%. In order to better understand the mechanisms behind the differential lipid accumulation in these two strains, tracer experiments with (13)C-glucose were performed with the growth of M. circinelloides and subsequent gas chromatography-mass spectrometric detection of (13)C-patterns in proteinogenic amino acids was carried out to identify the metabolic network topology and estimate intracellular fluxes. Our results showed that the high oleaginous strain WJ11 had higher flux of pentose phosphate pathway and malic enzyme, lower flux in tricarboxylic acid cycle, higher flux in glyoxylate cycle and ATP: citrate lyase, together, it might provide more NADPH and substrate acetyl-CoA for fatty acid synthesis.

  3. Central metabolic responses to the overproduction of fatty acids in Escherichia coli based on 13C-metabolic flux analysis.

    PubMed

    He, Lian; Xiao, Yi; Gebreselassie, Nikodimos; Zhang, Fuzhong; Antoniewiez, Maciek R; Tang, Yinjie J; Peng, Lifeng

    2014-03-01

    We engineered a fatty acid overproducing Escherichia coli strain through overexpressing tesA (“pull”) and fadR (“push”) and knocking out fadE (“block”). This “pull-push-block” strategy yielded 0.17 g of fatty acids (C12–C18) per gram of glucose (equivalent to 48% of the maximum theoretical yield) in batch cultures during the exponential growth phase under aerobic conditions. Metabolic fluxes were determined for the engineered E. coli and its control strain using tracer ([1,2-13C]glucose) experiments and 13C-metabolic flux analysis. Cofactor (NADPH) and energy (ATP) balances were also investigated for both strains based on estimated fluxes. Compared to the control strain, fatty acid overproduction led to significant metabolic responses in the central metabolism: (1) Acetic acid secretion flux decreased 10-fold; (2) Pentose phosphate pathway and Entner–Doudoroff pathway fluxes increased 1.5- and 2.0-fold, respectively; (3) Biomass synthesis flux was reduced 1.9-fold; (4) Anaplerotic phosphoenolpyruvate carboxylation flux decreased 1.7-fold; (5) Transhydrogenation flux converting NADH to NADPH increased by 1.7-fold. Real-time quantitative RT-PCR analysis revealed the engineered strain increased the transcription levels of pntA (encoding the membrane-bound transhydrogenase) by 2.1-fold and udhA (encoding the soluble transhydrogenase) by 1.4-fold, which is in agreement with the increased transhydrogenation flux. Cofactor and energy balances analyses showed that the fatty acid overproducing E. coli consumed significantly higher cellular maintenance energy than the control strain. We discussed the strategies to future strain development and process improvements for fatty acid production in E. coli.

  4. IsoDesign: a software for optimizing the design of 13C-metabolic flux analysis experiments.

    PubMed

    Millard, Pierre; Sokol, Serguei; Letisse, Fabien; Portais, Jean-Charles

    2014-01-01

    The growing demand for (13) C-metabolic flux analysis ((13) C-MFA) in the field of metabolic engineering and systems biology is driving the need to rationalize expensive and time-consuming (13) C-labeling experiments. Experimental design is a key step in improving both the number of fluxes that can be calculated from a set of isotopic data and the precision of flux values. We present IsoDesign, a software that enables these parameters to be maximized by optimizing the isotopic composition of the label input. It can be applied to (13) C-MFA investigations using a broad panel of analytical tools (MS, MS/MS, (1) H NMR, (13) C NMR, etc.) individually or in combination. It includes a visualization module to intuitively select the optimal label input depending on the biological question to be addressed. Applications of IsoDesign are described, with an example of the entire (13) C-MFA workflow from the experimental design to the flux map including important practical considerations. IsoDesign makes the experimental design of (13) C-MFA experiments more accessible to a wider biological community. IsoDesign is distributed under an open source license at http://metasys.insa-toulouse.fr/software/isodes/

  5. Comparative 13C metabolic flux analysis of pyruvate dehydrogenase complex-deficient, L-valine-producing Corynebacterium glutamicum.

    PubMed

    Bartek, Tobias; Blombach, Bastian; Lang, Siegmund; Eikmanns, Bernhard J; Wiechert, Wolfgang; Oldiges, Marco; Nöh, Katharina; Noack, Stephan

    2011-09-01

    L-Valine can be formed successfully using C. glutamicum strains missing an active pyruvate dehydrogenase enzyme complex (PDHC). Wild-type C. glutamicum and four PDHC-deficient strains were compared by (13)C metabolic flux analysis, especially focusing on the split ratio between glycolysis and the pentose phosphate pathway (PPP). Compared to the wild type, showing a carbon flux of 69% ± 14% through the PPP, a strong increase in the PPP flux was observed in PDHC-deficient strains with a maximum of 113% ± 22%. The shift in the split ratio can be explained by an increased demand of NADPH for l-valine formation. In accordance, the introduction of the Escherichia coli transhydrogenase PntAB, catalyzing the reversible conversion of NADH to NADPH, into an L-valine-producing C. glutamicum strain caused the PPP flux to decrease to 57% ± 6%, which is below the wild-type split ratio. Hence, transhydrogenase activity offers an alternative perspective for sufficient NADPH supply, which is relevant for most amino acid production systems. Moreover, as demonstrated for L-valine, this bypass leads to a significant increase of product yield due to a concurrent reduction in carbon dioxide formation via the PPP.

  6. Optimal tracers for parallel labeling experiments and (13)C metabolic flux analysis: A new precision and synergy scoring system.

    PubMed

    Crown, Scott B; Long, Christopher P; Antoniewicz, Maciek R

    2016-11-01

    (13)C-Metabolic flux analysis ((13)C-MFA) is a widely used approach in metabolic engineering for quantifying intracellular metabolic fluxes. The precision of fluxes determined by (13)C-MFA depends largely on the choice of isotopic tracers and the specific set of labeling measurements. A recent advance in the field is the use of parallel labeling experiments for improved flux precision and accuracy. However, as of today, no systemic methods exist for identifying optimal tracers for parallel labeling experiments. In this contribution, we have addressed this problem by introducing a new scoring system and evaluating thousands of different isotopic tracer schemes. Based on this extensive analysis we have identified optimal tracers for (13)C-MFA. The best single tracers were doubly (13)C-labeled glucose tracers, including [1,6-(13)C]glucose, [5,6-(13)C]glucose and [1,2-(13)C]glucose, which consistently produced the highest flux precision independent of the metabolic flux map (here, 100 random flux maps were evaluated). Moreover, we demonstrate that pure glucose tracers perform better overall than mixtures of glucose tracers. For parallel labeling experiments the optimal isotopic tracers were [1,6-(13)C]glucose and [1,2-(13)C]glucose. Combined analysis of [1,6-(13)C]glucose and [1,2-(13)C]glucose labeling data improved the flux precision score by nearly 20-fold compared to widely use tracer mixture 80% [1-(13)C]glucose +20% [U-(13)C]glucose.

  7. Investigate the Metabolic Reprogramming of Saccharomyces cerevisiae for Enhanced Resistance to Mixed Fermentation Inhibitors via 13C Metabolic Flux Analysis

    PubMed Central

    Guo, Weihua; Chen, Yingying; Wei, Na; Feng, Xueyang

    2016-01-01

    The fermentation inhibitors from the pretreatment of lignocellulosic materials, e.g., acetic acid and furfural, are notorious due to their negative effects on the cell growth and chemical production. However, the metabolic reprogramming of the cells under these stress conditions, especially metabolic response for resistance to mixed inhibitors, has not been systematically investigated and remains mysterious. Therefore, in this study, 13C metabolic flux analysis (13C-MFA), a powerful tool to elucidate the intracellular carbon flux distributions, has been applied to two Saccharomyces cerevisiae strains with different tolerances to the inhibitors under acetic acid, furfural, and mixed (i.e., acetic acid and furfural) stress conditions to unravel the key metabolic responses. By analyzing the intracellular carbon fluxes as well as the energy and cofactor utilization under different conditions, we uncovered varied metabolic responses to different inhibitors. Under acetate stress, ATP and NADH production was slightly impaired, while NADPH tended towards overproduction. Under furfural stress, ATP and cofactors (including both NADH and NADPH) tended to be overproduced. However, under dual-stress condition, production of ATP and cofactors was severely impaired due to synergistic stress caused by the simultaneous addition of two fermentation inhibitors. Such phenomenon indicated the pivotal role of the energy and cofactor utilization in resisting the mixed inhibitors of acetic acid and furfural. Based on the discoveries, valuable insights are provided to improve the tolerance of S. cerevisiae strain and further enhance lignocellulosic fermentation. PMID:27532329

  8. Integration of a constraint-based metabolic model of Brassica napus developing seeds with (13)C-metabolic flux analysis.

    PubMed

    Hay, Jordan O; Shi, Hai; Heinzel, Nicolas; Hebbelmann, Inga; Rolletschek, Hardy; Schwender, Jorg

    2014-01-01

    The use of large-scale or genome-scale metabolic reconstructions for modeling and simulation of plant metabolism and integration of those models with large-scale omics and experimental flux data is becoming increasingly important in plant metabolic research. Here we report an updated version of bna572, a bottom-up reconstruction of oilseed rape (Brassica napus L.; Brassicaceae) developing seeds with emphasis on representation of biomass-component biosynthesis. New features include additional seed-relevant pathways for isoprenoid, sterol, phenylpropanoid, flavonoid, and choline biosynthesis. Being now based on standardized data formats and procedures for model reconstruction, bna572+ is available as a COBRA-compliant Systems Biology Markup Language (SBML) model and conforms to the Minimum Information Requested in the Annotation of Biochemical Models (MIRIAM) standards for annotation of external data resources. Bna572+ contains 966 genes, 671 reactions, and 666 metabolites distributed among 11 subcellular compartments. It is referenced to the Arabidopsis thaliana genome, with gene-protein-reaction (GPR) associations resolving subcellular localization. Detailed mass and charge balancing and confidence scoring were applied to all reactions. Using B. napus seed specific transcriptome data, expression was verified for 78% of bna572+ genes and 97% of reactions. Alongside bna572+ we also present a revised carbon centric model for (13)C-Metabolic Flux Analysis ((13)C-MFA) with all its reactions being referenced to bna572+ based on linear projections. By integration of flux ratio constraints obtained from (13)C-MFA and by elimination of infinite flux bounds around thermodynamically infeasible loops based on COBRA loopless methods, we demonstrate improvements in predictive power of Flux Variability Analysis (FVA). Using this combined approach we characterize the difference in metabolic flux of developing seeds of two B. napus genotypes contrasting in starch and oil content.

  9. Integration of a constraint-based metabolic model of Brassica napus developing seeds with 13C-metabolic flux analysis

    PubMed Central

    Hay, Jordan O.; Shi, Hai; Heinzel, Nicolas; Hebbelmann, Inga; Rolletschek, Hardy; Schwender, Jorg

    2014-01-01

    The use of large-scale or genome-scale metabolic reconstructions for modeling and simulation of plant metabolism and integration of those models with large-scale omics and experimental flux data is becoming increasingly important in plant metabolic research. Here we report an updated version of bna572, a bottom-up reconstruction of oilseed rape (Brassica napus L.; Brassicaceae) developing seeds with emphasis on representation of biomass-component biosynthesis. New features include additional seed-relevant pathways for isoprenoid, sterol, phenylpropanoid, flavonoid, and choline biosynthesis. Being now based on standardized data formats and procedures for model reconstruction, bna572+ is available as a COBRA-compliant Systems Biology Markup Language (SBML) model and conforms to the Minimum Information Requested in the Annotation of Biochemical Models (MIRIAM) standards for annotation of external data resources. Bna572+ contains 966 genes, 671 reactions, and 666 metabolites distributed among 11 subcellular compartments. It is referenced to the Arabidopsis thaliana genome, with gene-protein-reaction (GPR) associations resolving subcellular localization. Detailed mass and charge balancing and confidence scoring were applied to all reactions. Using B. napus seed specific transcriptome data, expression was verified for 78% of bna572+ genes and 97% of reactions. Alongside bna572+ we also present a revised carbon centric model for 13C-Metabolic Flux Analysis (13C-MFA) with all its reactions being referenced to bna572+ based on linear projections. By integration of flux ratio constraints obtained from 13C-MFA and by elimination of infinite flux bounds around thermodynamically infeasible loops based on COBRA loopless methods, we demonstrate improvements in predictive power of Flux Variability Analysis (FVA). Using this combined approach we characterize the difference in metabolic flux of developing seeds of two B. napus genotypes contrasting in starch and oil content. PMID

  10. Integration of a constraint-based metabolic model of Brassica napus developing seeds with 13C-metabolic flux analysis

    DOE PAGES

    Hay, Jordan O.; Shi, Hai; Heinzel, Nicolas; ...

    2014-12-19

    The use of large-scale or genome-scale metabolic reconstructions for modeling and simulation of plant metabolism and integration of those models with large-scale omics and experimental flux data is becoming increasingly important in plant metabolic research. Here we report an updated version of bna572, a bottom-up reconstruction of oilseed rape (Brassica napus L.; Brassicaceae) developing seeds with emphasis on representation of biomass-component biosynthesis. New features include additional seed-relevant pathways for isoprenoid, sterol, phenylpropanoid, flavonoid, and choline biosynthesis. Being now based on standardized data formats and procedures for model reconstruction, bna572+ is available as a COBRA-compliant Systems Biology Markup Language (SBML) modelmore » and conforms to the Minimum Information Requested in the Annotation of Biochemical Models (MIRIAM) standards for annotation of external data resources. Bna572+ contains 966 genes, 671 reactions, and 666 metabolites distributed among 11 subcellular compartments. It is referenced to the Arabidopsis thaliana genome, with gene-protein-reaction (GPR) associations resolving subcellular localization. Detailed mass and charge balancing and confidence scoring were applied to all reactions. Using B. napus seed specific transcriptome data, expression was verified for 78% of bna572+ genes and 97% of reactions. Alongside bna572+ we also present a revised carbon centric model for 13C-Metabolic Flux Analysis (13C-MFA) with all its reactions being referenced to bna572+ based on linear projections. By integration of flux ratio constraints obtained from 13C-MFA and by elimination of infinite flux bounds around thermodynamically infeasible loops based on COBRA loopless methods, we demonstrate improvements in predictive power of Flux Variability Analysis (FVA). In conclusion, using this combined approach we characterize the difference in metabolic flux of developing seeds of two B. napus genotypes contrasting in starch

  11. (13)C metabolic flux analysis of the extremely thermophilic, fast growing, xylose-utilizing Geobacillus strain LC300.

    PubMed

    Cordova, Lauren T; Antoniewicz, Maciek R

    2016-01-01

    Thermophiles are increasingly used as versatile hosts in the biotechnology industry. One of the key advantages of thermophiles is the potential to achieve high rates of feedstock conversion at elevated temperatures. The recently isolated Geobacillus strain LC300 grows extremely fast on xylose, with a doubling time of less than 30 min. In the accompanying paper, the genome of Geobacillus LC300 was sequenced and annotated. In this work, we have experimentally validated the metabolic network model using parallel (13)C-labeling experiments and applied (13)C-metabolic flux analysis to quantify precise metabolic fluxes. Specifically, the complete set of singly labeled xylose tracers, [1-(13)C], [2-(13)C], [3-(13)C], [4-(13)C], and [5-(13)C]xylose, was used for the first time. Isotopic labeling of biomass amino acids was measured by gas chromatography mass spectrometry (GC-MS). Isotopic labeling of carbon dioxide in the off-gas was also measured by an on-line mass spectrometer. The (13)C-labeling data was then rigorously integrated for flux elucidation using the COMPLETE-MFA approach. The results provided important new insights into the metabolism of Geobacillus LC300, its efficient xylose utilization pathways, and the balance between carbon, redox and energy fluxes. The pentose phosphate pathway, glycolysis and TCA cycle were found to be highly active in Geobacillus LC300. The oxidative pentose phosphate pathway was also active and contributed significantly to NADPH production. No transhydrogenase activity was detected. Results from this work provide a solid foundation for future studies of this strain and its metabolic engineering and biotechnological applications.

  12. Metabolic network reconstruction, growth characterization and 13C-metabolic flux analysis of the extremophile Thermus thermophilus HB8.

    PubMed

    Swarup, Aditi; Lu, Jing; DeWoody, Kathleen C; Antoniewicz, Maciek R

    2014-07-01

    Thermus thermophilus is an extremely thermophilic bacterium with significant biotechnological potential. In this work, we have characterized aerobic growth characteristics of T. thermophilus HB8 at temperatures between 50 and 85°C, constructed a metabolic network model of its central carbon metabolism and validated the model using (13)C-metabolic flux analysis ((13)C-MFA). First, cells were grown in batch cultures in custom constructed mini-bioreactors at different temperatures to determine optimal growth conditions. The optimal temperature for T. thermophilus grown on defined medium with glucose was 81°C. The maximum growth rate was 0.25h(-1). Between 50 and 81°C the growth rate increased by 7-fold and the temperature dependence was described well by an Arrhenius model with an activation energy of 47kJ/mol. Next, we performed a (13)C-labeling experiment with [1,2-(13)C] glucose as the tracer and calculated intracellular metabolic fluxes using (13)C-MFA. The results provided support for the constructed network model and highlighted several interesting characteristics of T. thermophilus metabolism. We found that T. thermophilus largely uses glycolysis and TCA cycle to produce biosynthetic precursors, ATP and reducing equivalents needed for cells growth. Consistent with its proposed metabolic network model, we did not detect any oxidative pentose phosphate pathway flux or Entner-Doudoroff pathway activity. The biomass precursors erythrose-4-phosphate and ribose-5-phosphate were produced via the non-oxidative pentose phosphate pathway, and largely via transketolase, with little contribution from transaldolase. The high biomass yield on glucose that was measured experimentally was also confirmed independently by (13)C-MFA. The results presented here provide a solid foundation for future studies of T. thermophilus and its metabolic engineering applications.

  13. Computational analysis of storage synthesis in developing Brassica napus L. (oilseed rape) embryos: Flux variability analysis in relation to 13C-metabolic flux analysis

    SciTech Connect

    Hay, J.; Schwender, J.

    2011-08-01

    Plant oils are an important renewable resource, and seed oil content is a key agronomical trait that is in part controlled by the metabolic processes within developing seeds. A large-scale model of cellular metabolism in developing embryos of Brassica napus (bna572) was used to predict biomass formation and to analyze metabolic steady states by flux variability analysis under different physiological conditions. Predicted flux patterns are highly correlated with results from prior 13C metabolic flux analysis of B. napus developing embryos. Minor differences from the experimental results arose because bna572 always selected only one sugar and one nitrogen source from the available alternatives, and failed to predict the use of the oxidative pentose phosphate pathway. Flux variability, indicative of alternative optimal solutions, revealed alternative pathways that can provide pyruvate and NADPH to plastidic fatty acid synthesis. The nutritional values of different medium substrates were compared based on the overall carbon conversion efficiency (CCE) for the biosynthesis of biomass. Although bna572 has a functional nitrogen assimilation pathway via glutamate synthase, the simulations predict an unexpected role of glycine decarboxylase operating in the direction of NH4+ assimilation. Analysis of the light-dependent improvement of carbon economy predicted two metabolic phases. At very low light levels small reductions in CO2 efflux can be attributed to enzymes of the tricarboxylic acid cycle (oxoglutarate dehydrogenase, isocitrate dehydrogenase) and glycine decarboxylase. At higher light levels relevant to the 13C flux studies, ribulose-1,5-bisphosphate carboxylase activity is predicted to account fully for the light-dependent changes in carbon balance.

  14. Evidence for transketolase-like TKTL1 flux in CHO cells based on parallel labeling experiments and (13)C-metabolic flux analysis.

    PubMed

    Ahn, Woo Suk; Crown, Scott B; Antoniewicz, Maciek R

    2016-09-01

    The pentose phosphate pathway (PPP) is a fundamental component of cellular metabolism. It provides precursors for the biosynthesis of nucleotides and contributes to the production of reducing power in the form of NADPH. It has been hypothesized that mammalian cells may contain a hidden reaction in PPP catalyzed by transketolase-like protein 1 (TKTL1) that is closely related to the classical transketolase enzyme; however, until now there has been no direct experimental evidence for this reaction. In this work, we have applied state-of-the-art techniques in (13)C metabolic flux analysis ((13)C-MFA) based on parallel labeling experiments and integrated flux fitting to estimate the TKTL1 flux in CHO cells. We identified a set of three parallel labeling experiments with [1-(13)C]glucose+[4,5,6-(13)C]glucose, [2-(13)C]glucose+[4,5,6-(13)C]glucose, and [3-(13)C]glucose+[4,5,6-(13)C]glucose and developed a new method to measure (13)C-labeling of fructose 6-phosphate by GC-MS that allows intuitive interpretation of mass isotopomer distributions to determine key fluxes in the model, including glycolysis, oxidative PPP, non-oxidative PPP, and the TKTL1 flux. Using these tracers we detected a significant TKTL1 flux in CHO cells at the stationary phase. The flux results suggest that the main function of oxidative PPP in CHO cells at the stationary phase is to fuel the TKTL1 reaction. Overall, this study demonstrates for the first time that carbon atoms can be lost in the PPP, by means other than the oxidative PPP, and that this loss of carbon atoms is consistent with the hypothesized TKTL1 reaction in mammalian cells.

  15. (13)C Metabolic Flux Analysis for Systematic Metabolic Engineering of S. cerevisiae for Overproduction of Fatty Acids.

    PubMed

    Ghosh, Amit; Ando, David; Gin, Jennifer; Runguphan, Weerawat; Denby, Charles; Wang, George; Baidoo, Edward E K; Shymansky, Chris; Keasling, Jay D; García Martín, Héctor

    2016-01-01

    Efficient redirection of microbial metabolism into the abundant production of desired bioproducts remains non-trivial. Here, we used flux-based modeling approaches to improve yields of fatty acids in Saccharomyces cerevisiae. We combined (13)C labeling data with comprehensive genome-scale models to shed light onto microbial metabolism and improve metabolic engineering efforts. We concentrated on studying the balance of acetyl-CoA, a precursor metabolite for the biosynthesis of fatty acids. A genome-wide acetyl-CoA balance study showed ATP citrate lyase from Yarrowia lipolytica as a robust source of cytoplasmic acetyl-CoA and malate synthase as a desirable target for downregulation in terms of acetyl-CoA consumption. These genetic modifications were applied to S. cerevisiae WRY2, a strain that is capable of producing 460 mg/L of free fatty acids. With the addition of ATP citrate lyase and downregulation of malate synthase, the engineered strain produced 26% more free fatty acids. Further increases in free fatty acid production of 33% were obtained by knocking out the cytoplasmic glycerol-3-phosphate dehydrogenase, which flux analysis had shown was competing for carbon flux upstream with the carbon flux through the acetyl-CoA production pathway in the cytoplasm. In total, the genetic interventions applied in this work increased fatty acid production by ~70%.

  16. 13C Metabolic Flux Analysis for Systematic Metabolic Engineering of S. cerevisiae for Overproduction of Fatty Acids

    PubMed Central

    Ghosh, Amit; Ando, David; Gin, Jennifer; Runguphan, Weerawat; Denby, Charles; Wang, George; Baidoo, Edward E. K.; Shymansky, Chris; Keasling, Jay D.; García Martín, Héctor

    2016-01-01

    Efficient redirection of microbial metabolism into the abundant production of desired bioproducts remains non-trivial. Here, we used flux-based modeling approaches to improve yields of fatty acids in Saccharomyces cerevisiae. We combined 13C labeling data with comprehensive genome-scale models to shed light onto microbial metabolism and improve metabolic engineering efforts. We concentrated on studying the balance of acetyl-CoA, a precursor metabolite for the biosynthesis of fatty acids. A genome-wide acetyl-CoA balance study showed ATP citrate lyase from Yarrowia lipolytica as a robust source of cytoplasmic acetyl-CoA and malate synthase as a desirable target for downregulation in terms of acetyl-CoA consumption. These genetic modifications were applied to S. cerevisiae WRY2, a strain that is capable of producing 460 mg/L of free fatty acids. With the addition of ATP citrate lyase and downregulation of malate synthase, the engineered strain produced 26% more free fatty acids. Further increases in free fatty acid production of 33% were obtained by knocking out the cytoplasmic glycerol-3-phosphate dehydrogenase, which flux analysis had shown was competing for carbon flux upstream with the carbon flux through the acetyl-CoA production pathway in the cytoplasm. In total, the genetic interventions applied in this work increased fatty acid production by ~70%. PMID:27761435

  17. Photobioreactor design for isotopic non-stationary 13C-metabolic flux analysis (INST 13C-MFA) under photoautotrophic conditions.

    PubMed

    Martzolff, Arnaud; Cahoreau, Edern; Cogne, Guillaume; Peyriga, Lindsay; Portais, Jean-Charles; Dechandol, Emmanuel; Le Grand, Fabienne; Massou, Stéphane; Gonçalves, Olivier; Pruvost, Jérémy; Legrand, Jack

    2012-12-01

    Adaptive metabolic behavior of photoautotrophic microorganisms toward genetic and environmental perturbations can be interpreted in a quantitative depiction of carbon flow through a biochemical reaction network using isotopic non-stationary (13) C-metabolic flux analysis (INST (13) C-MFA). To evaluate (13) C-metabolic flux maps for Chlamydomonas reinhardtii, an original experimental framework was designed allowing rapid, reliable collection of high-quality isotopomer data against time. It involved (i) a short-time (13) C labeling injection device based on mixing control in a torus-shaped photobioreactor with plug-flow hydrodynamics allowing a sudden step-change in the (13) C proportion in the substrate feed and (ii) a rapid sampling procedure using an automatic fast filtration method coupled to a manual rapid liquid nitrogen quenching step. (13) C-substrate labeling enrichment was controlled through the total dissolved inorganic carbon concentration in the pulsed solution. First results were obtained from steady-state continuous culture measurements allowing the characterization of the kinetics of label incorporation into light-limited growing cells cultivated in a photobioreactor operating at the maximal biomass productivity for an incident photon flux density of 200 µmol m(-2) s(-1). (13)C label incorporation was measured for 21 intracellular metabolites using IC-MS/MS in 58 samples collected across a labeling experiment duration of 7 min. The fastest labeling rate was observed for 2/3-phosphoglycerate with an apparent isotopic stationary state reached after 300 s. The labeling rate was consistent with the optimized mixing time of about 4.9 s inside the reactor and the shortest reliable sampling period assessed at 5 s.

  18. Metabolomic and (13)C-metabolic flux analysis of a xylose-consuming Saccharomyces cerevisiae strain expressing xylose isomerase.

    PubMed

    Wasylenko, Thomas M; Stephanopoulos, Gregory

    2015-03-01

    Over the past two decades, significant progress has been made in the engineering of xylose-consuming Saccharomyces cerevisiae strains for production of lignocellulosic biofuels. However, the ethanol productivities achieved on xylose are still significantly lower than those observed on glucose for reasons that are not well understood. We have undertaken an analysis of central carbon metabolite pool sizes and metabolic fluxes on glucose and on xylose under aerobic and anaerobic conditions in a strain capable of rapid xylose assimilation via xylose isomerase in order to investigate factors that may limit the rate of xylose fermentation. We find that during xylose utilization the flux through the non-oxidative Pentose Phosphate Pathway (PPP) is high but the flux through the oxidative PPP is low, highlighting an advantage of the strain employed in this study. Furthermore, xylose fails to elicit the full carbon catabolite repression response that is characteristic of glucose fermentation in S. cerevisiae. We present indirect evidence that the incomplete activation of the fermentation program on xylose results in a bottleneck in lower glycolysis, leading to inefficient re-oxidation of NADH produced in glycolysis.

  19. Impacts of high β-galactosidase expression on central metabolism of recombinant Pichia pastoris GS115 using glucose as sole carbon source via (13)C metabolic flux analysis.

    PubMed

    Nie, Yongsheng; Huang, Mingzhi; Lu, Junjie; Qian, Jiangchao; Lin, Weilu; Chu, Ju; Zhuang, Yingping; Zhang, Siliang

    2014-10-10

    The yeast Pichia pastoris GS115 is a widely used microbial cell factory for the production of heterologous protein. In order to reveal the impacts of high heterologous protein expression on the central metabolism of Pichia pastoris GS115 using glucose as sole carbon source, we engineered a high β-galactosidase expression strain P. pastoris G1HL and a low expression control strain P. pastoris GHL through controlling the initiation strength of constitutive promoter pGAP. The carbon flux distributions in these two strains were quantified via (13)C metabolic flux analysis. Compared to the control strain, G1HL showed a lower growth rate, a higher flux through glycolysis pathway, a higher flux through pentose phosphate pathway, and a lower flux through by-products secretion pathway. The metabolic flux redistribution in G1HL was thought to compensate the increased redox cofactors and energy demands caused by the high protein expression. Although the fluxes through Krebs cycle in two engineered strains were almost the same, they were significantly lower than those in wild strain. The enhanced expression of β-galactosidase by glutamate supplementation demonstrated the potential of P. pastoris GS115 to catabolize more carbon through the Krebs cycle for even higher protein expression. In conclusion, our work indicates that P. pastoris GS115 can readjusts the central metabolism for higher heterologous protein expression and provides strategies for strain development or process optimization for enhancing production of heterologous protein.

  20. Metabolite channeling and compartmentation in the human cell line AGE1.HN determined by 13C labeling experiments and 13C metabolic flux analysis.

    PubMed

    Niklas, Jens; Sandig, Volker; Heinzle, Elmar

    2011-12-01

    This study focused on analyzing active pathways and the metabolic flux distribution in human neuronal AGE1.HN cells that is a desirable basis for a rational design and optimization of producing cell lines and production processes for biopharmaceuticals. (13)C-labeling experiments and (13)C metabolic flux analysis were conducted using glucose, glutamine, alanine and lactate tracers in parallel experiments. Connections between cytosolic and mitochondrial metabolite pools were verified, e.g., flux from TCA cycle metabolite (13)C to glycolytic metabolites. It was also found that lactate and alanine are produced from the same pyruvate pool and that consumed alanine is mainly directly metabolized and secreted as lactate. Activity of the pentose phosphate pathway was low being around 2.3% of the glucose uptake flux. This might be compensated in AGE1.HN by high mitochondrial malic enzyme flux producing NADPH. Mitochondrial pyruvate transport was almost zero. Instead pyruvate carbons were channeled via oxaloacetate into the TCA cycle which was mainly fed via α-ketoglutarate and oxaloacetate during the investigated phase. The data indicate that further optimization of this cell line should focus on the improved substrate usage which can be accomplished by an improved connectivity between glycolytic and mitochondrial pyruvate pools or by better control of the substrate uptake.

  1. 13C metabolic flux analysis shows that resistin impairs the metabolic response to insulin in L6E9 myotubes

    PubMed Central

    2014-01-01

    Background It has been suggested that the adipokine resistin links obesity and insulin resistance, although how resistin acts on muscle metabolism is controversial. We aimed to quantitatively analyse the effects of resistin on the glucose metabolic flux profile and on insulin response in L6E9 myotubes at the metabolic level using a tracer-based metabolomic approach and our in-house developed software, Isodyn. Results Resistin significantly increased glucose uptake and glycolysis, altering pyruvate utilisation by the cell. In the presence of resistin, insulin only slightly increased glucose uptake and glycolysis, and did not alter the flux profile around pyruvate induced by resistin. Resistin prevented the increase in gene expression in pyruvate dehydrogenase-E1 and the sharp decrease in gene expression in cytosolic phosphoenolpyruvate carboxykinase-1 induced by insulin. Conclusions These data suggest that resistin impairs the metabolic activation of insulin. This impairment cannot be explained by the activity of a single enzyme, but instead due to reorganisation of the whole metabolic flux distribution. PMID:25217974

  2. Comprehensive analysis of glucose and xylose metabolism in Escherichia coli under aerobic and anaerobic conditions by (13)C metabolic flux analysis.

    PubMed

    Gonzalez, Jacqueline E; Long, Christopher P; Antoniewicz, Maciek R

    2017-01-01

    Glucose and xylose are the two most abundant sugars derived from the breakdown of lignocellulosic biomass. While aerobic glucose metabolism is relatively well understood in E. coli, until now there have been only a handful of studies focused on anaerobic glucose metabolism and no (13)C-flux studies on xylose metabolism. In the absence of experimentally validated flux maps, constraint-based approaches such as MOMA and RELATCH cannot be used to guide new metabolic engineering designs. In this work, we have addressed this critical gap in current understanding by performing comprehensive characterizations of glucose and xylose metabolism under aerobic and anaerobic conditions, using recent state-of-the-art techniques in (13)C metabolic flux analysis ((13)C-MFA). Specifically, we quantified precise metabolic fluxes for each condition by performing parallel labeling experiments and analyzing the data through integrated (13)C-MFA using the optimal tracers [1,2-(13)C]glucose, [1,6-(13)C]glucose, [1,2-(13)C]xylose and [5-(13)C]xylose. We also quantified changes in biomass composition and confirmed turnover of macromolecules by applying [U-(13)C]glucose and [U-(13)C]xylose tracers. We demonstrated that under anaerobic growth conditions there is significant turnover of lipids and that a significant portion of CO2 originates from biomass turnover. Using knockout strains, we also demonstrated that β-oxidation is critical for anaerobic growth on xylose. Quantitative analysis of co-factor balances (NADH/FADH2, NADPH, and ATP) for different growth conditions provided new insights regarding the interplay of energy and redox metabolism and the impact on E. coli cell physiology.

  3. Exo-MFA - A 13C metabolic flux analysis framework to dissect tumor microenvironment-secreted exosome contributions towards cancer cell metabolism.

    PubMed

    Achreja, Abhinav; Zhao, Hongyun; Yang, Lifeng; Yun, Tae Hyun; Marini, Juan; Nagrath, Deepak

    2017-01-11

    Dissecting the pleiotropic roles of tumor micro-environment (TME) on cancer progression has been brought to the foreground of research on cancer pathology. Extracellular vesicles such as exosomes, transport proteins, lipids, and nucleic acids, to mediate intercellular communication between TME components and have emerged as candidates for anti-cancer therapy. We previously reported that cancer-associated fibroblast (CAF) derived exosomes (CDEs) contain metabolites in their cargo that are utilized by cancer cells for central carbon metabolism and promote cancer growth. However, the metabolic fluxes involved in donor cells towards packaging of metabolites in extracellular vesicles and exosome-mediated metabolite flux upregulation in recipient cells are still not known. Here, we have developed a novel empirical and computational technique, exosome-mediated metabolic flux analysis (Exo-MFA) to quantify flow of cargo from source cells to recipient cells via vesicular transport. Our algorithm, which is based on (13)C metabolic flux analysis, successfully predicts packaging fluxes to metabolite cargo in CAFs, dynamic changes in rate of exosome internalization by cancer cells, and flux of cargo release over time. We find that cancer cells internalize exosomes rapidly leading to depletion of extracellular exosomes within 24h. However, metabolite cargo significantly alters intracellular metabolism over the course of 24h by regulating glycolysis pathway fluxes via lactate supply. Furthermore, it can supply up to 35% of the TCA cycle fluxes by providing TCA intermediates and glutamine. Our algorithm will help gain insight into (i) metabolic interactions in multicellular systems (ii) biogenesis of extracellular vesicles and their differential packaging of cargo under changing environments, and (iii) regulation of cancer cell metabolism by its microenvironment.

  4. Non-stationary 13C metabolic flux analysis of Chinese hamster ovary cells in batch culture using extracellular labeling highlights metabolic reversibility and compartmentation

    PubMed Central

    2014-01-01

    Background Mapping the intracellular fluxes for established mammalian cell lines becomes increasingly important for scientific and economic reasons. However, this is being hampered by the high complexity of metabolic networks, particularly concerning compartmentation. Results Intracellular fluxes of the CHO-K1 cell line central carbon metabolism were successfully determined for a complex network using non-stationary 13C metabolic flux analysis. Mass isotopomers of extracellular metabolites were determined using [U-13C6] glucose as labeled substrate. Metabolic compartmentation and extracellular transport reversibility proved essential to successfully reproduce the dynamics of the labeling patterns. Alanine and pyruvate reversibility changed dynamically even if their net production fluxes remained constant. Cataplerotic fluxes of cytosolic phosphoenolpyruvate carboxykinase and mitochondrial malic enzyme and pyruvate carboxylase were successfully determined. Glycolytic pyruvate channeling to lactate was modeled by including a separate pyruvate pool. In the exponential growth phase, alanine, glycine and glutamate were excreted, and glutamine, aspartate, asparagine and serine were taken up; however, all these amino acids except asparagine were exchanged reversibly with the media. High fluxes were determined in the pentose phosphate pathway and the TCA cycle. The latter was fueled mainly by glucose but also by amino acid catabolism. Conclusions The CHO-K1 central metabolism in controlled batch culture proves to be robust. It has the main purpose to ensure fast growth on a mixture of substrates and also to mitigate oxidative stress. It achieves this by using compartmentation to control NADPH and NADH availability and by simultaneous synthesis and catabolism of amino acids. PMID:24773761

  5. Synergy between 13C-metabolic flux analysis and flux balance analysis for understanding metabolic adaption to anaerobiosis in e. coli

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genome-based Flux Balance Analysis (FBA, constraints based flux analysis) and steady state isotopic-labeling-based Metabolic Flux Analysis (MFA) are complimentary approaches to predicting and measuring the operation and regulation of metabolic networks. Here a genome-derived model of E. coli metabol...

  6. (13)C metabolic flux analysis in neurons utilizing a model that accounts for hexose phosphate recycling within the pentose phosphate pathway.

    PubMed

    Gebril, Hoda M; Avula, Bharathi; Wang, Yan-Hong; Khan, Ikhlas A; Jekabsons, Mika B

    2016-02-01

    Glycolysis, mitochondrial substrate oxidation, and the pentose phosphate pathway (PPP) are critical for neuronal bioenergetics and oxidation-reduction homeostasis, but quantitating their fluxes remains challenging, especially when processes such as hexose phosphate (i.e., glucose/fructose-6-phosphate) recycling in the PPP are considered. A hexose phosphate recycling model was developed which exploited the rates of glucose consumption, lactate production, and mitochondrial respiration to infer fluxes through the major glucose consuming pathways of adherent cerebellar granule neurons by replicating [(13)C]lactate labeling from metabolism of [1,2-(13)C2]glucose. Flux calculations were predicated on a steady-state system with reactions having known stoichiometries and carbon atom transitions. Non-oxidative PPP activity and consequent hexose phosphate recycling, as well as pyruvate production by cytoplasmic malic enzyme, were optimized by the model and found to account for 28 ± 2% and 7.7 ± 0.2% of hexose phosphate and pyruvate labeling, respectively. From the resulting fluxes, 52 ± 6% of glucose was metabolized by glycolysis, compared to 19 ± 2% by the combined oxidative/non-oxidative pentose cycle that allows for hexose phosphate recycling, and 29 ± 8% by the combined oxidative PPP/de novo nucleotide synthesis reactions. By extension, 62 ± 6% of glucose was converted to pyruvate, the metabolism of which resulted in 16 ± 1% of glucose oxidized by mitochondria and 46 ± 6% exported as lactate. The results indicate a surprisingly high proportion of glucose utilized by the pentose cycle and the reactions synthesizing nucleotides, and exported as lactate. While the in vitro conditions to which the neurons were exposed (high glucose, no lactate or other exogenous substrates) limit extrapolating these results to the in vivo state, the approach provides a means of assessing a number of metabolic fluxes within the context of hexose phosphate recycling in the PPP from a

  7. GC-MS/MS survey of collision-induced dissociation of tert-butyldimethylsilyl-derivatized amino acids and its application to (13)C-metabolic flux analysis of Escherichia coli central metabolism.

    PubMed

    Okahashi, Nobuyuki; Kawana, Shuichi; Iida, Junko; Shimizu, Hiroshi; Matsuda, Fumio

    2016-09-01

    Stable isotope labeling experiments using mass spectrometry have been employed to investigate carbon flow levels (metabolic flux) in mammalian, plant, and microbial cells. To achieve a more precise (13)C-metabolic flux analysis ((13)C-MFA), novel fragmentations of tert-butyldimethylsilyl (TBDMS)-amino acids were investigated by gas chromatography-tandem mass spectrometry (GC-MS/MS). The product ion scan analyses of 15 TBDMS-amino acids revealed 24 novel fragment ions. The amino acid-derived carbons included in the five fragment ions were identified by the analyses of (13)C-labeled authentic standards. The identification of the fragment ion at m/z 170 indicated that the isotopic abundance of S-methyl carbon in methionine could be determined from the cleavage of C5 in the precursor of [M-159](+) (m/z 218). It was also confirmed that the precision of (13)C-MFA in Escherichia coli central carbon metabolism could be improved by introducing (13)C-labeling data derived from novel fragmentations. Graphical Abstract Novel collision-induced dissociation fragmentations of tert-butyldimethylsilyl amino acids were investigated and identified by GC-MS/MS.

  8. Gas Chromatography-Quadrupole Time-of-Flight Mass Spectrometry-Based Determination of Isotopologue and Tandem Mass Isotopomer Fractions of Primary Metabolites for (13)C-Metabolic Flux Analysis.

    PubMed

    Mairinger, Teresa; Steiger, Matthias; Nocon, Justyna; Mattanovich, Diethard; Koellensperger, Gunda; Hann, Stephan

    2015-12-01

    For the first time an analytical work flow based on accurate mass gas chromatography-quadrupole time-of-flight mass spectrometry (GC-QTOFMS) with chemical ionization for analysis providing a comprehensive picture of (13)C distribution along the primary metabolism is elaborated. The method provides a powerful new toolbox for (13)C-based metabolic flux analysis, which is an emerging strategy in metabolic engineering. In this field, stable isotope tracer experiments based on, for example, (13)C are central for providing characteristic patterns of labeled metabolites, which in turn give insights into the regulation of metabolic pathway kinetics. The new method enables the analysis of isotopologue fractions of 42 free intracellular metabolites within biotechnological samples, while tandem mass isotopomer information is also accessible for a large number of analytes. Hence, the method outperforms previous approaches in terms of metabolite coverage, while also providing rich isotopomer information for a significant number of key metabolites. Moreover, the established work flow includes novel evaluation routines correcting for isotope interference of naturally distributed elements, which is crucial following derivatization of metabolites. Method validation in terms of trueness, precision, and limits of detection was performed, showing excellent analytical figures of merit with an overall maximum bias of 5.8%, very high precision for isotopologue and tandem mass isotopomer fractions representing >10% of total abundance, and absolute limits of detection in the femtomole range. The suitability of the developed method is demonstrated on a flux experiment of Pichia pastoris employing two different tracers, i.e., 1,6(13)C2-glucose and uniformly labeled (13)C-glucose.

  9. 13C-metabolic flux ratio and novel carbon path analyses confirmed that Trichoderma reesei uses primarily the respirative pathway also on the preferred carbon source glucose

    PubMed Central

    Jouhten, Paula; Pitkänen, Esa; Pakula, Tiina; Saloheimo, Markku; Penttilä, Merja; Maaheimo, Hannu

    2009-01-01

    Background The filamentous fungus Trichoderma reesei is an important host organism for industrial enzyme production. It is adapted to nutrient poor environments where it is capable of producing large amounts of hydrolytic enzymes. In its natural environment T. reesei is expected to benefit from high energy yield from utilization of respirative metabolic pathway. However, T. reesei lacks metabolic pathway reconstructions and the utilization of the respirative pathway has not been investigated on the level of in vivo fluxes. Results The biosynthetic pathways of amino acids in T. reesei supported by genome-level evidence were reconstructed with computational carbon path analysis. The pathway reconstructions were a prerequisite for analysis of in vivo fluxes. The distribution of in vivo fluxes in both wild type strain and cre1, a key regulator of carbon catabolite repression, deletion strain were quantitatively studied by performing 13C-labeling on both repressive carbon source glucose and non-repressive carbon source sorbitol. In addition, the 13C-labeling on sorbitol was performed both in the presence and absence of sophorose that induces the expression of cellulase genes. Carbon path analyses and the 13C-labeling patterns of proteinogenic amino acids indicated high similarity between biosynthetic pathways of amino acids in T. reesei and yeast Saccharomyces cerevisiae. In contrast to S. cerevisiae, however, mitochondrial rather than cytosolic biosynthesis of Asp was observed under all studied conditions. The relative anaplerotic flux to the TCA cycle was low and thus characteristic to respiratory metabolism in both strains and independent of the carbon source. Only minor differences were observed in the flux distributions of the wild type and cre1 deletion strain. Furthermore, the induction of the hydrolytic gene expression did not show altered flux distributions and did not affect the relative amino acid requirements or relative anabolic and respirative activities

  10. Metabolic flux analysis using 13C peptide label measurements

    Technology Transfer Automated Retrieval System (TEKTRAN)

    13C metabolic flux analysis (MFA) has become the experimental method of choice to investigate cellular metabolism. MFA has established flux maps of central metabolism for dozens of microbes, cell cultures, and plant seeds. Steady-state MFA utilizes isotopic labeling measurements of amino acids obtai...

  11. Hyperpolarized 13C Metabolic MRI of the Human Heart

    PubMed Central

    Lau, Justin Y.C.; Chen, Albert P.; Geraghty, Benjamin J.; Perks, William J.; Roifman, Idan; Wright, Graham A.; Connelly, Kim A.

    2016-01-01

    Rationale: Altered cardiac energetics is known to play an important role in the progression toward heart failure. A noninvasive method for imaging metabolic markers that could be used in longitudinal studies would be useful for understanding therapeutic approaches that target metabolism. Objective: To demonstrate the first hyperpolarized 13C metabolic magnetic resonance imaging of the human heart. Methods and Results: Four healthy subjects underwent conventional proton cardiac magnetic resonance imaging followed by 13C imaging and spectroscopic acquisition immediately after intravenous administration of a 0.1 mmol/kg dose of hyperpolarized [1-13C]pyruvate. All subjects tolerated the procedure well with no adverse effects reported ≤1 month post procedure. The [1-13C]pyruvate signal appeared within the chambers but not within the muscle. Imaging of the downstream metabolites showed 13C-bicarbonate signal mainly confined to the left ventricular myocardium, whereas the [1-13C]lactate signal appeared both within the chambers and in the myocardium. The mean 13C image signal:noise ratio was 115 for [1-13C]pyruvate, 56 for 13C-bicarbonate, and 53 for [1-13C]lactate. Conclusions: These results represent the first 13C images of the human heart. The appearance of 13C-bicarbonate signal after administration of hyperpolarized [1-13C]pyruvate was readily detected in this healthy cohort (n=4). This shows that assessment of pyruvate metabolism in vivo in humans is feasible using current technology. Clinical Trial Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT02648009. PMID:27635086

  12. Methods and advances in metabolic flux analysis: a mini-review.

    PubMed

    Antoniewicz, Maciek R

    2015-03-01

    Metabolic flux analysis (MFA) is one of the pillars of metabolic engineering. Over the past three decades, it has been widely used to quantify intracellular metabolic fluxes in both native (wild type) and engineered biological systems. Through MFA, changes in metabolic pathway fluxes are quantified that result from genetic and/or environmental interventions. This information, in turn, provides insights into the regulation of metabolic pathways and may suggest new targets for further metabolic engineering of the strains. In this mini-review, we discuss and classify the various methods of MFA that have been developed, which include stoichiometric MFA, (13)C metabolic flux analysis, isotopic non-stationary (13)C metabolic flux analysis, dynamic metabolic flux analysis, and (13)C dynamic metabolic flux analysis. For each method, we discuss key advantages and limitations and conclude by highlighting important recent advances in flux analysis approaches.

  13. Simultaneous imaging of 13C metabolism and 1H structure: technical considerations and potential applications.

    PubMed

    Gordon, Jeremy W; Fain, Sean B; Niles, David J; Ludwig, Kai D; Johnson, Kevin M; Peterson, Eric T

    2015-05-01

    Real-time imaging of (13)C metabolism in vivo has been enabled by recent advances in hyperpolarization. As a result of the inherently low natural abundance of endogenous (13)C nuclei, hyperpolarized (13)C images lack structural information that could be used to aid in motion detection and anatomical registration. Motion before or during the (13)C acquisition can therefore result in artifacts and misregistration that may obscure measures of metabolism. In this work, we demonstrate a method to simultaneously image both (1)H and (13)C nuclei using a dual-nucleus spectral-spatial radiofrequency excitation and a fully coincident readout for rapid multinuclear spectroscopic imaging. With the appropriate multinuclear hardware, and the means to simultaneously excite and receive on both channels, this technique is straightforward to implement requiring little to no increase in scan time. Phantom and in vivo experiments were performed with both Cartesian and spiral trajectories to validate and illustrate the utility of simultaneous acquisitions. Motion compensation of dynamic metabolic measurements acquired during free breathing was demonstrated using motion tracking derived from (1)H data. Simultaneous multinuclear imaging provides structural (1)H and metabolic (13)C images that are correlated both spatially and temporally, and are therefore amenable to joint (1)H and (13)C analysis and correction of structure-function images.

  14. Flux analysis in plant metabolic networks: increasing throughput and coverage.

    PubMed

    Junker, Björn H

    2014-04-01

    Quantitative information about metabolic networks has been mainly obtained at the level of metabolite contents, transcript abundance, and enzyme activities. However, the active process of metabolism is represented by the flow of matter through the pathways. These metabolic fluxes can be predicted by Flux Balance Analysis or determined experimentally by (13)C-Metabolic Flux Analysis. These relatively complicated and time-consuming methods have recently seen significant improvements at the level of coverage and throughput. Metabolic models have developed from single cell models into whole-organism dynamic models. Advances in lab automation and data handling have significantly increased the throughput of flux measurements. This review summarizes advances to increase coverage and throughput of metabolic flux analysis in plants.

  15. Flux analysis and metabolomics for systematic metabolic engineering of microorganisms.

    PubMed

    Toya, Yoshihiro; Shimizu, Hiroshi

    2013-11-01

    Rational engineering of metabolism is important for bio-production using microorganisms. Metabolic design based on in silico simulations and experimental validation of the metabolic state in the engineered strain helps in accomplishing systematic metabolic engineering. Flux balance analysis (FBA) is a method for the prediction of metabolic phenotype, and many applications have been developed using FBA to design metabolic networks. Elementary mode analysis (EMA) and ensemble modeling techniques are also useful tools for in silico strain design. The metabolome and flux distribution of the metabolic pathways enable us to evaluate the metabolic state and provide useful clues to improve target productivity. Here, we reviewed several computational applications for metabolic engineering by using genome-scale metabolic models of microorganisms. We also discussed the recent progress made in the field of metabolomics and (13)C-metabolic flux analysis techniques, and reviewed these applications pertaining to bio-production development. Because these in silico or experimental approaches have their respective advantages and disadvantages, the combined usage of these methods is complementary and effective for metabolic engineering.

  16. Metabolic flux analysis of Escherichia coli knockouts: lessons from the Keio collection and future outlook.

    PubMed

    Long, Christopher P; Antoniewicz, Maciek R

    2014-08-01

    Cellular metabolic and regulatory systems are of fundamental interest to biologists and engineers. Incomplete understanding of these complex systems remains an obstacle to progress in biotechnology and metabolic engineering. An established method for obtaining new information on network structure, regulation and dynamics is to study the cellular system following a perturbation such as a genetic knockout. The Keio collection of all viable Escherichia coli single-gene knockouts is facilitating a systematic investigation of the regulation and metabolism of E. coli. Of all omics measurements available, the metabolic flux profile (the fluxome) provides the most direct and relevant representation of the cellular phenotype. Recent advances in (13)C-metabolic flux analysis are now permitting highly precise and accurate flux measurements for investigating cellular systems and guiding metabolic engineering efforts.

  17. Carbon 13-Metabolic Flux Analysis derived constraint-based metabolic modelling of Clostridium acetobutylicum in stressed chemostat conditions.

    PubMed

    Wallenius, Janne; Maaheimo, Hannu; Eerikäinen, Tero

    2016-11-01

    The metabolism of butanol producing bacteria Clostridium acetobutylicum was studied in chemostat with glucose limited conditions, butanol stimulus, and as a reference cultivation. COnstraint-Based Reconstruction and Analysis (COBRA) was applied using additional constraints from (13)C Metabolic Flux Analysis ((13)C-MFA) and experimental measurement results. A model consisting of 451 metabolites and 604 reactions was utilized in flux balance analysis (FBA). The stringency of the flux spaces considering different optimization objectives, i.e. growth rate maximization, ATP maintenance, and NADH/NADPH formation, for flux variance analysis (FVA) was studied in the different modelled conditions. Also a previously uncharacterized exopolysaccharide (EPS) produced by C. acetobutylicum was characterized on monosaccharide level. The major monosaccharide components of the EPS were 40n-% rhamnose, 34n-% glucose, 13n-% mannose, 10n-% galactose, and 2n-% arabinose. The EPS was studied to have butanol adsorbing property, 70(butanol)mg(EPS)g(-1) at 37°C.

  18. Radiative Flux Analysis

    DOE Data Explorer

    Long, Chuck [NOAA

    2008-05-14

    The Radiative Flux Analysis is a technique for using surface broadband radiation measurements for detecting periods of clear (i.e. cloudless) skies, and using the detected clear-sky data to fit functions which are then used to produce continuous clear-sky estimates. The clear-sky estimates and measurements are then used in various ways to infer cloud macrophysical properties.

  19. Metabolic flux analysis of Escherichia coli MG1655 under octanoic acid (C8) stress.

    PubMed

    Fu, Yanfen; Yoon, Jong Moon; Jarboe, Laura; Shanks, Jacqueline V

    2015-05-01

    Systems metabolic engineering has made the renewable production of industrial chemicals a feasible alternative to modern operations. One major example of a renewable process is the production of carboxylic acids, such as octanoic acid (C8), from Escherichia coli, engineered to express thioesterase enzymes. C8, however, is toxic to E. coli above a certain concentration, which limits the final titer. (13)C metabolic flux analysis of E. coli was performed for both C8 stress and control conditions using NMR2Flux with isotopomer balancing. A mixture of labeled and unlabeled glucose was used as the sole carbon source for bacterial growth for (13)C flux analysis. By comparing the metabolic flux maps of the control condition and C8 stress condition, pathways that were altered under the stress condition were identified. C8 stress was found to reduce carbon flux in several pathways: the tricarboxylic acid (TCA) cycle, the CO2 production, and the pyruvate dehydrogenase pathway. Meanwhile, a few pathways became more active: the pyruvate oxidative pathway, and the extracellular acetate production. These results were statistically significant for three biological replicates between the control condition and C8 stress. As a working hypothesis, the following causes are proposed to be the main causes for growth inhibition and flux alteration for a cell under stress: membrane disruption, low activity of electron transport chain, and the activation of the pyruvate dehydrogenase regulator (PdhR).

  20. OpenMebius: an open source software for isotopically nonstationary 13C-based metabolic flux analysis.

    PubMed

    Kajihata, Shuichi; Furusawa, Chikara; Matsuda, Fumio; Shimizu, Hiroshi

    2014-01-01

    The in vivo measurement of metabolic flux by (13)C-based metabolic flux analysis ((13)C-MFA) provides valuable information regarding cell physiology. Bioinformatics tools have been developed to estimate metabolic flux distributions from the results of tracer isotopic labeling experiments using a (13)C-labeled carbon source. Metabolic flux is determined by nonlinear fitting of a metabolic model to the isotopic labeling enrichment of intracellular metabolites measured by mass spectrometry. Whereas (13)C-MFA is conventionally performed under isotopically constant conditions, isotopically nonstationary (13)C metabolic flux analysis (INST-(13)C-MFA) has recently been developed for flux analysis of cells with photosynthetic activity and cells at a quasi-steady metabolic state (e.g., primary cells or microorganisms under stationary phase). Here, the development of a novel open source software for INST-(13)C-MFA on the Windows platform is reported. OpenMebius (Open source software for Metabolic flux analysis) provides the function of autogenerating metabolic models for simulating isotopic labeling enrichment from a user-defined configuration worksheet. Analysis using simulated data demonstrated the applicability of OpenMebius for INST-(13)C-MFA. Confidence intervals determined by INST-(13)C-MFA were less than those determined by conventional methods, indicating the potential of INST-(13)C-MFA for precise metabolic flux analysis. OpenMebius is the open source software for the general application of INST-(13)C-MFA.

  1. Comparative metabolic flux analysis of an Ashbya gossypii wild type strain and a high riboflavin-producing mutant strain.

    PubMed

    Jeong, Bo-Young; Wittmann, Christoph; Kato, Tatsuya; Park, Enoch Y

    2015-01-01

    In the present study, we analyzed the central metabolic pathway of an Ashbya gossypii wild type strain and a riboflavin over-producing mutant strain developed in a previous study in order to characterize the riboflavin over-production pathway. (13)C-Metabolic flux analysis ((13)C-MFA) was carried out in both strains, and the resulting data were fit to a steady-state flux isotopomer model using OpenFLUX. Flux to pentose-5-phosphate (P5P) via the pentose phosphate pathway (PPP) was 9% higher in the mutant strain compared to the wild type strain. The flux from purine synthesis to riboflavin in the mutant strain was 1.6%, while that of the wild type strain was only 0.1%, a 16-fold difference. In addition, the flux from the cytoplasmic pyruvate pool to the extracellular metabolites, pyruvate, lactate, and alanine, was 2-fold higher in the mutant strain compared to the wild type strain. This result demonstrates that increased guanosine triphosphate (GTP) flux through the PPP and purine synthesis pathway (PSP) increased riboflavin production in the mutant strain. The present study provides the first insight into metabolic flux through the central carbon pathway in A. gossypii and sets the foundation for development of a quantitative and functional model of the A. gossypii metabolic network.

  2. Metabolic flux analysis of Cyanothece sp. ATCC 51142 under mixotrophic conditions.

    PubMed

    Alagesan, Swathi; Gaudana, Sandeep B; Sinha, Avinash; Wangikar, Pramod P

    2013-11-01

    Cyanobacteria are a group of photosynthetic prokaryotes capable of utilizing solar energy to fix atmospheric carbon dioxide to biomass. Despite several "proof of principle" studies, low product yield is an impediment in commercialization of cyanobacteria-derived biofuels. Estimation of intracellular reaction rates by (13)C metabolic flux analysis ((13)C-MFA) would be a step toward enhancing biofuel yield via metabolic engineering. We report (13)C-MFA for Cyanothece sp. ATCC 51142, a unicellular nitrogen-fixing cyanobacterium, known for enhanced hydrogen yield under mixotrophic conditions. Rates of reactions in the central carbon metabolism under nitrogen-fixing and -non-fixing conditions were estimated by monitoring the competitive incorporation of (12)C and (13)C from unlabeled CO2 and uniformly labeled glycerol, respectively, into terminal metabolites such as amino acids. The observed labeling patterns suggest mixotrophic growth under both the conditions, with a larger fraction of unlabeled carbon in nitrate-sufficient cultures asserting a greater contribution of carbon fixation by photosynthesis and an anaplerotic pathway. Indeed, flux analysis complements the higher growth observed under nitrate-sufficient conditions. On the other hand, the flux through the oxidative pentose phosphate pathway and tricarboxylic acid cycle was greater in nitrate-deficient conditions, possibly to supply the precursors and reducing equivalents needed for nitrogen fixation. In addition, an enhanced flux through fructose-6-phosphate phosphoketolase possibly suggests the organism's preferred mode under nitrogen-fixing conditions. The (13)C-MFA results complement the reported predictions by flux balance analysis and provide quantitative insight into the organism's distinct metabolic features under nitrogen-fixing and -non-fixing conditions.

  3. Metabolic flux analysis of CHO cells at growth and non-growth phases using isotopic tracers and mass spectrometry.

    PubMed

    Ahn, Woo Suk; Antoniewicz, Maciek R

    2011-09-01

    Chinese hamster ovary (CHO) cells are the main platform for production of biotherapeutics in the biopharmaceutical industry. However, relatively little is known about the metabolism of CHO cells in cell culture. In this work, metabolism of CHO cells was studied at the growth phase and early stationary phase using isotopic tracers and mass spectrometry. CHO cells were grown in fed-batch culture over a period of six days. On days 2 and 4, [1,2-(13)C] glucose was introduced and the labeling of intracellular metabolites was measured by gas chromatography-mass spectrometry (GC-MS) at 6, 12 and 24h following the introduction of tracer. Intracellular metabolic fluxes were quantified from measured extracellular rates and (13)C-labeling dynamics of intracellular metabolites using non-stationary (13)C-metabolic flux analysis ((13)C-MFA). The flux results revealed significant rewiring of intracellular metabolic fluxes in the transition from growth to non-growth, including changes in energy metabolism, redox metabolism, oxidative pentose phosphate pathway and anaplerosis. At the exponential phase, CHO cell metabolism was characterized by a high flux of glycolysis from glucose to lactate, anaplerosis from pyruvate to oxaloacetate and from glutamate to α-ketoglutarate, and cataplerosis though malic enzyme. At the stationary phase, the flux map was characterized by a reduced flux of glycolysis, net lactate uptake, oxidative pentose phosphate pathway flux, and reduced rate of anaplerosis. The fluxes of pyruvate dehydrogenase and TCA cycle were similar at the exponential and stationary phase. The results presented here provide a solid foundation for future studies of CHO cell metabolism for applications such as cell line development and medium optimization for high-titer production of recombinant proteins.

  4. Experimental flux measurements on a network scale

    SciTech Connect

    Schwender, J.

    2011-10-11

    Metabolic flux is a fundamental property of living organisms. In recent years, methods for measuring metabolic flux in plants on a network scale have evolved further. One major challenge in studying flux in plants is the complexity of the plant's metabolism. In particular, in the presence of parallel pathways in multiple cellular compartments, the core of plant central metabolism constitutes a complex network. Hence, a common problem with the reliability of the contemporary results of {sup 13}C-Metabolic Flux Analysis in plants is the substantial reduction in complexity that must be included in the simulated networks; this omission partly is due to limitations in computational simulations. Here, I discuss recent emerging strategies that will better address these shortcomings.

  5. Genome-based metabolic mapping and 13C flux analysis reveal systematic properties of an oleaginous microalga Chlorella protothecoides.

    PubMed

    Wu, Chao; Xiong, Wei; Dai, Junbiao; Wu, Qingyu

    2015-02-01

    Integrated and genome-based flux balance analysis, metabolomics, and (13)C-label profiling of phototrophic and heterotrophic metabolism in Chlorella protothecoides, an oleaginous green alga for biofuel. The green alga Chlorella protothecoides, capable of autotrophic and heterotrophic growth with rapid lipid synthesis, is a promising candidate for biofuel production. Based on the newly available genome knowledge of the alga, we reconstructed the compartmentalized metabolic network consisting of 272 metabolic reactions, 270 enzymes, and 461 encoding genes and simulated the growth in different cultivation conditions with flux balance analysis. Phenotype-phase plane analysis shows conditions achieving theoretical maximum of the biomass and corresponding fatty acid-producing rate for phototrophic cells (the ratio of photon uptake rate to CO2 uptake rate equals 8.4) and heterotrophic ones (the glucose uptake rate to O2 consumption rate reaches 2.4), respectively. Isotope-assisted liquid chromatography-mass spectrometry/mass spectrometry reveals higher metabolite concentrations in the glycolytic pathway and the tricarboxylic acid cycle in heterotrophic cells compared with autotrophic cells. We also observed enhanced levels of ATP, nicotinamide adenine dinucleotide (phosphate), reduced, acetyl-Coenzyme A, and malonyl-Coenzyme A in heterotrophic cells consistently, consistent with a strong activity of lipid synthesis. To profile the flux map in experimental conditions, we applied nonstationary (13)C metabolic flux analysis as a complementing strategy to flux balance analysis. The result reveals negligible photorespiratory fluxes and a metabolically low active tricarboxylic acid cycle in phototrophic C. protothecoides. In comparison, high throughput of amphibolic reactions and the tricarboxylic acid cycle with no glyoxylate shunt activities were measured for heterotrophic cells. Taken together, the metabolic network modeling assisted by experimental metabolomics and (13)C

  6. Genome-Based Metabolic Mapping and 13C Flux Analysis Reveal Systematic Properties of an Oleaginous Microalga Chlorella protothecoides

    DOE PAGES

    Wu, Chao; Xiong, Wei; Dai, Junbiao; ...

    2014-12-15

    We report that integrated and genome-based flux balance analysis, metabolomics, and 13C-label profiling of phototrophic and heterotrophic metabolism in Chlorella protothecoides, an oleaginous green alga for biofuel. The green alga Chlorella protothecoides, capable of autotrophic and heterotrophic growth with rapid lipid synthesis, is a promising candidate for biofuel production. Based on the newly available genome knowledge of the alga, we reconstructed the compartmentalized metabolic network consisting of 272 metabolic reactions, 270 enzymes, and 461 encoding genes and simulated the growth in different cultivation conditions with flux balance analysis. Phenotype-phase plane analysis shows conditions achieving theoretical maximum of the biomass andmore » corresponding fatty acid-producing rate for phototrophic cells (the ratio of photon uptake rate to CO2 uptake rate equals 8.4) and heterotrophic ones (the glucose uptake rate to O2 consumption rate reaches 2.4), respectively. Isotope-assisted liquid chromatography-mass spectrometry/mass spectrometry reveals higher metabolite concentrations in the glycolytic pathway and the tricarboxylic acid cycle in heterotrophic cells compared with autotrophic cells. We also observed enhanced levels of ATP, nicotinamide adenine dinucleotide (phosphate), reduced, acetyl-Coenzyme A, and malonyl-Coenzyme A in heterotrophic cells consistently, consistent with a strong activity of lipid synthesis. To profile the flux map in experimental conditions, we applied nonstationary 13C metabolic flux analysis as a complementing strategy to flux balance analysis. We found that the result reveals negligible photorespiratory fluxes and a metabolically low active tricarboxylic acid cycle in phototrophic C. protothecoides. In comparison, high throughput of amphibolic reactions and the tricarboxylic acid cycle with no glyoxylate shunt activities were measured for heterotrophic cells. Lastly, taken together, the metabolic network modeling assisted

  7. Analysis of metabolic flux phenotypes for two Arabidopsis mutants with severe impairment in seed storage lipid synthesis

    SciTech Connect

    Lonien, J.; Schwender, J.

    2009-11-01

    Major storage reserves of Arabidopsis (Arabidopsis thaliana) seeds are triacylglycerols (seed oils) and proteins. Seed oil content is severely reduced for the regulatory mutant wrinkled1 (wri1-1; At3g54320) and for a double mutant in two isoforms of plastidic pyruvate kinase (pkp{beta}{sub 1}pkp{alpha}; At5g52920 and At3g22960). Both already biochemically well-characterized mutants were now studied by {sup 13}C metabolic flux analysis of cultured developing embryos based on comparison with their respective genetic wild-type backgrounds. For both mutations, in seeds as well as in cultured embryos, the oil fraction was strongly reduced while the fractions of proteins and free metabolites increased. Flux analysis in cultured embryos revealed changes in nutrient uptakes and fluxes into biomass as well as an increase in tricarboxylic acid cycle activity for both mutations. While in both wild types plastidic pyruvate kinase (PK{sub p}) provides most of the pyruvate for plastidic fatty acid synthesis, the flux through PK{sub p} is reduced in pkp{beta}{sub 1}pkp{alpha} by 43% of the wild-type value. In wri1-1, PK{sub p} flux is even more reduced (by 82%), although the genes PKp{beta}{sub 1} and PKp{alpha} are still expressed. Along a common paradigm of metabolic control theory, it is hypothesized that a large reduction in PK{sub p} enzyme activity in pkp{beta}{sub 1}pkp{alpha} has less effect on PK{sub p} flux than multiple smaller reductions in glycolytic enzymes in wri1-1. In addition, only in the wri1-1 mutant is the large reduction in PK{sub p} flux compensated in part by an increased import of cytosolic pyruvate and by plastidic malic enzyme. No such limited compensatory bypass could be observed in pkp{beta}{sub 1}pkp{alpha}.

  8. Strategies for investigating the plant metabolic network with steady-state metabolic flux analysis: lessons from an Arabidopsis cell culture and other systems.

    PubMed

    Kruger, N J; Masakapalli, S K; Ratcliffe, R G

    2012-03-01

    Steady-state (13)C metabolic flux analysis (MFA) is currently the experimental method of choice for generating flux maps of the compartmented network of primary metabolism in heterotrophic and mixotrophic plant tissues. While statistically robust protocols for the application of steady-state MFA to plant tissues have been developed by several research groups, the implementation of the method is still far from routine. The effort required to produce a flux map is more than justified by the information that it contains about the metabolic phenotype of the system, but it remains the case that steady-state MFA is both analytically and computationally demanding. This article provides an overview of principles that underpin the implementation of steady-state MFA, focusing on the definition of the metabolic network responsible for redistribution of the label, experimental considerations relating to data collection, the modelling process that allows a set of metabolic fluxes to be deduced from the labelling data, and the interpretation of flux maps. The article draws on published studies of Arabidopsis cell cultures and other systems, including developing oilseeds, with the aim of providing practical guidance and strategies for handling the issues that arise when applying steady-state MFA to the complex metabolic networks encountered in plants.

  9. Constrained Allocation Flux Balance Analysis

    PubMed Central

    Mori, Matteo; Hwa, Terence; Martin, Olivier C.

    2016-01-01

    New experimental results on bacterial growth inspire a novel top-down approach to study cell metabolism, combining mass balance and proteomic constraints to extend and complement Flux Balance Analysis. We introduce here Constrained Allocation Flux Balance Analysis, CAFBA, in which the biosynthetic costs associated to growth are accounted for in an effective way through a single additional genome-wide constraint. Its roots lie in the experimentally observed pattern of proteome allocation for metabolic functions, allowing to bridge regulation and metabolism in a transparent way under the principle of growth-rate maximization. We provide a simple method to solve CAFBA efficiently and propose an “ensemble averaging” procedure to account for unknown protein costs. Applying this approach to modeling E. coli metabolism, we find that, as the growth rate increases, CAFBA solutions cross over from respiratory, growth-yield maximizing states (preferred at slow growth) to fermentative states with carbon overflow (preferred at fast growth). In addition, CAFBA allows for quantitatively accurate predictions on the rate of acetate excretion and growth yield based on only 3 parameters determined by empirical growth laws. PMID:27355325

  10. Multi-objective experimental design for (13)C-based metabolic flux analysis.

    PubMed

    Bouvin, Jeroen; Cajot, Simon; D'Huys, Pieter-Jan; Ampofo-Asiama, Jerry; Anné, Jozef; Van Impe, Jan; Geeraerd, Annemie; Bernaerts, Kristel

    2015-10-01

    (13)C-based metabolic flux analysis is an excellent technique to resolve fluxes in the central carbon metabolism but costs can be significant when using specialized tracers. This work presents a framework for cost-effective design of (13)C-tracer experiments, illustrated on two different networks. Linear and non-linear optimal input mixtures are computed for networks for Streptomyces lividans and a carcinoma cell line. If only glucose tracers are considered as labeled substrate for a carcinoma cell line or S. lividans, the best parameter estimation accuracy is obtained by mixtures containing high amounts of 1,2-(13)C2 glucose combined with uniformly labeled glucose. Experimental designs are evaluated based on a linear (D-criterion) and non-linear approach (S-criterion). Both approaches generate almost the same input mixture, however, the linear approach is favored due to its low computational effort. The high amount of 1,2-(13)C2 glucose in the optimal designs coincides with a high experimental cost, which is further enhanced when labeling is introduced in glutamine and aspartate tracers. Multi-objective optimization gives the possibility to assess experimental quality and cost at the same time and can reveal excellent compromise experiments. For example, the combination of 100% 1,2-(13)C2 glucose with 100% position one labeled glutamine and the combination of 100% 1,2-(13)C2 glucose with 100% uniformly labeled glutamine perform equally well for the carcinoma cell line, but the first mixture offers a decrease in cost of $ 120 per ml-scale cell culture experiment. We demonstrated the validity of a multi-objective linear approach to perform optimal experimental designs for the non-linear problem of (13)C-metabolic flux analysis. Tools and a workflow are provided to perform multi-objective design. The effortless calculation of the D-criterion can be exploited to perform high-throughput screening of possible (13)C-tracers, while the illustrated benefit of multi

  11. From elementary flux modes to elementary flux vectors: Metabolic pathway analysis with arbitrary linear flux constraints.

    PubMed

    Klamt, Steffen; Regensburger, Georg; Gerstl, Matthias P; Jungreuthmayer, Christian; Schuster, Stefan; Mahadevan, Radhakrishnan; Zanghellini, Jürgen; Müller, Stefan

    2017-04-01

    Elementary flux modes (EFMs) emerged as a formal concept to describe metabolic pathways and have become an established tool for constraint-based modeling and metabolic network analysis. EFMs are characteristic (support-minimal) vectors of the flux cone that contains all feasible steady-state flux vectors of a given metabolic network. EFMs account for (homogeneous) linear constraints arising from reaction irreversibilities and the assumption of steady state; however, other (inhomogeneous) linear constraints, such as minimal and maximal reaction rates frequently used by other constraint-based techniques (such as flux balance analysis [FBA]), cannot be directly integrated. These additional constraints further restrict the space of feasible flux vectors and turn the flux cone into a general flux polyhedron in which the concept of EFMs is not directly applicable anymore. For this reason, there has been a conceptual gap between EFM-based (pathway) analysis methods and linear optimization (FBA) techniques, as they operate on different geometric objects. One approach to overcome these limitations was proposed ten years ago and is based on the concept of elementary flux vectors (EFVs). Only recently has the community started to recognize the potential of EFVs for metabolic network analysis. In fact, EFVs exactly represent the conceptual development required to generalize the idea of EFMs from flux cones to flux polyhedra. This work aims to present a concise theoretical and practical introduction to EFVs that is accessible to a broad audience. We highlight the close relationship between EFMs and EFVs and demonstrate that almost all applications of EFMs (in flux cones) are possible for EFVs (in flux polyhedra) as well. In fact, certain properties can only be studied with EFVs. Thus, we conclude that EFVs provide a powerful and unifying framework for constraint-based modeling of metabolic networks.

  12. Genome-Based Metabolic Mapping and 13C Flux Analysis Reveal Systematic Properties of an Oleaginous Microalga Chlorella protothecoides1[OPEN

    PubMed Central

    Wu, Chao; Xiong, Wei; Dai, Junbiao; Wu, Qingyu

    2015-01-01

    Integrated and genome-based flux balance analysis, metabolomics, and 13C-label profiling of phototrophic and heterotrophic metabolism in Chlorella protothecoides, an oleaginous green alga for biofuel. The green alga Chlorella protothecoides, capable of autotrophic and heterotrophic growth with rapid lipid synthesis, is a promising candidate for biofuel production. Based on the newly available genome knowledge of the alga, we reconstructed the compartmentalized metabolic network consisting of 272 metabolic reactions, 270 enzymes, and 461 encoding genes and simulated the growth in different cultivation conditions with flux balance analysis. Phenotype-phase plane analysis shows conditions achieving theoretical maximum of the biomass and corresponding fatty acid-producing rate for phototrophic cells (the ratio of photon uptake rate to CO2 uptake rate equals 8.4) and heterotrophic ones (the glucose uptake rate to O2 consumption rate reaches 2.4), respectively. Isotope-assisted liquid chromatography-mass spectrometry/mass spectrometry reveals higher metabolite concentrations in the glycolytic pathway and the tricarboxylic acid cycle in heterotrophic cells compared with autotrophic cells. We also observed enhanced levels of ATP, nicotinamide adenine dinucleotide (phosphate), reduced, acetyl-Coenzyme A, and malonyl-Coenzyme A in heterotrophic cells consistently, consistent with a strong activity of lipid synthesis. To profile the flux map in experimental conditions, we applied nonstationary 13C metabolic flux analysis as a complementing strategy to flux balance analysis. The result reveals negligible photorespiratory fluxes and a metabolically low active tricarboxylic acid cycle in phototrophic C. protothecoides. In comparison, high throughput of amphibolic reactions and the tricarboxylic acid cycle with no glyoxylate shunt activities were measured for heterotrophic cells. Taken together, the metabolic network modeling assisted by experimental metabolomics and 13C labeling

  13. Metabolic flux pattern of glucose utilization by Xanthomonas campestris pv. campestris: prevalent role of the Entner-Doudoroff pathway and minor fluxes through the pentose phosphate pathway and glycolysis.

    PubMed

    Schatschneider, Sarah; Huber, Claudia; Neuweger, Heiko; Watt, Tony Francis; Pühler, Alfred; Eisenreich, Wolfgang; Wittmann, Christoph; Niehaus, Karsten; Vorhölter, Frank-Jörg

    2014-10-01

    The well-studied plant pathogenic bacterium Xanthomonas campestris pv. campestris (Xcc) synthesizes the biotechnologically important polysaccharide xanthan gum, which is also regarded as a virulence factor in plant interactions. In Xcc, sugars like glucose are utilized as a source to generate energy and biomass for growth and pathogenicity. In this study, we used [1-(13)C]glucose as a tracer to analyze the fluxes in the central metabolism of the bacterium growing in a minimal medium. (13)C-Metabolic flux analysis based on gas chromatography-mass spectrometry (GC-MS) confirmed the prevalent catabolic role of the Entner-Doudoroff pathway. Comparative nuclear magnetic resonance (NMR)-based isotopologue profiling of a mutant deficient in glycolysis gave evidence for a moderate flux via glycolysis in the wild-type. In addition to reconfirming the Entner-Doudoroff pathway as a catabolic main route, this approach affirmed a numerically minor but important flux via the pentose phosphate pathway.

  14. Recurrence Analysis of Eddy Covariance Fluxes

    NASA Astrophysics Data System (ADS)

    Lange, Holger; Flach, Milan; Foken, Thomas; Hauhs, Michael

    2015-04-01

    The eddy covariance (EC) method is one key method to quantify fluxes in biogeochemical cycles in general, and carbon and energy transport across the vegetation-atmosphere boundary layer in particular. EC data from the worldwide net of flux towers (Fluxnet) have also been used to validate biogeochemical models. The high resolution data are usually obtained at 20 Hz sampling rate but are affected by missing values and other restrictions. In this contribution, we investigate the nonlinear dynamics of EC fluxes using Recurrence Analysis (RA). High resolution data from the site DE-Bay (Waldstein-Weidenbrunnen) and fluxes calculated at half-hourly resolution from eight locations (part of the La Thuile dataset) provide a set of very long time series to analyze. After careful quality assessment and Fluxnet standard gapfilling pretreatment, we calculate properties and indicators of the recurrent structure based both on Recurrence Plots as well as Recurrence Networks. Time series of RA measures obtained from windows moving along the time axis are presented. Their interpretation is guided by three different questions: (1) Is RA able to discern periods where the (atmospheric) conditions are particularly suitable to obtain reliable EC fluxes? (2) Is RA capable to detect dynamical transitions (different behavior) beyond those obvious from visual inspection? (3) Does RA contribute to an understanding of the nonlinear synchronization between EC fluxes and atmospheric parameters, which is crucial for both improving carbon flux models as well for reliable interpolation of gaps? (4) Is RA able to recommend an optimal time resolution for measuring EC data and for analyzing EC fluxes? (5) Is it possible to detect non-trivial periodicities with a global RA? We will demonstrate that the answers to all five questions is affirmative, and that RA provides insights into EC dynamics not easily obtained otherwise.

  15. Lithium metaborate flux in silicate analysis

    USGS Publications Warehouse

    Ingamells, C.O.

    1970-01-01

    Lithium metaborate is an effective flux for silicates and other rock-forming minerals. The glass resulting from fusion is mechanically strong, reasonably nonhygroscopic, and is readily soluble in dilute acids. These characteristics lead to its use in X-ray spectrography and in methods which require whole-rock solutions, such as atomic absorption and emission spectrometry. Difficulties have been encountered in the use of such techniques : a high-quality reagent has been difficult to obtain ; fusion conditions must be rather closely controlled; graphite crucibles used in the fusions need special treatment. Methods for overcoming these difficulties are outlined. Selected procedures for various instrumental methods of analysis are described. ?? 1970.

  16. Comparison and analysis of objective functions in flux balance analysis.

    PubMed

    García Sánchez, Carlos Eduardo; Torres Sáez, Rodrigo Gonzalo

    2014-01-01

    Flux balance analysis (FBA) is currently one of the most important and used techniques for estimation of metabolic reaction rates (fluxes). This mathematical approach utilizes an optimization criterion in order to select a distribution of fluxes from the feasible space delimited by the metabolic reactions and some restrictions imposed over them, assuming that cellular metabolism is in steady state. Therefore, the obtained flux distribution depends on the specific objective function used. Multiple studies have been aimed to compare distinct objective functions at given conditions, in order to determine which of those functions produces values of fluxes closer to real data when used as objective in the FBA; in other words, what is the best objective function for modeling cell metabolism at a determined environmental condition. However, these comparative studies have been designed in very dissimilar ways, and in general, several factors that can change the ideal objective function in a cellular condition have not been adequately considered. Additionally, most of them have used only one dataset for representing one condition of cell growth, and different measuring techniques have been used. For these reasons, a rigorous study on the effect of factors such as the quantity of used data, the number and type of fluxes utilized as input data, and the selected classification of growth conditions, are required in order to obtain useful conclusions for these comparative studies, allowing limiting clearly the application range on any of those results.

  17. Mathematical modeling of isotope labeling experiments for metabolic flux analysis.

    PubMed

    Nargund, Shilpa; Sriram, Ganesh

    2014-01-01

    Isotope labeling experiments (ILEs) offer a powerful methodology to perform metabolic flux analysis. However, the task of interpreting data from these experiments to evaluate flux values requires significant mathematical modeling skills. Toward this, this chapter provides background information and examples to enable the reader to (1) model metabolic networks, (2) simulate ILEs, and (3) understand the optimization and statistical methods commonly used for flux evaluation. A compartmentalized model of plant glycolysis and pentose phosphate pathway illustrates the reconstruction of a typical metabolic network, whereas a simpler example network illustrates the underlying metabolite and isotopomer balancing techniques. We also discuss the salient features of commonly used flux estimation software 13CFLUX2, Metran, NMR2Flux+, FiatFlux, and OpenFLUX. Furthermore, we briefly discuss methods to improve flux estimates. A graphical checklist at the end of the chapter provides a reader a quick reference to the mathematical modeling concepts and resources.

  18. Flux-coupled genes and their use in metabolic flux analysis.

    PubMed

    Kim, Hyun Uk; Kim, Won Jun; Lee, Sang Yup

    2013-09-01

    As large volumes of omics data have become available, systems biology is playing increasingly important roles in elucidating new biological phenomena, especially through genome-scale metabolic network modeling and simulation. Much effort has been exerted on integrating omics data with metabolic flux simulation, but further development is necessary for more accurate flux estimation. To move one step forward, we adopted the concept of flux-coupled genes (FCGs), which show that their expression transition patterns upon perturbations are correlated with their corresponding flux values, as additional constraints in metabolic flux analysis. It was found that gnd, pfkB, rpe, sdhB, sdhD, sucA, and zwf genes, mostly associated with pentose phosphate pathway and TCA cycle, were the most consistent FCGs in Escherichia coli based on its transcriptome and (13) C-flux data obtained from the chemostat cultivation at five different dilution rates. Consequently, constraints-based flux analyses with FCGs as additional constraints were conducted for the seven single-gene knockout mutants, compared with those obtained without using FCGs. This strategy of constraining the metabolic flux analysis with FCGs is expected to be useful due to the relative ease in obtaining transcriptional information in the functional genomics era.

  19. Statistical analysis of micrometeoroids flux on Mercury

    NASA Astrophysics Data System (ADS)

    Borin, P.; Cremonese, G.; Marzari, F.; Bruno, M.; Marchi, S.

    2009-08-01

    Context: Meteoroid impacts are an important source of neutral atoms in the exosphere of Mercury. Impacting particles of size smaller than 1 cm have been proposed to be the major contribution to exospheric gases. However, our knowledge of the fluxes and impact velocities of different sizes is based on old extrapolations of similar quantities on Earth. Aims: We compute by means of N-body numerical integrations the orbital evolution of a large number of dust particles supposedly produced in the Main Belt. They migrate inward under the effect of drag forces until they encounter a terrestrial planet or eventually fall into the Sun. From our numerical simulations, we compute the flux of particles hitting Mercury's surface and the corresponding distribution of impact velocities. Methods: The orbital evolution of dust particles of different sizes is computed with a numerical code based on a physical model developed previously by Marzari & Vanzani (1994, A&A, 283, 275). It includes the effects of Poynting-Robertson drag, solar wind drag, and planetary perturbations. A precise calibration of the particle flux on Mercury has been performed by comparing our model predictions for dust infall on to Earth with observational data. Results: We provide predictions of the flux to different size particles impacting Mercury and their collisional velocity distribution. We compare our results with previous estimates and we find that these collisional velocities are lower but that the fluxes are significantly higher.

  20. Flux frequency analysis of seasonally dry ecosystem fluxes in two unique biomes of Sonora Mexico

    NASA Astrophysics Data System (ADS)

    Verduzco, V. S.; Yepez, E. A.; Robles-Morua, A.; Garatuza, J.; Rodriguez, J. C.; Watts, C.

    2013-05-01

    Complex dynamics from the interactions of ecosystems processes makes difficult to model the behavior of ecosystems fluxes of carbon and water in response to the variation of environmental and biological drivers. Although process oriented ecosystem models are critical tools for studying land-atmosphere fluxes, its validity depends on the appropriate parameterization of equations describing temporal and spatial changes of model state variables and their interactions. This constraint often leads to discrepancies between model simulations and observed data that reduce models reliability especially in arid and semiarid ecosystems. In the semiarid north western Mexico, ecosystem processes are fundamentally controlled by the seasonality of water and the intermittence of rain pulses which are conditions that require calibration of specific fitting functions to describe the response of ecosystem variables (i.e. NEE, GPP, ET, respiration) to these wetting and drying periods. The goal is to find functions that describe the magnitude of ecosystem fluxes during individual rain pulses and the seasonality of the ecosystem. Relaying on five years of eddy covariance flux data of a tropical dry forest and a subtropical shrubland we present a flux frequency analysis that describe the variation of net ecosystem exchange (NEE) of CO2 to highlight the relevance of pulse driven dynamics controlling this flux. Preliminary results of flux frequency analysis of NEE indicate that these ecosystems are strongly controlled by the frequency distribution of rain. Also, the output of fitting functions for NEE, GPP, ET and respiration using semi-empirical functions applied at specific rain pulses compared with season-long statistically generated simulations do not agree. Seasonality and the intrinsic nature of individual pulses have different effects on ecosystem flux responses. This suggests that relationships between the nature of seasonality and individual pulses can help improve the

  1. Software applications toward quantitative metabolic flux analysis and modeling.

    PubMed

    Dandekar, Thomas; Fieselmann, Astrid; Majeed, Saman; Ahmed, Zeeshan

    2014-01-01

    Metabolites and their pathways are central for adaptation and survival. Metabolic modeling elucidates in silico all the possible flux pathways (flux balance analysis, FBA) and predicts the actual fluxes under a given situation, further refinement of these models is possible by including experimental isotopologue data. In this review, we initially introduce the key theoretical concepts and different analysis steps in the modeling process before comparing flux calculation and metabolite analysis programs such as C13, BioOpt, COBRA toolbox, Metatool, efmtool, FiatFlux, ReMatch, VANTED, iMAT and YANA. Their respective strengths and limitations are discussed and compared to alternative software. While data analysis of metabolites, calculation of metabolic fluxes, pathways and their condition-specific changes are all possible, we highlight the considerations that need to be taken into account before deciding on a specific software. Current challenges in the field include the computation of large-scale networks (in elementary mode analysis), regulatory interactions and detailed kinetics, and these are discussed in the light of powerful new approaches.

  2. Thermal-structural finite element analysis using linear flux formulation

    NASA Technical Reports Server (NTRS)

    Pandey, Ajay K.; Dechaumphai, Pramote; Wieting, Allan R.

    1990-01-01

    A linear flux approach is developed for a finite element thermal-structural analysis of steady state thermal and structural problems. The element fluxes are assumed to vary linearly in the same form as the element unknown variables, and the finite element matrices are evaluated in closed form. Since numerical integration is avoided, significant computational time saving is achieved. Solution accuracy and computational speed improvements are demonstrated by solving several two and three dimensional thermal-structural examples.

  3. 13C-based metabolic flux analysis: fundamentals and practice.

    PubMed

    Yang, Tae Hoon

    2013-01-01

    Isotope-based metabolic flux analysis is one of the emerging technologies applied to system level metabolic phenotype characterization in metabolic engineering. Among the developed approaches, (13)C-based metabolic flux analysis has been established as a standard tool and has been widely applied to quantitative pathway characterization of diverse biological systems. To implement (13)C-based metabolic flux analysis in practice, comprehending the underlying mathematical and computational modeling fundamentals is of importance along with carefully conducted experiments and analytical measurements. Such knowledge is also crucial when designing (13)C-labeling experiments and properly acquiring key data sets essential for in vivo flux analysis implementation. In this regard, the modeling fundamentals of (13)C-labeling systems and analytical data processing are the main topics we will deal with in this chapter. Along with this, the relevant numerical optimization techniques are addressed to help implementation of the entire computational procedures aiming at (13)C-based metabolic flux analysis in vivo.

  4. Flux tube analysis of L-band ionospheric scintillation

    NASA Astrophysics Data System (ADS)

    Shume, E. B.; Mannucci, A. J.; Butala, M. D.; Pi, X.; Valladares, C. E.

    2013-06-01

    This manuscript presents magnetic flux tube analysis of L-band signal scintillation in the nighttime equatorial and low-latitude ionosphere. Residues of the scintillation index S4 estimated from the L-band signals received from Geostationary Earth Orbit (GEO) satellites are employed in the analysis. The S4 estimates have been shown to be associated with simultaneous GPS VTEC variations derived from JPL's GIPSY-GIM package. We have applied the wavelet decomposition technique simultaneously on the S4 time series in a flux tube over the equatorial and low-latitude regions. The technique decomposes the S4 signal to identify the dominant mode of variabilities and the temporal variations of scintillation-producing irregularities in the context of a flux tube. Statistically significant regions of the wavelet power spectra considered in our study have mainly shown that (a) dominant plasma irregularities associated with S4 variabilities in a flux tube have periods of about 4 to 15 minutes (horizontal irregularity scales of about 24 to 90 km). These periods match short period gravity waves, (b) scintillation-producing irregularities are anisotropic along the flux tube and in the east-west direction, and (c) the occurrences of scintillation-producing irregularities along the flux tube indicate that the entire flux tube became unstable. However, plasma instability occurrences were not simultaneous in most cases along the flux tube, there were time delays of various orders. Understanding the attributes of L-band scintillation-producing irregularities could be important for developing measures to mitigate L-band signal degradation.

  5. Dynamic Modeling of Aerobic Growth of Shewanella oneidensis. Predicting Triauxic Growth, Flux Distributions and Energy Requirement for Growth

    SciTech Connect

    Song, Hyun-Seob; Ramkrishna, Doraiswami; Pinchuk, Grigoriy E.; Beliaev, Alex S.; Konopka, Allan; Fredrickson, Jim K.

    2013-01-01

    A model-based analysis is conducted to investigate metabolism of Shewanella oneidensis MR-1 strain in aerobic batch culture, which exhibits an intriguing growth pattern by sequentially consuming substrate (i.e., lactate) and by-products (i.e., pyruvate and acetate). A general protocol is presented for developing a detailed network-based dynamic model for S. oneidensis based on the Lumped Hybrid Cybernetic Model (LHCM) framework. The L-HCM, although developed from only limited data, is shown to accurately reproduce exacting dynamic metabolic shifts, and provide reasonable estimates of energy requirement for growth. Flux distributions in S. oneidensis predicted by the L-HCM compare very favorably with 13C-metabolic flux analysis results reported in the literature. Predictive accuracy is enhanced by incorporating measurements of only a few intracellular fluxes, in addition to extracellular metabolites. The L-HCM developed here for S. oneidensis is consequently a promising tool for the analysis of intracellular flux distribution and metabolic engineering.

  6. An Analysis of Moisture Fluxes into the Gulf of California

    NASA Technical Reports Server (NTRS)

    Wu, Man-Li C.; Schubert, Siegfried D.; Suarez, Max J.; Huang, Norden E.

    2009-01-01

    This study examines the nature of episodes of enhanced warm-season moisture flux into the Gulf of California. Both spatial structure and primary time scales of the fluxes are examined using the 40-yr ECMWF Re-Analysis data for the period 1980-2001. The analysis approach consists of a compositing technique that is keyed on the low-level moisture fluxes into the Gulf of California. The results show that the fluxes have a rich spectrum of temporal variability, with periods of enhanced transport over the gulf linked to African easterly waves on subweekly (3-8 day) time scales, the Madden-Julian oscillation (MJO) at intraseasonal time scales (20-90 day), and intermediate (10-15 day) time-scale disturbances that appear to originate primarily in the Caribbean Sea-western Atlantic Ocean. In the case of the MJO, enhanced low-level westerlies and large-scale rising motion provide an environment that favors large-scale cyclonic development near the west coast of Central America that, over the course of about 2 weeks, expands northward along the coast eventually reaching the mouth of the Gulf of California where it acts to enhance the southerly moisture flux in that region. On a larger scale, the development includes a northward shift in the eastern Pacific ITCZ, enhanced precipitation over much of Mexico and the southwestern United States, and enhanced southerly/southeasterly fluxes from the Gulf of Mexico into Mexico and the southwestern and central United States. In the case of the easterly waves, the systems that reach Mexico appear to redevelop/reorganize on the Pacific coast and then move rapidly to the northwest to contribute to the moisture flux into the Gulf of California. The most intense fluxes into the gulf on these time scales appear to be synchronized with a midlatitude short-wave trough over the U.S. West Coast and enhanced low-level southerly fluxes over the U.S. Great Plains. The intermediate (10-15 day) time-scale systems have zonal wavelengths roughly twice

  7. Noise analysis of DC SQUIDs with damped superconducting flux transformers

    NASA Astrophysics Data System (ADS)

    Faley, M. I.; Poppe, U.; Urban, K.; Fagaly, R. L.

    2010-06-01

    An analysis was performed of intrinsic noise for high-Tc DC SQUID with superconducting flux transformer (FT) containing resistive elements. For a SQUID with a loop inductance of about 40 pH we observed voltage swings of ~55 μV and a flux noise of ~4 μΦ0/√Hz at 77 K. Inductive coupling of an 8-mm multilayer superconducting FT to the SQUID increased voltage swings to ~70 μV due to effective reduction of the SQUID loop inductance. This also increased the flux noise to ~6μΦ0/√Hz, corresponding to a field resolution of ~18 fT/√Hz at 77 K with a white noise spectrum down to frequency ~10 Hz. The main sources of white flux noise were the Nyquist noise in the Josephson junctions and the FT, as well as the suppression of the DC SQUID voltage swings caused by parasitic capacitance between the FT and the SQUID. An ultra-low-ohmic resistor with resistance value between the flux-creep-induced resistances of superconductors (below ~0.1 nΩ) and resistances of conventional resistors (above ~0.1 mΩ) was developed. An RL-circuit based high-pass filter (HPF) with time constant ~7 sec was realized and integrated in the superconducting FT. The contribution of the HPF to the noise of the sensors was measured and compared with calculated values.

  8. Structural analysis of airborne flux estimates over a region

    NASA Technical Reports Server (NTRS)

    Caramori, Paulo; Schuepp, Peter; Desjardins, Raymond; Macpherson, Ian

    1994-01-01

    Aircraft-based observations of turbulence fields of velocity, moisture, and temperature are used to study coherent turbulent structures that dominate turbulent transfer of moisture and heat above three different eco-systems. Flux traces are defragmented, to reconstruct the presumed full size (along the sampled transect) of these structures, and flux traces are simplified by elimination of those that contribute negligibly to the flux estimate. Structures are analyzed in terms of size, spatial distribution, and contribution to the flux, in the four 'quadrant' modes of eddy-covariance transfer (excess up/down and deficit up/down). The effect of nonlinear detrending of moisture and temperature data on this 'structural analysis,' over surfaces with heterogeneous surface wetness, is also examined. Results over grassland, wetland, and moist and dry agricultural land, show that nonlinear detrending may provide a more physically realistic description of structures. Significant differences are observed between structure size and associated relative flux contribution, between moist and dry areas, with smaller structures playing a more important role over the moist areas. Structure size generally increases with height, as spatial reorganization from smaller structures into larger ones takes place. This coincides with a gradual loss of surface 'signature' (position and clustering of plumes above localized source areas). The data are expected to provide a basis for an eventual statistical description of boundary-layer transfer events , and help to interpret the link between boundary-layer transfer and hydrological surface conditions.

  9. E-Flux2 and SPOT: Validated Methods for Inferring Intracellular Metabolic Flux Distributions from Transcriptomic Data

    PubMed Central

    Kim, Min Kyung; Lane, Anatoliy; Kelley, James J.; Lun, Desmond S.

    2016-01-01

    Background Several methods have been developed to predict system-wide and condition-specific intracellular metabolic fluxes by integrating transcriptomic data with genome-scale metabolic models. While powerful in many settings, existing methods have several shortcomings, and it is unclear which method has the best accuracy in general because of limited validation against experimentally measured intracellular fluxes. Results We present a general optimization strategy for inferring intracellular metabolic flux distributions from transcriptomic data coupled with genome-scale metabolic reconstructions. It consists of two different template models called DC (determined carbon source model) and AC (all possible carbon sources model) and two different new methods called E-Flux2 (E-Flux method combined with minimization of l2 norm) and SPOT (Simplified Pearson cOrrelation with Transcriptomic data), which can be chosen and combined depending on the availability of knowledge on carbon source or objective function. This enables us to simulate a broad range of experimental conditions. We examined E. coli and S. cerevisiae as representative prokaryotic and eukaryotic microorganisms respectively. The predictive accuracy of our algorithm was validated by calculating the uncentered Pearson correlation between predicted fluxes and measured fluxes. To this end, we compiled 20 experimental conditions (11 in E. coli and 9 in S. cerevisiae), of transcriptome measurements coupled with corresponding central carbon metabolism intracellular flux measurements determined by 13C metabolic flux analysis (13C-MFA), which is the largest dataset assembled to date for the purpose of validating inference methods for predicting intracellular fluxes. In both organisms, our method achieves an average correlation coefficient ranging from 0.59 to 0.87, outperforming a representative sample of competing methods. Easy-to-use implementations of E-Flux2 and SPOT are available as part of the open

  10. Acetate fluxes in Escherichia coli are determined by the thermodynamic control of the Pta-AckA pathway

    PubMed Central

    Enjalbert, Brice; Millard, Pierre; Dinclaux, Mickael; Portais, Jean-Charles; Létisse, Fabien

    2017-01-01

    Escherichia coli excretes acetate upon growth on fermentable sugars, but the regulation of this production remains elusive. Acetate excretion on excess glucose is thought to be an irreversible process. However, dynamic 13C-metabolic flux analysis revealed a strong bidirectional exchange of acetate between E. coli and its environment. The Pta-AckA pathway was found to be central for both flux directions, while alternative routes (Acs or PoxB) play virtually no role in glucose consumption. Kinetic modelling of the Pta-AckA pathway predicted that its flux is thermodynamically controlled by the extracellular acetate concentration in vivo. Experimental validations confirmed that acetate production can be reduced and even reversed depending solely on its extracellular concentration. Consistently, the Pta-AckA pathway can rapidly switch from acetate production to consumption. Contrary to current knowledge, E. coli is thus able to co-consume glucose and acetate under glucose excess. These metabolic capabilities were confirmed on other glycolytic substrates which support the growth of E. coli in the gut. These findings highlight the dual role of the Pta-AckA pathway in acetate production and consumption during growth on glycolytic substrates, uncover a novel regulatory mechanism that controls its flux in vivo, and significantly expand the metabolic capabilities of E. coli. PMID:28186174

  11. Evolutionary engineered Saccharomyces cerevisiae wine yeast strains with increased in vivo flux through the pentose phosphate pathway.

    PubMed

    Cadière, Axelle; Ortiz-Julien, Anne; Camarasa, Carole; Dequin, Sylvie

    2011-05-01

    Amplification of the flux toward the pentose phosphate (PP) pathway might be of interest for various S. cerevisiae based industrial applications. We report an evolutionary engineering strategy based on a long-term batch culture on gluconate, a substrate that is poorly assimilated by S. cerevisiae cells and is metabolized by the PP pathway. After adaptation for various periods of time, we selected strains that had evolved a greater consumption capacity for gluconate. (13)C metabolic flux analysis on glucose revealed a redirection of carbon flux from glycolysis towards the PP pathway and a greater synthesis of lipids. The relative flux into the PP pathway was 17% for the evolved strain (ECA5) versus 11% for the parental strain (EC1118). During wine fermentation, the evolved strains displayed major metabolic changes, such as lower levels of acetate production, higher fermentation rates and enhanced production of aroma compounds. These represent a combination of novel traits, which are of great interest in the context of modern winemaking.

  12. A reliable simulator for dynamic flux balance analysis.

    PubMed

    Höffner, K; Harwood, S M; Barton, P I

    2013-03-01

    Dynamic flux balance analysis (DFBA) provides a platform for detailed design, control and optimization of biochemical process technologies. It is a promising modeling framework that combines genome-scale metabolic network analysis with dynamic simulation of the extracellular environment. Dynamic flux balance analysis assumes that the intracellular species concentrations are in equilibrium with the extracellular environment. The resulting underdetermined stoichiometric model is solved under the assumption of a biochemical objective such as growth rate maximization. The model of the metabolism is coupled with the dynamic mass balance equations of the extracellular environment via expressions for the rates of substrate uptake and product excretion, which imposes additional constraints on the linear program (LP) defined by growth rate maximization of the metabolism. The linear program is embedded into the dynamic model of the bioreactor, and together with the additional constraints this provides an accurate model of the substrate consumption, product secretion, and biomass production during operation. A DFBA model consists of a system of ordinary differential equations for which the evaluation of the right-hand side requires not only function evaluations, but also the solution of one or more linear programs. The numerical tool presented here accurately and efficiently simulates large-scale dynamic flux balance models. The main advantages that this approach has over existing implementation are that the integration scheme has a variable step size, that the linear program only has to be solved when qualitative changes in the optimal flux distribution of the metabolic network occur, and that it can reliably simulate behavior near the boundary of the domain where the model is defined. This is illustrated through large-scale examples taken from the literature.

  13. Geoneutrinos at Jinping: Flux prediction and oscillation analysis

    NASA Astrophysics Data System (ADS)

    Wan, Linyan; Hussain, Ghulam; Wang, Zhe; Chen, Shaomin

    2017-03-01

    Geoneutrinos are electron antineutrinos (ν¯e) generated by the beta decays of radionuclides naturally occurring inside the Earth, in particular U 238 , Th 232 , and K 40 . Measurement of these neutrinos provides powerful constraints on the radiogenic heat of the Earth and tests on the Earth models. Since the prediction of ν¯e's in geoneutrino flux is subject to neutrino oscillation effects, we performed a calculation including detailed oscillation analysis in the propagation of geoneutrinos and reactor neutrinos generated around the Earth. The expected geoneutrino signal, the reactor neutrino background rates, and the systematic error budget are provided for a proposed 3 kt neutrino detector at the Jinping underground lab in Sichuan, China. In addition, we evaluated sensitivities for the geoneutrino flux, Th/U ratio, and power of a possible fission reactor in the interior of the Earth.

  14. Analysis of metabolic flux using dynamic labelling and metabolic modelling.

    PubMed

    Fernie, A R; Morgan, J A

    2013-09-01

    Metabolic fluxes and the capacity to modulate them are a crucial component of the ability of the plant cell to react to environmental perturbations. Our ability to quantify them and to attain information concerning the regulatory mechanisms that control them is therefore essential to understand and influence metabolic networks. For all but the simplest of flux measurements labelling methods have proven to be the most informative. Both steady-state and dynamic labelling approaches have been adopted in the study of plant metabolism. Here the conceptual basis of these complementary approaches, as well as their historical application in microbial, mammalian and plant sciences, is reviewed, and an update on technical developments in label distribution analyses is provided. This is supported by illustrative cases studies involving the kinetic modelling of secondary metabolism. One issue that is particularly complex in the analysis of plant fluxes is the extensive compartmentation of the plant cell. This problem is discussed from both theoretical and experimental perspectives, and the current approaches used to address it are assessed. Finally, current limitations and future perspectives of kinetic modelling of plant metabolism are discussed.

  15. Robustness analysis of elementary flux modes generated by column generation.

    PubMed

    Oddsdóttir, Hildur Æsa; Hagrot, Erika; Chotteau, Véronique; Forsgren, Anders

    2016-03-01

    Elementary flux modes (EFMs) are vectors defined from a metabolic reaction network, giving the connections between substrates and products. EFMs-based metabolic flux analysis (MFA) estimates the flux over each EFM from external flux measurements through least-squares data fitting. The measurements used in the data fitting are subject to errors. A robust optimization problem includes information on errors and gives a way to examine the sensitivity of the solution of the EFMs-based MFA to these errors. In general, formulating a robust optimization problem may make the problem significantly harder. We show that in the case of the EFMs-based MFA, when the errors are only in measurements and bounded by an interval, the robust problem can be stated as a convex quadratic programming (QP) problem. We have previously shown how the data fitting problem may be solved in a column-generation framework. In this paper, we show how column generation may be applied also to the robust problem, thereby avoiding explicit enumeration of EFMs. Furthermore, the option to indicate intervals on metabolites that are not measured is introduced in this column generation framework. The robustness of the data is evaluated in a case-study, which indicates that the solutions of our non-robust problems are in fact near-optimal also when robustness is considered, implying that the errors in measurement do not have a large impact on the optimal solution. Furthermore, we showed that the addition of intervals on unmeasured metabolites resulted in a change in the optimal solution.

  16. 13C labeling analysis of sugars by high resolution-mass spectrometry for metabolic flux analysis.

    PubMed

    Acket, Sébastien; Degournay, Anthony; Merlier, Franck; Thomasset, Brigitte

    2017-02-14

    Metabolic flux analysis is particularly complex in plant cells because of highly compartmented metabolism. Analysis of free sugars is interesting because it provides data to define fluxes around hexose, pentose, and triose phosphate pools in different compartment. In this work, we present a method to analyze the isotopomer distribution of free sugars labeled with carbon 13 using a liquid chromatography-high resolution mass spectrometry, without derivatized procedure, adapted for Metabolic flux analysis. Our results showed a good sensitivity, reproducibility and better accuracy to determine isotopic enrichments of free sugars compared to our previous methods [5, 6].

  17. Fast Flux Test Facility final safety analysis report. Amendment 73

    SciTech Connect

    Gantt, D.A.

    1993-08-01

    This report provides Final Safety Analysis Report (FSAR) Amendment 73 for incorporation into the Fast Flux Test Facility (FFTR) FSAR set. This page change incorporates Engineering Change Notices (ECNs) issued subsequent to Amendment 72 and approved for incorparoration before May 6, 1993. These changes include: Chapter 3, design criteria structures, equipment, and systems; chapter 5B, reactor coolant system; chapter 7, instrumentation and control systems; chapter 9, auxiliary systems; chapter 11, reactor refueling system; chapter 12, radiation protection and waste management; chapter 13, conduct of operations; chapter 17, technical specifications; chapter 20, FFTF criticality specifications; appendix C, local fuel failure events; and appendix Fl, operation at 680{degrees}F inlet temperature.

  18. In vivo NMR for ¹³C Metabolic Flux Analysis.

    PubMed

    Roscher, Albrecht; Troufflard, Stéphanie; Taghki, Abdelghani Idrissi

    2014-01-01

    The use of in vivo NMR within the framework of Metabolic Flux Analysis in plants is presented. In vivo NMR allows to visualize the active metabolic network, to determine metabolic and isotopic steady state and to measure metabolic fluxes which are not necessarily accessible by isotopic steady state (stationary) Metabolic Flux Analysis. The kinetic data can be used as input for dynamic (nonstationary) Metabolic Flux Analysis. Both 1D and 2D NMR methods are employed.

  19. Dynamic analysis of CO₂ labeling and cell respiration using membrane-inlet mass spectrometry.

    PubMed

    Yang, Tae Hoon

    2014-01-01

    Here, we introduce a mass spectrometry-based analytical method and relevant technical details for dynamic cell respiration and CO2 labeling analysis. Such measurements can be utilized as additional information and constraints for model-based (13)C metabolic flux analysis. Dissolved dynamics of oxygen consumption and CO2 mass isotopomer evolution from (13)C-labeled tracer substrates through different cellular processes can be precisely measured on-line using a miniaturized reactor system equipped with a membrane-inlet mass spectrometer. The corresponding specific rates of physiologically relevant gases and CO2 mass isotopomers can be quantified within a short-term range based on the liquid-phase dynamics of dissolved fermentation gases.

  20. Investigation of the central carbon metabolism of Sorangium cellulosum: metabolic network reconstruction and quantification of pathway fluxes.

    PubMed

    Bolten, Christoph J; Heinzle, Elmar; Müller, Rolf; Wittmann, Christoph

    2009-01-01

    In the present work, the metabolic network of primary metabolism of the slow-growing myxobacterium Sorangium cellulosum was reconstructed from the annotated genome sequence of the type strain So ce56. During growth on glucose as the carbon source and asparagine as the nitrogen source, So ce56 showed a very low growth rate of 0.23 d-(1), equivalent to a doubling time of 3 days. Based on a complete stoichiometric and isotopomer model of the central metabolism, 13C metabolic flux analysis was carried out for growth with glucose as carbon and asparagine as nitrogen sources. Normalized to the uptake flux for glucose (100%), cells recruited glycolysis (51%) and the pentose phosphate pathway (48%) as major catabolic pathways. The Entner-Doudoroff pathway and glyoxylate shunt were not active. A high flux through the TCA cycle (118%) enabled a strong formation of ATP, but cells revealed a rather low yield for biomass. Inspection of fluxes linked to energy metabolism revealed that S. cellulosum utilized only 10% of the ATP formed for growth, whereas 90% is required for maintenance. This explains the apparent discrepancy between the relatively low biomass yield and the high flux through the energy-delivering TCA cycle. The total flux of NADPH supply (216%) was higher than the demand for anabolism (156%), indicating additional reactions for balancing of NADPH. The cells further exhibited a highly active metabolic cycle, interconverting C3 and C4 metabolites of glycolysis and the TCA cycle. The present work provides the first insight into fluxes of the primary metabolism of myxobacteria, especially for future investigation on the supply of cofactors, building blocks, and energy in myxobacteria, producing natural compounds of biotechnological interest.

  1. Dynamic metabolic flux analysis using a convex analysis approach: Application to hybridoma cell cultures in perfusion.

    PubMed

    Fernandes de Sousa, Sofia; Bastin, Georges; Jolicoeur, Mario; Vande Wouwer, Alain

    2016-05-01

    In recent years, dynamic metabolic flux analysis (DMFA) has been developed in order to evaluate the dynamic evolution of the metabolic fluxes. Most of the proposed approaches are dedicated to exactly determined or overdetermined systems. When an underdetermined system is considered, the literature suggests the use of dynamic flux balance analysis (DFBA). However the main challenge of this approach is to determine an appropriate objective function, which remains valid over the whole culture. In this work, we propose an alternative dynamic metabolic flux analysis based on convex analysis, DMFCA, which allows the determination of bounded intervals for the fluxes using the available knowledge of the metabolic network and information provided by the time evolution of extracellular component concentrations. Smoothing splines and mass balance differential equations are used to estimate the time evolution of the uptake and excretion rates from this experimental data. The main advantage of the proposed procedure is that it does not require additional constraints or objective functions, and provides relatively narrow intervals for the intracellular metabolic fluxes. DMFCA is applied to experimental data from hybridoma HB58 cell perfusion cultures, in order to investigate the influence of the operating mode (batch and perfusion) on the metabolic flux distribution.

  2. Combining pathway analysis with flux balance analysis for the comprehensive study of metabolic systems.

    PubMed

    Schilling, C H; Edwards, J S; Letscher, D; Palsson, B Ø

    The elucidation of organism-scale metabolic networks necessitates the development of integrative methods to analyze and interpret the systemic properties of cellular metabolism. A shift in emphasis from single metabolic reactions to systemically defined pathways is one consequence of such an integrative analysis of metabolic systems. The constraints of systemic stoichiometry, and limited thermodynamics have led to the definition of the flux space within the context of convex analysis. The flux space of the metabolic system, containing all allowable flux distributions, is constrained to a convex polyhedral cone in a high-dimensional space. From metabolic pathway analysis, the edges of the high-dimensional flux cone are vectors that correspond to systemically defined "extreme pathways" spanning the capabilities of the system. The addition of maximum flux capacities of individual metabolic reactions serves to further constrain the flux space and has led to the development of flux balance analysis using linear optimization to calculate optimal flux distributions. Here we provide the precise theoretical connections between pathway analysis and flux balance analysis allowing for their combined application to study integrated metabolic function. Shifts in metabolic behavior are calculated using linear optimization and are then interpreted using the extreme pathways to demonstrate the concept of pathway utilization. Changes to the reaction network, such as the removal of a reaction, can lead to the generation of suboptimal phenotypes that can be directly attributed to the loss of pathway function and capabilities. Optimal growth phenotypes are calculated as a function of environmental variables, such as the availability of substrate and oxygen, leading to the definition of phenotypic phase planes. It is illustrated how optimality properties of the computed flux distributions can be interpreted in terms of the extreme pathways. Together these developments are applied to an

  3. Using flux balance analysis to guide microbial metabolic engineering.

    PubMed

    Curran, Kathleen A; Crook, Nathan C; Alper, Hal S

    2012-01-01

    Metabolic engineers modify biological systems through the use of modern molecular biology tools in order to obtain desired phenotypes. However, due to the extreme complexity and interconnectedness of metabolism in all organisms, it is often difficult to a priori predict which changes will yield the optimal results. Flux balance analysis (FBA) is a mathematical approach that uses a genomic-scale metabolic network models to afford in silico prediction and optimization of metabolic changes. In particular, a genome-scale approach can help select gene targets for knockout and overexpression. This approach can be used to help expedite the strain engineering process. Here, we give an introduction to the use of FBA and provide details for its implementation in a microbial metabolic engineering context.

  4. Finding elementary flux modes in metabolic networks based on flux balance analysis and flux coupling analysis: application to the analysis of Escherichia coli metabolism.

    PubMed

    Tabe-Bordbar, Shayan; Marashi, Sayed-Amir

    2013-12-01

    Elementary modes (EMs) are steady-state metabolic flux vectors with minimal set of active reactions. Each EM corresponds to a metabolic pathway. Therefore, studying EMs is helpful for analyzing the production of biotechnologically important metabolites. However, memory requirements for computing EMs may hamper their applicability as, in most genome-scale metabolic models, no EM can be computed due to running out of memory. In this study, we present a method for computing randomly sampled EMs. In this approach, a network reduction algorithm is used for EM computation, which is based on flux balance-based methods. We show that this approach can be used to recover the EMs in the medium- and genome-scale metabolic network models, while the EMs are sampled in an unbiased way. The applicability of such results is shown by computing “estimated” control-effective flux values in Escherichia coli metabolic network.

  5. Flux balance analysis of myocardial mitochondrial metabolic network

    NASA Astrophysics Data System (ADS)

    Luo, Ruoyu; Liao, Sha; Liu, Bifeng; Liu, Manxi; Zhang, Hongming; Luo, Qingming

    2005-03-01

    A large number of biological information has been available from genome sequencing and bioinformatics. To further understand the qualities of the biological networks (such as metabolic network) in the complex biological system, representations of integrated function in silico have been widely investigated, and various modeling approaches have been designed, most of which are based on detailed kinetic information except flux balance analysis (FBA). FBA, just based on stoichimetrical information of reactions, is a suitable method for the study of metabolic pathways, and it analyzes the behaviors of the network from the viewpoint of the whole system. Herein, this modeling approach has been utilized to reconstruct the mitochondrial metabolic network to integrate and analyze its capability of producing energy. Besides, extreme pathways analysis (EPA) and shadow prices analysis have also been integrated to study the interior characters of the network. Our modeling results have indicated for the first time that the covalent regulative property of pyruvate dehydrogenase is restrained by the feedback of acetyl-CoA. Combined with the biological experiments, these simulations in silico could be pretty useful for the further understanding of functions and characters of the biological network as a complex system.

  6. Dynamic metabolic flux analysis of plant cell wall synthesis.

    PubMed

    Chen, Xuewen; Alonso, Ana P; Shachar-Hill, Yair

    2013-07-01

    The regulation of plant cell wall synthesis pathways remains poorly understood. This has become a bottleneck in designing bioenergy crops. The goal of this study was to analyze the regulation of plant cell wall precursor metabolism using metabolic flux analysis based on dynamic labeling experiments. Arabidopsis T87 cells were cultured heterotrophically with (13)C labeled sucrose. The time course of ¹³C labeling patterns in cell wall precursors and related sugar phosphates was monitored using liquid chromatography tandem mass spectrometry until steady state labeling was reached. A kinetic model based on mass action reaction mechanisms was developed to simulate the carbon flow in the cell wall synthesis network. The kinetic parameters of the model were determined by fitting the model to the labeling time course data, cell wall composition, and synthesis rates. A metabolic control analysis was performed to predict metabolic regulations that may improve plant biomass composition for biofuel production. Our results describe the routes and rates of carbon flow from sucrose to cell wall precursors. We found that sucrose invertase is responsible for the entry of sucrose into metabolism and UDP-glucose-4-epimerase plays a dominant role in UDP-Gal synthesis in heterotrophic Aradidopsis cells under aerobic conditions. We also predicted reactions that exert strong regulatory influence over carbon flow to cell wall synthesis and its composition.

  7. Metabolomics integrated elementary flux mode analysis in large metabolic networks.

    PubMed

    Gerstl, Matthias P; Ruckerbauer, David E; Mattanovich, Diethard; Jungreuthmayer, Christian; Zanghellini, Jürgen

    2015-03-10

    Elementary flux modes (EFMs) are non-decomposable steady-state pathways in metabolic networks. They characterize phenotypes, quantify robustness or identify engineering targets. An EFM analysis (EFMA) is currently restricted to medium-scale models, as the number of EFMs explodes with the network's size. However, many topologically feasible EFMs are biologically irrelevant. We present thermodynamic EFMA (tEFMA), which calculates only the small(er) subset of thermodynamically feasible EFMs. We integrate network embedded thermodynamics into EFMA and show that we can use the metabolome to identify and remove thermodynamically infeasible EFMs during an EFMA without losing biologically relevant EFMs. Calculating only the thermodynamically feasible EFMs strongly reduces memory consumption and program runtime, allowing the analysis of larger networks. We apply tEFMA to study the central carbon metabolism of E. coli and find that up to 80% of its EFMs are thermodynamically infeasible. Moreover, we identify glutamate dehydrogenase as a bottleneck, when E. coli is grown on glucose and explain its inactivity as a consequence of network embedded thermodynamics. We implemented tEFMA as a Java package which is available for download at https://github.com/mpgerstl/tEFMA.

  8. Fast Flux Test Facility final safety analysis report. Amendment 72

    SciTech Connect

    Gantt, D. A.

    1992-08-01

    This document provides the Final Safety Analysis Report (FSAR) Amendment 72 for incorporation into the Fast Flux Test Facility (FFTF) FSAR set. This amendment change incorporates Engineering Change Notices issued subsequent to Amendment 71 and approved for incorporation before June 24, 1992. These include changes in: Chapter 2, Site Characteristics; Chapter 3, Design Criteria Structures, Equipment, and Systems; Chapter 5B, Reactor Coolant System; Chapter 7, Instrumentation and Control Systems; Chapter 8, Electrical Systems - The description of the Class 1E, 125 Vdc systems is updated for the higher capacity of the newly installed, replacement batteries; Chapter 9, Auxiliary Systems - The description of the inert cell NASA systems is corrected to list the correct number of spare sample points; Chapter 11, Reactor Refueling System; Chapter 12, Radiation Protection and Waste Management; Chapter 13, Conduct of Operations; Chapter 16, Quality Assurance; Chapter 17, Technical Specifications; Chapter 19, FFTF Fire Specifications for Fire Detection, Alarm, and Protection Systems; Chapter 20, FFTF Criticality Specifications; and Appendix B, Primary Piping Integrity Evaluation.

  9. Shortwave flux profile analysis at the Cabauw BSRN site

    NASA Astrophysics Data System (ADS)

    Wang, P.; Baltink, H. Klein; Knap, W. H.; Stammes, P.

    2013-05-01

    The vertical distribution of the shortwave flux in the atmosphere is important for understanding the energy budget and the validation of climate models. We calculated shortwave flux profiles for cloudy cases by using the Doubling-Adding KNMI radiative transfer model with water vapour and cloud liquid water profiles derived from the Integrated Profiling Technique (IPT). As an example, we will show the approach for 3 March 2012. The calculated downward flux at the surface for this day compares well with measurements made at the Cabauw Baseline Surface Radiation Network (BSRN) site (51.97°N, 4.93°E), the Netherlands.

  10. Flux effect analysis in WWER-440 reactor pressure vessel steels

    NASA Astrophysics Data System (ADS)

    Kryukov, A.; Blagoeva, D.; Debarberis, L.

    2013-11-01

    The results of long term research programme concerning the determination of irradiation embrittlement dependence on fast neutron flux for WWER-440 reactor pressure vessel steels before and after annealing are presented in this paper. The study of flux effect was carried out on commercial WWER-440 steels which differ significantly in phosphorous (0.013-0.036 wt%) and copper (0.08-0.20 wt%) contents. All specimens were irradiated in surveillance channel positions under similar conditions at high ˜4 × 1012 сm-2 s-1 and low ˜6 × 1011 сm-2 s-1 fluxes (E > 0.5 MeV) at a temperature of 270 °С. The radiation embrittlement was evaluated by transition temperature shift on the basis of Charpy specimens test results. In case of low flux, the measured Tk shifts could be 25-50 °C bigger than the Tk shifts obtained from high flux data. A significant flux effect is observed in WWER-440 reactor pressure vessel steels with higher copper content (>0.13 wt%).

  11. Quantitative flux analysis reveals folate-dependent NADPH production

    NASA Astrophysics Data System (ADS)

    Fan, Jing; Ye, Jiangbin; Kamphorst, Jurre J.; Shlomi, Tomer; Thompson, Craig B.; Rabinowitz, Joshua D.

    2014-06-01

    ATP is the dominant energy source in animals for mechanical and electrical work (for example, muscle contraction or neuronal firing). For chemical work, there is an equally important role for NADPH, which powers redox defence and reductive biosynthesis. The most direct route to produce NADPH from glucose is the oxidative pentose phosphate pathway, with malic enzyme sometimes also important. Although the relative contribution of glycolysis and oxidative phosphorylation to ATP production has been extensively analysed, similar analysis of NADPH metabolism has been lacking. Here we demonstrate the ability to directly track, by liquid chromatography-mass spectrometry, the passage of deuterium from labelled substrates into NADPH, and combine this approach with carbon labelling and mathematical modelling to measure NADPH fluxes. In proliferating cells, the largest contributor to cytosolic NADPH is the oxidative pentose phosphate pathway. Surprisingly, a nearly comparable contribution comes from serine-driven one-carbon metabolism, in which oxidation of methylene tetrahydrofolate to 10-formyl-tetrahydrofolate is coupled to reduction of NADP+ to NADPH. Moreover, tracing of mitochondrial one-carbon metabolism revealed complete oxidation of 10-formyl-tetrahydrofolate to make NADPH. As folate metabolism has not previously been considered an NADPH producer, confirmation of its functional significance was undertaken through knockdown of methylenetetrahydrofolate dehydrogenase (MTHFD) genes. Depletion of either the cytosolic or mitochondrial MTHFD isozyme resulted in decreased cellular NADPH/NADP+ and reduced/oxidized glutathione ratios (GSH/GSSG) and increased cell sensitivity to oxidative stress. Thus, although the importance of folate metabolism for proliferating cells has been long recognized and attributed to its function of producing one-carbon units for nucleic acid synthesis, another crucial function of this pathway is generating reducing power.

  12. Partial inhibition and bilevel optimization in flux balance analysis

    PubMed Central

    2013-01-01

    Motivation Within Flux Balance Analysis, the investigation of complex subtasks, such as finding the optimal perturbation of the network or finding an optimal combination of drugs, often requires to set up a bilevel optimization problem. In order to keep the linearity and convexity of these nested optimization problems, an ON/OFF description of the effect of the perturbation (i.e. Boolean variable) is normally used. This restriction may not be realistic when one wants, for instance, to describe the partial inhibition of a reaction induced by a drug. Results In this paper we present a formulation of the bilevel optimization which overcomes the oversimplified ON/OFF modeling while preserving the linear nature of the problem. A case study is considered: the search of the best multi-drug treatment which modulates an objective reaction and has the minimal perturbation on the whole network. The drug inhibition is described and modulated through a convex combination of a fixed number of Boolean variables. The results obtained from the application of the algorithm to the core metabolism of E.coli highlight the possibility of finding a broader spectrum of drug combinations compared to a simple ON/OFF modeling. Conclusions The method we have presented is capable of treating partial inhibition inside a bilevel optimization, without loosing the linearity property, and with reasonable computational performances also on large metabolic networks. The more fine-graded representation of the perturbation allows to enlarge the repertoire of synergistic combination of drugs for tasks such as selective perturbation of cellular metabolism. This may encourage the use of the approach also for other cases in which a more realistic modeling is required. PMID:24286232

  13. Observational & modeling analysis of surface heat and moisture fluxes

    SciTech Connect

    Smith, E.

    1995-09-01

    An observational and modeling study was conducted to help assess how well current GCMs are predicting surface fluxes under the highly variable cloudiness and flow conditions characteristic of the real atmosphere. The observational data base for the study was obtained from a network of surface flux stations operated during the First ISLSCP Field Experiment (FIFE). The study included examination of a surface-driven secondary circulation in the boundary layer resulting from a persistent cross-site gradient in soil moisture, to demonstrate the sensitivity of boundary layer dynamics to heterogeneous surface fluxes, The performance of a biosphere model in reproducing the measured surface fluxes was evaluated with and without the use of satellite retrieval of three key canopy variables with RMS uncertainties commensurate with those of the measurements themselves. Four sensible heat flux closure schemes currently being used in GCMs were then evaluated against the FIFE observations. Results indicate that the methods by which closure models are calibrated lead to exceedingly large errors when the schemes are applied to variable boundary layer conditions. 4 refs., 2 figs.

  14. Numerical Analysis of a Radiant Heat Flux Calibration System

    NASA Technical Reports Server (NTRS)

    Jiang, Shanjuan; Horn, Thomas J.; Dhir, V. K.

    1998-01-01

    A radiant heat flux gage calibration system exists in the Flight Loads Laboratory at NASA's Dryden Flight Research Center. This calibration system must be well understood if the heat flux gages calibrated in it are to provide useful data during radiant heating ground tests or flight tests of high speed aerospace vehicles. A part of the calibration system characterization process is to develop a numerical model of the flat plate heater element and heat flux gage, which will help identify errors due to convection, heater element erosion, and other factors. A 2-dimensional mathematical model of the gage-plate system has been developed to simulate the combined problem involving convection, radiation and mass loss by chemical reaction. A fourth order finite difference scheme is used to solve the steady state governing equations and determine the temperature distribution in the gage and plate, incident heat flux on the gage face, and flat plate erosion. Initial gage heat flux predictions from the model are found to be within 17% of experimental results.

  15. Analysis of uncertainty in the nitrous oxide flux values measured using a coupled eddy covariance - flux gradient technique

    NASA Astrophysics Data System (ADS)

    Mukherjee, S.; Sturman, A.; Mcmillan, A. M.; Harvey, M.; Zawar-Reza, P.

    2012-12-01

    An error analysis has been performed for a typical flux gradient - eddy covariance system setup for N2O flux measurement using data from two field campaigns. The diffusivity parameters were estimated using both a thermal and parameterization approach. Algebraic relationships of random relative error have been derived between the diffusivity parameters and the surface layer stability and turbulence terms. Based on these relationships, a Monte Carlo type analysis was performed to explore the dependency of the diffusivity terms on their contributing factors during the stable and unstable atmospheric conditions in the surface layer. The total relative uncertainty in the flux values due to errors in diffusivity terms and concentration gradients were then estimated. It was found that the mean uncertainty in the diffusivity parameter derived using the thermal method is higher (11%) than the parameterization method (≈7%), irrespective of stability. However, depending on the initial uncertainty among the surface layer variables, the maximum uncertainty can vary between 0-80% and 0-37% for the thermal and parameterization methods irrespective of stability. These maximum variations were obtained from the synthetic population of the random errors of the diffusivity parameters. The probability density function of the error anomaly of the diffusivity term from parameterization method was found to have higher kurtosis during unstable atmosphere, whereas marginally higher positive skewness was observed in the relative error term of the same diffusivity parameter during the unstable condition. Errors in the concentration gradients were estimated based on the minimum resolvable estimates from the gas analyzer and the associated random errors were found to be 6% and 8% for unstable and stable conditions. Finally, the mean total error in the N2O flux values was found to be approximately of the order of 9% and 11% for the parameterization method for unstable and stable conditions

  16. Dimensional Analysis of Thermoelectric Modules Under Constant Heat Flux

    NASA Astrophysics Data System (ADS)

    Suzuki, Ryosuke O.; Fujisaka, Takeyuki; Ito, Keita O.; Meng, Xiangning; Sui, Hong-Tao

    2015-01-01

    Thermoelectric power generation is examined in the case of radiative heating. A constant heat flux is assumed in addition to consideration of the Seebeck effect, Peltier effect, and Joule heating with temperature-dependent material properties. Numerical evaluations are conducted using a combination of the finite-volume method and an original simultaneous solver for the heat transfer, thermoelectric, and electric transportation phenomena. Comparison with experimental results shows that the new solver could work well in the numerical calculations. The calculations predict that the Seebeck effect becomes larger for longer thermoelectric elements because of the larger temperature difference. The heat transfer to the cold surface is critical to determine the junction temperatures under a constant heat flux from the hot surface. The negative contribution from Peltier cooling and heating can be minimized when the current is smaller for longer elements. Therefore, a thicker TE module can generate more electric power even under a constant heat flux.

  17. COMPLETE-MFA: complementary parallel labeling experiments technique for metabolic flux analysis.

    PubMed

    Leighty, Robert W; Antoniewicz, Maciek R

    2013-11-01

    We have developed a novel approach for measuring highly accurate and precise metabolic fluxes in living cells, termed COMPLETE-MFA, short for complementary parallel labeling experiments technique for metabolic flux analysis. The COMPLETE-MFA method is based on combined analysis of multiple isotopic labeling experiments, where the synergy of using complementary tracers greatly improves the precision of estimated fluxes. In this work, we demonstrate the COMPLETE-MFA approach using all singly labeled glucose tracers, [1-(13)C], [2-(13)C], [3-(13)C], [4-(13)C], [5-(13)C], and [6-(13)C]glucose to determine precise metabolic fluxes for wild-type Escherichia coli. Cells were grown in six parallel cultures on defined medium with glucose as the only carbon source. Mass isotopomers of biomass amino acids were measured by gas chromatography-mass spectrometry (GC-MS). The data from all six experiments were then fitted simultaneously to a single flux model to determine accurate intracellular fluxes. We obtained a statistically acceptable fit with more than 300 redundant measurements. The estimated flux map is the most precise flux result obtained thus far for E. coli cells. To our knowledge, this is the first time that six isotopic labeling experiments have been successfully integrated for high-resolution (13)C-flux analysis.

  18. Analysis of Surface Fluxes at Eureka Climate Observatory in Arctic

    NASA Astrophysics Data System (ADS)

    Grachev, Andrey; Albee, Robert; Fairall, Christopher; Hare, Jeffrey; Persson, Ola; Uttal, Taneil

    2010-05-01

    The Arctic region is experiencing unprecedented changes associated with increasing average temperatures (faster than the pace of the globally-averaged increase) and significant decreases in both the areal extent and thickness of the Arctic pack ice. These changes are early warning signs of shifts in the global climate system that justifies increased scientific focus on this region. The increase in atmospheric carbon dioxide has raised concerns worldwide about future climate change. Recent studies suggest that huge stores of carbon dioxide (and other climate relevant compounds) locked up in Arctic soils could be unexpectedly released due to global warming. Observational evidence suggests that atmospheric energy fluxes are a major contributor to the decrease of the Arctic pack ice, seasonal land snow cover and the warming of the surrounding land areas and permafrost layers. To better understand the atmosphere-surface exchange mechanisms, improve models, and to diagnose climate variability in the Arctic, accurate measurements are required of all components of the net surface energy budget and the carbon dioxide cycle over representative areas and over multiple years. In this study we analyze variability of turbulent fluxes including water vapor and carbon dioxide transfer based on long-term measurements made at Eureka observatory (80.0 N, 85.9 W) located near the coast of the Arctic Ocean (Canadian territory of Nunavut). Turbulent fluxes and mean meteorological data are continuously measured and reported hourly at various levels on a 10-m flux tower. Sonic anemometers are located at 3 and 8 m heights while high-speed Licor 7500 infrared gas analyzer (water moisture and carbon dioxide measurements) at 7.5 m height. According to our data, that the sensible heat flux, carbon dioxide and water vapor fluxes exhibited clear diurnal cycles in Arctic summer. This behavior is similar to the diurnal variation of the fluxes in mid-latitudes during the plants growing season, with

  19. Synthesis: comparing effects of resource and consumer fluxes into recipient food webs using meta-analysis.

    PubMed

    Allen, Daniel C; Wesner, Jeff S

    2016-03-01

    Here we synthesize empirical research using meta-analysis to compare how consumer and resource fluxes affect recipient food webs. We tested the following hypotheses: (H1) The direct effects of resource fluxes (bottom-up) should be stronger than the direct effects of consumer fluxes (top-down), because resource fluxes are permanent (do not return to the food web in which they were produced) but consumer fluxes may not be (consumers can leave). (H2) Following H1, the indirect effects should attenuate (weaken) more quickly for consumer fluxes than for resource fluxes due to their direct effects being weaker, (H3) The effects of resource fluxes should be stronger when recipient food webs are in different ecosystems than donor food webs due to differences in elevation that accompany cross-ecosystem food web interfaces, often increasing flux quantity due to gravity, while the effects of consumer fluxes should be stronger when donor and recipient food webs are in the same ecosystem as they should more easily assimilate into the recipient food web. We found no differences in the magnitude of bottom-up and top-down direct effects for resource and consumer fluxes, but top-down direct effects were 122% stronger than top-down indirect effects. Indirect effects of prey and predator fluxes quickly attenuated while indirect effects of non-prey resource and herbivore fluxes did not, as the overall direct effects of prey and predator fluxes were 123% and 163% stronger than their indirect effects, respectively. This result suggests that the magnitude of indirect effects decrease as the trophic level of resource and consumer fluxes increases, and also contrasts with results from studies showing in situ top-down indirect effects are stronger than in situ bottom-up indirect effects. We found that resource and consumer flux effect sizes were similar when they occurred between ecosystems, but when they occurred within ecosystems predator flux effects were 107% stronger than nutrient flux

  20. Quantifying plant phenotypes with isotopic labeling and metabolic flux analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Analyses of metabolic flux using stable isotopes in plants have traditionally been restricted to tissues with presumed homogeneous cell populations such as developing seeds, cell suspensions, or cultured roots and root tips. It is now possible to describe these and other more complex tissues such a...

  1. An analysis of flux-split algorithms for Euler's equations with real gases

    NASA Technical Reports Server (NTRS)

    Grossman, B.; Walters, R. W.

    1987-01-01

    An analysis of flux-splitting procedures for the solution of Euler's equations with real gas effects is presented. An alternative real-gas flux-splitting is derived which can easily be implemented into existing codes. This approach, which takes the form of an 'equivalent' gamma representation is not an ad hoc model, but is based on theoretical considerations. Details of this method with the Steger-Warming and Van Leer flux vector splittings and the Roe flux-difference splitting are given. Applications of the method to several high Mach number, high temperature flows are presented for one and two space dimensions.

  2. Theoretical analysis of integral neutron transport equation using collision probability method with quadratic flux approach

    SciTech Connect

    Shafii, Mohammad Ali Meidianti, Rahma Wildian, Fitriyani, Dian; Tongkukut, Seni H. J.; Arkundato, Artoto

    2014-09-30

    Theoretical analysis of integral neutron transport equation using collision probability (CP) method with quadratic flux approach has been carried out. In general, the solution of the neutron transport using the CP method is performed with the flat flux approach. In this research, the CP method is implemented in the cylindrical nuclear fuel cell with the spatial of mesh being conducted into non flat flux approach. It means that the neutron flux at any point in the nuclear fuel cell are considered different each other followed the distribution pattern of quadratic flux. The result is presented here in the form of quadratic flux that is better understanding of the real condition in the cell calculation and as a starting point to be applied in computational calculation.

  3. Hierarchical organization of fluxes in Escherichia coli metabolic network: using flux coupling analysis for understanding the physiological properties of metabolic genes.

    PubMed

    Hosseini, Zhaleh; Marashi, Sayed-Amir

    2015-05-01

    Flux coupling analysis is a method for investigating the connections between reactions of metabolic networks. Here, we construct the hierarchical flux coupling graph for the reactions of the Escherichia coli metabolic network model to determine the level of each reaction in the graph. This graph is constructed based on flux coupling analysis of metabolic network: if zero flux through reaction a results in zero flux through reaction b (and not vice versa), then reaction a is located at the top of reaction b in the flux coupling graph. We show that in general, more important, older and essential reactions are located at the top of the graph. Strikingly, genes corresponding to these reactions are found to be the genes which are most regulated.

  4. Analysis of microfiltration performance with constant flux processing of secondary effluent.

    PubMed

    Parameshwaran, K; Fane, A G; Cho, B D; Kim, K J

    2001-12-01

    This study involves the microfiltration (MF) of secondary effluent from a sequencing batch reactor processing industrial waste. The MF unit was a hollow fibre module with gas backwash capability, and operated with pumped permeate (controlled flux) and dead-end, crossflow or intermittent feed. The results showed that crossflow had no effect on flux and that intermittent dead-end filtration was less productive than non-intermittent operation. For dead-end filtration the cycle-time between gas backwashes depends very significantly on the imposed flux (varying from about 100 min at 30 L/m2 h to about 5 min at 90 L, m2 h) and the feed solids content. Optimal operation has to balance operating (energy for backwash) costs and the capital (membrane area) costs. Cost analysis based on capital and energy costs indicates that for lower energy cost the unit needs to be operated at lower imposed flux but to minimise total cost it is necessary to operate the unit above 60 L/m2 h imposed flux depending on the maximum transmembrane pressure (TMP) allowed before back washing. Further analysis of TMP profiles showed that membrane resistance increased over time towards a maximum, which tended to increase with imposed flux. This implies more frequent chemical cleaning for high flux operation. Specific cake resistances were deduced from the profiles and indicated cake compression at higher flux and larger maximum TMP. Results of long-term trials are also reported. Water quality analysis shows consistent quality of permeate

  5. Error analysis of flux limiter schemes at extrema

    NASA Astrophysics Data System (ADS)

    Kriel, A. J.

    2017-01-01

    Total variation diminishing (TVD) schemes have been an invaluable tool for the solution of hyperbolic conservation laws. One of the major shortcomings of commonly used TVD methods is the loss of accuracy near extrema. Although large amounts of anti-diffusion usually benefit the resolution of discontinuities, a balanced limiter such as Van Leer's performs better at extrema. Reliable criteria, however, for the performance of a limiter near extrema are not readily apparent. This work provides theoretical quantitative estimates for the local truncation errors of flux limiter schemes at extrema for a uniform grid. Moreover, the component of the error attributed to the flux limiter was obtained. This component is independent of the problem and grid spacing, and may be considered a property of the limiter that reflects the performance at extrema. Numerical test problems validate the results.

  6. Insights into primary metabolism in oilseeds from labeling and flux analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Labeling investigations along with metabolic flux analysis have enabled quantification of important cellular phenotypes. These descriptions have documented uses of enzymes in unique ways and characterized the contributions of pathways to oil, protein and carbohydrate compositions in seeds. The diffe...

  7. Analysis of surface currents on the CTX mesh flux conserver

    SciTech Connect

    Wright, B.L.

    1984-02-01

    The use of discharge cleaning and a mesh structure for the flux conserver have led to hotter, less resistive, spheromak configurations in the CTX experiment. Achievement of these conditions has been accompanied by the appearance of oscillations - most notably seen on magnetic probe signals - that were previously not present. These oscillations are observed both during the sustainment (V/sub gun/ not equal to 0) and the decaying (V/sub gun/ = 0) phases of the discharge and are attributed to the presence of internal kink modes driven by the departure of the plasma current distribution from the Taylor state: j/B = const. Computations suggest that an n-1 mode becomes unstable when j/B increases away from the magnetic axis (sustained spheromak) whereas an n=2 mode becomes unstable for radially decreasing j/B (decaying spheromak). The results reported are from an 80-cm-diameter mesh flux conserver constructed form 0.5''-diameter copper stock. It is of the same oblate rooftop design as used for previous work with a dimension of 40 cm from front to back.

  8. Integrated metabolic flux and omics analysis of Synechocystis sp. PCC 6803 under mixotrophic and photoheterotrophic conditions.

    PubMed

    Nakajima, Tsubasa; Kajihata, Shuichi; Yoshikawa, Katsunori; Matsuda, Fumio; Furusawa, Chikara; Hirasawa, Takashi; Shimizu, Hiroshi

    2014-09-01

    Cyanobacteria have flexible metabolic capability that enables them to adapt to various environments. To investigate their underlying metabolic regulation mechanisms, we performed an integrated analysis of metabolic flux using transcriptomic and metabolomic data of a cyanobacterium Synechocystis sp. PCC 6803, under mixotrophic and photoheterotrophic conditions. The integrated analysis indicated drastic metabolic flux changes, with much smaller changes in gene expression levels and metabolite concentrations between the conditions, suggesting that the flux change was not caused mainly by the expression levels of the corresponding genes. Under photoheterotrophic conditions, created by the addition of the photosynthesis inhibitor atrazine in mixotrophic conditions, the result of metabolic flux analysis indicated the significant repression of carbon fixation and the activation of the oxidative pentose phosphate pathway (PPP). Moreover, we observed gluconeogenic activity of upstream of glycolysis, which enhanced the flux of the oxidative PPP to compensate for NADPH depletion due to the inhibition of the light reaction of photosynthesis. 'Omics' data suggested that these changes were probably caused by the repression of the gap1 gene, which functions as a control valve in the metabolic network. Since metabolic flux is the outcome of a complicated interplay of cellular components, integrating metabolic flux with other 'omics' layers can identify metabolic changes and narrow down these regulatory mechanisms more effectively.

  9. Multiple Triangulation Analysis: another approach to determine the orientation of magnetic flux ropes

    NASA Astrophysics Data System (ADS)

    Zhou, X.-Z.; Zong, Q.-G.; Pu, Z. Y.; Fritz, T. A.; Dunlop, M. W.; Shi, Q. Q.; Wang, J.; Wei, Y.

    2006-07-01

    Another approach (Multiple Triangulation Analysis, MTA) is presented to determine the orientation of magnetic flux rope, based on 4-point measurements. A 2-D flux rope model is used to examine the accuracy of the MTA technique in a theoretical way. It is found that the precision of the estimated orientation is dependent on both the spacecraft separation and the constellation path relative to the flux rope structure. However, the MTA error range can be shown to be smaller than that of the traditional MVA technique. As an application to real Cluster data, several flux rope events on 26 January 2001 are analyzed using MTA, to obtain their orientations. The results are compared with the ones obtained by several other methods which also yield flux rope orientation. The estimated axis orientations are shown to be fairly close, suggesting the reliability of the MTA method.

  10. Genome-scale metabolic flux analysis of Streptomyces lividans growing on a complex medium.

    PubMed

    D'Huys, Pieter-Jan; Lule, Ivan; Vercammen, Dominique; Anné, Jozef; Van Impe, Jan F; Bernaerts, Kristel

    2012-09-15

    Constraint-based metabolic modeling comprises various excellent tools to assess experimentally observed phenotypic behavior of micro-organisms in terms of intracellular metabolic fluxes. In combination with genome-scale metabolic networks, micro-organisms can be investigated in much more detail and under more complex environmental conditions. Although complex media are ubiquitously applied in industrial fermentations and are often a prerequisite for high protein secretion yields, such multi-component conditions are seldom investigated using genome-scale flux analysis. In this paper, a systematic and integrative approach is presented to determine metabolic fluxes in Streptomyces lividans TK24 grown on a nutritious and complex medium. Genome-scale flux balance analysis and randomized sampling of the solution space are combined to extract maximum information from exometabolome profiles. It is shown that biomass maximization cannot predict the observed metabolite production pattern as such. Although this cellular objective commonly applies to batch fermentation data, both input and output constraints are required to reproduce the measured biomass production rate. Rich media hence not necessarily lead to maximum biomass growth. To eventually identify a unique intracellular flux vector, a hierarchical optimization of cellular objectives is adopted. Out of various tested secondary objectives, maximization of the ATP yield per flux unit returns the closest agreement with the maximum frequency in flux histograms. This unique flux estimation is hence considered as a reasonable approximation for the biological fluxes. Flux maps for different growth phases show no active oxidative part of the pentose phosphate pathway, but NADPH generation in the TCA cycle and NADPH transdehydrogenase activity are most important in fulfilling the NADPH balance. Amino acids contribute to biomass growth by augmenting the pool of available amino acids and by boosting the TCA cycle, particularly

  11. Metabolic flux and nodes control analysis of brewer's yeasts under different fermentation temperature during beer brewing.

    PubMed

    Yu, Zhimin; Zhao, Haifeng; Zhao, Mouming; Lei, Hongjie; Li, Huiping

    2012-12-01

    The aim of this work was to further investigate the glycolysis performance of lager and ale brewer's yeasts under different fermentation temperature using a combined analysis of metabolic flux, glycolytic enzyme activities, and flux control. The results indicated that the fluxes through glycolytic pathway decreased with the change of the fermentation temperature from 15 °C to 10 °C, which resulted in the prolonged fermentation times. The maximum activities (V (max)) of hexokinase (HK), phosphofructokinase (PFK), and pyruvate kinase (PK) at key nodes of glycolytic pathway decreased with decreasing fermentation temperature, which was estimated to have different control extent (22-84 %) on the glycolytic fluxes in exponential or flocculent phase. Moreover, the decrease of V (max) of PFK or PK displayed the crucial role in down-regulation of flux in flocculent phase. In addition, the metabolic state of ale strain was more sensitive to the variation of temperature than that of lager strain. The results of the metabolic flux and nodes control analysis in brewer's yeasts under different fermentation temperature may provide an alternative approach to regulate glycolytic flux by changing V (max) and improve the production efficiency and beer quality.

  12. [Breeding of Actinobacillus succiniogenes mutants with improved succinate production based on metabolic flux analysis].

    PubMed

    Pan, Lijun; Li, Xingjiang; Jiang, Shaotong; Wei, Zhaojun; Chen, Xiaohui; Cai, Licheng; Wang, Hefeng; Jiang, Jijun

    2008-09-01

    It is very important to obtain high yield mutant strains on the base of metabolic flux analysis of Actinobacillus succinogenes S.JST for the industrial bioconversion of succinic acid. The metabolic pathway was analized at first and the flux of the metabolic networks was calculated by matrix. In order to decrease acetic acid flux, the strains mutated by soft X-ray of synchronous radiation were screened on the plates with high concentration of fluoroacetic acid. For decreasing the metabolic flux of ethanol the site-directed mutagenesis was carried out for the reduction of alcohol dehydrogenase(Adh) specific activity. Then the enzyme activity determination and the gene sequence analysis of the mutant strain was compared with those of the parent strain. Metabolic flux analysis of the parent strain indicated that the flux of succinic acid was 1.78(mmol/g/h) and that the flux of acetic acid and ethanol were 0.60 (mmol/g/h) and 1.04( mmol/g/h), respectively. Meanwhile the metabolic pathway analysis showed that the ethanol metabolism enhanced the lacking of H electron donor during the synthesis of succinic acid and that the succinic acid flux was weakened by the metabolism of byproducts ethanol and acetic acid. Compared with the parent strain, the acetic acid flux of anti-fluoroacetic mutant strain S.JST1 was 0.024 (mmol/g/h), decreasing by 96%. Then the enzyme determination showed that the specific activity unit of phosphotransacetylase(Pta) decreased from 602 to 74 and a mutated site was founded in the pta gene of the mutant strain S.JST1. Compared with that of the parent strain S.JST1 the ethanol flux of adh-site-directed mutant strain S.JST2 was 0.020 (mmol/g/h), decreasing by 98%. Then the enzyme determination showed that the specific activity unit of Adh decreased from 585 to 62 and the yield of end product succinic acid was 65.7 (g/L). The interdiction of Adh and Pta decreased the metabolism of byproducts and the H electron donor was well balanced, thus the succinic

  13. RELATIONSHIP BETWEEN MASS FLUX REDUCTION AND SOURCE-ZONE MASS REMOVAL: ANALYSIS OF FIELD DATA

    PubMed Central

    DiFilippo, Erica L.

    2010-01-01

    The magnitude of contaminant mass flux reduction associated with a specific amount of contaminant mass removed is a key consideration for evaluating the effectiveness of a source-zone remediation effort. Thus, there is great interest in characterizing, estimating, and predicting relationships between mass flux reduction and mass removal. Published data collected for several field studies were examined to evaluate relationships between mass flux reduction and source-zone mass removal. The studies analyzed herein represent a variety of source-zone architectures, immiscible-liquid compositions, and implemented remediation technologies. There are two general approaches to characterizing the mass-flux-reduction/mass-removal relationship, end-point analysis and time-continuous analysis. End-point analysis, based on comparing masses and mass fluxes measured before and after a source-zone remediation effort, was conducted for 21 remediation projects. Mass removals were greater than 60% for all but three of the studies. Mass flux reductions ranging from slightly less than to slightly greater than one-to-one were observed for the majority of the sites. However, these single-snapshot characterizations are limited in that the antecedent behavior is indeterminate. Time-continuous analysis, based on continuous monitoring of mass removal and mass flux, was performed for two sites, both for which data were obtained under water-flushing conditions. The reductions in mass flux were significantly different for the two sites (90% vs. ~8%) for similar mass removals (~40%). These results illustrate the dependence of the mass-flux-reduction/mass-removal relationship on source-zone architecture and associated mass-transfer processes. Minimal mass flux reduction was observed for a system wherein mass removal was relatively efficient (ideal mass transfer and displacement). Conversely, a significant degree of mass flux reduction was observed for a site wherein mass removal was inefficient

  14. INCA: a computational platform for isotopically non-stationary metabolic flux analysis.

    PubMed

    Young, Jamey D

    2014-05-01

    13C flux analysis studies have become an essential component of metabolic engineering research. The scope of these studies has gradually expanded to include both isotopically steady-state and transient labeling experiments, the latter of which are uniquely applicable to photosynthetic organisms and slow-to-label mammalian cell cultures. Isotopomer network compartmental analysis (INCA) is the first publicly available software package that can perform both steady-state metabolic flux analysis and isotopically non-stationary metabolic flux analysis. The software provides a framework for comprehensive analysis of metabolic networks using mass balances and elementary metabolite unit balances. The generation of balance equations and their computational solution is completely automated and can be performed on networks of arbitrary complexity.

  15. Dynamic metabolic flux analysis--tools for probing transient states of metabolic networks.

    PubMed

    Antoniewicz, Maciek R

    2013-12-01

    Computational approaches for analyzing dynamic states of metabolic networks provide a practical framework for design, control, and optimization of biotechnological processes. In recent years, two promising modeling approaches have emerged for characterizing transients in cellular metabolism, dynamic metabolic flux analysis (DMFA), and dynamic flux balance analysis (DFBA). Both approaches combine metabolic network analysis based on pseudo steady-state (PSS) assumption for intracellular metabolism with dynamic models for extracellular environment. One strategy to capture dynamics is by combining network analysis with a kinetic model. Predictive models are thus established that can be used to optimize bioprocessing conditions and identify useful genetic manipulations. Alternatively, by combining network analysis with methods for analyzing extracellular time-series data, transients in intracellular metabolic fluxes can be determined and applied for process monitoring and control.

  16. Subchannel void fraction prediction via drift-flux analysis

    SciTech Connect

    Khan, H.J.; Kosaly, G.

    1986-10-01

    Void fraction calculations have been performed using the subchannel drift-flux code CANAL. Using void and flow distributions in rod bundle geometry, a value of C/sub 0/ has been estimated for bundle-averaged void fraction calculation in one-dimensional approximations. Successful prediction of the average void fraction is observed for the annular rod bundle geometry of the FRIGG experiment. In order to perform subchannel void fraction calculation, a C/sub 0/ model has been developed for one-dimensional subchannel geometry. The implicit form of the C/sub 0/ model developed accounts for void and flow conditions in the adjacent subchannels existing at the common interfaces, i.e., at the gap spacing between the subchannels. It appears that the magnitude of C/sub 0/ varies between subchannels (annular rings of FRIGG geometry) but remains almost constant within each subchannel. Good agreement is observed between prediction and data for subchannel void fractions in axially uniform and nonuniform heated rod bundles.

  17. Efficient Modeling of MS/MS Data for Metabolic Flux Analysis.

    PubMed

    Tepper, Naama; Shlomi, Tomer

    2015-01-01

    Metabolic flux analysis (MFA) is a widely used method for quantifying intracellular metabolic fluxes. It works by feeding cells with isotopic labeled nutrients, measuring metabolite isotopic labeling, and computationally interpreting the measured labeling data to estimate flux. Tandem mass-spectrometry (MS/MS) has been shown to be useful for MFA, providing positional isotopic labeling data. Specifically, MS/MS enables the measurement of a metabolite tandem mass-isotopomer distribution, representing the abundance in which certain parent and product fragments of a metabolite have different number of labeled atoms. However, a major limitation in using MFA with MS/MS data is the lack of a computationally efficient method for simulating such isotopic labeling data. Here, we describe the tandemer approach for efficiently computing metabolite tandem mass-isotopomer distributions in a metabolic network, given an estimation of metabolic fluxes. This approach can be used by MFA to find optimal metabolic fluxes, whose induced metabolite labeling patterns match tandem mass-isotopomer distributions measured by MS/MS. The tandemer approach is applied to simulate MS/MS data in a small-scale metabolic network model of mammalian methionine metabolism and in a large-scale metabolic network model of E. coli. It is shown to significantly improve the running time by between two to three orders of magnitude compared to the state-of-the-art, cumomers approach. We expect the tandemer approach to promote broader usage of MS/MS technology in metabolic flux analysis.

  18. Elucidating acetogenic H2 consumption in dark fermentation using flux balance analysis.

    PubMed

    Lalman, Jerald A; Chaganti, Subba Rao; Moon, Chungman; Kim, Dong-Hoon

    2013-10-01

    In this study, a flux balance analysis (FBA) was adopted to estimate the activity of acetogenic H2-consuming reaction. Experimental data at different substrate concentrations of 10, 20, and 30 g COD/L showing the lowest, medium, and highest H2 yields, respectively, were used in the FBA to calculate the fluxes. It was interesting to note that the hydrogenase activity based on R12 (2Fd(+)+2H(+)→2Fd(2+)+H2, ferredoxin (Fd)) flux was most active at 10 g COD/L. The flux of R17 (4H2+2CO2→CH3COOH), a mechanism for reutilizing produced H2, increased in steps of 0.030, 0.119, and 0.467 as the substrate concentration decreased. Contradictory to our general understanding, acetate production found to have a negligible or even negative effect on the final H2 yield in dark fermentation.

  19. Analysis of field measurements of carbon dioxide and water vapor fluxes

    NASA Technical Reports Server (NTRS)

    Verma, Shashi B.

    1991-01-01

    Analysis of the field measurements of carbon dioxide and water vapor fluxes is discussed. These data were examined in conjunction with reflectance obtained from helicopter mounted Modular Multiband Radiometer. These measurements are representative of the canopy scale (10 to 100 m)(exp 2) and provide a good basis for investigating the hypotheses/relationship potentially useful in remote sensing applications. All the micrometeorological data collected during FIFE-89 were processed and fluxes of CO2, water vapor, and sensible heat were calculated. Soil CO2 fluxes were also estimated. Employing these soil CO2 flux values, in conjunction with micrometeorological measurements, canopy photosynthesis is being estimated. A biochemical model of leaf photosynthesis was adapted to the prairie vegetation. The modeled leaf photosynthesis rates were scaled up to the canopy level. This model and a multiplicative stomatal conductance model are also used to calculate canopy conductance.

  20. An improved path flux analysis with multi generations method for mechanism reduction

    NASA Astrophysics Data System (ADS)

    Wang, Wei; Gou, Xiaolong

    2016-03-01

    An improved path flux analysis with a multi generations (IMPFA) method is proposed to eliminate unimportant species and reactions, and to generate skeletal mechanisms. The production and consumption path fluxes of each species at multiple reaction paths are calculated and analysed to identify the importance of the species and of the elementary reactions. On the basis of the indexes of each reaction path of the first, second, and third generations, the improved path flux analysis with two generations (IMPFA2) and improved path flux analysis with three generations (IMPFA3) are used to generate skeletal mechanisms that contain different numbers of species. The skeletal mechanisms are validated in the case of homogeneous autoignition and perfectly stirred reactor of methane and n-decane/air mixtures. Simulation results of the skeletal mechanisms generated by IMPFA2 and IMPFA3 are compared with those obtained by path flux analysis (PFA) with two and three generations, respectively. The comparisons of ignition delay times, final temperatures, and temperature dependence on flow residence time show that the skeletal mechanisms generated by the present IMPFA method are more accurate than those obtained by the PFA method, with almost the same number of species under a range of initial conditions. By considering the accuracy and computational efficiency, when using the IMPFA (or PFA) method, three generations may be the best choice for the reduction of large-scale detailed chemistry.

  1. The relative importance of head, flux, and prior information in hydraulic tomography analysis

    NASA Astrophysics Data System (ADS)

    Michael Tso, Chak-Hau; Zha, Yuanyuan; Jim Yeh, Tian-Chyi; Wen, Jet-Chau

    2016-01-01

    Using cross-correlation analysis, we demonstrate that flux measurements at observation locations during hydraulic tomography (HT) surveys carry nonredundant information about heterogeneity that are complementary to head measurements at the same locations. We then hypothesize that a joint interpretation of head and flux data, even when the same observation network as head has been used, can enhance the resolution of HT estimates. Subsequently, we use numerical experiments to test this hypothesis and investigate the impact of flux conditioning and prior information (such as correlation lengths and initial mean models (i.e., uniform mean or distributed means)) on the HT estimates of a nonstationary, layered medium. We find that the addition of flux conditioning to HT analysis improves the estimates in all of the prior models tested. While prior information on geologic structures could be useful, its influence on the estimates reduces as more nonredundant data (i.e., flux) are used in the HT analysis. Lastly, recommendations for conducting HT surveys and analysis are presented.

  2. Bayesian Statistical Analysis Applied to NAA Data for Neutron Flux Spectrum Determination

    NASA Astrophysics Data System (ADS)

    Chiesa, D.; Previtali, E.; Sisti, M.

    2014-04-01

    In this paper, we present a statistical method, based on Bayesian statistics, to evaluate the neutron flux spectrum from the activation data of different isotopes. The experimental data were acquired during a neutron activation analysis (NAA) experiment [A. Borio di Tigliole et al., Absolute flux measurement by NAA at the Pavia University TRIGA Mark II reactor facilities, ENC 2012 - Transactions Research Reactors, ISBN 978-92-95064-14-0, 22 (2012)] performed at the TRIGA Mark II reactor of Pavia University (Italy). In order to evaluate the neutron flux spectrum, subdivided in energy groups, we must solve a system of linear equations containing the grouped cross sections and the activation rate data. We solve this problem with Bayesian statistical analysis, including the uncertainties of the coefficients and the a priori information about the neutron flux. A program for the analysis of Bayesian hierarchical models, based on Markov Chain Monte Carlo (MCMC) simulations, is used to define the problem statistical model and solve it. The energy group fluxes and their uncertainties are then determined with great accuracy and the correlations between the groups are analyzed. Finally, the dependence of the results on the prior distribution choice and on the group cross section data is investigated to confirm the reliability of the analysis.

  3. Global analysis of seasonality in the shell flux of extant planktonic foraminifera

    NASA Astrophysics Data System (ADS)

    Jonkers, L.; Kučera, M.

    2015-01-01

    Shell fluxes of planktonic foraminifera species vary intra-annually in a pattern that appears to follow the seasonal cycle. However, the variation in the timing and prominence of seasonal flux maxima in space and among species remain poorly constrained. Thus, although changing seasonality may result in a flux-weighted temperature offset of more than 5 °C within a species, this effect is often ignored in the interpretation of foraminifera-based paleoceanographic records. Here we present an analysis of the intra-annual pattern of shell flux variability in 37 globally distributed time series. The existence of a seasonal component in flux variability was objectively characterised using periodic regression. This analysis yielded estimates of the number, timing and prominence of seasonal flux maxima. Over 80% of the flux series across all species showed a statistically significant periodic component, indicating that a considerable part of the intra-annual flux variability is predictable. Temperature appears to be a powerful predictor of flux seasonality, but its effect differs among species. Three different modes of seasonality are distinguishable. Tropical and subtropical species (G. ruber (white and pink varieties), N. dutertrei, G. sacculifer, O. universa, G. siphonifera, P. obliquiloculata, G. menardii, G. rubescens, G. tenella and G. conglobatus) appear to have a less predictable flux pattern with random peak timing in warm waters. In colder waters, seasonality is more prevalent: peak fluxes occur shortly after summer temperature maxima and peak prominence increases. This tendency is stronger in species with a narrower temperature range, implying that warm-adapted species find it increasingly difficult to reproduce outside their optimum temperature range and that with decreasing mean temperature, their flux is progressively focussed into the warm season. The second group includes the temperate to cold-water species Globigerina bulloides, G. glutinata, N. incompta

  4. Global analysis of seasonality in the shell flux of extant planktonic Foraminifera

    NASA Astrophysics Data System (ADS)

    Jonkers, L.; Kučera, M.

    2015-04-01

    Shell fluxes of planktonic Foraminifera species vary intra-annually in a pattern that appears to follow the seasonal cycle. However, the variation in the timing and prominence of seasonal flux maxima in space and among species remains poorly constrained. Thus, although changing seasonality may result in a flux-weighted temperature offset of more than 5° C within a species, this effect is often ignored in the interpretation of Foraminifera-based paleoceanographic records. To address this issue we present an analysis of the intra-annual pattern of shell flux variability in 37 globally distributed time series. The existence of a seasonal component in flux variability was objectively characterised using periodic regression. This analysis yielded estimates of the number, timing and prominence of seasonal flux maxima. Over 80% of the flux series across all species showed a statistically significant periodic component, indicating that a considerable part of the intra-annual flux variability is predictable. Temperature appears to be a powerful predictor of flux seasonality, but its effect differs among species. Three different modes of seasonality are distinguishable. Tropical and subtropical species (Globigerinoides ruber (white and pink varieties), Neogloboquadrina dutertrei, Globigerinoides sacculifer, Orbulina universa, Globigerinella siphonifera, Pulleniatina obliquiloculata, Globorotalia menardii, Globoturborotalita rubescens, Globoturborotalita tenella and Globigerinoides conglobatus) appear to have a less predictable flux pattern, with random peak timing in warm waters. In colder waters, seasonality is more prevalent: peak fluxes occur shortly after summer temperature maxima and peak prominence increases. This tendency is stronger in species with a narrower temperature range, implying that warm-adapted species find it increasingly difficult to reproduce outside their optimum temperature range and that, with decreasing mean temperature, their flux is progressively

  5. Analysis of Atmosphere-Ocean Surface Flux Feedbacks in Recent Satellite and Model Reanalysis Products

    NASA Technical Reports Server (NTRS)

    Roberts, J. Brent; Robertson, F. R.; Clayson, C. A.

    2010-01-01

    Recent investigations have examined observations in an attempt to determine when and how the ocean forces the atmosphere, and vice versa. These studies focus primarily on relationships between sea surface temperature anomalies and the turbulent and radiative surface heat fluxes. It has been found that both positive and negative feedbacks, which enhance or reduce sea surface temperature anomaly amplitudes, can be generated through changes in the surface boundary layer. Consequent changes in sea surface temperature act to change boundary layer characteristics through changes in static stability or turbulent fluxes. Previous studies over the global oceans have used coarse-resolution observational and model products such as ICOADS and the NCEP Reanalysis. This study focuses on documenting the atmosphere ocean feedbacks that exist in recently produced higher resolution products, namely the SeaFlux v1.0 product and the NASA Modern Era Retrospective-Analysis for Research and Applications (MERRA). It has been noted in recent studies that evidence of oceanic forcing of the atmosphere exists on smaller scales than the usually more dominant atmospheric forcing of the ocean, particularly in higher latitudes. It is expected that use of these higher resolution products will allow for a more comprehensive description of these small-scale ocean-atmosphere feedbacks. The SeaFlux intercomparisons have revealed large scatter between various surface flux climatologies. This study also investigates the uncertainty in surface flux feedbacks based on several of these recent satellite based climatologies

  6. Metabolic flux analysis using ¹³C peptide label measurements.

    PubMed

    Mandy, Dominic E; Goldford, Joshua E; Yang, Hong; Allen, Doug K; Libourel, Igor G L

    2014-02-01

    ¹³C metabolic flux analysis (MFA) has become the experimental method of choice to investigate the cellular metabolism of microbes, cell cultures and plant seeds. Conventional steady-state MFA utilizes isotopic labeling measurements of amino acids obtained from protein hydrolysates. To retain spatial information in conventional steady-state MFA, tissues or subcellular fractions must be dissected or biochemically purified. In contrast, peptides retain their identity in complex protein extracts, and may therefore be associated with a specific time of expression, tissue type and subcellular compartment. To enable 'single-sample' spatially and temporally resolved steady-state flux analysis, we investigated the suitability of peptide mass distributions (PMDs) as an alternative to amino acid label measurements. PMDs are the discrete convolution of the mass distributions of the constituent amino acids of a peptide. We investigated the requirements for the unique deconvolution of PMDs into amino acid mass distributions (AAMDs), the influence of peptide sequence length on parameter sensitivity, and how AAMD and flux estimates that are determined through deconvolution compare to estimates from a conventional GC-MS measurement-based approach. Deconvolution of PMDs of the storage protein β-conglycinin of soybean (Glycine max) resulted in good AAMD and flux estimates if fluxes were directly fitted to PMDs. Unconstrained deconvolution resulted in inferior AAMD and flux estimates. PMD measurements do not include amino acid backbone fragments, which increase the information content in GC-MS-derived analyses. Nonetheless, the resulting flux maps were of comparable quality due to the precision of Orbitrap quantification and the larger number of peptide measurements.

  7. Development of high flux thermal neutron generator for neutron activation analysis

    NASA Astrophysics Data System (ADS)

    Vainionpaa, Jaakko H.; Chen, Allan X.; Piestrup, Melvin A.; Gary, Charles K.; Jones, Glenn; Pantell, Richard H.

    2015-05-01

    The new model DD110MB neutron generator from Adelphi Technology produces thermal (<0.5 eV) neutron flux that is normally achieved in a nuclear reactor or larger accelerator based systems. Thermal neutron fluxes of 3-5 · 107 n/cm2/s are measured. This flux is achieved using four ion beams arranged concentrically around a target chamber containing a compact moderator with a central sample cylinder. Fast neutron yield of ∼2 · 1010 n/s is created at the titanium surface of the target chamber. The thickness and material of the moderator is selected to maximize the thermal neutron flux at the center. The 2.5 MeV neutrons are quickly thermalized to energies below 0.5 eV and concentrated at the sample cylinder. The maximum flux of thermal neutrons at the target is achieved when approximately half of the neutrons at the sample area are thermalized. In this paper we present simulation results used to characterize performance of the neutron generator. The neutron flux can be used for neutron activation analysis (NAA) prompt gamma neutron activation analysis (PGNAA) for determining the concentrations of elements in many materials. Another envisioned use of the generator is production of radioactive isotopes. DD110MB is small enough for modest-sized laboratories and universities. Compared to nuclear reactors the DD110MB produces comparable thermal flux but provides reduced administrative and safety requirements and it can be run in pulsed mode, which is beneficial in many neutron activation techniques.

  8. AN ANALYSIS OF MAGNETOHYDRODYNAMIC INVARIANTS OF MAGNETIC FLUCTUATIONS WITHIN INTERPLANETARY FLUX ROPES

    SciTech Connect

    Telloni, D.; Perri, S.; Carbone, V.; Bruno, R.; D Amicis, R.

    2013-10-10

    A statistical analysis of magnetic flux ropes, identified by large-amplitude, smooth rotations of the magnetic field vector and a low level of both proton density and temperature, has been performed by computing the invariants of the ideal magnetohydrodynamic (MHD) equations, namely the magnetic helicity, the cross-helicity, and the total energy, via magnetic field and plasma fluctuations in the interplanetary medium. A technique based on the wavelet spectrograms of the MHD invariants allows the localization and characterization of those structures in both scales and time: it has been observed that flux ropes show, as expected, high magnetic helicity states (|σ{sub m}| in [0.6: 1]), but extremely variable cross-helicity states (|σ{sub c}| in [0: 0.8]), which, however, are not independent of the magnetic helicity content of the flux rope itself. The two normalized MHD invariants observed within the flux ropes tend indeed to distribute, neither trivially nor automatically, along the √(σ{sub m}{sup 2}+σ{sub c}{sup 2})=1 curve, thus suggesting that some constraint should exist between the magnetic and cross-helicity content of the structures. The analysis carried out has further showed that the flux rope properties are totally independent of their time duration and that they are detected either as a sort of interface between different portions of solar wind or as isolated structures embedded in the same stream.

  9. Analysis and Engineering of Metabolic Pathway Fluxes in Corynebacterium glutamicum

    NASA Astrophysics Data System (ADS)

    Wittmann, Christoph

    The Gram-positive soil bacterium Corynebacterium glutamicum was discovered as a natural overproducer of glutamate about 50 years ago. Linked to the steadily increasing economical importance of this microorganism for production of glutamate and other amino acids, the quest for efficient production strains has been an intense area of research during the past few decades. Efficient production strains were created by applying classical mutagenesis and selection and especially metabolic engineering strategies with the advent of recombinant DNA technology. Hereby experimental and computational approaches have provided fascinating insights into the metabolism of this microorganism and directed strain engineering. Today, C. glutamicum is applied to the industrial production of more than 2 million tons of amino acids per year. The huge achievements in recent years, including the sequencing of the complete genome and efficient post genomic approaches, now provide the basis for a new, fascinating era of research - analysis of metabolic and regulatory properties of C. glutamicum on a global scale towards novel and superior bioprocesses.

  10. Discussion about modeling the effects of neutron flux exposure for nuclear reactor core analysis

    SciTech Connect

    Vondy, D.R.

    1986-04-01

    Methods used to calculate the effects of exposure to a neutron flux are described. The modeling of the nuclear-reactor core history presents an analysis challenge. The nuclide chain equations must be solved, and some of the methods in use for this are described. Techniques for treating reactor-core histories are discussed and evaluated.

  11. Tools for the analysis of metabolic flux through the sphingolipid pathway.

    PubMed

    Martínez-Montañés, Fernando; Schneiter, Roger

    2016-11-01

    Discerning the complex regulation of the enzymatic steps necessary for sphingolipid biosynthesis is facilitated by the utilization of tracers that allow a time-resolved analysis of the pathway dynamics without affecting the metabolic flux. Different strategies have been used and new tools are continuously being developed to probe the various enzymatic conversions that occur within this complex pathway. Here, we provide a short overview of the divergent fungal and mammalian sphingolipid biosynthetic routes, and of the tracers and methods that are frequently employed to follow the flux of intermediates throughout these pathways.

  12. Theoretical analysis of interferometer wave front tilt and fringe radiant flux on a rectangular photodetector.

    PubMed

    Smith, Robert; Fuss, Franz Konstantin

    2013-09-06

    This paper is a theoretical analysis of mirror tilt in a Michelson interferometer and its effect on the radiant flux over the active area of a rectangular photodetector or image sensor pixel. It is relevant to sensor applications using homodyne interferometry where these opto-electronic devices are employed for partial fringe counting. Formulas are derived for radiant flux across the detector for variable location within the fringe pattern and with varying wave front angle. The results indicate that the flux is a damped sine function of the wave front angle, with a decay constant of the ratio of wavelength to detector width. The modulation amplitude of the dynamic fringe pattern reduces to zero at wave front angles that are an integer multiple of this ratio and the results show that the polarity of the radiant flux changes exclusively at these multiples. Varying tilt angle causes radiant flux oscillations under an envelope curve, the frequency of which is dependent on the location of the detector with the fringe pattern. It is also shown that a fringe count of zero can be obtained for specific photodetector locations and wave front angles where the combined effect of fringe contraction and fringe tilt can have equal and opposite effects. Fringe tilt as a result of a wave front angle of 0.05° can introduce a phase measurement difference of 16° between a photodetector/pixel located 20 mm and one located 100 mm from the optical origin.

  13. Two-stage flux balance analysis of metabolic networks for drug target identification

    PubMed Central

    2011-01-01

    Background Efficient identification of drug targets is one of major challenges for drug discovery and drug development. Traditional approaches to drug target identification include literature search-based target prioritization and in vitro binding assays which are both time-consuming and labor intensive. Computational integration of different knowledge sources is a more effective alternative. Wealth of omics data generated from genomic, proteomic and metabolomic techniques changes the way researchers view drug targets and provides unprecedent opportunities for drug target identification. Results In this paper, we develop a method based on flux balance analysis (FBA) of metabolic networks to identify potential drug targets. This method consists of two linear programming (LP) models, which first finds the steady optimal fluxes of reactions and the mass flows of metabolites in the pathologic state and then determines the fluxes and mass flows in the medication state with the minimal side effect caused by the medication. Drug targets are identified by comparing the fluxes of reactions in both states and examining the change of reaction fluxes. We give an illustrative example to show that the drug target identification problem can be solved effectively by our method, then apply it to a hyperuricemia-related purine metabolic pathway. Known drug targets for hyperuricemia are correctly identified by our two-stage FBA method, and the side effects of these targets are also taken into account. A number of other promising drug targets are found to be both effective and safe. Conclusions Our method is an efficient procedure for drug target identification through flux balance analysis of large-scale metabolic networks. It can generate testable predictions, provide insights into drug action mechanisms and guide experimental design of drug discovery. PMID:21689470

  14. Combined analysis of neutron and photon flux measurements for the Jules Horowitz reactor core mapping

    SciTech Connect

    Fourmentel, D.; Villard, J. F.; Lyoussi, A.; Reynard-Carette, C.; Bignan, G.; Chauvin, J. P.; Gonnier, C.; Guimbal, P.; Malo, J. Y.; Carette, M.; Janulyte, A.; Merroun, O.; Brun, J.; Zerega, Y.; Andre, J.

    2011-07-01

    We study the combined analysis of nuclear measurements to improve the knowledge of the irradiation conditions in the experimental locations of the future Jules Horowitz Reactor (JHR). The goal of the present work is to measure more accurately neutron flux, photon flux and nuclear heating in the reactor. In a Material Testing Reactor (MTR), nuclear heating is a crucial parameter to design the experimental devices to be irradiated in harsh nuclear conditions. This parameter drives the temperature of the devices and of the samples. The numerical codes can predict this parameter but in-situ measurements are necessary to reach the expected accuracy. For this reason, one objective of the IN-CORE program [1] is to study the combined measurements of neutron and photon flux and their cross advanced interpretation. It should be reminded that both neutron and photon sensors are not totally selective as their signals are due to neutron and photon interactions. We intend to measure the neutron flux by three different kinds of sensors (Uranium Fission chamber, Plutonium Fission chamber and Self Powered Neutron Detector), the photon flux by two different sensors (Ionization chamber and Self Powered Gamma Detector) and the nuclear heating by two different ones (Differential calorimeter and Gamma Thermometer). For the same parameter, we expect that the use of different kinds of sensors will allow a better estimation of the aimed parameter by mixing different spectrum responses and different neutron and gamma contributions. An experimental test called CARMEN-1 is scheduled in OSIRIS reactor (CEA Saclay - France) at the end of 2011, with the goal to map irradiation locations in the reactor reflector to get a first validation of the analysis model. This article focuses on the sensor selection for CARMEN-1 experiment and to the way to link neutron and photon flux measurements in view to reduce their uncertainties but also to better assess the neutron and photon contributions to nuclear

  15. Extreme relativistic electron fluxes at geosynchronous orbit: Analysis of GOES E > 2 MeV electrons

    NASA Astrophysics Data System (ADS)

    Meredith, Nigel P.; Horne, Richard B.; Isles, John D.; Rodriguez, Juan V.

    2015-03-01

    Relativistic electrons (E > 1 MeV) cause internal charging on satellites and are an important space weather hazard. A key requirement in space weather research concerns extreme events and knowledge of the largest flux expected to be encountered over the lifetime of a satellite mission. This is interesting both from scientific and practical points of view since satellite operators, engineers, and the insurance industry need this information to better evaluate the effects of extreme events on their spacecraft. Here we conduct an extreme value analysis of daily averaged E > 2 MeV electron fluxes from the Geostationary Operational Environmental Satellites (GOES) during the 19.5 year period from 1 January 1995 to 30 June 2014. We find that the daily averaged flux measured at GOES West is typically a factor of about 2.5 higher than that measured at GOES East, and we conduct independent analyses for these two locations. The 1 in 10, 1 in 50, and 1 in 100 year daily averaged E > 2 MeV electron fluxes at GOES West are 1.84 ×105, 5.00 ×105, and 7.68 ×105 cm-2 s-1 sr-1, respectively. The corresponding fluxes at GOES East are 6.53 ×104, 1.98 ×105, and 3.25 ×105 cm-2 s-1 sr-1, respectively. The largest fluxes seen during the 19.5 year period on 29 July 2004 were particularly extreme and were seen by satellites at GOES West and GOES East. The extreme value analysis suggests that this event was a 1 in 50 year event.

  16. Leonid Storm Flux Analysis From One Leonid MAC Video AL50R

    NASA Technical Reports Server (NTRS)

    Gural, Peter S.; Jenniskens, Peter; DeVincenzi, Donald L. (Technical Monitor)

    2000-01-01

    A detailed meteor flux analysis is presented of a seventeen-minute portion of one videotape, collected on November 18, 1999, during the Leonid Multi-instrument Aircraft Campaign. The data was recorded around the peak of the Leonid meteor storm using an intensified CCD camera pointed towards the low southern horizon. Positions of meteors on the sky were measured. These measured meteor distributions were compared to a Monte Carlo simulation, which is a new approach to parameter estimation for mass ratio and flux. Comparison of simulated flux versus observed flux levels, seen between 1:50:00 and 2:06:41 UT, indicate a magnitude population index of r = 1.8 +/- 0.1 and mass ratio of s = 1.64 +/- 0.06. The average spatial density of the material contributing to the Leonid storm peak is measured at 0.82 +/- 0.19 particles per square kilometer per hour for particles of at least absolute visual magnitude +6.5. Clustering analysis of the arrival times of Leonids impacting the earth's atmosphere over the total observing interval shows no enhancement or clumping down to time scales of the video frame rate. This indicates a uniformly random temporal distribution of particles in the stream encountered during the 1999 epoch. Based on the observed distribution of meteors on the sky and the model distribution, recommendations am made for the optimal pointing directions for video camera meteor counts during future ground and airborne missions.

  17. Leonid Storm Flux Analysis from One Leonid Mac Video AL50R

    NASA Astrophysics Data System (ADS)

    Gural, Peter S.; Jenniskens, Peter

    A detailed meteor flux analysis is presented of a seventeen-minute portion of one videotape, collected on November 18, 1999, during the Leonid Multi-instrument Aircraft Campaign. The data was recorded around the peak of the Leonid meteor storm using an intensified CCD camera pointed towards the low southern horizon. Positions of meteors on the sky were measured. These measured meteor distributions were compared to a Monte Carlo simulation, which is a new approach to parameter estimation for mass ratio and flux. Comparison of simulated flux versus observed flux levels, seen between 1:50:00 and 2:06:41 UT, indicate a magnitude population index of r = 1.8 +/- 0.1 and mass ratio of s = 1.64 +/- 0.06. The average spatial density of the material contributing to the Leonid storm peak is measured at 0.82 +/- 0.19 particles per square kilometer per hour for particles of at least absolute visual magnitude +6.5. Clustering analysis of the arrival times of Leonids impacting the earth's atmosphere over the total observing interval shows no enhancement or clumping down to time scales of the video frame rate. This indicates a uniformly random temporal distribution of particles in the stream encountered during the 1999 epoch. Based on the observed distribution of meteors on the sky and the model distribution, recommendations are made for the optimal pointing directions for video camera meteor counts during future ground and airborne missions.

  18. Metabolic Flux Analysis of Mitochondrial Uncoupling in 3T3-L1 Adipocytes

    PubMed Central

    Si, Yaguang; Shi, Hai; Lee, Kyongbum

    2009-01-01

    Background Increasing energy expenditure at the cellular level offers an attractive option to limit adiposity and improve whole body energy balance. In vivo and in vitro observations have correlated mitochondrial uncoupling protein-1 (UCP1) expression with reduced white adipose tissue triglyceride (TG) content. The metabolic basis for this correlation remains unclear. Methodology/Principal Findings This study tested the hypothesis that mitochondrial uncoupling requires the cell to compensate for the decreased oxidation phosphorylation efficiency by up-regulating lactate production, thus redirecting carbon flux away from TG synthesis. Metabolic flux analysis was used to characterize the effects of non-lethal, long-term mitochondrial uncoupling (up to 18 days) on the pathways of intermediary metabolism in differentiating 3T3-L1 adipocytes. Uncoupling was induced by forced expression of UCP1 and chemical (FCCP) treatment. Chemical uncoupling significantly decreased TG content by ca. 35%. A reduction in the ATP level suggested diminished oxidative phosphorylation efficiency in the uncoupled adipocytes. Flux analysis estimated significant up-regulation of glycolysis and down-regulation of fatty acid synthesis, with chemical uncoupling exerting quantitatively larger effects. Conclusions/Significance The results of this study support our hypothesis regarding uncoupling-induced redirection of carbon flux into glycolysis and lactate production, and suggest mitochondrial proton translocation as a potential target for controlling adipocyte lipid metabolism. PMID:19746157

  19. Hands-on metabolism analysis of complex biochemical networks using elementary flux modes.

    PubMed

    Schäuble, Sascha; Schuster, Stefan; Kaleta, Christoph

    2011-01-01

    The aim of this chapter is to discuss the basic principles and reasoning behind elementary flux mode analysis (EFM analysis)--an important tool for the analysis of metabolic networks. We begin with a short introduction into metabolic pathway analysis and subsequently outline in detail fundamentals of EFM analysis by way of a small example network. We discuss issues arising in the reconstruction of metabolic networks required for EFM analysis and how they can be circumvented. Subsequently, we analyze a more elaborate example network representing photosynthate metabolism. Finally, we give an overview of applications of EFM analysis in biotechnology and other fields and discuss issues arising when applying methods from metabolic pathway analysis to genome-scale metabolic networks.

  20. Impact of water use efficiency on eddy covariance flux partitioning using correlation structure analysis

    NASA Astrophysics Data System (ADS)

    Anderson, Ray; Skaggs, Todd; Alfieri, Joseph; Kustas, William; Wang, Dong; Ayars, James

    2016-04-01

    Partitioned land surfaces fluxes (e.g. evaporation, transpiration, photosynthesis, and ecosystem respiration) are needed as input, calibration, and validation data for numerous hydrological and land surface models. However, one of the most commonly used techniques for measuring land surface fluxes, Eddy Covariance (EC), can directly measure net, combined water and carbon fluxes (evapotranspiration and net ecosystem exchange/productivity). Analysis of the correlation structure of high frequency EC time series (hereafter flux partitioning or FP) has been proposed to directly partition net EC fluxes into their constituent components using leaf-level water use efficiency (WUE) data to separate stomatal and non-stomatal transport processes. FP has significant logistical and spatial representativeness advantages over other partitioning approaches (e.g. isotopic fluxes, sap flow, microlysimeters), but the performance of the FP algorithm is reliant on the accuracy of the intercellular CO2 (ci) concentration used to parameterize WUE for each flux averaging interval. In this study, we tested several parameterizations for ci as a function of atmospheric CO2 (ca), including (1) a constant ci/ca ratio for C3 and C4 photosynthetic pathway plants, (2) species-specific ci/ca-Vapor Pressure Deficit (VPD) relationships (quadratic and linear), and (3) generalized C3 and C4 photosynthetic pathway ci/ca-VPD relationships. We tested these ci parameterizations at three agricultural EC towers from 2011-present in C4 and C3 crops (sugarcane - Saccharum officinarum L. and peach - Prunus persica), and validated again sap-flow sensors installed at the peach site. The peach results show that the species-specific parameterizations driven FP algorithm came to convergence significantly more frequently (~20% more frequently) than the constant ci/ca ratio or generic C3-VPD relationship. The FP algorithm parameterizations with a generic VPD relationship also had slightly higher transpiration (5 Wm-2

  1. Power Law Regression Analysis of Heat Flux Width in Type I ELMs

    NASA Astrophysics Data System (ADS)

    Stephens, C. D.; Makowski, M. A.; Leonard, A. W.; Osborne, T. H.

    2014-10-01

    In this project, a database of Type I ELM characteristics has been assembled and will be used to investigate possible dependencies of the heat flux width on physics and engineering parameters. At the edge near the divertor, high impulsive heat loads are imparted onto the surface. The impact of these ELMs can cause a reduction in divertor lifetime if the heat flux is great enough due to material erosion. A program will be used to analyze data, extract relevant, measurable quantities, and record the quantities in the table. Care is taken to accurately capture the complex space/time structure of the ELM. Then correlations between discharge and equilibrium parameters will be investigated. Power law regression analysis will be used to help determine the dependence of the heat flux width on these various measurable quantities and parameters. This will enable us to better understand the physics of heat flux at the edge. Work supported in part by the National Undergraduate Fellowship Program in Plasma Physics and Fusion Energy Sciences and the US DOE under DE-FG02-04ER54761, DE-AC52-07NA27344, DE-FC02-04ER54698.

  2. Metabolic Flux Analysis -application in plant metabolic modelling for advanced life support systems

    NASA Astrophysics Data System (ADS)

    Sasidharan L, Swathy; Hezard, Pauline; Poughon, Laurent; Dussap, Claude-Gilles

    Plants have an important role in providing food and fresh oxygen for humans in a closed environment during long duration missions to Mars or Moon. Also, plants play an important role for recycling water. Thus, plant modelling (crop composition, yield prediction and the responses to its environment within the closed loop) gets much attention in the development of closed ecological life support systems. In order to achieve this, metabolic flux computation methods accounting for reactions stoichiometry and chemical energy conservation obtained from metabolic pathways description of different plant parts are required. The basic ideas of metabolic modelling and their application to various plant parts will be discussed. Metabolic systems consist of a set of metabolites and reactions that consume or produce them. The metabolic pathways within a metabolic network for each plant part or sub level are characterised and the metabolic fluxes, defined as the amount of converted metabolite per unit time and per unit mass of tissue (or per plant part), can be calculated. MBA (Metabolic flux analysis) which is a constraint based approach is effective at calculating flux distributions through bio-chemical networks. This methodology can be applied to several plants' growth situations. In terms of space appli-cations, it is shown how this approach could bring valuable tools for assessing and quantifying the effects of the environment of a close system on growth rate and conversion yields.

  3. Analysis of a multi-machine database on divertor heat fluxes

    SciTech Connect

    Makowski, M. A.; Lasnier, C. J.; Elder, D.; Stangeby, P. C.; Gray, T. K.; Maingi, R.; LaBombard, B.; Terry, J. L.; Leonard, A. W.; Osborne, T. H.; Watkins, J.

    2012-05-15

    A coordinated effort to measure divertor heat flux characteristics in fully attached, similarly shaped H-mode plasmas on C-Mod, DIII-D, and NSTX was carried out in 2010 in order to construct a predictive scaling relation applicable to next step devices including ITER, FNSF, and DEMO. Few published scaling laws are available and those that have been published were obtained under widely varying conditions and divertor geometries, leading to conflicting predictions for this critically important quantity. This study was designed to overcome these deficiencies. Analysis of the combined data set reveals that the primary dependence of the parallel heat flux width is robustly inverse with I{sub p}, which all three tokamaks independently demonstrate. An improved Thomson scattering system on DIII-D has yielded very accurate scrape off layer (SOL) profile measurements from which tests of parallel transport models have been made. It is found that a flux-limited model agrees best with the data at all collisionalities, while a Spitzer resistivity model agrees at higher collisionality where it is more valid. The SOL profile measurements and divertor heat flux scaling are consistent with a heuristic drift based model as well as a critical gradient model.

  4. Extraction, purification, methylation and GC-MS analysis of short-chain carboxylic acids for metabolic flux analysis.

    PubMed

    Tivendale, Nathan D; Jewett, Erin M; Hegeman, Adrian D; Cohen, Jerry D

    2016-08-15

    Dynamic metabolic flux analysis requires efficient and effective methods for extraction, purification and analysis of a plethora of naturally-occurring compounds. One area of metabolism that would be highly informative to study using metabolic flux analysis is the tricarboxylic acid (TCA) cycle, which consists of short-chain carboxylic acids. Here, we describe a newly-developed method for extraction, purification, derivatization and analysis of short-chain carboxylic acids involved in the TCA cycle. The method consists of snap-freezing the plant material, followed by maceration and a 12-15h extraction at -80 °C. The extracts are then subject to reduction (to stabilize β-keto acids), purified by strong anion exchange solid phase extraction and methylated with methanolic HCl. This method could also be readily adapted to quantify many other short-chain carboxylic acids.

  5. Uncertainty analysis of the Operational Simplified Surface Energy Balance (SSEBop) model at multiple flux tower sites

    NASA Astrophysics Data System (ADS)

    Chen, Mingshi; Senay, Gabriel B.; Singh, Ramesh K.; Verdin, James P.

    2016-05-01

    Evapotranspiration (ET) is an important component of the water cycle - ET from the land surface returns approximately 60% of the global precipitation back to the atmosphere. ET also plays an important role in energy transport among the biosphere, atmosphere, and hydrosphere. Current regional to global and daily to annual ET estimation relies mainly on surface energy balance (SEB) ET models or statistical and empirical methods driven by remote sensing data and various climatological databases. These models have uncertainties due to inevitable input errors, poorly defined parameters, and inadequate model structures. The eddy covariance measurements on water, energy, and carbon fluxes at the AmeriFlux tower sites provide an opportunity to assess the ET modeling uncertainties. In this study, we focused on uncertainty analysis of the Operational Simplified Surface Energy Balance (SSEBop) model for ET estimation at multiple AmeriFlux tower sites with diverse land cover characteristics and climatic conditions. The 8-day composite 1-km MODerate resolution Imaging Spectroradiometer (MODIS) land surface temperature (LST) was used as input land surface temperature for the SSEBop algorithms. The other input data were taken from the AmeriFlux database. Results of statistical analysis indicated that the SSEBop model performed well in estimating ET with an R2 of 0.86 between estimated ET and eddy covariance measurements at 42 AmeriFlux tower sites during 2001-2007. It was encouraging to see that the best performance was observed for croplands, where R2 was 0.92 with a root mean square error of 13 mm/month. The uncertainties or random errors from input variables and parameters of the SSEBop model led to monthly ET estimates with relative errors less than 20% across multiple flux tower sites distributed across different biomes. This uncertainty of the SSEBop model lies within the error range of other SEB models, suggesting systematic error or bias of the SSEBop model is within the

  6. Uncertainty analysis of the Operational Simplified Surface Energy Balance (SSEBop) model at multiple flux tower sites

    USGS Publications Warehouse

    Chen, Mingshi; Senay, Gabriel B.; Singh, Ramesh K.; Verdin, James P.

    2016-01-01

    Evapotranspiration (ET) is an important component of the water cycle – ET from the land surface returns approximately 60% of the global precipitation back to the atmosphere. ET also plays an important role in energy transport among the biosphere, atmosphere, and hydrosphere. Current regional to global and daily to annual ET estimation relies mainly on surface energy balance (SEB) ET models or statistical and empirical methods driven by remote sensing data and various climatological databases. These models have uncertainties due to inevitable input errors, poorly defined parameters, and inadequate model structures. The eddy covariance measurements on water, energy, and carbon fluxes at the AmeriFlux tower sites provide an opportunity to assess the ET modeling uncertainties. In this study, we focused on uncertainty analysis of the Operational Simplified Surface Energy Balance (SSEBop) model for ET estimation at multiple AmeriFlux tower sites with diverse land cover characteristics and climatic conditions. The 8-day composite 1-km MODerate resolution Imaging Spectroradiometer (MODIS) land surface temperature (LST) was used as input land surface temperature for the SSEBop algorithms. The other input data were taken from the AmeriFlux database. Results of statistical analysis indicated that the SSEBop model performed well in estimating ET with an R2 of 0.86 between estimated ET and eddy covariance measurements at 42 AmeriFlux tower sites during 2001–2007. It was encouraging to see that the best performance was observed for croplands, where R2 was 0.92 with a root mean square error of 13 mm/month. The uncertainties or random errors from input variables and parameters of the SSEBop model led to monthly ET estimates with relative errors less than 20% across multiple flux tower sites distributed across different biomes. This uncertainty of the SSEBop model lies within the error range of other SEB models, suggesting systematic error or bias of the SSEBop model is within

  7. Some advance on the comprehension of SR analysis for estimating the flux of a scalar

    NASA Astrophysics Data System (ADS)

    Castellví, Dr

    2009-04-01

    In agronomy, the eddy covariance, EC, method likely is the preferred for measuring surface scalar fluxes. For latent heat flux, however, weighing lysimeters maybe preferred in agriculture, but they are rarely affordable and not portable. The dissipation method, DM, is considered the most reliable technique for measuring scalar fluxes over open water because instrument motion contaminates the EC measurements. The main advantage of DM over EC is that it is less sensitive to low frequency instrument platform motions (such as ship and buoys), sensor alignment, precise orientation and stringent steadiness in the mean meteorological conditions (Fairall and Larsen, 1986; Kader, 1992; Edson and Fairall, 1998). Over land, keeping in mind that the EC and DM methods require the same measurements for scalar flux measurement, the DM has several disadvantages versus the EC. Direct measurement of the scalar variance dissipation rate, VDR, requires to capture eddies in the Kolmogorov's microscale (thus scalar time series measured at frequencies in the order of kHz are needed). Therefore, it is not practical. Indirect methods to estimate VDR (such as spectral analysis and second or third order structure functions) requires implementing iterative methods involving similarity relationships that are not well established (Hsieh and Katul, 1997; Castellvi and Snyder, 2008). Currently, there is ample evidence that the DM as explained in traditional micrometeorological books (such as, Panofsky and Dutton, 1984; Brutsaert, 1988; Kaimal and Finnigan; 1994) is, in general, not correct. Accordingly, it likely explains why DM is typically omitted in revisits of micrometeorological methods for estimating scalar fluxes in agronomy. Within the last decade, over some agricultural surfaces, evidence has been shown on the advantages over other micrometeorological methods and the reliability (i.e., close performance to the EC method) of Surface Renewal, SR, theory in conjunction with the Analysis of

  8. Quantitative analysis of bidirectional electron fluxes within coronal mass ejections at 1 AU

    NASA Technical Reports Server (NTRS)

    Phillips, J. L.; Gosling, J. T.; Mccomas, D. J.; Bame, S. J.; Feldman, W. C.

    1992-01-01

    The solar wind electron heat flux is carried primarily by suprathermal electrons beamed antisunward along the interplanetary magnetic field. However, analysis of electron observations at 1 AU has shown that counterstreaming electron beams, suggesting closed magnetic structures, prevail within coronal mass ejections (CMEs). These structures might be magnetic 'tongues', magnetically detached plasmoids, or complex flux ropes. Here we show results of analysis of ISEE-3 observations within 39 CMEs, including the asymmetry between the two beams, its control by magnetic field orientation, and the variation of the electron distributions as CMEs convect past the spacecraft. We find that some CMEs are strongly asymmetric, with the antisunward beam generally dominant, while others contain nearly symmetric beams. The beam asymmetries, and the magnetic field orientations, exhibit characteristic trends as CMEs pass over the spacecraft. We present an example of a distinctive 'strahl-on-strahl' distribution, suggesting continued magnetic connection to the corona, in which a narrow antisunward beam is superimposed on a broader beam. Our results favor continuing magnetic connection to the Sun in a tongue or flux rope geometry rather than a fully detached plasmoid.

  9. #FluxFlow: Visual Analysis of Anomalous Information Spreading on Social Media.

    PubMed

    Zhao, Jian; Cao, Nan; Wen, Zhen; Song, Yale; Lin, Yu-Ru; Collins, Christopher

    2014-12-01

    We present FluxFlow, an interactive visual analysis system for revealing and analyzing anomalous information spreading in social media. Everyday, millions of messages are created, commented, and shared by people on social media websites, such as Twitter and Facebook. This provides valuable data for researchers and practitioners in many application domains, such as marketing, to inform decision-making. Distilling valuable social signals from the huge crowd's messages, however, is challenging, due to the heterogeneous and dynamic crowd behaviors. The challenge is rooted in data analysts' capability of discerning the anomalous information behaviors, such as the spreading of rumors or misinformation, from the rest that are more conventional patterns, such as popular topics and newsworthy events, in a timely fashion. FluxFlow incorporates advanced machine learning algorithms to detect anomalies, and offers a set of novel visualization designs for presenting the detected threads for deeper analysis. We evaluated FluxFlow with real datasets containing the Twitter feeds captured during significant events such as Hurricane Sandy. Through quantitative measurements of the algorithmic performance and qualitative interviews with domain experts, the results show that the back-end anomaly detection model is effective in identifying anomalous retweeting threads, and its front-end interactive visualizations are intuitive and useful for analysts to discover insights in data and comprehend the underlying analytical model.

  10. Atmospheric neutrino oscillation analysis using Fluka three-dimensional flux and Super-Kamiokande data

    NASA Astrophysics Data System (ADS)

    Kibayashi, Atsuko

    A new neutrino oscillation analysis was carried out with the atmospheric neutrino data taken by the Super-Kamiokande detector. The updated 1489 live-days of data, or 91.6 kton-yr exposure, has been compared to a new 3-dimensional atmospheric flux calculation. The resulting best-fit oscillation parameters are (sin2 2theta, Delta m2) = (1.00, 2.51 x 10-3 eV2). At 90% C.L., the allowed region parameters are sin2(2theta) > 0.90 and 1.7 x 10-3 < Deltam2 < 4.2 x 10 -3. No flux model dependence on the results were seen. Results are considered with muon-tau neutrino mixing alone. Some admixture of sterile and electron neutrinos remains permissible, but oscillations of muon to tau neutrinos, as opposed to alternative solutions, seems inescapable.

  11. Metabolic distance estimation based on principle component analysis of metabolic turnover.

    PubMed

    Nakayama, Yasumune; Putri, Sastia P; Bamba, Takeshi; Fukusaki, Eiichiro

    2014-09-01

    Visualization of metabolic dynamism is important for various types of metabolic studies including studies on optimization of bio-production processes and studies of metabolism-related diseases. Many methodologies have been developed for metabolic studies. Among these, metabolic turnover analysis (MTA) is often used to analyze metabolic dynamics. MTA involves observation of changes in the isotopomer ratio of metabolites over time following introduction of isotope-labeled substrates. MTA has several advantages compared with (13)C-metabolic flux analysis, including the diversity of applicable samples, the variety of isotope tracers, and the wide range of target pathways. However, MTA produces highly complex data from which mining useful information becomes difficult. For easy understanding of MTA data, a new approach was developed using principal component analysis (PCA). The resulting PCA score plot visualizes the metabolic distance, which is defined as distance between metabolites on the real metabolic map. And the score plot gives us some hints of interesting metabolism for further study. We used this method to analyze the central metabolism of Saccharomyces cerevisiae under moderated aerobic conditions, and time course data for 77 isotopomers of 14 metabolites were obtained. The PCA score plot for this dataset represented a metabolic map and indicated interesting phenomena such as activity of fumarate reductase under aerated condition. These findings show the importance of a multivariate analysis to MTA. In addition, because the approach is not biased, this method has potential application for analysis of less-studied pathways and organisms.

  12. 13C MRS and LC–MS Flux Analysis of Tumor Intermediary Metabolism

    PubMed Central

    Shestov, Alexander A.; Lee, Seung-Cheol; Nath, Kavindra; Guo, Lili; Nelson, David S.; Roman, Jeffrey C.; Leeper, Dennis B.; Wasik, Mariusz A.; Blair, Ian A.; Glickson, Jerry D.

    2016-01-01

    We present the first validated metabolic network model for analysis of flux through key pathways of tumor intermediary metabolism, including glycolysis, the oxidative and non-oxidative arms of the pentose pyrophosphate shunt, the TCA cycle as well as its anaplerotic pathways, pyruvate–malate shuttling, glutaminolysis, and fatty acid biosynthesis and oxidation. The model that is called Bonded Cumomer Analysis for application to 13C magnetic resonance spectroscopy (13C MRS) data and Fragmented Cumomer Analysis for mass spectrometric data is a refined and efficient form of isotopomer analysis that can readily be expanded to incorporate glycogen, phospholipid, and other pathways thereby encompassing all the key pathways of tumor intermediary metabolism. Validation was achieved by demonstrating agreement of experimental measurements of the metabolic rates of oxygen consumption, glucose consumption, lactate production, and glutamate pool size with independent measurements of these parameters in cultured human DB-1 melanoma cells. These cumomer models have been applied to studies of DB-1 melanoma and DLCL2 human diffuse large B-cell lymphoma cells in culture and as xenografts in nude mice at 9.4 T. The latter studies demonstrate the potential translation of these methods to in situ studies of human tumor metabolism by MRS with stable 13C isotopically labeled substrates on instruments operating at high magnetic fields (≥7 T). The melanoma studies indicate that this tumor line obtains 51% of its ATP by mitochondrial metabolism and 49% by glycolytic metabolism under both euglycemic (5 mM glucose) and hyperglycemic conditions (26 mM glucose). While a high level of glutamine uptake is detected corresponding to ~50% of TCA cycle flux under hyperglycemic conditions, and ~100% of TCA cycle flux under euglycemic conditions, glutaminolysis flux and its contributions to ATP synthesis were very small. Studies of human lymphoma cells demonstrated that inhibition of

  13. Metabolic flux distribution analysis by 13C-tracer experiments using the Markov chain-Monte Carlo method.

    PubMed

    Yang, J; Wongsa, S; Kadirkamanathan, V; Billings, S A; Wright, P C

    2005-12-01

    Metabolic flux analysis using 13C-tracer experiments is an important tool in metabolic engineering since intracellular fluxes are non-measurable quantities in vivo. Current metabolic flux analysis approaches are fully based on stoichiometric constraints and carbon atom balances, where the over-determined system is iteratively solved by a parameter estimation approach. However, the unavoidable measurement noises involved in the fractional enrichment data obtained by 13C-enrichment experiment and the possible existence of unknown pathways prevent a simple parameter estimation method for intracellular flux quantification. The MCMC (Markov chain-Monte Carlo) method, which obtains intracellular flux distributions through delicately constructed Markov chains, is shown to be an effective approach for deep understanding of the intracellular metabolic network. Its application is illustrated through the simulation of an example metabolic network.

  14. Design and analysis of a new axial flux coreless PMSG with three rotors and double stators

    NASA Astrophysics Data System (ADS)

    Minaz, Mehmet Recep; Çelebi, Mehmet

    In this study, axial flux coreless permanent magnet synchronous generator (PMSG) is designed as double stators and three rotors and its electromagnetic and structural characteristics are analyzed. Designing aimed the axial flux generator is placed into the single end of the side rotor in the machine and permanent magnets are placed into the double ends of the middle rotor. One more rotor than the number of stators here is used. Core is not used in the stator of the machine intended to be designed. Aim of this study is to provide both reduction of iron loss and making the machine become lighter by reducing the number of the rotors to be used. Moreover, easiness in the production stage of the machine is provided. Three-dimensioned electromagnetic analysis of the designed machine has been done through the finite element method and transient solutions are suggested based on this. Within this study, arrangements have been made depending on certain standards in order that permanent magnets and coils obtain direct alternating current. The designed new axial flux generator move as permanent speed of 500 rpm and so maximum voltage of approximately 120 V per phase is obtained. Furthermore, this PMSG does not need a gear system due to its design structure.

  15. ¹³C-based metabolic flux analysis of Saccharomyces cerevisiae with a reduced Crabtree effect.

    PubMed

    Kajihata, Shuichi; Matsuda, Fumio; Yoshimi, Mika; Hayakawa, Kenshi; Furusawa, Chikara; Kanda, Akihisa; Shimizu, Hiroshi

    2015-08-01

    Saccharomyces cerevisiae shows a Crabtree effect that produces ethanol in a high glucose concentration even under fully aerobic condition. For efficient production of cake yeast or compressed yeast for baking, ethanol by-production is not desired since glucose limited chemostat or fed-batch cultivations are performed to suppress the Crabtree effect. In this study, the (13)C-based metabolic flux analysis ((13)C-MFA) was performed for the S288C derived S. cerevisiae strain to characterize a metabolic state under the reduced Crabtree effect. S. cerevisiae cells were cultured at a low dilution rate (0.1 h(-1)) under the glucose-limited chemostat condition. The estimated metabolic flux distribution showed that the acetyl-CoA in mitochondria was mainly produced from pyruvate by pyruvate dehydrogenase (PDH) reaction and that the level of the metabolic flux through the pentose phosphate pathway was much higher than that of the Embden-Meyerhof-Parnas pathway, which contributes to high biomass yield at low dilution rate by supplying NADPH required for cell growth.

  16. Metabolic flux analysis during the exponential growth phase of Saccharomyces cerevisiae in wine fermentations.

    PubMed

    Quirós, Manuel; Martínez-Moreno, Rubén; Albiol, Joan; Morales, Pilar; Vázquez-Lima, Felícitas; Barreiro-Vázquez, Antonio; Ferrer, Pau; Gonzalez, Ramon

    2013-01-01

    As a consequence of the increase in global average temperature, grapes with the adequate phenolic and aromatic maturity tend to be overripe by the time of harvest, resulting in increased sugar concentrations and imbalanced C/N ratios in fermenting musts. This fact sets obvious additional hurdles in the challenge of obtaining wines with reduced alcohols levels, a new trend in consumer demands. It would therefore be interesting to understand Saccharomyces cerevisiae physiology during the fermentation of must with these altered characteristics. The present study aims to determine the distribution of metabolic fluxes during the yeast exponential growth phase, when both carbon and nitrogen sources are in excess, using continuous cultures. Two different sugar concentrations were studied under two different winemaking temperature conditions. Although consumption and production rates for key metabolites were severely affected by the different experimental conditions studied, the general distribution of fluxes in central carbon metabolism was basically conserved in all cases. It was also observed that temperature and sugar concentration exerted a higher effect on the pentose phosphate pathway and glycerol formation than on glycolysis and ethanol production. Additionally, nitrogen uptake, both quantitatively and qualitatively, was strongly influenced by environmental conditions. This work provides the most complete stoichiometric model used for Metabolic Flux Analysis of S. cerevisiae in wine fermentations employed so far, including the synthesis and release of relevant aroma compounds and could be used in the design of optimal nitrogen supplementation of wine fermentations.

  17. Core Fueling and Edge Particle Flux Analysis in Ohmically and Auxiliary Heated NSTX Plasmas

    SciTech Connect

    V.A. Soukhanovskii; R. Maingi; R. Raman; H.W. Kugel; B.P. LeBlanc; L. Roquemore; C.H. Skinner; NSTX Research Team

    2002-06-12

    The Boundary Physics program of the National Spherical Torus Experiment (NSTX) is focusing on optimization of the edge power and particle flows in b * 25% L- and H-mode plasmas of t {approx} 0.8 s duration heated by up to 6 MW of high harmonic fast wave and up to 5 MW of neutral beam injection. Particle balance and core fueling efficiencies of low and high field side gas fueling of L-mode homic and NBI heated plasmas have been compared using an analytical zero dimensional particle balance model and measured ion and neutral fluxes. Gas fueling efficiencies are in the range of 0.05-0.20 and do not depend on discharge magnetic configuration, density or poloidal location of the injector. The particle balance modeling indicates that the addition of HFS fueling results in a reversal of the wall loading rate and higher wall inventories. Initial particle source estimates obtained from neutral pressure and spectroscopic measurements indicate that ion flux into the divertor greatly exceeds midplane ion flux from the main plasma, suggesting that the scrape-off cross-field transport plays a minor role in diverted plasmas. Present analysis provides the basis for detailed fluid modeling of core and edge particle flows and particle confinement properties of NSTX plasmas. This research was supported by the U.S. Department of Energy under contracts No. DE-AC02-76CH03073, DE-AC05-00OR22725, and W-7405-ENG-36.

  18. Quantitative analysis of bidirectional electron fluxes within coronal mass ejections at 1 AU

    NASA Astrophysics Data System (ADS)

    Phillips, J. L.; Gosling, J. T.; McComas, D. J.; Bame, S. J.; Feldman, W. C.

    The solar wind electron heat flux is carried primarily by suprathermal 'halo' electrons beamed antisunward along the interplanetary magnetic field (IMF), indicating magnetic connection to the Sun only in one direction. However, electron observations at 1 AU show that counterstreaming halo beams, suggesting closed magnetic structures, prevail within coronal mass ejections (CME's). These structures might be magnetic 'tongues', tied to the Sun at both ends, magnetically detached plasmoids, or complex flux rope structures. Here, we present first results of analysis of ISEE-3 observations within 39 CME's, including the asymmetry between the counterstreaming beams and its control by the IMF orientation, and the variation of the electron distributions as CME's convect past the spacecraft. We find that some CME's contain nearly symmetric electron beams, while others are strongly asymmetric, and that the antisunward beam is generally dominant. The more nearly radial the IMF, the greater the asymmetry between outward and inward beams. We present an example of a distinctive 'strahl-on-strahl' distribution, suggesting continued magnetic connection to the corona, in which a narrow antisunward beam is superimposed on a broader beam. Taken as a whole, our results appear to favor a tongue or flux rope scenario rather than a fully detached plasmoid.

  19. Metabolic Flux Analysis during the Exponential Growth Phase of Saccharomyces cerevisiae in Wine Fermentations

    PubMed Central

    Quirós, Manuel; Martínez-Moreno, Rubén; Albiol, Joan; Morales, Pilar; Vázquez-Lima, Felícitas; Barreiro-Vázquez, Antonio; Ferrer, Pau; Gonzalez, Ramon

    2013-01-01

    As a consequence of the increase in global average temperature, grapes with the adequate phenolic and aromatic maturity tend to be overripe by the time of harvest, resulting in increased sugar concentrations and imbalanced C/N ratios in fermenting musts. This fact sets obvious additional hurdles in the challenge of obtaining wines with reduced alcohols levels, a new trend in consumer demands. It would therefore be interesting to understand Saccharomyces cerevisiae physiology during the fermentation of must with these altered characteristics. The present study aims to determine the distribution of metabolic fluxes during the yeast exponential growth phase, when both carbon and nitrogen sources are in excess, using continuous cultures. Two different sugar concentrations were studied under two different winemaking temperature conditions. Although consumption and production rates for key metabolites were severely affected by the different experimental conditions studied, the general distribution of fluxes in central carbon metabolism was basically conserved in all cases. It was also observed that temperature and sugar concentration exerted a higher effect on the pentose phosphate pathway and glycerol formation than on glycolysis and ethanol production. Additionally, nitrogen uptake, both quantitatively and qualitatively, was strongly influenced by environmental conditions. This work provides the most complete stoichiometric model used for Metabolic Flux Analysis of S. cerevisiae in wine fermentations employed so far, including the synthesis and release of relevant aroma compounds and could be used in the design of optimal nitrogen supplementation of wine fermentations. PMID:23967264

  20. Lidar and Hyperspectral Remote Sensing for the Analysis of Coniferous Biomass Stocks and Fluxes

    NASA Astrophysics Data System (ADS)

    Halligan, K. Q.; Roberts, D. A.

    2006-12-01

    Airborne lidar and hyperspectral data can improve estimates of aboveground carbon stocks and fluxes through their complimentary responses to vegetation structure and biochemistry. While strong relationships have been demonstrated between lidar-estimated vegetation structural parameters and field data, research is needed to explore the portability of these methods across a range of topographic conditions, disturbance histories, vegetation type and climate. Additionally, research is needed to evaluate contributions of hyperspectral data in refining biomass estimates and determination of fluxes. To address these questions we are a conducting study of lidar and hyperspectral remote sensing data across sites including coniferous forests, broadleaf deciduous forests and a tropical rainforest. Here we focus on a single study site, Yellowstone National Park, where tree heights, stem locations, above ground biomass and basal area were mapped using first-return small-footprint lidar data. A new method using lidar intensity data was developed for separating the terrain and vegetation components in lidar data using a two-scale iterative local minima filter. Resulting Digital Terrain Models (DTM) and Digital Canopy Models (DCM) were then processed to retrieve a diversity of vertical and horizontal structure metrics. Univariate linear models were used to estimate individual tree heights while stepwise linear regression was used to estimate aboveground biomass and basal area. Three small-area field datasets were compared for their utility in model building and validation of vegetation structure parameters. All structural parameters were linearly correlated with lidar-derived metrics, with higher accuracies obtained where field and imagery data were precisely collocated . Initial analysis of hyperspectral data suggests that vegetation health metrics including measures of live and dead vegetation and stress indices may provide good indicators of carbon flux by mapping vegetation

  1. Exploiting diurnal variations to evaluate the ISCCP-FD flux calculations and radiative-flux-analysis-processed surface observations from BSRN, ARM, and SURFRAD

    SciTech Connect

    Zhang, Yuanchong; Long, Charles N.; Rossow, William B.; Dutton, Ellsworth G.

    2010-01-01

    Based on monthly-3-hourly and 3-hourly mean surface radiative fluxes and their associated meteorological parameters for 2004 from the International Satellite Cloud Climatology Project-FD (ISCCP-FD) and the Radiative Flux Analysis method-Produced Surface Observations (RFA-PSO) for 15 high-quality-controlled surface stations, operated by the Baseline Surface Radiation Network (BSRN), the Atmospheric Radiation Measurement (ARM) and the National Oceanic and Atmospheric Administration's Surface Radiation budget network (SURFRAD), this work, goes beyond the previous validation for FD against surface observation by introducing the Meteorological Similarity Comparison Method (MSCM) to make a more precise, mutual evaluation of both FD and PSO products. The comparison results in substantial uncertainty reduction and provides reasonable physical explanations for the flux differences. This approach compares fluxes for cases where the atmospheric and surface physical properties (specifically, the input parameters for radiative transfer model) are as close as possible to the values determined at the observational sites by matching the RFA-produced cloud fraction (CF) and/or optical thickness (Tau), etc., or alternatively, by directly changing the model input variables for FD to match PSO values, and using such-produced matched sub-datasets to make more accurate comparisons based on more similar meteorological environments between FD and PSO. The crucial part is the availability of flux-associated meteorological parameters from RFA-PSO, which was only recently made available that makes this work possible. For surface downwelling shortwave(SW) flux (SWdn) and its two components, diffuse (Dif) and direct (Dir), uncertainty for monthly mean is 15, 15 and 17 W/m2, respectively, smaller than the separately estimated uncertainty values from both FD and PSO. When applying MSCM by reducing their CF difference, the differences can be reduced by a factor of 2. The strength of

  2. Some advance on the comprehension of SR analysis for estimating the flux of a scalar

    NASA Astrophysics Data System (ADS)

    Castellví, Dr

    2009-04-01

    In agronomy, the eddy covariance, EC, method likely is the preferred for measuring surface scalar fluxes. For latent heat flux, however, weighing lysimeters maybe preferred in agriculture, but they are rarely affordable and not portable. The dissipation method, DM, is considered the most reliable technique for measuring scalar fluxes over open water because instrument motion contaminates the EC measurements. The main advantage of DM over EC is that it is less sensitive to low frequency instrument platform motions (such as ship and buoys), sensor alignment, precise orientation and stringent steadiness in the mean meteorological conditions (Fairall and Larsen, 1986; Kader, 1992; Edson and Fairall, 1998). Over land, keeping in mind that the EC and DM methods require the same measurements for scalar flux measurement, the DM has several disadvantages versus the EC. Direct measurement of the scalar variance dissipation rate, VDR, requires to capture eddies in the Kolmogorov's microscale (thus scalar time series measured at frequencies in the order of kHz are needed). Therefore, it is not practical. Indirect methods to estimate VDR (such as spectral analysis and second or third order structure functions) requires implementing iterative methods involving similarity relationships that are not well established (Hsieh and Katul, 1997; Castellvi and Snyder, 2008). Currently, there is ample evidence that the DM as explained in traditional micrometeorological books (such as, Panofsky and Dutton, 1984; Brutsaert, 1988; Kaimal and Finnigan; 1994) is, in general, not correct. Accordingly, it likely explains why DM is typically omitted in revisits of micrometeorological methods for estimating scalar fluxes in agronomy. Within the last decade, over some agricultural surfaces, evidence has been shown on the advantages over other micrometeorological methods and the reliability (i.e., close performance to the EC method) of Surface Renewal, SR, theory in conjunction with the Analysis of

  3. High-throughput hyperpolarized 13C metabolic investigations using a multi-channel acquisition system

    NASA Astrophysics Data System (ADS)

    Lee, Jaehyuk; Ramirez, Marc S.; Walker, Christopher M.; Chen, Yunyun; Yi, Stacey; Sandulache, Vlad C.; Lai, Stephen Y.; Bankson, James A.

    2015-11-01

    Magnetic resonance imaging and spectroscopy of hyperpolarized (HP) compounds such as [1-13C]-pyruvate have shown tremendous potential for offering new insight into disease and response to therapy. New applications of this technology in clinical research and care will require extensive validation in cells and animal models, a process that may be limited by the high cost and modest throughput associated with dynamic nuclear polarization. Relatively wide spectral separation between [1-13C]-pyruvate and its chemical endpoints in vivo are conducive to simultaneous multi-sample measurements, even in the presence of a suboptimal global shim. Multi-channel acquisitions could conserve costs and accelerate experiments by allowing acquisition from multiple independent samples following a single dissolution. Unfortunately, many existing preclinical MRI systems are equipped with only a single channel for broadband acquisitions. In this work, we examine the feasibility of this concept using a broadband multi-channel digital receiver extension and detector arrays that allow concurrent measurement of dynamic spectroscopic data from ex vivo enzyme phantoms, in vitro anaplastic thyroid carcinoma cells, and in vivo in tumor-bearing mice. Throughput and the cost of consumables were improved by up to a factor of four. These preliminary results demonstrate the potential for efficient multi-sample studies employing hyperpolarized agents.

  4. Robust hyperpolarized (13)C metabolic imaging with selective non-excitation of pyruvate (SNEP).

    PubMed

    Chen, Way Cherng; Teo, Xing Qi; Lee, Man Ying; Radda, George K; Lee, Philip

    2015-08-01

    In vivo metabolic imaging using hyperpolarized [1-(13)C]pyruvate provides localized biochemical information and is particularly useful in detecting early disease changes, as well as monitoring disease progression and treatment response. However, a major limitation of hyperpolarized magnetization is its unrecoverable decay, due not only to T1 relaxation but also to radio-frequency (RF) excitation. RF excitation schemes used in metabolic imaging must therefore be able to utilize available hyperpolarized magnetization efficiently and robustly for the optimal detection of substrate and metabolite activities. In this work, a novel RF excitation scheme called selective non-excitation of pyruvate (SNEP) is presented. This excitation scheme involves the use of a spectral selective RF pulse to specifically exclude the excitation of [1-(13)C]pyruvate, while uniformly exciting the key metabolites of interest (namely [1-(13)C]lactate and [1-(13)C]alanine) and [1-(13)C]pyruvate-hydrate. By eliminating the loss of hyperpolarized [1-(13)C]pyruvate magnetization due to RF excitation, the signal from downstream metabolite pools is increased together with enhanced dynamic range. Simulation results, together with phantom measurements and in vivo experiments, demonstrated the improvement in signal-to-noise ratio (SNR) and the extension of the lifetime of the [1-(13)C]lactate and [1-(13)C]alanine pools when compared with conventional non-spectral selective (NS) excitation. SNEP has also been shown to perform comparably well with multi-band (MB) excitation, yet SNEP possesses distinct advantages, including ease of implementation, less stringent demands on gradient performance, increased robustness to frequency drifts and B0 inhomogeneity as well as easier quantification involving the use of [1-(13)C]pyruvate-hydrate as a proxy for the actual [1-(13)C] pyruvate signal. SNEP is therefore a promising alternative for robust hyperpolarized [1-(13)C]pyruvate metabolic imaging with high fidelity.

  5. Analysis of the influence of rainfall variables on urban effluents concentrations and fluxes in wet weather

    NASA Astrophysics Data System (ADS)

    Gooré Bi, Eustache; Monette, Frédéric; Gasperi, Johnny

    2015-04-01

    Urban rainfall runoff has been a topic of increasing importance over the past years, a result of both the increase in impervious land area arising from constant urban growth and the effects of climate change on urban drainage. The main goal of the present study is to assess and analyze the correlations between rainfall variables and common indicators of urban water quality, namely event mean concentrations (EMCs) and event fluxes (EFs), in order to identify and explain the impacts of each of the main rainfall variables on the generation process of urban pollutants during wet periods. To perform this analysis, runoff from eight summer rainfall events that resulted in combined sewer overflow (CSO) was sampled simultaneously from two distinct catchment areas in order to quantify discharges at the respective outfalls. Pearson statistical analysis of total suspended solids (TSS), chemical oxygen demand (COD), carbonaceous biochemical oxygen demand at 5 days (CBOD5), total phosphorus (Ptot) and total kedjal nitrogen (N-TKN) showed significant correlations (ρ = 0.05) between dry antecedent time (DAT) and EMCs on one hand, and between total rainfall (TR) and the volume discharged (VD) during EFs, on the other. These results show that individual rainfall variables strongly affect either EMCs or EFs and are good predictors to consider when selecting variables for statistical modeling of urban runoff quality. The results also show that in a combined sewer network, there is a linear relationship between TSS event fluxes and COD, CBOD5, Ptot, and N-TKN event fluxes; this explains 97% of the variability of these pollutants which adsorb onto TSS during wet weather, which therefore act as tracers. Consequently, the technological solution selected for TSS removal will also lead to a reduction of these pollutants. Given the huge volumes involved, urban runoffs contribute substantially to pollutant levels in receiving water bodies, a situation which, in a climate change context, may

  6. Job/task analysis for I C (Instrumentation and Controls) instrument technicians at the High Flux Isotope Reactor

    SciTech Connect

    Duke, L.L.

    1989-09-01

    To comply with Department of Energy Order 5480.XX (Draft), a job/task analysis was initiated by the Maintenance Management Department at Oak Ridge National Laboratory (ORNL). The analysis was applicable to instrument technicians working at the ORNL High Flux Isotope Reactor (HFIR). This document presents the procedures and results of that analysis. 2 refs., 2 figs.

  7. Arrangement Analysis of Leaves Optimized on Photon Flux Density or Photosynthetic Rate

    NASA Astrophysics Data System (ADS)

    Obara, Shin'ya; Tanno, Itaru

    By clarifying a plant evolutive process, useful information may be obtained on engineering. Consequently, an analysis algorithm that investigates the optimal arrangement of plant leaves was developed. In the developed algorithm, the Monte Carlo method is introduced and sunlight is simulated. Moreover, the arrangement optimization of leaves is analyzed using a Genetic Algorithm (GA). The number of light quanta (photon flux density) that reaches leaves, or the average photosynthetic rate of the same was set as the objective function, and leaf models of a dogwood and a ginkgo tree were analyzed. The number of leaf models was set between two to four, and the position of the leaf was expressed in terms of the angle of direction, elevation angle, rotation angle, and the representative length of the branch of a leaf. The chromosome model introduced into GA consists of information concerning the position of the leaf. Based on the analysis results, the characteristics of the leaf of an actual plant could be simulated by ensuring the algorithm had multiple constrained conditions. The optimal arrangement of leaves differs in maximization of the photon flux density, and that of the average value of a photosynthetic rate. Furthermore, the leaf form affecting the optimal arrangement of leave and also having a significant influence also on a photosynthetic rate was shown.

  8. Application of (13)C flux analysis to identify high-productivity CHO metabolic phenotypes.

    PubMed

    Templeton, Neil; Smith, Kevin D; McAtee-Pereira, Allison G; Dorai, Haimanti; Betenbaugh, Michael J; Lang, Steven E; Young, Jamey D

    2017-01-23

    Industrial bioprocesses place high demands on the energy metabolism of host cells to meet biosynthetic requirements for maximal protein expression. Identifying metabolic phenotypes that promote high expression is therefore a major goal of the biotech industry. We conducted a series of (13)C flux analysis studies to examine the metabolic response to IgG expression during early stationary phase of CHO cell cultures grown in 3L fed-batch bioreactors. We examined eight clones expressing four different IgGs and compared with three non-expressing host-cell controls. Some clones were genetically manipulated to be apoptosis-resistant by expressing Bcl-2Δ, which correlated with increased IgG production and elevated glucose metabolism. The metabolic phenotypes of the non-expressing, IgG-expressing, and Bcl-2Δ/IgG-expressing clones were fully segregated by hierarchical clustering analysis. Lactate consumption and citric acid cycle fluxes were most strongly associated with specific IgG productivity. These studies indicate that enhanced oxidative metabolism is a characteristic of high-producing CHO cell lines.

  9. Insights into pH-induced metabolic switch by flux balance analysis.

    PubMed

    Ivarsson, Marija; Noh, Heeju; Morbidelli, Massimo; Soos, Miroslav

    2015-01-01

    Lactate accumulation in mammalian cell culture is known to impede cellular growth and productivity. The control of lactate formation and consumption in a hybridoma cell line was achieved by pH alteration during the early exponential growth phase. In particular, lactate consumption was induced even at high glucose concentrations at pH 6.8, whereas highly increased production of lactate was obtained at pH 7.8. Consequently, constraint-based metabolic flux analysis was used to examine pH-induced metabolic states in the same growth state. We demonstrated that lactate influx at pH 6.8 led cells to maintain high fluxes in the TCA cycle and malate-aspartate shuttle resulting in a high ATP production rate. In contrast, under increased pH conditions, less ATP was generated and different ATP sources were utilized. Gene expression analysis led to the conclusion that lactate formation at high pH was enabled by gluconeogenic pathways in addition to facilitated glucose uptake. The obtained results provide new insights into the influence of pH on cellular metabolism, and are of importance when considering pH heterogeneities typically present in large scale industrial bioreactors.

  10. Elementary Flux Mode Analysis of Acetyl-CoA Pathway in Carboxydothermus hydrogenoformans Z-2901

    PubMed Central

    Chinnasamy Perumal, Rajadurai; Selvaraj, Ashok; Ramesh Kumar, Gopal

    2014-01-01

    Carboxydothermus hydrogenoformans is a carboxydotrophic hydrogenogenic bacterium species that produces hydrogen molecule by utilizing carbon monoxide (CO) or pyruvate as a carbon source. To investigate the underlying biochemical mechanism of hydrogen production, an elementary mode analysis of acetyl-CoA pathway was performed to determine the intermediate fluxes by combining linear programming (LP) method available in CellNetAnalyzer software. We hypothesized that addition of enzymes necessary for carbon monoxide fixation and pyruvate dissimilation would enhance the theoretical yield of hydrogen. An in silico gene knockout of pyk, pykC, and mdh genes of modeled acetyl-CoA pathway allows the maximum theoretical hydrogen yield of 47.62 mmol/gCDW/h for 1 mole of carbon monoxide (CO) uptake. The obtained hydrogen yield is comparatively two times greater than the previous experimental data. Therefore, it could be concluded that this elementary flux mode analysis is a crucial way to achieve efficient hydrogen production through acetyl-CoA pathway and act as a model for strain improvement. PMID:24822064

  11. Differential Bees Flux Balance Analysis with OptKnock for In Silico Microbial Strains Optimization

    PubMed Central

    Choon, Yee Wen; Mohamad, Mohd Saberi; Deris, Safaai; Illias, Rosli Md.; Chong, Chuii Khim; Chai, Lian En; Omatu, Sigeru; Corchado, Juan Manuel

    2014-01-01

    Microbial strains optimization for the overproduction of desired phenotype has been a popular topic in recent years. The strains can be optimized through several techniques in the field of genetic engineering. Gene knockout is a genetic engineering technique that can engineer the metabolism of microbial cells with the objective to obtain desirable phenotypes. However, the complexities of the metabolic networks have made the process to identify the effects of genetic modification on the desirable phenotypes challenging. Furthermore, a vast number of reactions in cellular metabolism often lead to the combinatorial problem in obtaining optimal gene deletion strategy. Basically, the size of a genome-scale metabolic model is usually large. As the size of the problem increases, the computation time increases exponentially. In this paper, we propose Differential Bees Flux Balance Analysis (DBFBA) with OptKnock to identify optimal gene knockout strategies for maximizing the production yield of desired phenotypes while sustaining the growth rate. This proposed method functions by improving the performance of a hybrid of Bees Algorithm and Flux Balance Analysis (BAFBA) by hybridizing Differential Evolution (DE) algorithm into neighborhood searching strategy of BAFBA. In addition, DBFBA is integrated with OptKnock to validate the results for improving the reliability the work. Through several experiments conducted on Escherichia coli, Bacillus subtilis, and Clostridium thermocellum as the model organisms, DBFBA has shown a better performance in terms of computational time, stability, growth rate, and production yield of desired phenotypes compared to the methods used in previous works. PMID:25047076

  12. Metabolome, transcriptome and metabolic flux analysis of arabinose fermentation by engineered Saccharomyces cerevisiae.

    PubMed

    Wisselink, H Wouter; Cipollina, Chiara; Oud, Bart; Crimi, Barbara; Heijnen, Joseph J; Pronk, Jack T; van Maris, Antonius J A

    2010-11-01

    One of the challenges in strain improvement by evolutionary engineering is to subsequently determine the molecular basis of the improved properties that were enriched from the natural genetic variation during the selective conditions. This study focuses on Saccharomyces cerevisiae IMS0002 which, after metabolic and evolutionary engineering, ferments the pentose sugar arabinose. Glucose- and arabinose-limited anaerobic chemostat cultures of IMS0002 and its non-evolved ancestor were subjected to transcriptome analysis, intracellular metabolite measurements and metabolic flux analysis. Increased expression of the GAL-regulon and deletion of GAL2 in IMS0002 confirmed that the galactose transporter is essential for growth on arabinose. Elevated intracellular concentrations of pentose-phosphate-pathway intermediates and upregulation of TKL2 and YGR043c (encoding transketolase and transaldolase isoenzymes) suggested an involvement of these genes in flux-controlling reactions in arabinose fermentation. Indeed, deletion of these genes in IMS0002 caused a 21% reduction of the maximum specific growth rate on arabinose.

  13. Isotopically non-stationary metabolic flux analysis: complex yet highly informative.

    PubMed

    Wiechert, Wolfgang; Nöh, Katharina

    2013-12-01

    Metabolic flux analysis (MFA) using isotopic tracers aims at the experimental determination of in vivo reaction rates (fluxes). In recent years, the well-established 13C-MFA method based on metabolic and isotopic steady state was extended to INST-MFA (isotopically non-stationary MFA), which is performed in a transient labeling state. INST-MFA offers short-time experiments with a maximal information gain, and can moreover be applied to a wider range of growth conditions or organisms. Some of these conditions are not accessible by conventional methods. This comes at the price of significant methodological complexity involving high-frequency sampling and quenching, precise analysis of many samples and an extraordinary computational effort. This review gives a brief overview of basic principles, experimental workflows, and recent progress in this field. Special emphasis is laid on the trade-off between total effort and information gain, particularly on the suitability of INST-MFA for certain types of biological questions. In order to integrate INST-MFA as a viable method into the toolbox of MFA, some major challenges must be addressed in the coming years. These are discussed in the outlook.

  14. Metabolic engineering and flux analysis of Corynebacterium glutamicum for L-serine production.

    PubMed

    Lai, Shujuan; Zhang, Yun; Liu, Shuwen; Liang, Yong; Shang, Xiuling; Chai, Xin; Wen, Tingyi

    2012-04-01

    L-Serine plays a critical role as a building block for cell growth, and thus it is difficult to achieve the direct fermentation of L-serine from glucose. In this study, Corynebacterium glutamicum ATCC 13032 was engineered de novo by blocking and attenuating the conversion of L-serine to pyruvate and glycine, releasing the feedback inhibition by L-serine to 3-phosphoglycerate dehydrogenase (PGDH), in combination with the co-expression of 3-phosphoglycerate kinase (PGK) and feedback-resistant PGDH (PGDH(r)). The resulting strain, SER-8, exhibited a lower specific growth rate and significant differences in L-serine levels from Phase I to Phase V as determined for fed-batch fermentation. The intracellular L-serine pool reached (14.22 ± 1.41) μmol g(CDM) (-1), which was higher than glycine pool, contrary to fermentation with the wild-type strain. Furthermore, metabolic flux analysis demonstrated that the over-expression of PGK directed the flux of the pentose phosphate pathway (PPP) towards the glycolysis pathway (EMP), and the expression of PGDH(r) improved the L-serine biosynthesis pathway. In addition, the flux from L-serine to glycine dropped by 24%, indicating that the deletion of the activator GlyR resulted in down-regulation of serine hydroxymethyltransferase (SHMT) expression. Taken together, our findings imply that L-serine pool management is fundamental for sustaining the viability of C. glutamicum, and improvement of C(1) units generation by introducing the glycine cleavage system (GCV) to degrade the excessive glycine is a promising target for L-serine production in C. glutamicum.

  15. Substrate cycles in Penicillium chrysogenum quantified by isotopic non-stationary flux analysis

    PubMed Central

    2012-01-01

    Background Penicillium chrysogenum, the main production strain for penicillin-G, has a high content of intracellular carbohydrates, especially reduced sugars such as mannitol, arabitol, erythritol, as well as trehalose and glycogen. In previous steady state 13C wash-in experiments a delay of labeling enrichments in glycolytic intermediates was observed, which suggests turnover of storage carbohydrates. The turnover of storage pools consumes ATP which is expected to reduce the product yield for energy demanding production pathways like penicillin-G. Results In this study, a 13C labeling wash-in experiment of 1 hour was performed to systematically quantify the intracellular flux distribution including eight substrate cycles. The experiments were performed using a mixed carbon source of 85% CmolGlc/CmolGlc+EtOH labeled glucose (mixture of 90% [1-13C1] and 10% [U-13C6]) and 15% ethanol [U-13C2]. It was found, that (1) also several extracellular pools are enriched with 13C labeling rapidly (trehalose, mannitol, and others), (2) the intra- to extracellular metabolite concentration ratios were comparable for a large set of metabolites while for some carbohydrates (mannitol, trehalose, and glucose) the measured ratios were much higher. Conclusions The fast enrichment of several extracellular carbohydrates and a concentration ratio higher than the ratio expected from cell lysis (2%) indicate active (e.g. ATP consuming) transport cycles over the cellular membrane. The flux estimation indicates, that substrate cycles account for about 52% of the gap in the ATP balance based on metabolic flux analysis. PMID:23098235

  16. LES of large wind farm during a diurnal cycle: Analysis of Energy and Scalar flux budgets

    NASA Astrophysics Data System (ADS)

    Sharma, V.; Calaf, M.; Parlange, M. B.

    2014-12-01

    With an expanding role of wind energy in satisfying energy demands around the world, wind farms are covering increasingly larger surfaces to the point where interaction between wind farms and the atmospheric boundary layer (ABL) might have significant implications. Furthermore, many wind farm sites lie over existing farmland for which water is a precious resource. In this context, a relevant question yet to be fully understood, is whether large wind farms alter near surface temperatures and evaporation rates and if so, by how much. In the present study, Large Eddy Simulation (LES) of a geostrophic wind driven ABL with two active scalars, temperature and specific humidity, in the presence of Coriolis forces with an embedded wind farm are performed. Multiple 'synthetic' diurnal cycles are simulated by imposing a time-varying surface temperature and specific humidity. Wind turbines are modeled using the "actuator disk" approach along with the flexibility to reorient according to varying flow directions. LES is performed using the "pseudo-spectral" approach implying that an infinitely large wind farm is simulated. Comparison of simulations with and without wind farms show clear differences in vertical profiles of horizontal velocity magnitude and direction, turbulent kinetic energy and scalar fluxes. To better understand these differences, a detailed analysis of the constituent terms of budget equations of mean and turbulent kinetic energy and sensible and latent heat fluxes has been performed for different stratification regimes as the ABL evolves during the diurnal cycle. The analyses help explain the effect of wind farms on the characteristics of the low-level jet, depth of the stable boundary layer, formation and growth of the convective boundary layer (CBL) and scalar fluxes at the surface.

  17. Isotopically nonstationary 13C flux analysis of changes in Arabidopsis thaliana leaf metabolism due to high light acclimation

    DOE PAGES

    Ma, Fangfang; Jazmin, Lara J.; Young, Jamey D.; ...

    2014-11-03

    Improving plant productivity is an important aim for metabolic engineering. There are few comprehensive methods that quantitatively describe leaf metabolism, although such information would be valuable for increasing photosynthetic capacity, enhancing biomass production, and rerouting carbon flux toward desirable end products. Isotopically nonstationary metabolic flux analysis (INST-MFA) has been previously applied to map carbon fluxes in photoautotrophic bacteria, which involves model-based regression of transient 13C-labeling patterns of intracellular metabolites. However, experimental and computational difficulties have hindered its application to terrestrial plant systems. Here, we performed in vivo isotopic labeling of Arabidopsis thaliana rosettes with 13CO2 and estimated fluxes throughout leafmore » photosynthetic metabolism by INST-MFA. Plants grown at 200 µmol m$-$2s$-$1 light were compared with plants acclimated for 9 d at an irradiance of 500 µmol∙m$-$2∙s$-$1. Approximately 1,400 independent mass isotopomer measurements obtained from analysis of 37 metabolite fragment ions were regressed to estimate 136 total fluxes (54 free fluxes) under each condition. The results provide a comprehensive description of changes in carbon partitioning and overall photosynthetic flux after long-term developmental acclimation of leaves to high light. Despite a doubling in the carboxylation rate, the photorespiratory flux increased from 17 to 28% of net CO2 assimilation with high-light acclimation (Vc/Vo: 3.5:1 vs. 2.3:1, respectively). In conclusion, this study highlights the potential of 13C INST-MFA to describe emergent flux phenotypes that respond to environmental conditions or plant physiology and cannot be obtained by other complementary approaches.« less

  18. Application of metabolic flux analysis for the identification of metabolic bottlenecks in the biosynthesis of penicillin-G.

    PubMed

    van Gulik, W M; de Laat, W T; Vinke, J L; Heijnen, J J

    2000-06-20

    A detailed stoichiometric model was developed for growth and penicillin-G production in Penicillium chrysogenum. From an a priori metabolic flux analysis using this model it appeared that penicillin production requires significant changes in fluxes through the primary metabolic pathways. This is brought about by the biosynthesis of carbon precursors for the beta-lactan nucleus and an increased demand for NADPH, mainly for sulfate reduction. As a result, significant changes in flux partitioning occur around four principal nodes in primary metabolism. These are located at: (1) glucose-6-phosphate; (2) 3-phosphoglycerate; (3) mitochondrial pyruvate; and (4) mitochondrial isocitrate. These nodes should be regarded as potential bottlenecks for increased productivity. The flexibility of these principal nodes was investigated by experimental manipulation of the fluxes through the central metabolic pathways using a high-producing strain of P. chrysogenum. Metabolic fluxes were manipulated through growth of the cells on different substrates in carbon-limited chemostat culture. Metabolic flux analysis, based on measured input and output fluxes, was used to calculate the fluxes around the principal nodes. It was found that, for growth on glucose, ethanol, and acetate, the flux partitioning around these nodes differed significantly. However, this had hardly any effect on penicillin productivity, showing that primary carbon metabolism is not likely to contain potential bottlenecks. Further experiments were performed to manipulate the total metabolic demand for the cofactor nicotinamide adenine dinucleotide phosphate (NADPH). NADPH demand was increased stepwise by cultivating the cells on glucose or xylose as the carbon source combined with either ammonia or nitrate as the nitrogen source, which resulted in a stepwise decrease of penicillin production. This clearly shows that, in penicillin fermentation, possible limitations in primary metabolism reside in the supply

  19. Coordinated analysis and quantification of sedimentary fluxes and budgets in cold environments: The SEDIBUD Programme

    NASA Astrophysics Data System (ADS)

    Beylich, Achim A.; Lamoureux, Scott F.

    2010-05-01

    Amplified climate change and ecological sensitivity of polar and high-altitude cold environments has been highlighted as a key global environmental issue. Projected climate change in cold climate environments is expected to alter melt season duration and intensity, along with the number of extreme rainfall events, total annual precipitation and the balance between snowfall and rainfall. Similarly, changes to the thermal balance are expected to reduce the extent of permafrost and seasonal ground frost and increase active layer depth. These effects will undoubtedly change surface environments in cold environments and alter fluxes of sediments, nutrients and solutes, but the absense of data and coordinated analysis to understand the sensitivity of the surface environment are acute in cold climate environments. The SEDIBUD (Sediment Budgets in Cold Environments) Programme of the International Association of Geomorphologists (I.A.G./A.I.G.) was formed in 2005 to address this key knowledge gap. SEDIBUD has currently about 400 members worldwide and the Steering Committee of this international programme is composed of ten scientists from nine different countries. The central research question of this global group of scientists is to Assess the contemporary sedimentary fluxes in cold climates, with emphasis on both particulate and dissolved components. Research carried out at currently 38 defined SEDIBUD Key Test Sites varies by programme, logistics and available ressources, but typically represent interdisciplinary collaborations of geomorphologists, hydrologists, ecologists, and permafrost scientists and glaciologists with different levels of detail. SEDIBUD key test sites provide data on annual climate conditions, total runoff and particulate and dissolved fluxes as well as information on other relevant surface processes. A number of selected key test sites are providing high-resolution data on climatic conditions, runoff and fluvial fluxes, which in addition to the

  20. Metabolic flux analysis of Saccharomyces cerevisiae in a sealed winemaking fermentation system.

    PubMed

    Li, Hua; Su, Jing; Ma, Wen; Guo, Anque; Shan, Zuhua; Wang, Hua

    2015-03-01

    A sealed fermentation (SF) system and an anaerobic fermentation (AF) system (under normal atmospheric pressure conditions) were employed to study the influence of endogenous carbon dioxide (CO2) on the metabolism of Saccharomyces cerevisiae. The results showed that the fermentation stopped when 82.0 g L(-1) glucose was consumed and the endogenously produced CO2: pressure reached to 14.3 MPa in SF system, while the sugar was used up during AF. The total yeast viable count in the end of AF was higher than that of SF. It was also observed that the ethanol yield in AF and SF was similar, the glycerol yield in AF was 1.26 times higher than that in SF, while the succinic acid and acetic acid yields in SF were 24.7 and 26 times higher than that in AF, respectively. Additionally, this work provides a stoichiometric model used for metabolic flux analysis of S. cerevisiae to compare the flux distribution in SF and AF. The results showed that CO2 had an important effect on the pathways of oxaloacetic acid formation from pyruvic acid and ribose-5-phosphate formation from glucose-6-phosphate. However, the pathway of ethanol formation from pyruvic acid (decarboxylation reaction), catalyzed by pyruvate decarboxylase, was insensitive to CO2.

  1. Two-Flux Green's Function Analysis for Transient Spectral Radiation in a Composite

    NASA Technical Reports Server (NTRS)

    Siegel, Robert

    1996-01-01

    An analysis is developed for obtaining transient temperatures in a two-layer semitransparent composite with spectrally dependent properties. Each external boundary of the composite is subjected to radiation and convection. The two-flux radiative transfer equations are solved by deriving a Green's function. This yields the local radiative heat source needed to numerically solve the transient energy equation. An advantage of the two-flux method is that isotropic scattering is included without added complexity. The layer refractive indices are larger than one. This produces internal reflections at the boundaries and the internal interface; the reflections are assumed diffuse. Spectral results using the Green's function method are verified by comparing with numerical solutions using the exact radiative transfer equations. Transient temperature distributions are given to illustrate the effect of radiative heating on one side of a composite with external convective cooling. The protection of a material from incident radiation is illustrated by adding scattering to the layer adjacent to the radiative source.

  2. Quantification of Metabolic Rearrangements During Neural Stem Cells Differentiation into Astrocytes by Metabolic Flux Analysis.

    PubMed

    Sá, João V; Kleiderman, Susanne; Brito, Catarina; Sonnewald, Ursula; Leist, Marcel; Teixeira, Ana P; Alves, Paula M

    2017-01-01

    Proliferation and differentiation of neural stem cells (NSCs) have a crucial role to ensure neurogenesis and gliogenesis in the mammalian brain throughout life. As there is growing evidence for the significance of metabolism in regulating cell fate, knowledge on the metabolic programs in NSCs and how they evolve during differentiation into somatic cells may provide novel therapeutic approaches to address brain diseases. In this work, we applied a quantitative analysis to assess how the central carbon metabolism evolves upon differentiation of NSCs into astrocytes. Murine embryonic stem cell (mESC)-derived NSCs and astrocytes were incubated with labelled [1-(13)C]glucose and the label incorporation into intracellular metabolites was followed by GC-MS. The obtained (13)C labelling patterns, together with uptake/secretion rates determined from supernatant analysis, were integrated into an isotopic non-stationary metabolic flux analysis ((13)C-MFA) model to estimate intracellular flux maps. Significant metabolic differences between NSCs and astrocytes were identified, with a general downregulation of central carbon metabolism during astrocytic differentiation. While glucose uptake was 1.7-fold higher in NSCs (on a per cell basis), a high lactate-secreting phenotype was common to both cell types. Furthermore, NSCs consumed glutamine from the medium; the highly active reductive carboxylation of alpha-ketoglutarate indicates that this was converted to citrate and used for biosynthetic purposes. In astrocytes, pyruvate entered the TCA cycle mostly through pyruvate carboxylase (81%). This pathway supported glutamine and citrate secretion, recapitulating well described metabolic features of these cells in vivo. Overall, this fluxomics study allowed us to quantify the metabolic rewiring accompanying astrocytic lineage specification from NSCs.

  3. Advances in the quantification of mitochondrial function in primary human immune cells through extracellular flux analysis

    PubMed Central

    Ip, Blanche C.; Habib, Chloe; Ritou, Eleni; Grammatopoulos, Tom N.; Steenkamp, Devin; Dooms, Hans; Apovian, Caroline M.; Lauffenburger, Douglas A.

    2017-01-01

    Numerous studies show that mitochondrial energy generation determines the effectiveness of immune responses. Furthermore, changes in mitochondrial function may regulate lymphocyte function in inflammatory diseases like type 2 diabetes. Analysis of lymphocyte mitochondrial function has been facilitated by introduction of 96-well format extracellular flux (XF96) analyzers, but the technology remains imperfect for analysis of human lymphocytes. Limitations in XF technology include the lack of practical protocols for analysis of archived human cells, and inadequate data analysis tools that require manual quality checks. Current analysis tools for XF outcomes are also unable to automatically assess data quality and delete untenable data from the relatively high number of biological replicates needed to power complex human cell studies. The objectives of work presented herein are to test the impact of common cellular manipulations on XF outcomes, and to develop and validate a new automated tool that objectively analyzes a virtually unlimited number of samples to quantitate mitochondrial function in immune cells. We present significant improvements on previous XF analyses of primary human cells that will be absolutely essential to test the prediction that changes in immune cell mitochondrial function and fuel sources support immune dysfunction in chronic inflammatory diseases like type 2 diabetes. PMID:28178278

  4. Advances in the quantification of mitochondrial function in primary human immune cells through extracellular flux analysis.

    PubMed

    Nicholas, Dequina; Proctor, Elizabeth A; Raval, Forum M; Ip, Blanche C; Habib, Chloe; Ritou, Eleni; Grammatopoulos, Tom N; Steenkamp, Devin; Dooms, Hans; Apovian, Caroline M; Lauffenburger, Douglas A; Nikolajczyk, Barbara S

    2017-01-01

    Numerous studies show that mitochondrial energy generation determines the effectiveness of immune responses. Furthermore, changes in mitochondrial function may regulate lymphocyte function in inflammatory diseases like type 2 diabetes. Analysis of lymphocyte mitochondrial function has been facilitated by introduction of 96-well format extracellular flux (XF96) analyzers, but the technology remains imperfect for analysis of human lymphocytes. Limitations in XF technology include the lack of practical protocols for analysis of archived human cells, and inadequate data analysis tools that require manual quality checks. Current analysis tools for XF outcomes are also unable to automatically assess data quality and delete untenable data from the relatively high number of biological replicates needed to power complex human cell studies. The objectives of work presented herein are to test the impact of common cellular manipulations on XF outcomes, and to develop and validate a new automated tool that objectively analyzes a virtually unlimited number of samples to quantitate mitochondrial function in immune cells. We present significant improvements on previous XF analyses of primary human cells that will be absolutely essential to test the prediction that changes in immune cell mitochondrial function and fuel sources support immune dysfunction in chronic inflammatory diseases like type 2 diabetes.

  5. Sensitivity Analysis of the Land Surface Model NOAH-MP for Different Model Fluxes

    NASA Astrophysics Data System (ADS)

    Thober, S.; Mai, J.; Samaniego, L. E.; Clark, M. P.; Mendoza, P. A.; Wulfmeyer, V. G.; Branch, O.; Attinger, S.; Kumar, R.; Cuntz, M.

    2014-12-01

    The land-atmosphere fluxes of water, energy and carbon, as computed by the Land Surface Model (LSM), are a critical component of Earth System Models and Numerical Weather Prediction models. Processes and parameters of LSMs are validated mostly against point measurements, for example from Eddy-covariance towers, with much attention given to biophysical processes and vegetation parameters. River discharge on the other hand is not considered very often although it provides an integrated signal of the hydrologic cycle over a catchment. Sensitivity analyses of hydrologic models have shown that soil parameters have then the largest impact on modeled river discharge. In this study, we quantify parametric sensitivities of the land surface model NOAH-MP simultaneously for model outputs at different spatial resolutions. NOAH-MP is a state-of-the-art LSM, which is used at regional scale as the land surface scheme of the atmospheric Weather Research and Forecasting Model (WRF). NOAH-MP contains multiple process parameterizations (hence MP), yielding a considerable amount of parameters (> 500). Standard methods for sensitivity analysis such as Sobol indexes require too many model evaluations in case of many parameters. We therefore use first a recently developed inexpensive screening method based on Elementary Effects that has proven to identify the same informative parameters as the Sobol method but requires only 1% of model evaluations. This reduces the number of parameters to a feasible amount for a thorough sensitivity analysis. The study is conducted on twelve Model Parameter Estimation Experiment (MOPEX) catchments. This allows investigation of parametric sensitivities for distinct hydro-climatic characteristics, emphasizing different land-surface processes. The river basins range in size from 1020 to 4421 km^2, allowing fast model evaluation. The screening and sensitivity analysis identifies the most informative parameters of NOAH-MP for different model output variables

  6. Metabolic network reconstruction and flux variability analysis of storage synthesis in developing oilseed rape (Brassica napus L.) embryos

    SciTech Connect

    Hay, J.; Schwender, J.

    2011-08-01

    Computational simulation of large-scale biochemical networks can be used to analyze and predict the metabolic behavior of an organism, such as a developing seed. Based on the biochemical literature, pathways databases and decision rules defining reaction directionality we reconstructed bna572, a stoichiometric metabolic network model representing Brassica napus seed storage metabolism. In the highly compartmentalized network about 25% of the 572 reactions are transport reactions interconnecting nine subcellular compartments and the environment. According to known physiological capabilities of developing B. napus embryos, four nutritional conditions were defined to simulate heterotrophy or photoheterotrophy, each in combination with the availability of inorganic nitrogen (ammonia, nitrate) or amino acids as nitrogen sources. Based on mathematical linear optimization the optimal solution space was comprehensively explored by flux variability analysis, thereby identifying for each reaction the range of flux values allowable under optimality. The range and variability of flux values was then categorized into flux variability types. Across the four nutritional conditions, approximately 13% of the reactions have variable flux values and 10-11% are substitutable (can be inactive), both indicating metabolic redundancy given, for example, by isoenzymes, subcellular compartmentalization or the presence of alternative pathways. About one-third of the reactions are never used and are associated with pathways that are suboptimal for storage synthesis. Fifty-seven reactions change flux variability type among the different nutritional conditions, indicating their function in metabolic adjustments. This predictive modeling framework allows analysis and quantitative exploration of storage metabolism of a developing B. napus oilseed.

  7. Radiation flux and spectral analysis of the multi-temperature Z dynamic hohlraum

    SciTech Connect

    Lockard, T. E.; Idzorek, G. C.; Tierney, T. E. IV; Watt, R. G.

    2008-10-15

    Experiments performed at the Sandia National Laboratories (SNL) Z-machine, located in Albuquerque, New Mexico produce hot ({approx}220 eV) plasmas. X-ray emission from the plasma is used to drive radiation flow experiments. Our standard plasma diagnostic suite consists of x-ray diodes (XRDs), silicon photodiodes, and nickel thin film bolometers. Small diagnostic holes allow us to view the hot plasma from the side, top axial anode side, and bottom axial cathode side. Computer software has been written to process the raw data to calculate data quality, fold in detector spectral response and experiment geometry for emitted flux, calculate a multidetector spectral unfold, and yield an equivalent time-dependent Planckian temperature profile. Spectral unfolds of our XRD data generally yield a Planckian-like spectrum. In our presentation we will compare our diagnostic techniques, analysis, and results to more accurately characterize spectral unfolds in order to establish better drive conditions for our experiments.

  8. Equilibrium analysis of carbon pools and fluxes of forest biomes in the former Soviet Union

    SciTech Connect

    Kolchugina, T.P.; Vinson, T.S.

    1993-01-01

    Forests are an important component of the biosphere and sequestration of carbon in boreal forests may represent one of the few realistic alternatives to ameliorate changes in atmospheric chemistry. The former Soviet Union has the greatest expanse of boreal forests in the world; however, the role of these forests in the terrestrial carbon cycle is not fully understood because the carbon budget of the Soviet forest sector has not been established. In recognition of the need to determine the role of these forests in the global carbon cycle, the carbon budget of forest biomes in the former Soviet Union was assessed based on an equilibrium analysis of carbon cycle pools and fluxes. Net primary productivity was used to identify the rate of carbon turnover in the forest biomes.

  9. Flux control coefficients determined by inhibitor titration: the design and analysis of experiments to minimize errors.

    PubMed Central

    Small, J R

    1993-01-01

    This paper is a study into the effects of experimental error on the estimated values of flux control coefficients obtained using specific inhibitors. Two possible techniques for analysing the experimental data are compared: a simple extrapolation method (the so-called graph method) and a non-linear function fitting method. For these techniques, the sources of systematic errors are identified and the effects of systematic and random errors are quantified, using both statistical analysis and numerical computation. It is shown that the graph method is very sensitive to random errors and, under all conditions studied, that the fitting method, even under conditions where the assumptions underlying the fitted function do not hold, outperformed the graph method. Possible ways of designing experiments to minimize the effects of experimental errors are analysed and discussed. PMID:8257434

  10. The May 17, 2012 Solar Event: Back-Tracing Analysis and Flux Reconstruction with PAMELA

    NASA Technical Reports Server (NTRS)

    Bruno, A.; Adriani, O.; Barbarino, G. C.; Bazilevskaya, G. A.; Bellotti, R.; Boezio, M.; Bogomolov, E. A.; Bongi, M.; Bonvicini, V.; Bottai, S.; Bravar, U.; Cafagna, F.; Campana, D.; Carbone, R.; Carlson, P.; Casolino, M.; Castellini, G.; Christian, Eric R.

    2016-01-01

    The PAMELA space experiment is providing first direct observations of Solar Energetic Particles (SEPs) with energies from about 80 MeV to several GeV in near-Earth orbit, bridging the low energy measurements by other spacecrafts and the Ground Level Enhancement (GLE) data by the worldwide network of neutron monitors. Its unique observational capabilities include the possibility of measuring the flux angular distribution and thus investigating possible anisotropies associated to SEP events. The analysis is supported by an accurate back-tracing simulation based on a realistic description of the Earth's magnetosphere, which is exploited to estimate the SEP energy spectra as a function of the asymptotic direction of arrival with respect to the Interplanetary Magnetic Field (IMF). In this work we report the results for the May 17, 2012 event.

  11. Radiation flux and spectral analysis of the multi-temperature Z dynamic hohlraum.

    PubMed

    Lockard, T E; Idzorek, G C; Tierney, T E; Watt, R G

    2008-10-01

    Experiments performed at the Sandia National Laboratories (SNL) Z-machine, located in Albuquerque, New Mexico produce hot (approximately 220 eV) plasmas. X-ray emission from the plasma is used to drive radiation flow experiments. Our standard plasma diagnostic suite consists of x-ray diodes (XRDs), silicon photodiodes, and nickel thin film bolometers. Small diagnostic holes allow us to view the hot plasma from the side, top axial anode side, and bottom axial cathode side. Computer software has been written to process the raw data to calculate data quality, fold in detector spectral response and experiment geometry for emitted flux, calculate a multidetector spectral unfold, and yield an equivalent time-dependent Planckian temperature profile. Spectral unfolds of our XRD data generally yield a Planckian-like spectrum. In our presentation we will compare our diagnostic techniques, analysis, and results to more accurately characterize spectral unfolds in order to establish better drive conditions for our experiments.

  12. The Infrared Astronomical Satellite /IRAS/ Scientific Data Analysis System /SDAS/ sky flux subsystem

    NASA Technical Reports Server (NTRS)

    Stagner, J. R.; Girard, M. A.

    1980-01-01

    The sky flux subsystem of the Infrared Astronomical Satellite Scientific Data Analysis System is described. Its major output capabilities are (1) the all-sky lune maps (8-arcminute pixel size), (2) galactic plane maps (2-arcminute pixel size) and (3) regional maps of small areas such as extended sources greater than 1-degree in extent. The major processing functions are to (1) merge the CRDD and pointing data, (2) phase the detector streams, (3) compress the detector streams in the in-scan and cross-scan directions, and (4) extract data. Functional diagrams of the various capabilities of the subsystem are given. Although this device is inherently nonimaging, various calibrated and geometrically controlled imaging products are created, suitable for quantitative and qualitative scientific interpretation.

  13. The May 17, 2012 solar event: back-tracing analysis and flux reconstruction with PAMELA

    NASA Astrophysics Data System (ADS)

    Bruno, A.; Adriani, O.; Barbarino, G. C.; Bazilevskaya, G. A.; Bellotti, R.; Boezio, M.; Bogomolov, E. A.; Bongi, M.; Bonvicini, V.; Bottai, S.; Bravar, U.; Cafagna, F.; Campana, D.; Carbone, R.; Carlson, P.; Casolino, M.; Castellini, G.; Christian, E. C.; De Donato, C.; de Nolfo, G. A.; De Santis, C.; De Simone, N.; Di Felice, V.; Formato, V.; Galper, A. M.; Karelin, A. V.; Koldashov, S. V.; Koldobskiy, S.; Krutkov, S. Y.; Kvashnin, A. N.; Lee, M.; Leonov, A.; Malakhov, V.; Marcelli, L.; Martucci, M.; Mayorov, A. G.; Menn, W.; Mergè, M.; Mikhailov, V. V.; Mocchiutti, E.; Monaco, A.; Mori, N.; Munini, R.; Osteria, G.; Palma, F.; Panico, B.; Papini, P.; Pearce, M.; Picozza, P.; Ricci, M.; Ricciarini, S. B.; Ryan, J. M.; Sarkar, R.; Scotti, V.; Simon, M.; Sparvoli, R.; Spillantini, P.; Stochaj, S.; Stozhkov, Y. I.; Vacchi, A.; Vannuccini, E.; Vasilyev, G. I.; Voronov, S. A.; Yurkin, Y. T.; Zampa, G.; Zampa, N.; Zverev, V. G.

    2016-02-01

    The PAMELA space experiment is providing first direct observations of Solar Energetic Particles (SEPs) with energies from about 80 MeV to several GeV in near-Earth orbit, bridging the low energy measurements by other spacecrafts and the Ground Level Enhancement (GLE) data by the worldwide network of neutron monitors. Its unique observational capabilities include the possibility of measuring the flux angular distribution and thus investigating possible anisotropies associated to SEP events. The analysis is supported by an accurate back-tracing simulation based on a realistic description of the Earth's magnetosphere, which is exploited to estimate the SEP energy spectra as a function of the asymptotic direction of arrival with respect to the Interplanetary Magnetic Field (IMF). In this work we report the results for the May 17, 2012 event.

  14. OM-FBA: Integrate Transcriptomics Data with Flux Balance Analysis to Decipher the Cell Metabolism.

    PubMed

    Guo, Weihua; Feng, Xueyang

    2016-01-01

    Constraint-based metabolic modeling such as flux balance analysis (FBA) has been widely used to simulate cell metabolism. Thanks to its simplicity and flexibility, numerous algorithms have been developed based on FBA and successfully predicted the phenotypes of various biological systems. However, their phenotype predictions may not always be accurate in FBA because of using the objective function that is assumed for cell metabolism. To overcome this challenge, we have developed a novel computational framework, namely omFBA, to integrate multi-omics data (e.g. transcriptomics) into FBA to obtain omics-guided objective functions with high accuracy. In general, we first collected transcriptomics data and phenotype data from published database (e.g. GEO database) for different microorganisms such as Saccharomyces cerevisiae. We then developed a "Phenotype Match" algorithm to derive an objective function for FBA that could lead to the most accurate estimation of the known phenotype (e.g. ethanol yield). The derived objective function was next correlated with the transcriptomics data via regression analysis to generate the omics-guided objective function, which was next used to accurately simulate cell metabolism at unknown conditions. We have applied omFBA in studying sugar metabolism of S. cerevisiae and found that the ethanol yield could be accurately predicted in most of the cases tested (>80%) by using transcriptomics data alone, and revealed valuable metabolic insights such as the dynamics of flux ratios. Overall, omFBA presents a novel platform to potentially integrate multi-omics data simultaneously and could be incorporated with other FBA-derived tools by replacing the arbitrary objective function with the omics-guided objective functions.

  15. Soil carbonyl sulfide fluxes in a Mediterranean ecosystem: insights from model-data fusion analysis

    NASA Astrophysics Data System (ADS)

    Sun, W.; Seibt, U. H.; Maseyk, K. S.; Lett, C.

    2013-12-01

    Carbonyl sulfide (COS) is linked to biosphere components of the carbon cycle, due in large part to its hydrolysis by the enzyme carbonic anhydrase (CA). Stomatal diffusion models and observations at leaf and ecosystem scales have demonstrated the potential of COS as a tracer for Gross Primary Production (GPP). Although considered small relative to canopy COS fluxes, accurate knowledge of soil COS fluxes is required for the use of net ecosystem COS fluxes in carbon flux partitioning. However, extensive field measurements of soil COS fluxes are rare and process-based modeling is limited. Here we report continuous chamber measurements of soil COS fluxes in a Mediterranean ecosystem in the Santa Monica Mountains, California during April and early May 2013. Both COS uptake and emissions were observed, but the soil acted as a net sink in most conditions and was a net source only when soil temperatures were above 22 C. COS sink fluxes were positively correlated with soil water content and CO2 fluxes. COS uptake had a maximum at a temperature around 15 C. However, no single environmental variable could be correlated to COS fluxes with an r-square > 0.6. COS fluxes from soil chambers ranged from -9 to 2.5 pmol m-2 s-1. Leaf litter appeared to increase soil COS metabolic activity. We observed huge bursts of soil COS uptake induced by a precipitation event, probably due to enhanced soil microbial activity resulting from alleviated water limitation and a decrease in soil temperature towards the optimum. We used a soil gas exchange model coupled with CA enzyme kinetics to simulate the soil COS fluxes. Micrometeorological and soil data were used to drive the soil flux model. Model simulations indicated that diurnal and synoptic variations of COS fluxes were driven by soil temperature and water content, controlling both CA activity and diffusion. We suggest that multiple parameters need to be optimized to reduce uncertainties in models of soil COS fluxes at larger scales.

  16. Flux balance analysis reveals acetate metabolism modulates cyclic electron flow and alternative glycolytic pathways in Chlamydomonas reinhardtii.

    PubMed

    Chapman, Stephen P; Paget, Caroline M; Johnson, Giles N; Schwartz, Jean-Marc

    2015-01-01

    Cells of the green alga Chlamydomonas reinhardtii cultured in the presence of acetate perform mixotrophic growth, involving both photosynthesis and organic carbon assimilation. Under such conditions, cells exhibit a reduced capacity for photosynthesis but a higher growth rate, compared to phototrophic cultures. Better understanding of the down regulation of photosynthesis would enable more efficient conversion of carbon into valuable products like biofuels. In this study, Flux Balance Analysis (FBA) and Flux Variability Analysis (FVA) have been used with a genome scale model of C. reinhardtii to examine changes in intracellular flux distribution in order to explain their changing physiology. Additionally, a reaction essentiality analysis was performed to identify which reaction subsets are essential for a given growth condition. Our results suggest that exogenous acetate feeds into a modified tricarboxylic acid (TCA) cycle, which bypasses the CO2 evolution steps, explaining increases in biomass, consistent with experimental data. In addition, reactions of the oxidative pentose phosphate and glycolysis pathways, inactive under phototrophic conditions, show substantial flux under mixotrophic conditions. Importantly, acetate addition leads to an increased flux through cyclic electron flow (CEF), but results in a repression of CO2 fixation via Rubisco, explaining the down regulation of photosynthesis. However, although CEF enhances growth on acetate, it is not essential-impairment of CEF results in alternative metabolic pathways being increased. We have demonstrated how the reactions of photosynthesis interconnect with carbon metabolism on a global scale, and how systems approaches play a viable tool in understanding complex relationships at the scale of the organism.

  17. Flux balance analysis reveals acetate metabolism modulates cyclic electron flow and alternative glycolytic pathways in Chlamydomonas reinhardtii

    PubMed Central

    Chapman, Stephen P.; Paget, Caroline M.; Johnson, Giles N.; Schwartz, Jean-Marc

    2015-01-01

    Cells of the green alga Chlamydomonas reinhardtii cultured in the presence of acetate perform mixotrophic growth, involving both photosynthesis and organic carbon assimilation. Under such conditions, cells exhibit a reduced capacity for photosynthesis but a higher growth rate, compared to phototrophic cultures. Better understanding of the down regulation of photosynthesis would enable more efficient conversion of carbon into valuable products like biofuels. In this study, Flux Balance Analysis (FBA) and Flux Variability Analysis (FVA) have been used with a genome scale model of C. reinhardtii to examine changes in intracellular flux distribution in order to explain their changing physiology. Additionally, a reaction essentiality analysis was performed to identify which reaction subsets are essential for a given growth condition. Our results suggest that exogenous acetate feeds into a modified tricarboxylic acid (TCA) cycle, which bypasses the CO2 evolution steps, explaining increases in biomass, consistent with experimental data. In addition, reactions of the oxidative pentose phosphate and glycolysis pathways, inactive under phototrophic conditions, show substantial flux under mixotrophic conditions. Importantly, acetate addition leads to an increased flux through cyclic electron flow (CEF), but results in a repression of CO2 fixation via Rubisco, explaining the down regulation of photosynthesis. However, although CEF enhances growth on acetate, it is not essential—impairment of CEF results in alternative metabolic pathways being increased. We have demonstrated how the reactions of photosynthesis interconnect with carbon metabolism on a global scale, and how systems approaches play a viable tool in understanding complex relationships at the scale of the organism. PMID:26175742

  18. Effects of measurement resolution on the analysis of temperature time series for stream-aquifer flux estimation

    NASA Astrophysics Data System (ADS)

    Soto-López, Carlos D.; Meixner, Thomas; Ferré, Ty P. A.

    2011-12-01

    From its inception in the mid-1960s, the use of temperature time series (thermographs) to estimate vertical fluxes has found increasing use in the hydrologic community. Beginning in 2000, researchers have examined the impacts of measurement and parameter uncertainty on the estimates of vertical fluxes. To date, the effects of temperature measurement discretization (resolution), a characteristic of all digital temperature loggers, on the determination of vertical fluxes has not been considered. In this technical note we expand the analysis of recently published work to include the effects of temperature measurement resolution on estimates of vertical fluxes using temperature amplitude and phase shift information. We show that errors in thermal front velocity estimation introduced by discretizing thermographs differ when amplitude or phase shift data are used to estimate vertical fluxes. We also show that under similar circumstances sensor resolution limits the range over which vertical velocities are accurately reproduced more than uncertainty in temperature measurements, uncertainty in sensor separation distance, and uncertainty in the thermal diffusivity combined. These effects represent the baseline error present and thus the best-case scenario when discrete temperature measurements are used to infer vertical fluxes. The errors associated with measurement resolution can be minimized by using the highest-resolution sensors available. But thoughtful experimental design could allow users to select the most cost-effective temperature sensors to fit their measurement needs.

  19. Flux balance analysis of ammonia assimilation network in E. coli predicts preferred regulation point.

    PubMed

    Wang, Lu; Lai, Luhua; Ouyang, Qi; Tang, Chao

    2011-01-25

    Nitrogen assimilation is a critical biological process for the synthesis of biomolecules in Escherichia coli. The central ammonium assimilation network in E. coli converts carbon skeleton α-ketoglutarate and ammonium into glutamate and glutamine, which further serve as nitrogen donors for nitrogen metabolism in the cell. This reaction network involves three enzymes: glutamate dehydrogenase (GDH), glutamine synthetase (GS) and glutamate synthase (GOGAT). In minimal media, E. coli tries to maintain an optimal growth rate by regulating the activity of the enzymes to match the availability of the external ammonia. The molecular mechanism and the strategy of the regulation in this network have been the research topics for many investigators. In this paper, we develop a flux balance model for the nitrogen metabolism, taking into account of the cellular composition and biosynthetic requirements for nitrogen. The model agrees well with known experimental results. Specifically, it reproduces all the (15)N isotope labeling experiments in the wild type and the two mutant (ΔGDH and ΔGOGAT) strains of E. coli. Furthermore, the predicted catalytic activities of GDH, GS and GOGAT in different ammonium concentrations and growth rates for the wild type, ΔGDH and ΔGOGAT strains agree well with the enzyme concentrations obtained from western blots. Based on this flux balance model, we show that GS is the preferred regulation point among the three enzymes in the nitrogen assimilation network. Our analysis reveals the pattern of regulation in this central and highly regulated network, thus providing insights into the regulation strategy adopted by the bacteria. Our model and methods may also be useful in future investigations in this and other networks.

  20. Observations and analysis of small-scale magnetic flux ropes in the solar wind

    NASA Astrophysics Data System (ADS)

    Zheng, Jinlei; Hu, Qiang

    2016-11-01

    The small-scale magnetic flux ropes (of duration ranging from a few minutes to a few hours) in the solar wind have the typical topology of winding field lines around a central axis, which is similar to the large-scale flux ropes, i.e., magnetic clouds. However, accumulating evidence suggests that their plasma characteristics, origin, formation mechanism and evolution are different from those of large-scale flux ropes. The small-scale flux ropes are intensively studied in recent years, since they affect particle transport and energization, and are considered as the potential source of local acceleration. The Grad-Shafranov reconstruction technique is a tool to reconstruct the two and a half dimensional field structure based on in-situ measurements captured by an observing platform moving past it. In this study, we reconstruct the flux rope structures in two events using the Grad-Shafranov reconstruction approach. In one event, a twin flux rope structure at 1 AU occurring on 2002 February 1 and two following single flux rope structures are identified behind an interplanetary shock. In the other event, we reconstruct the flux rope structures occurring on 1998 March 25 and 26 at 1 AU in the ambient solar wind. The associated energetic particle signatures and the possible origin of these flux rope structures are discussed.

  1. Does the Blazar Gamma-ray Spectrum Harden with Increasing Flux? - Analysis of Nine Years of EGRET Data

    NASA Technical Reports Server (NTRS)

    Nandikotkur, Giridhar; Jahoda, Keith M.; Hartman, R. C.; Mukherjee, R.; Sreekumar, P.; Boettcher, M.

    2007-01-01

    The Energetic Gamma Ray Experiment Telescope (EGRET) on the Compton Gamma Ray Observatory (CGRO) discovered gamma-ray emission from more than 67 blazars during its nine-year lifetime. We conducted an exhaustive search of the EGRET archives and selected all the blazars that were observed multiple times and were bright enough to enable a spectral analysis using standard powerlaw models. The sample consists of 18 flat-spectrum radio quasars (FSRQs), 6 low-frequency-peaked BL Lacs (LBLs) and 2 high-frequency-peaked BL Lacs (HBLs). We do not detect any clear pattern in'the variation of spectral index with flux. Some of the blazars do not show any statistical evidence for spectral variability. The spectrum hardens with increasing flux in a few cases. There is also evidence for a flux-hardness anticorrelation at lo\\v fluxes in five blazars. The well observed blazars (3C 279,3C 273, PKS 0528-i-134, PKS 1622-297, PKS 0208- 512) do not show any overall trend in the long-term spectral dependence on flux, but the sample shows a mixture of hard and soft states. We observed spectral hysteresis at weekly timescales in all the three FSRQs for which data from flares lasting for 3 approx. 4 weeks were available. All three sources show a counterclockwise rotation despite the widely different flux profiles. Hysteresis in the spectral index vs. flux space has never been observed in FSRQs in gamma-rays at weekly timescales. itre analyze the observed spectral behavior in the context of various inverse-Compton mechanisms believed to be responsible for emission in the EGRET energy range. Our analysis uses the EGRET skymaps that were regenerated to include the changes in performance during the mission.

  2. Partial Safety Analysis for a Reduced Uranium Enrichment Core for the High Flux Isotope Reactor

    SciTech Connect

    Primm, Trent; Gehin, Jess C

    2009-04-01

    A computational model of the reactor core of the High Flux Isotope Rector (HFIR) was developed in order to analyze non-destructive accidents caused by transients during reactor operation. The reactor model was built for the latest version of the nuclear analysis software package called Program for the Analysis of Reactor Transients (PARET). Analyses performed with the model constructed were compared with previous data obtained with other tools in order to benchmark the code. Finally, the model was used to analyze the behavior of the reactor under transients using a different nuclear fuel with lower enrichment of uranium (LEU) than the fuel currently used, which has a high enrichment of uranium (HEU). The study shows that the presence of fertile isotopes in LEU fuel, which increases the neutron resonance absorption, reduces the impact of transients on the fuel and enhances the negative reactivity feedback, thus, within the limitations of this study, making LEU fuel appear to be a safe alternative fuel for the reactor core.

  3. Integrated, step-wise, mass-isotopomeric flux analysis of the TCA Cycle

    PubMed Central

    Alves, Tiago C.; Pongratz, Rebecca L.; Zhao, Xiaojian; Yarborough, Orlando; Sereda, Sam; Shirihai, Orian; Cline, Gary W.; Mason, Graeme; Kibbey, Richard G.

    2015-01-01

    Summary Mass isotopomer multi-ordinate spectral analysis (MIMOSA) is a step-wise flux analysis platform to measure discrete glycolytic and mitochondrial metabolic rates. Importantly, direct citrate synthesis rates were obtained by deconvolving the mass spectra generated from [U-13C6]-D-glucose labeling for position-specific enrichments of mitochondrial acetyl-CoA, oxaloacetate and citrate. Comprehensive steady-state and dynamic analyses of key metabolic rates (pyruvate dehydrogenase, β-oxidation, pyruvate carboxylase, isocitrate dehydrogenase and PEP/pyruvate cycling) were calculated from the position-specific transfer of 13C from sequential precursors to their products. Important limitations of previous techniques were identified. In INS-1 cells, citrate synthase rates correlated with both insulin secretion and oxygen consumption. Pyruvate carboxylase rates were substantially lower than previously reported but showed the highest fold change in response to glucose stimulation. In conclusion, MIMOSA measures key metabolic rates from the precursor/product position-specific transfer of 13C label between metabolites and has broad applicability to any glucose-oxidizing cell. PMID:26411341

  4. Snubber reduction analysis of secondary hot leg in Fast Flux Test Facility

    SciTech Connect

    Chen, W.W.; Lindquist, M.R.

    1995-01-01

    This paper describes analyses to qualify the structural integrity of the secondary hot leg (SHL) dump heat exchanger(DHX) piping system of the Fast Flux Test Facility (FFTF) under seismic loading after the deletion of a number of seismic snubbers. A baseline geometric model was developed, with ADLPIPE computer code, Version 2ElA, 1984. Several seismic analyses were performed by iteration with the maximum number of snubbers reduced to obtain a support configuration having acceptable anchor and support loads as well as acceptable piping stresses. A thermal analysis was performed at an operating temperature of 965{degrees}F (518{degrees}C) to qualify the piping system following the replacement of some seismic snubbers with rigid supports. A series of displacements from flow- or pump-induced vibration were measured. These vibrations, on the order of tens of seconds or minutes apart, were not continuous and steady, but rather of a series of forced displacements that rapidly damped out to zero. A scoping evaluation of the effect of the vibration found that the calculated stress is within limits. Results of the stress analysis of the piping with snubber reduction are qualified in accordance with ASME, Section III, Class 1, requirements. Nearly 67% of the snubbers in loop 1 of the SHL piping system may be eliminated or replaced with rigid struts at the same location and orientation. The effects of inelastic strain accumulation, creep-fatigue damage and interaction, and elastic follow-up are evaluated by comparison to the results of inelastic analysis. Revised anchor and hanger loads also were evaluated by comparison to those from the original seismic analysis.

  5. ANALYSIS OF SOIL VAPOR EXTRACTION DATA TO EVALUATE MASS-TRANSFER CONSTRAINTS AND ESTIMATE SOURCE-ZONE MASS FLUX

    PubMed Central

    Rohay, Virginia; Truex, Michael J.

    2013-01-01

    Methods are developed to use data collected during cyclic operation of soil vapor extraction (SVE) systems to help characterize the magnitudes and timescales of mass flux associated with vadose zone contaminant sources. Operational data collected at the Department of Energy’s Hanford site are used to illustrate the use of such data. An analysis was conducted of carbon tetrachloride vapor concentrations collected during and between SVE operations. The objective of the analysis was to evaluate changes in concentrations measured during periods of operation and non-operation of SVE, with a focus on quantifying temporal dynamics of the vadose zone contaminant mass flux, and associated source strength. Three mass-flux terms, representing mass flux during the initial period of a SVE cycle, during the asymptotic period of a cycle, and during the rebound period, were calculated and compared. It was shown that it is possible to use the data to estimate time frames for effective operation of an SVE system if a sufficient set of historical cyclic operational data exists. This information could then be used to help evaluate changes in SVE operations, including system closure. The mass-flux data would also be useful for risk assessments of the impact of vadose-zone sources on groundwater contamination or vapor intrusion. PMID:23516336

  6. Elementary Flux Mode Analysis Revealed Cyclization Pathway as a Powerful Way for NADPH Regeneration of Central Carbon Metabolism.

    PubMed

    Rui, Bin; Yi, Yin; Shen, Tie; Zheng, Meijuan; Zhou, Wenwei; Du, Honglin; Fan, Yadong; Wang, Yongkang; Zhang, Zhengdong; Xu, Shengsheng; Liu, Zhijie; Wen, Han; Xie, Xiaoyao

    2015-01-01

    NADPH regeneration capacity is attracting growing research attention due to its important role in resisting oxidative stress. Besides, NADPH availability has been regarded as a limiting factor in production of industrially valuable compounds. The central carbon metabolism carries the carbon skeleton flux supporting the operation of NADPH-regenerating enzyme and offers flexibility in coping with NADPH demand for varied intracellular environment. To acquire an insightful understanding of its NADPH regeneration capacity, the elementary mode method was employed to compute all elementary flux modes (EFMs) of a network representative of central carbon metabolism. Based on the metabolic flux distributions of these modes, a cluster analysis of EFMs with high NADPH regeneration rate was conducted using the self-organizing map clustering algorithm. The clustering results were used to study the relationship between the flux of total NADPH regeneration and the flux in each NADPH producing enzyme. The results identified several reaction combinations supporting high NADPH regeneration, which are proven to be feasible in cells via thermodynamic analysis and coincident with a great deal of previous experimental report. Meanwhile, the reaction combinations showed some common characteristics: there were one or two decarboxylation oxidation reactions in the combinations that produced NADPH and the combination constitution included certain gluconeogenesis pathways. These findings suggested cyclization pathways as a powerful way for NADPH regeneration capacity of bacterial central carbon metabolism.

  7. Accuracy and Precision Analysis of Chamber-Based Nitrous Oxide Gas Flux Estimates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Chamber-based estimates of soil-to-atmosphere nitrous oxide (N2O) gas flux tend to underestimate actual emission rates due to inherently non-linear time series data. In theory, this limitation can be minimized by adjusting measurement conditions to reduce non-linearity and/or by using flux-calculati...

  8. Impact of water use efficiency on eddy covariance flux partitioning using correlation structure analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Partitioned land surfaces fluxes (e.g. evaporation, transpiration, photosynthesis, and ecosystem respiration) are needed as input, calibration, and validation data for numerous hydrological and land surface models. However, one of the most commonly used techniques for measuring land surface fluxes,...

  9. Entropy Analysis of Kinetic Flux Vector Splitting Schemes for the Compressible Euler Equations

    NASA Technical Reports Server (NTRS)

    Shiuhong, Lui; Xu, Jun

    1999-01-01

    Flux Vector Splitting (FVS) scheme is one group of approximate Riemann solvers for the compressible Euler equations. In this paper, the discretized entropy condition of the Kinetic Flux Vector Splitting (KFVS) scheme based on the gas-kinetic theory is proved. The proof of the entropy condition involves the entropy definition difference between the distinguishable and indistinguishable particles.

  10. Combining Flux Balance and Energy Balance Analysis for Large-Scale Metabolic Network: Biochemical Circuit Theory for Analysis of Large-Scale Metabolic Networks

    NASA Technical Reports Server (NTRS)

    Beard, Daniel A.; Liang, Shou-Dan; Qian, Hong; Biegel, Bryan (Technical Monitor)

    2001-01-01

    Predicting behavior of large-scale biochemical metabolic networks represents one of the greatest challenges of bioinformatics and computational biology. Approaches, such as flux balance analysis (FBA), that account for the known stoichiometry of the reaction network while avoiding implementation of detailed reaction kinetics are perhaps the most promising tools for the analysis of large complex networks. As a step towards building a complete theory of biochemical circuit analysis, we introduce energy balance analysis (EBA), which compliments the FBA approach by introducing fundamental constraints based on the first and second laws of thermodynamics. Fluxes obtained with EBA are thermodynamically feasible and provide valuable insight into the activation and suppression of biochemical pathways.

  11. Advanced flux leakage and eddy current signal analysis for casing and pipe inspection. Final report, September 1994-October 1996

    SciTech Connect

    Merchant, G.A.

    1997-01-01

    The objective of the project was to develop signal analysis methods and algorithms which will extract from the magnetic flux leakage and eddy current (FLEC) signals all the quantitative information they inherently contain and format that information in a manner which facilitates its interpretation in terms of casing condition.

  12. Validation Test Report for the NRL Ocean Surface Flux (NFLUX) Quality Control and 2D Variational Analysis System

    DTIC Science & Technology

    2014-06-11

    Test Report for the NRL Ocean Surface Flux (NFLUX) Quality Control and 2D Variational Analysis System Jackie May Neil VaN de Voorde QinetiQ North...OF RESPONSIBLE PERSON 19b. TELEPHONE NUMBER (include area code) b. ABSTRACT c. THIS PAGE 18. NUMBER OF PAGES 17. LIMITATION OF ABSTRACT Validation Test ...1 2.0 VALIDATION TEST DESIGN

  13. Quantitative analysis of autophagic flux by confocal pH-imaging of autophagic intermediates

    PubMed Central

    Maulucci, Giuseppe; Chiarpotto, Michela; Papi, Massimiliano; Samengo, Daniela; Pani, Giovambattista; De Spirito, Marco

    2015-01-01

    Although numerous techniques have been developed to monitor autophagy and to probe its cellular functions, these methods cannot evaluate in sufficient detail the autophagy process, and suffer limitations from complex experimental setups and/or systematic errors. Here we developed a method to image, contextually, the number and pH of autophagic intermediates by using the probe mRFP-GFP-LC3B as a ratiometric pH sensor. This information is expressed functionally by AIPD, the pH distribution of the number of autophagic intermediates per cell. AIPD analysis reveals how intermediates are characterized by a continuous pH distribution, in the range 4.5–6.5, and therefore can be described by a more complex set of states rather than the usual biphasic one (autophagosomes and autolysosomes). AIPD shape and amplitude are sensitive to alterations in the autophagy pathway induced by drugs or environmental states, and allow a quantitative estimation of autophagic flux by retrieving the concentrations of autophagic intermediates. PMID:26506895

  14. The flux of organic matter through a peatland ecosystem - evidence from thermogravimetric analysis

    NASA Astrophysics Data System (ADS)

    Worrall, Fred; Moody, Catherine; Clay, Gareth

    2016-04-01

    Carbon budgets of peatlands are now common and studies have considered nitrogen, oxygen and energy budgets, but no study has considered the whole composition of the organic matter as it transfers through and into a peatland. Organic matter samples were taken from each organic matter reservoir found in and each fluvial flux from a peatland and analysed the samples by thermogravimetric analysis. The samples analysed were: aboveground, belowground, heather, mosses and sedges, litter layer, a peat core, and monthly samples of particulate and dissolved organic matter. All organic matter samples were taken from a 100% peat catchment within Moor House National Nature Reserve in the North Pennines, UK, and collected samples were compared to standards of lignin, cellulose, humic acid and plant protein. Results showed that the thermogravimetric trace of the sampled organic matter were distinctive with the DOM traces being marked out by very low thermal stability relative other organic matter types. The peat profile shows a significant trend with depth from vegetation- to lignin-like composition. When all traces are weighted according to the observed dry matter and carbon budgets for the catchment then it is possible to judge what has been lost in the transition through and into the ecosystem. By plotting this "lost" trace it possible to assess its composition which is either 97% cellulose and 3% humic acid or 92% and 8% lignin. This has important implications for what controls the organic matter balance of peatlands and it suggests that the oxidation state (OR) of peatland is less than 1.

  15. Quantitative analysis of autophagic flux by confocal pH-imaging of autophagic intermediates.

    PubMed

    Maulucci, Giuseppe; Chiarpotto, Michela; Papi, Massimiliano; Samengo, Daniela; Pani, Giovambattista; De Spirito, Marco

    2015-01-01

    Although numerous techniques have been developed to monitor autophagy and to probe its cellular functions, these methods cannot evaluate in sufficient detail the autophagy process, and suffer limitations from complex experimental setups and/or systematic errors. Here we developed a method to image, contextually, the number and pH of autophagic intermediates by using the probe mRFP-GFP-LC3B as a ratiometric pH sensor. This information is expressed functionally by AIPD, the pH distribution of the number of autophagic intermediates per cell. AIPD analysis reveals how intermediates are characterized by a continuous pH distribution, in the range 4.5-6.5, and therefore can be described by a more complex set of states rather than the usual biphasic one (autophagosomes and autolysosomes). AIPD shape and amplitude are sensitive to alterations in the autophagy pathway induced by drugs or environmental states, and allow a quantitative estimation of autophagic flux by retrieving the concentrations of autophagic intermediates.

  16. Gene Knockout Identification Using an Extension of Bees Hill Flux Balance Analysis

    PubMed Central

    Choon, Yee Wen; Mohamad, Mohd Saberi; Deris, Safaai; Chong, Chuii Khim; Omatu, Sigeru; Corchado, Juan Manuel

    2015-01-01

    Microbial strain optimisation for the overproduction of a desired phenotype has been a popular topic in recent years. Gene knockout is a genetic engineering technique that can modify the metabolism of microbial cells to obtain desirable phenotypes. Optimisation algorithms have been developed to identify the effects of gene knockout. However, the complexities of metabolic networks have made the process of identifying the effects of genetic modification on desirable phenotypes challenging. Furthermore, a vast number of reactions in cellular metabolism often lead to a combinatorial problem in obtaining optimal gene knockout. The computational time increases exponentially as the size of the problem increases. This work reports an extension of Bees Hill Flux Balance Analysis (BHFBA) to identify optimal gene knockouts to maximise the production yield of desired phenotypes while sustaining the growth rate. This proposed method functions by integrating OptKnock into BHFBA for validating the results automatically. The results show that the extension of BHFBA is suitable, reliable, and applicable in predicting gene knockout. Through several experiments conducted on Escherichia coli, Bacillus subtilis, and Clostridium thermocellum as model organisms, extension of BHFBA has shown better performance in terms of computational time, stability, growth rate, and production yield of desired phenotypes. PMID:25874200

  17. Implicit flux-split Euler schemes for unsteady aerodynamic analysis involving unstructured dynamic meshes

    NASA Technical Reports Server (NTRS)

    Batina, John T.

    1990-01-01

    Improved algorithm for the solution of the time-dependent Euler equations are presented for unsteady aerodynamic analysis involving unstructured dynamic meshes. The improvements were developed recently to the spatial and temporal discretizations used by unstructured grid flow solvers. The spatial discretization involves a flux-split approach which is naturally dissipative and captures shock waves sharply with at most one grid point within the shock structure. The temporal discretization involves an implicit time-integration scheme using a Gauss-Seidel relaxation procedure which is computationally efficient for either steady or unsteady flow problems. For example, very large time steps may be used for rapid convergence to steady state, and the step size for unsteady cases may be selected for temporal accuracy rather than for numerical stability. Steady and unsteady flow results are presented for the NACA 0012 airfoil to demonstrate applications of the new Euler solvers. The unsteady results were obtained for the airfoil pitching harmonically about the quarter chord. The resulting instantaneous pressure distributions and lift and moment coefficients during a cycle of motion compare well with experimental data. A description of the Euler solvers is presented along with results and comparisons which assess the capability.

  18. Statistical analysis of the ion flux to the JET outer wall

    NASA Astrophysics Data System (ADS)

    Walkden, N. R.; Wynn, A.; Militello, F.; Lipschultz, B.; Matthews, G.; Guillemaut, C.; Harrison, J.; Moulton, D.; Contributors, JET

    2017-03-01

    Statistical analysis of the ion flux to the JET outer-wall is conducted in outer-wall limiter mounted Langmuir probe (OLP) time-series across a wide range of plasma current and line-averaged density during Ohmically heated horizontal target L-mode plasmas. The mean, μ, and the standard deviation, σ, of the ion-saturation current measured by the OLP show systematic variation with plasma current and density. Both increase as either plasma current decreases and/or density increases. Upon renormalization, achieved by subtraction of μ and rescaling by σ, the probability distribution functions (PDFs) of each signal collapse approximately onto a single curve. The shape of the curve deviates from a Γ distribution in the tail of the PDF and is better described by a log-normal distribution. The invariance in the shape of the PDF, which occurs over approximately four decades of the ordinate, is shown to be the result of a balance between the duration time of the average burst wave-form, {τd} and the waiting time between bursts, {τw} . This implies that the intermittency parameter, {τd}/{τw} , can be considered constant at the JET outer wall during horizontal target Ohmic L-mode operation. This result may be important both for model validation and prediction.

  19. Constrictor: Flux Balance Analysis Constraint Modification Provides Insight for Design of Biochemical Networks

    NASA Astrophysics Data System (ADS)

    Erickson, Keesha; Chatterjee, Anushree

    2014-03-01

    The use of in silico methods has become standard practice to correlate the structure of a biochemical network to the expression of a desired phenotype. Flux balance analysis (FBA) is one of the most prevalent techniques for modeling metabolism. FBA models have been successfully applied to obtain growth predictions, theoretical product yields from heterologous pathways, and genome engineering targets. We take inspiration from high-throughput recombineering techniques, which show that combinatorial exploration can reveal optimal mutants, and apply the advantages of computational techniques to analyze these combinations. We introduce Constrictor, an in silico tool for FBA that allows gene mutations to be analyzed in a combinatorial fashion, by applying simulated constraints accounting for regulation of gene expression. We apply this algorithm to study ethylene production in E. coli through the addition of the heterologous ethylene-forming enzyme from P. syringae. Targeting individual reactions as well as sets of reactions results in theoretical ethylene yields that are as much 65% greater than yields calculated using typical FBA. Constrictor is an adaptable technique that can be used to generate and analyze disparate populations of in silico mutants & select gene expression levels.

  20. Flux Balance Analysis of Plant Metabolism: The Effect of Biomass Composition and Model Structure on Model Predictions

    PubMed Central

    Yuan, Huili; Cheung, C. Y. Maurice; Hilbers, Peter A. J.; van Riel, Natal A. W.

    2016-01-01

    The biomass composition represented in constraint-based metabolic models is a key component for predicting cellular metabolism using flux balance analysis (FBA). Despite major advances in analytical technologies, it is often challenging to obtain a detailed composition of all major biomass components experimentally. Studies examining the influence of the biomass composition on the predictions of metabolic models have so far mostly been done on models of microorganisms. Little is known about the impact of varying biomass composition on flux prediction in FBA models of plants, whose metabolism is very versatile and complex because of the presence of multiple subcellular compartments. Also, the published metabolic models of plants differ in size and complexity. In this study, we examined the sensitivity of the predicted fluxes of plant metabolic models to biomass composition and model structure. These questions were addressed by evaluating the sensitivity of predictions of growth rates and central carbon metabolic fluxes to varying biomass compositions in three different genome-/large-scale metabolic models of Arabidopsis thaliana. Our results showed that fluxes through the central carbon metabolism were robust to changes in biomass composition. Nevertheless, comparisons between the predictions from three models using identical modeling constraints and objective function showed that model predictions were sensitive to the structure of the models, highlighting large discrepancies between the published models. PMID:27200014

  1. Pool size measurements facilitate the determination of fluxes at branching points in non-stationary metabolic flux analysis: the case of Arabidopsis thaliana.

    PubMed

    Heise, Robert; Fernie, Alisdair R; Stitt, Mark; Nikoloski, Zoran

    2015-01-01

    Pool size measurements are important for the estimation of absolute intracellular fluxes in particular scenarios based on data from heavy carbon isotope experiments. Recently, steady-state fluxes estimates were obtained for central carbon metabolism in an intact illuminated rosette of Arabidopsis thaliana grown photoautotrophically (Szecowka et al., 2013; Heise et al., 2014). Fluxes were estimated therein by integrating mass-spectrometric data of the dynamics of the unlabeled metabolic fraction, data on metabolic pool sizes, partitioning of metabolic pools between cellular compartments and estimates of photosynthetically inactive pools, with a simplified model of plant central carbon metabolism. However, the fluxes were determined by treating the pool sizes as fixed parameters. Here we investigated whether and, if so, to what extent the treatment of pool sizes as parameters to be optimized in three scenarios may affect the flux estimates. The results are discussed in terms of benchmark values for canonical pathways and reactions, including starch and sucrose synthesis as well as the ribulose-1,5-bisphosphate carboxylation and oxygenation reactions. In addition, we discuss pathways emerging from a divergent branch point for which pool sizes are required for flux estimation, irrespective of the computational approach used for the simulation of the observable labeling pattern. Therefore, our findings indicate the necessity for development of techniques for accurate pool size measurements to improve the quality of flux estimates from non-stationary flux estimates in intact plant cells in the absence of alternative flux measurements.

  2. Pool size measurements facilitate the determination of fluxes at branching points in non-stationary metabolic flux analysis: the case of Arabidopsis thaliana

    PubMed Central

    Heise, Robert; Fernie, Alisdair R.; Stitt, Mark; Nikoloski, Zoran

    2015-01-01

    Pool size measurements are important for the estimation of absolute intracellular fluxes in particular scenarios based on data from heavy carbon isotope experiments. Recently, steady-state fluxes estimates were obtained for central carbon metabolism in an intact illuminated rosette of Arabidopsis thaliana grown photoautotrophically (Szecowka et al., 2013; Heise et al., 2014). Fluxes were estimated therein by integrating mass-spectrometric data of the dynamics of the unlabeled metabolic fraction, data on metabolic pool sizes, partitioning of metabolic pools between cellular compartments and estimates of photosynthetically inactive pools, with a simplified model of plant central carbon metabolism. However, the fluxes were determined by treating the pool sizes as fixed parameters. Here we investigated whether and, if so, to what extent the treatment of pool sizes as parameters to be optimized in three scenarios may affect the flux estimates. The results are discussed in terms of benchmark values for canonical pathways and reactions, including starch and sucrose synthesis as well as the ribulose-1,5-bisphosphate carboxylation and oxygenation reactions. In addition, we discuss pathways emerging from a divergent branch point for which pool sizes are required for flux estimation, irrespective of the computational approach used for the simulation of the observable labeling pattern. Therefore, our findings indicate the necessity for development of techniques for accurate pool size measurements to improve the quality of flux estimates from non-stationary flux estimates in intact plant cells in the absence of alternative flux measurements. PMID:26082786

  3. Metabolic network reconstruction and flux variability analysis of storage synthesis in developing oilseed rape (Brassica napus L.) embryos.

    PubMed

    Hay, Jordan; Schwender, Jörg

    2011-08-01

    Computational simulation of large-scale biochemical networks can be used to analyze and predict the metabolic behavior of an organism, such as a developing seed. Based on the biochemical literature, pathways databases and decision rules defining reaction directionality we reconstructed bna572, a stoichiometric metabolic network model representing Brassica napus seed storage metabolism. In the highly compartmentalized network about 25% of the 572 reactions are transport reactions interconnecting nine subcellular compartments and the environment. According to known physiological capabilities of developing B. napus embryos, four nutritional conditions were defined to simulate heterotrophy or photoheterotrophy, each in combination with the availability of inorganic nitrogen (ammonia, nitrate) or amino acids as nitrogen sources. Based on mathematical linear optimization the optimal solution space was comprehensively explored by flux variability analysis, thereby identifying for each reaction the range of flux values allowable under optimality. The range and variability of flux values was then categorized into flux variability types. Across the four nutritional conditions, approximately 13% of the reactions have variable flux values and 10-11% are substitutable (can be inactive), both indicating metabolic redundancy given, for example, by isoenzymes, subcellular compartmentalization or the presence of alternative pathways. About one-third of the reactions are never used and are associated with pathways that are suboptimal for storage synthesis. Fifty-seven reactions change flux variability type among the different nutritional conditions, indicating their function in metabolic adjustments. This predictive modeling framework allows analysis and quantitative exploration of storage metabolism of a developing B. napus oilseed.

  4. [Effects of culture conditions on coenzyme Q10 production by Rhizobium radiobacter by metabolic flux analysis].

    PubMed

    Wu, Zu-fang; Du, Guo-cheng; Chen, Jian

    2005-04-01

    Metabolic pathway network of CoQ10 synthesis by R. radiobacter WSH2601 were instructed. The metabolic flux and its changes were determined under the conditions of changing DO concentration and addition of 1% CSL in the medium. The results illustrated that the Ru5P flux (r7) increased by 26.6 when increasing the DO concentration, r7 increased by 17.2 when addition of 1% CSL. The ratio of EMP and HMP flux as well as TCA flux decreased at these two conditions. DPP flux had a little change at these two conditions. Therefore, the CoQ10 accumulation is greatly determined by two key enzymes activities of condensation reaction between p-hydroxybenzoate acid (PHB) and decaprenyl diphosphate (DPP). The nodes of G6P, pyruvate and PEP are principal nodes in primary metabolism of CoQ10 fermentation. The flexibility of principal nodes was evaluated that the G6P node is elastic, while pyruvate node is weakly flexibility, at the condition of changing culture conditions. The increase of DCW is associated with the improvement of HMP pathway flux.

  5. Reconstruction and flux analysis of coupling between metabolic pathways of astrocytes and neurons: application to cerebral hypoxia

    PubMed Central

    Çakιr, Tunahan; Alsan, Selma; Saybaşιlι, Hale; Akιn, Ata; Ülgen, Kutlu Ö

    2007-01-01

    Background It is a daunting task to identify all the metabolic pathways of brain energy metabolism and develop a dynamic simulation environment that will cover a time scale ranging from seconds to hours. To simplify this task and make it more practicable, we undertook stoichiometric modeling of brain energy metabolism with the major aim of including the main interacting pathways in and between astrocytes and neurons. Model The constructed model includes central metabolism (glycolysis, pentose phosphate pathway, TCA cycle), lipid metabolism, reactive oxygen species (ROS) detoxification, amino acid metabolism (synthesis and catabolism), the well-known glutamate-glutamine cycle, other coupling reactions between astrocytes and neurons, and neurotransmitter metabolism. This is, to our knowledge, the most comprehensive attempt at stoichiometric modeling of brain metabolism to date in terms of its coverage of a wide range of metabolic pathways. We then attempted to model the basal physiological behaviour and hypoxic behaviour of the brain cells where astrocytes and neurons are tightly coupled. Results The reconstructed stoichiometric reaction model included 217 reactions (184 internal, 33 exchange) and 216 metabolites (183 internal, 33 external) distributed in and between astrocytes and neurons. Flux balance analysis (FBA) techniques were applied to the reconstructed model to elucidate the underlying cellular principles of neuron-astrocyte coupling. Simulation of resting conditions under the constraints of maximization of glutamate/glutamine/GABA cycle fluxes between the two cell types with subsequent minimization of Euclidean norm of fluxes resulted in a flux distribution in accordance with literature-based findings. As a further validation of our model, the effect of oxygen deprivation (hypoxia) on fluxes was simulated using an FBA-derivative approach, known as minimization of metabolic adjustment (MOMA). The results show the power of the constructed model to simulate

  6. Analysis of solar receiver flux distributions for US/Russian solar dynamic system demonstration on the MIR Space Station

    NASA Technical Reports Server (NTRS)

    Kerslake, Thomas W.; Fincannon, James

    1995-01-01

    The United States and Russia have agreed to jointly develop a solar dynamic (SD) system for flight demonstration on the Russian MIR space station starting in late 1997. Two important components of this SD system are the solar concentrator and heat receiver provided by Russia and the U.S., respectively. This paper describes optical analysis of the concentrator and solar flux predictions on target receiver surfaces. The optical analysis is performed using the code CIRCE2. These analyses account for finite sun size with limb darkening, concentrator surface slope and position errors, concentrator petal thermal deformation, gaps between petals, and the shading effect of the receiver support struts. The receiver spatial flux distributions are then combined with concentrator shadowing predictions. Geometric shadowing patterns are traced from the concentrator to the target receiver surfaces. These patterns vary with time depending on the chosen MIR flight attitude and orbital mechanics of the MIR spacecraft. The resulting predictions provide spatial and temporal receiver flux distributions for any specified mission profile. The impact these flux distributions have on receiver design and control of the Brayton engine are discussed.

  7. Analysis of solar receiver flux distributions for US/Russian solar dynamic system demonstration on the MIR Space Station

    NASA Astrophysics Data System (ADS)

    Kerslake, Thomas W.; Fincannon, James

    1995-06-01

    The United States and Russia have agreed to jointly develop a solar dynamic (SD) system for flight demonstration on the Russian MIR space station starting in late 1997. Two important components of this SD system are the solar concentrator and heat receiver provided by Russia and the U.S., respectively. This paper describes optical analysis of the concentrator and solar flux predictions on target receiver surfaces. The optical analysis is performed using the code CIRCE2. These analyses account for finite sun size with limb darkening, concentrator surface slope and position errors, concentrator petal thermal deformation, gaps between petals, and the shading effect of the receiver support struts. The receiver spatial flux distributions are then combined with concentrator shadowing predictions. Geometric shadowing patterns are traced from the concentrator to the target receiver surfaces. These patterns vary with time depending on the chosen MIR flight attitude and orbital mechanics of the MIR spacecraft. The resulting predictions provide spatial and temporal receiver flux distributions for any specified mission profile. The impact these flux distributions have on receiver design and control of the Brayton engine are discussed.

  8. A meta-analysis of soil salinization effects on nitrogen pools, cycles and fluxes in coastal ecosystems.

    PubMed

    Zhou, Minghua; Butterbach-Bahl, Klaus; Vereecken, Harry; Brüggemann, Nicolas

    2017-03-01

    Salinity intrusion caused by land subsidence resulting from increasing groundwater abstraction, decreasing river sediment loads and increasing sea level because of climate change has caused widespread soil salinization in coastal ecosystems. Soil salinization may greatly alter nitrogen (N) cycling in coastal ecosystems. However, a comprehensive understanding of the effects of soil salinization on ecosystem N pools, cycling processes and fluxes is not available for coastal ecosystems. Therefore, we compiled data from 551 observations from 21 peer-reviewed papers and conducted a meta-analysis of experimental soil salinization effects on 19 variables related to N pools, cycling processes and fluxes in coastal ecosystems. Our results showed that the effects of soil salinization varied across different ecosystem types and salinity levels. Soil salinization increased plant N content (18%), soil NH4(+) (12%) and soil total N (210%), although it decreased soil NO3(-) (2%) and soil microbial biomass N (74%). Increasing soil salinity stimulated soil N2 O fluxes as well as hydrological NH4(+) and NO2(-) fluxes more than threefold, although it decreased the hydrological dissolved organic nitrogen (DON) flux (59%). Soil salinization also increased the net N mineralization by 70%, although salinization effects were not observed on the net nitrification, denitrification and dissimilatory nitrate reduction to ammonium in this meta-analysis. Overall, this meta-analysis improves our understanding of the responses of ecosystem N cycling to soil salinization, identifies knowledge gaps and highlights the urgent need for studies on the effects of soil salinization on coastal agro-ecosystem and microbial N immobilization. Additional increases in knowledge are critical for designing sustainable adaptation measures to the predicted intrusion of salinity intrusion so that the productivity of coastal agro-ecosystems can be maintained or improved and the N losses and pollution of the natural

  9. Simulating cyanobacterial phenotypes by integrating flux balance analysis, kinetics, and a light distribution function

    SciTech Connect

    He, Lian; Wu, Stephen G.; Wan, Ni; Reding, Adrienne C.; Tang, Yinjie J.

    2015-12-24

    In this study, genome-scale models (GSMs) are widely used to predict cyanobacterial phenotypes in photobioreactors (PBRs). However, stoichiometric GSMs mainly focus on fluxome that result in maximal yields. Cyanobacterial metabolism is controlled by both intracellular enzymes and photobioreactor conditions. To connect both intracellular and extracellular information and achieve a better understanding of PBRs productivities, this study integrates a genome-scale metabolic model of Synechocystis 6803 with growth kinetics, cell movements, and a light distribution function. The hybrid platform not only maps flux dynamics in cells of sub-populations but also predicts overall production titer and rate in PBRs. Analysis of the integrated GSM demonstrates several results. First, cyanobacteria are capable of reaching high biomass concentration (>20 g/L in 21 days) in PBRs without light and CO2 mass transfer limitations. Second, fluxome in a single cyanobacterium may show stochastic changes due to random cell movements in PBRs. Third, insufficient light due to cell self-shading can activate the oxidative pentose phosphate pathway in subpopulation cells. Fourth, the model indicates that the removal of glycogen synthesis pathway may not improve cyanobacterial bio-production in large-size PBRs, because glycogen can support cell growth in the dark zones. Based on experimental data, the integrated GSM estimates that Synechocystis 6803 in shake flask conditions has a photosynthesis efficiency of ~2.7 %. Conclusions: The multiple-scale integrated GSM, which examines both intracellular and extracellular domains, can be used to predict production yield/rate/titer in large-size PBRs. More importantly, genetic engineering strategies predicted by a traditional GSM may work well only in optimal growth conditions. In contrast, the integrated GSM may reveal mutant physiologies in diverse bioreactor conditions, leading to the design of robust strains with high

  10. Simulating cyanobacterial phenotypes by integrating flux balance analysis, kinetics, and a light distribution function

    DOE PAGES

    He, Lian; Wu, Stephen G.; Wan, Ni; ...

    2015-12-24

    In this study, genome-scale models (GSMs) are widely used to predict cyanobacterial phenotypes in photobioreactors (PBRs). However, stoichiometric GSMs mainly focus on fluxome that result in maximal yields. Cyanobacterial metabolism is controlled by both intracellular enzymes and photobioreactor conditions. To connect both intracellular and extracellular information and achieve a better understanding of PBRs productivities, this study integrates a genome-scale metabolic model of Synechocystis 6803 with growth kinetics, cell movements, and a light distribution function. The hybrid platform not only maps flux dynamics in cells of sub-populations but also predicts overall production titer and rate in PBRs. Analysis of the integratedmore » GSM demonstrates several results. First, cyanobacteria are capable of reaching high biomass concentration (>20 g/L in 21 days) in PBRs without light and CO2 mass transfer limitations. Second, fluxome in a single cyanobacterium may show stochastic changes due to random cell movements in PBRs. Third, insufficient light due to cell self-shading can activate the oxidative pentose phosphate pathway in subpopulation cells. Fourth, the model indicates that the removal of glycogen synthesis pathway may not improve cyanobacterial bio-production in large-size PBRs, because glycogen can support cell growth in the dark zones. Based on experimental data, the integrated GSM estimates that Synechocystis 6803 in shake flask conditions has a photosynthesis efficiency of ~2.7 %. Conclusions: The multiple-scale integrated GSM, which examines both intracellular and extracellular domains, can be used to predict production yield/rate/titer in large-size PBRs. More importantly, genetic engineering strategies predicted by a traditional GSM may work well only in optimal growth conditions. In contrast, the integrated GSM may reveal mutant physiologies in diverse bioreactor conditions, leading to the design of robust strains with high chances of success in

  11. ANALYSIS OF WATER AND ENERGY FLUXES USING SATELLITE, ENERGY BALANCE MODELING AND OBSERVATIONS (Invited)

    NASA Astrophysics Data System (ADS)

    Irmak, A.

    2009-12-01

    Surface energy fluxes, including net radiation (Rn), sensible heat (H), latent heat (LE), and soil heat flux (G) are critical in surface energy balance of any terrain or landscapes. Estimation or measurement of these energy fluxes is important for completing the water balance in terrestrial ecosystems, and therefore accurately predicting the effects of global climate and land use change. The objectives of this study were to (1) use METRICtm (Mapping Evapotranspiration at high Resolution using Internalized Calibration) model for estimating land surface energy fluxes in Nebraska (NE) by utilizing satellite remote sensing data, (2) identify model bias in energy balance components compared with measurements from Bowen Ratio Energy Balance System (BREBS) in a subsurface drip-irrigated maize field in South-central Nebraska, and (3) understand the partitioning of available energy into latent heat for corn and soybean cropping systems at large scale. A total of 15 Landsat images were processed to estimate instantaneous surface energy fluxes at Landsat overpasses with METRIC model. Results showed that the model predictions of the surface energy fluxes and daily evapotranspiration were correlated well with the BREBS measurements. There is a need, however, to test the performance of the model with in-situ observations in other locations with different dataset before utilizing it for crucial water regulatory and policy decisions. The METRICtm approach illustrated how an ‘off-the-shelf’ model can be applied operationally over a significant time period and how that model behaves. The findings makes considerable contribution to our understanding of estimating land surface energy fluxes using remote sensing approach and experimentally describes the operational characteristics of METRICtm and presents its limitations.

  12. Mathematical analysis for internal filtration of convection-enhanced high-flux hemodialyzer.

    PubMed

    Lee, Jung Chan; Lee, Kyungsoo; Kim, Hee Chan

    2012-10-01

    Structural modifications using a conventional hemodialyzer improved the internal filtration and clearance of middle molecular weight wastes by enhanced convection effect. In this study, we employed a mathematical model describing the internal filtration rate as well as the hemodynamic and hematologic parameters in highflux dialyzer to interpret the previous reported experimental results. Conventional high-flux hemodialysis and convection-enhanced high-flux hemodialysis were configured in the mathematical forms and integrated into the iterative numerical method to predict the internal filtration phenomena inside the dialyzers during dialysis. The distributions of blood pressure, dialysate pressure, oncotic pressure, blood flow rates, dialysate flow rates, local ultrafiltration, hematocrit, protein concentration and blood viscosity along the axial length of dialyzer were calculated in order to estimate the internal filtration volume. The results show that the filtration volumes by internal filtration is two times higher in a convection-enhanced high-flux hemodialyzer than in a conventional high-flux hemodialzer and explains the experimental result of improved clearance of middle molecular size waste in convection-enhanced high-flux hemodialyzer.

  13. Thin film heat flux sensors for accurate transient and unidirectional heat transfer analysis

    NASA Astrophysics Data System (ADS)

    Azerou, B.; Garnier, B.; Lahmar, J.

    2012-11-01

    Heat flux measurement is needed in many heat transfer studies. For the best unbiased heat flux sensors (HFS), the heat flux is obtained using temperature measurements at different locations and also an inverse heat conduction method (function specification...) in order to calculate the heat flux. Systematic errors can come from the uncertainty in the wire thermocouples locations and from errors in the knowledge of distances between two consecutive wire thermocouples. The main idea in this work is to use thin film thermoresistances deposited on a flexible thin polymer substrate instead of wire thermocouples welded on metallic sample. The interest of using thin film thermoresistances instead of wire thermocouples is a lower disturbance due to the smaller thickness of the thin film sensors (typically less than 1μm) and a much better knowledge of the distances between the different thin film thermoresistances which are precisely defined in the mask used for the metallic thin film pattern fabrication. In this paper, we present the fabrication of the new heat flux sensor with thin film thermoresistances, the study of the effect of the self heating (due to Joule effect in thermoresistances) and the performances of this new HFS with the comparison with classical HFS using wire thermocouples. For this study, a symmetric experimental setup is used with metallic samples equipped with an etched foil heater and both classical and new HFS. For several heating conditions, it appears that a better accuracy is always obtained with the new HFS using thin film thermoresistances.

  14. Metabolic Flux Analysis of Shewanella spp. Reveals Evolutionary Robustness in Central Carbon Metabolism

    SciTech Connect

    Tang, Yinjie J.; Martin, Hector Garcia; Dehal, Paramvir S.; Deutschbauer, Adam; Llora, Xavier; Meadows, Adam; Arkin, Adam; Keasling, Jay D.

    2009-08-19

    Shewanella spp. are a group of facultative anaerobic bacteria widely distributed in marine and fresh-water environments. In this study, we profiled the central metabolic fluxes of eight recently sequenced Shewanella species grown under the same condition in minimal med-ium with [3-13C] lactate. Although the tested Shewanella species had slightly different growth rates (0.23-0.29 h31) and produced different amounts of acetate and pyruvate during early exponential growth (pseudo-steady state), the relative intracellular metabolic flux distributions were remarkably similar. This result indicates that Shewanella species share similar regulation in regard to central carbon metabolic fluxes under steady growth conditions: the maintenance of metabolic robustness is not only evident in a single species under genetic perturbations (Fischer and Sauer, 2005; Nat Genet 37(6):636-640), but also observed through evolutionary related microbial species. This remarkable conservation of relative flux profiles through phylogenetic differences prompts us to introduce the concept of metabotype as an alternative scheme to classify microbial fluxomics. On the other hand, Shewanella spp. display flexibility in the relative flux profiles when switching their metabolism from consuming lactate to consuming pyruvate and acetate.

  15. As-Run Thermal Analysis of the GTL-1 Experiment Irradiated in the ATR South Flux Trap

    SciTech Connect

    Donna P. Guillen

    2011-05-01

    The GTL-1 experiment was conducted to assess corrosion the performance of the proposed Boosted Fast Flux Loop booster fuel at heat flux levels {approx}30% above the design operating condition. Sixteen miniplates fabricated from 25% enriched, high-density U3Si2/Al dispersion fuel with 6061 aluminum cladding were subjected to peak beginning of cycle (BOC) heat fluxes ranging from 411 W/cm2 to 593 W/cm2. Miniplates fabricated with three different fuel variations (without fines, annealed, and with standard powder) performed equally well, with negligible irradiation-induced swelling and a normal fission density gradient. Both the standard and the modified prefilm procedures produced hydroxide films that adequately protected the miniplates from failure. A detailed finite element model was constructed to calculate temperatures and heat flux for an as-run cycle average effective south lobe power of 25.4 MW(t). Results of the thermal analysis are given at four times during the cycle: BOC at 0 effective full power days (EFPD), middle of cycle (MOC) at 18 EFPD, MOC at 36 EFPD, and end of cycle at 48.9 EFPD. The highest temperatures and heat fluxes occur at the BOC and decrease in a linear manner throughout the cycle. Miniplate heat flux levels and fuel, cladding, hydroxide, and coolant-hydroxide interface temperatures were calculated using the average measured hydroxide thickness on each miniplate. The hydroxide layers are the largest on miniplates nearest to the core midplane, where heat flux and temperature are highest. The hydroxide layer thickness averages 20.4 {mu}m on the six hottest miniplates (B3, B4, C1, C2, C3, and C4). This tends to exacerbate the heating of these miniplates, since a thicker hydroxide layer reduces the heat transfer from the fuel to the coolant. These six hottest miniplates have the following thermal characteristics at BOC: (1) Peak fuel centerline temperature >300 C; (2) Peak cladding temperature >200 C; (3) Peak hydroxide temperature >190 C; (4

  16. Dynamic vacuum analysis for APS high heat flux beamline front ends using optical ray-tracing simulation methods

    SciTech Connect

    Xu, S.; Nielsen, R.W.

    1992-01-01

    The high-power and high-flux x-ray beams produced by third generation synchrotron radiation sources such as the Advanced Photon Source (APS) can cause significantly high gas desorption rates on beamline front-end components if beam missteering occurs. The effect of this gas desorption needs to be understood for dynamic vacuum analysis. To simulate beam missteering conditions, optical ray-tracing methods have been employed. The results of the ray-tracing analysis have been entered into a system-oriented vacuum program to provide dynamic vacuum calculations for determination of pumping requirements for the beamline front-ends. The APS will provide several types of synchrotron radiation sources, for example, undulators, wigglers, and bending magnets. For the purpose of this study, the wiggler source was chosen as a worst case'' scenario due to its high photon flux, high beam power, and relatively large beam cross section.

  17. Dynamic vacuum analysis for APS high heat flux beamline front ends using optical ray-tracing simulation methods

    SciTech Connect

    Xu, S.; Nielsen, R.W.

    1992-09-01

    The high-power and high-flux x-ray beams produced by third generation synchrotron radiation sources such as the Advanced Photon Source (APS) can cause significantly high gas desorption rates on beamline front-end components if beam missteering occurs. The effect of this gas desorption needs to be understood for dynamic vacuum analysis. To simulate beam missteering conditions, optical ray-tracing methods have been employed. The results of the ray-tracing analysis have been entered into a system-oriented vacuum program to provide dynamic vacuum calculations for determination of pumping requirements for the beamline front-ends. The APS will provide several types of synchrotron radiation sources, for example, undulators, wigglers, and bending magnets. For the purpose of this study, the wiggler source was chosen as a ``worst case`` scenario due to its high photon flux, high beam power, and relatively large beam cross section.

  18. Hierarchical flux-based thermal-structural finite element analysis method

    NASA Technical Reports Server (NTRS)

    Polesky, Sandra P.

    1992-01-01

    A hierarchical flux-based finite element method is developed for both a one and two dimensional thermal structural analyses. Derivation of the finite element equations is presented. The resulting finite element matrices associated with the flux based formulation are evaluated in a closed form. The hierarchical finite elements include additional degrees of freedom in the approximation of the element variable distributions by the use of nodeless variables. The nodeless variables offer increased solution accuracy without the need for defining actual nodes and rediscretizing the finite element model. Thermal and structural responses are obtained from a conventional linear finite element method and exact solutions. Results show that the hierarchical flux-based method can provide improved thermal and structural solution accuracy with fewer elements when compared to results for the conventional linear element method.

  19. An Analysis of Turbulent Heat Fluxes and the Energy Balance During the REFLEX Campaign

    NASA Astrophysics Data System (ADS)

    Tol, Christiaan van der; Timmermans, Wim; Corbari, Chiara; Carrara, Arnaud; Timmermans, Joris; Su, Zhongbo

    2015-12-01

    Three eddy covariance stations were installed at the Barrax experimental farm during the Land-Atmosphere Exchanges (REFLEX) airborne training and measurement campaign to provide ground truth data of energy balance fluxes and vertical temperature and wind profiles. The energy balance closure ratio (EBR) was 105% for a homogeneous camelina site, 86% at a sparse reforestation site, and 73% for a vineyard. We hypothesize that the lower closure in the last site was related to the limited fetch. Incorporating a vertical gradient of soil thermal properties decreased the RMSE of the energy balance at the camelina site by 16 W m-2. At the camelina site, eddy covariance estimates of sensible and latent heat fluxes could be reproduced well using mean vertical profiles of wind and temperature, provided that the Monin-Obukhov length is known. Measured surface temperature and sensible heat fluxes suggested high excess resistance for heat (kB-1 = 17).

  20. An Analysis of Turbulent Heat Fluxes and the Energy Balance During the REFLEX Campaign

    NASA Astrophysics Data System (ADS)

    van der Tol, Christiaan; Timmermans, Wim; Corbari, Chiara; Carrara, Arnaud; Timmermans, Joris; Su, Zhongbo

    2015-12-01

    Three eddy covariance stations were installed at the Barrax experimental farm during the Land-Atmosphere Exchanges (REFLEX) airborne training and measurement campaign to provide ground truth data of energy balance fluxes and vertical temperature and wind profiles. The energy balance closure ratio (EBR) was 105% for a homogeneous camelina site, 86% at a sparse reforestation site, and 73% for a vineyard. We hypothesize that the lower closure in the last site was related to the limited fetch. Incorporating a vertical gradient of soil thermal properties decreased the RMSE of the energy balance at the camelina site by 16 W m-2. At the camelina site, eddy covariance estimates of sensible and latent heat fluxes could be reproduced well using mean vertical profiles of wind and temperature, provided that the Monin—Obukhov length is known. Measured surface temperature and sensible heat fluxes suggested high excess resistance for heat (kB-1 = 17).

  1. Computational Platform for Flux Analysis Using 13C-Label Tracing- Phase I SBIR Final Report

    SciTech Connect

    Van Dien, Stephen J.

    2005-04-12

    Isotopic label tracing is a powerful experimental technique that can be combined with metabolic models to quantify metabolic fluxes in an organism under a particular set of growth conditions. In this work we constructed a genome-scale metabolic model of Methylobacterium extorquens, a facultative methylotroph with potential application in the production of useful chemicals from methanol. A series of labeling experiments were performed using 13C-methanol, and the resulting distribution of labeled carbon in the proteinogenic amino acids was determined by mass spectrometry. Algorithms were developed to analyze this data in context of the metabolic model, yielding flux distributions for wild-type and several engineered strains of M. extorquens. These fluxes were compared to those predicted by model simulation alone, and also integrated with microarray data to give an improved understanding of the metabolic physiology of this organism.

  2. A benchmark analysis of radiation flux distribution for Boron Neutron Capture Therapy of canine brain tumors

    SciTech Connect

    Moran, J.M.

    1992-02-01

    Calculations of radiation flux and dose distributions for Boron Neutron Capture Therapy (BNCT) of brain tumors are typically performed using sophisticated three-dimensional analytical models based on either a homogeneous approximation or a simplified few-region approximation to the actual highly-heterogeneous geometry of the irradiation volume. Such models should be validated by comparison with calculations using detailed models in which all significant macroscopic tissue heterogeneities and geometric structures are explicitly represented as faithfully as possible. This work describes a validation exercise for BNCT of canine brain tumors. Geometric measurements of the canine anatomical structures of interest for this work were performed by dissecting and examining two essentially identical Labrador Retriever heads. Chemical analyses of various tissue samples taken during the dissections were conducted to obtain measurements of elemental compositions for tissues of interest. The resulting geometry and tissue composition data were then used to construct a detailed heterogeneous calculational model of the Labrador Retriever head. Calculations of three-dimensional radiation flux distributions pertinent to BNCT were performed for the model using the TORT discrete-ordinates radiation transport code. The calculations were repeated for a corresponding volume-weighted homogeneous tissue model. Comparison of the results showed that the peak neutron and photon flux magnitudes were quite similar for the two models (within 5%), but that the spatial flux profiles were shifted in the heterogeneous model such that the fluxes in some locations away from the peak differed from the corresponding fluxes in the homogeneous model by as much as 10-20%. Differences of this magnitude can be therapeutically significant, emphasizing the need for proper validation of simplified treatment planning models.

  3. Suspended sediment fluxes in a tidal wetland: Measurement, controlling factors, and error analysis

    USGS Publications Warehouse

    Ganju, N.K.; Schoellhamer, D.H.; Bergamaschi, B.A.

    2005-01-01

    Suspended sediment fluxes to and from tidal wetlands are of increasing concern because of habitat restoration efforts, wetland sustainability as sea level rises, and potential contaminant accumulation. We measured water and sediment fluxes through two channels on Browns Island, at the landward end of San Francisco Bay, United States, to determine the factors that control sediment fluxes on and off the island. In situ instrumentation was deployed between October 10 and November 13, 2003. Acoustic Doppler current profilers and the index velocity method were employed to calculate water fluxes. Suspended sediment concentrations (SSC) were determined with optical sensors and cross-sectional water sampling. All procedures were analyzed for their contribution to total error in the flux measurement. The inability to close the water balance and determination of constituent concentration were identified as the main sources of error; total error was 27% for net sediment flux. The water budget for the island was computed with an unaccounted input of 0.20 m 3 s-1 (22% of mean inflow), after considering channel flow, change in water storage, evapotranspiration, and precipitation. The net imbalance may be a combination of groundwater seepage, overland flow, and flow through minor channels. Change of island water storage, caused by local variations in water surface elevation, dominated the tidalty averaged water flux. These variations were mainly caused by wind and barometric pressure change, which alter regional water levels throughout the Sacramento-San Joaquin River Delta. Peak instantaneous ebb flow was 35% greater than peak flood flow, indicating an ebb-dominant system, though dominance varied with the spring-neap cycle. SSC were controlled by wind-wave resuspension adjacent to the island and local tidal currents that mobilized sediment from the channel bed. During neap tides sediment was imported onto the island but during spring tides sediment was exported because the main

  4. An analysis of return flux from the Space Shuttle Orbiter RCS engines

    NASA Technical Reports Server (NTRS)

    Ehlers, H. K. F.

    1984-01-01

    The return flux from the Space Shuttle Orbiter reaction control system (RCS) engines to sensors in the open payload bay has been analyzed on the basis of Shuttle/Payload Contamination Evaluation (SPACE II) model predictions and orbital flight measurements. Model data are presented showing the variations of molecular return flux values with Orbiter orientation and instrument direction. The effects of multiple molecular collisions within RCS engine plumes and in their vicinity are discussed. These collisions significantly influence the amount of plume molecules returning to payload instruments and, therefore, the amount of contaminants received.

  5. Analysis of the Flux Growth Rate in Emerging Active Regions on the Sun

    NASA Astrophysics Data System (ADS)

    Abramenko, V. I.; Kutsenko, A. S.; Tikhonova, O. I.; Yurchyshyn, V. B.

    2017-04-01

    We studied the emergence process of 42 active regions (ARs) by analyzing the time derivative, R(t), of the total unsigned flux. Line-of-sight magnetograms acquired by the Helioseismic and Magnetic Imager (HMI) onboard the Solar Dynamics Observatory (SDO) were used. A continuous piecewise linear fitting to the R(t)-profile was applied to detect an interval, Δ t2, of nearly constant R(t) covering one or several local maxima. The magnitude of R(t) averaged over Δ t2 was accepted as an estimate of the maximum value of the flux growth rate, R_{MAX}, which varies in a range of (0.5 - 5)×10^{20} Mx hour^{-1} for ARs with a maximum total unsigned flux of (0.5 - 3)× 10^{22} Mx. The normalized flux growth rate, RN, was defined under the assumption that the saturated total unsigned flux, F_{MAX}, equals unity. Out of 42 ARs in our initial list, 36 events were successfully fitted, and they form two subsets (with a small overlap of eight events): the ARs with a short (<13 hours) interval Δ t2 and a high (>0.024 hour^{-1}) normalized flux emergence rate, RN, form the "rapid" emergence event subset. The second subset consists of "gradual" emergence events, and it is characterized by a long (>13 hours) interval Δ t2 and a low RN (<0.024 hour^{-1}). In diagrams of R_{MAX} plotted versus F_{MAX}, the events from different subsets do not overlap, and each subset displays an individual power law. The power-law index derived from the entire ensemble of 36 events is 0.69 ± 0.10. The rapid emergence is consistent with a two-step emergence process of a single twisted flux tube. The gradual emergence is possibly related to a consecutive rising of several flux tubes emerging at nearly the same location in the photosphere.

  6. Recent advances in elementary flux modes and yield space analysis as useful tools in metabolic network studies.

    PubMed

    Horvat, Predrag; Koller, Martin; Braunegg, Gerhart

    2015-09-01

    A review of the use of elementary flux modes (EFMs) and their applications in metabolic engineering covered with yield space analysis (YSA) is presented. EFMs are an invaluable tool in mathematical modeling of biochemical processes. They are described from their inception in 1994, followed by various improvements of their computation in later years. YSA constitutes another precious tool for metabolic network modeling, and is presented in details along with EFMs in this article. The application of these techniques is discussed for several case studies of metabolic network modeling provided in respective original articles. The article is concluded by some case studies in which the application of EFMs and YSA turned out to be most useful, such as the analysis of intracellular polyhydroxyalkanoate (PHA) formation and consumption in Cupriavidus necator, including the constraint-based description of the steady-state flux cone of the strain's metabolic network, the profound analysis of a continuous five-stage bioreactor cascade for PHA production by C. necator using EFMs and, finally, the study of metabolic fluxes in the metabolic network of C. necator cultivated on glycerol.

  7. Heat loss analysis-based design of a 12 MW wind power generator module having an HTS flux pump exciter

    NASA Astrophysics Data System (ADS)

    Sung, Hae-Jin; Go, Byeong-Soo; Jiang, Zhenan; Park, Minwon; Yu, In-Keun

    2016-11-01

    The development of an effective high-temperature superconducting (HTS) generator is currently a research focus; however, the reduction of heat loss of a large-scale HTS generator is a challenge. This study deals with a heat loss analysis-based design of a 12 MW wind power generator module having an HTS flux pump exciter. The generator module consists of an HTS rotor of the generator and an HTS flux pump exciter. The specifications of the module were described, and the detailed configuration of the module was illustrated. For the heat loss analysis of the module, the excitation loss of the flux pump exciter, eddy current loss of all of the structures in the module, radiation loss, and conduction loss of an HTS coil supporter were assessed using a 3D finite elements method program. In the case of the conduction loss, different types of the supporters were compared to find out the supporter of the lowest conduction loss in the module. The heat loss analysis results of the module were reflected in the design of the generator module and discussed in detail. The results will be applied to the design of large-scale superconducting generators for wind turbines including a cooling system.

  8. Analysis of autophagic flux in response to sulforaphane in metastatic prostate cancer cells

    PubMed Central

    Watson, Gregory W; Wickramasekara, Samanthi; Fang, Yufeng; Palomera-Sanchez, Zoraya; Maier, Claudia S; Williams, David E; Dashwood, Roderick H; Perez, Viviana I; Ho, Emily

    2015-01-01

    Scope The phytochemical sulforaphane has been shown to decrease prostate cancer metastases in a genetic mouse model of prostate carcinogenesis, though the mechanism of action is not fully known. Sulforaphane has been reported to stimulate autophagy, and modulation of autophagy has been proposed to influence sulforaphane cytotoxicity; however, no conclusions about autophagy can be drawn without assessing autophagic flux, which has not been characterized in prostate cancer cells following sulforaphane treatment. Methods and Results We conducted an investigation to assess the impact of sulforaphane on autophagic flux in two metastatic prostate cancer cell lines at a concentration shown to decrease metastasis in vivo. Autophagic flux was assessed by multiple autophagy related proteins and substrates. We found that sulforaphane can stimulate autophagic flux and cell death only at high concentrations, above what has been observed in vivo. Conclusion These results suggest that sulforaphane does not directly stimulate autophagy or cell death in metastatic prostate cancer cells under physiologically relevant conditions, but instead supports the involvement of in vivo factors as important effectors of sulforaphane- mediated prostate cancer suppression. PMID:26108801

  9. Failure analysis of beryllium tile assembles following high heat flux testing for the ITER program

    SciTech Connect

    B. C. Odegard, Jr.; C. H. Cadden; N. Y. C. Yang

    2000-05-01

    The following document describes the processing, testing and post-test analysis of two Be-Cu assemblies that have successfully met the heat load requirements for the first wall and dome sections for the ITER (International Thermonuclear Experimental Reactor) fusion reactor. Several different joint assemblies were evaluated in support of a manufacturing technology investigation aimed at diffusion bonding or brazing a beryllium armor tile to a copper alloy heat sink for fusion reactor applications. Judicious selection of materials and coatings for these assemblies was essential to eliminate or minimize interactions with the highly reactive beryllium armor material. A thin titanium layer was used as a diffusion barrier to isolate the copper heat sink from the beryllium armor. To reduce residual stresses produced by differences in the expansion coefficients between the beryllium and copper, a compliant layer of aluminum or aluminum-beryllium (AlBeMet-150) was used. Aluminum was chosen because it does not chemically react with, and exhibits limited volubility in, beryllium. Two bonding processes were used to produce the assemblies. The primary process was a diffusion bonding technique. In this case, undesirable metallurgical reactions were minimized by keeping the materials in a solid state throughout the fabrication cycle. The other process employed an aluminum-silicon layer as a brazing filler material. In both cases, a hot isostatic press (HIP) furnace was used in conjunction with vacuum-canned assemblies in order to minimize oxidation and provide sufficient pressure on the assemblies for full metal-to-metal contact and subsequent bonding. The two final assemblies were subjected to a suite of tests including: tensile tests and electron and optical metallography. Finally, high heat flux testing was conducted at the electron beam testing system (EBTS) at Sandia National Laboratories, New Mexico. Here, test mockups were fabricated and subjected to normal heat loads to

  10. Uncertainty analysis of a coupled ecosystem response model simulating greenhouse gas fluxes from a temperate grassland

    NASA Astrophysics Data System (ADS)

    Liebermann, Ralf; Kraft, Philipp; Houska, Tobias; Breuer, Lutz; Müller, Christoph; Kraus, David; Haas, Edwin; Klatt, Steffen

    2015-04-01

    Among anthropogenic greenhouse gas emissions, CO2 is the dominant driver of global climate change. Next to its direct impact on the radiation budget, it also affects the climate system by triggering feedback mechanisms in terrestrial ecosystems. Such mechanisms - like stimulated photosynthesis, increased root exudations and reduced stomatal transpiration - influence both the input and the turnover of carbon and nitrogen compounds in the soil. The stabilization and decomposition of these compounds determines how increasing CO2 concentrations change the terrestrial trace gas emissions, especially CO2, N2O and CH4. To assess the potential reaction of terrestrial greenhouse gas emissions to rising tropospheric CO2 concentration, we make use of a comprehensive ecosystem model integrating known processes and fluxes of the carbon-nitrogen cycle in soil, vegetation and water. We apply a state-of-the-art ecosystem model with measurements from a long term field experiment of CO2 enrichment. The model - a grassland realization of LandscapeDNDC - simulates soil chemistry coupled with plant physiology, microclimate and hydrology. The data - comprising biomass, greenhouse gas emissions, management practices and soil properties - has been attained from a FACE (Free Air Carbon dioxide Enrichment) experiment running since 1997 on a temperate grassland in Giessen, Germany. Management and soil data, together with weather records, are used to drive the model, while cut biomass as well as CO2 and N2O emissions are used for calibration and validation. Starting with control data from installations without CO2 enhancement, we begin with a GLUE (General Likelihood Uncertainty Estimation) assessment using Latin Hypercube to reduce the range of the model parameters. This is followed by a detailed sensitivity analysis, the application of DREAM-ZS for model calibration, and an estimation of the effect of input uncertainty on the simulation results. Since first results indicate problems with

  11. Predictive Potential of Flux Balance Analysis of Saccharomyces cerevisiae Using as Optimization Function Combinations of Cell Compartmental Objectives

    PubMed Central

    García Sánchez, Carlos Eduardo; Vargas García, César Augusto; Torres Sáez, Rodrigo Gonzalo

    2012-01-01

    Background The main objective of flux balance analysis (FBA) is to obtain quantitative predictions of metabolic fluxes of an organism, and it is necessary to use an appropriate objective function to guarantee a good estimation of those fluxes. Methodology In this study, the predictive performance of FBA was evaluated, using objective functions arising from the linear combination of different cellular objectives. This approach is most suitable for eukaryotic cells, owing to their multiplicity of cellular compartments. For this reason, Saccharomyces cerevisiae was used as model organism, and its metabolic network was represented using the genome-scale metabolic model iMM904. As the objective was to evaluate the predictive performance from the FBA using the kind of objective function previously described, substrate uptake and oxygen consumption were the only input data used for the FBA. Experimental information about microbial growth and exchange of metabolites with the environment was used to assess the quality of the predictions. Conclusions The quality of the predictions obtained with the FBA depends greatly on the knowledge of the oxygen uptake rate. For the most of studied classifications, the best predictions were obtained with “maximization of growth”, and with some combinations that include this objective. However, in the case of exponential growth with unknown oxygen exchange flux, the objective function “maximization of growth, plus minimization of NADH production in cytosol, plus minimization of NAD(P)H consumption in mitochondrion” gave much more accurate estimations of fluxes than the obtained with any other objective function explored in this study. PMID:22912775

  12. Analysis of Metabolic Pathways and Fluxes in a Newly Discovered Thermophilic and Ethanol-Tolerant Geobacillus Strain

    SciTech Connect

    Tang, Yinjie J.; Sapra, Rajat; Joyner, Dominique; Hazen, Terry C.; Myers, Samuel; Reichmuth, David; Blanch, Harvey; Keasling, Jay D.

    2009-01-20

    A recently discovered thermophilic bacterium, Geobacillus thermoglucosidasius M10EXG, ferments a range of C5 (e.g., xylose) and C6 sugars (e.g., glucose) and istolerant to high ethanol concentrations (10percent, v/v). We have investigated the central metabolism of this bacterium using both in vitro enzyme assays and 13C-based flux analysis to provide insights into the physiological properties of this extremophile and explore its metabolism for bio-ethanol or other bioprocess applications. Our findings show that glucose metabolism in G. thermoglucosidasius M10EXG proceeds via glycolysis, the pentose phosphate pathway, and the TCA cycle; the Entner?Doudoroff pathway and transhydrogenase activity were not detected. Anaplerotic reactions (including the glyoxylate shunt, pyruvate carboxylase, and phosphoenolpyruvate carboxykinase) were active, but fluxes through those pathways could not be accuratelydetermined using amino acid labeling. When growth conditions were switched from aerobic to micro-aerobic conditions, fluxes (based on a normalized glucose uptake rate of 100 units (g DCW)-1 h-1) through the TCA cycle and oxidative pentose phosphate pathway were reduced from 64+-3 to 25+-2 and from 30+-2 to 19+-2, respectively. The carbon flux under micro-aerobic growth was directed formate. Under fully anerobic conditions, G. thermoglucosidasius M10EXG used a mixed acid fermentation process and exhibited a maximum ethanol yield of 0.38+-0.07 mol mol-1 glucose. In silico flux balance modeling demonstrates that lactate and acetate production from G. thermoglucosidasius M10EXG reduces the maximum ethanol yieldby approximately threefold, thus indicating that both pathways should be modified to maximize ethanol production.

  13. Flux balance analysis of CHO cells before and after a metabolic switch from lactate production to consumption.

    PubMed

    Martínez, Verónica S; Dietmair, Stefanie; Quek, Lake-Ee; Hodson, Mark P; Gray, Peter; Nielsen, Lars K

    2013-02-01

    Mammalian cell cultures typically exhibit an energy inefficient phenotype characterized by the consumption of large quantities of glucose and the concomitant production of large quantities of lactate. Under certain conditions, mammalian cells can switch to a more energy efficient state during which lactate is consumed. Using a metabolic model derived from a mouse genome scale model we performed flux balance analysis of Chinese hamster ovary cells before and after a metabolic switch from lactate production (in the presence of glucose) to lactate consumption (after glucose depletion). Despite a residual degree of freedom after accounting for measurements, the calculated flux ranges and associated errors were narrow enough to enable investigation of metabolic changes across the metabolic switch. Surprisingly, the fluxes through the lower part of the TCA cycle from oxoglutarate to malate were very similar (around 60 µmol/gDW/h) for both phases. A detailed analysis of the energy metabolism showed that cells consuming lactate have an energy efficiency (total ATP produced per total C-mol substrate consumed) six times greater than lactate producing cells.

  14. Regional flux analysis for discovering and quantifying anatomical changes: An application to the brain morphometry in Alzheimer's disease.

    PubMed

    Lorenzi, M; Ayache, N; Pennec, X

    2015-07-15

    In this study we introduce the regional flux analysis, a novel approach to deformation based morphometry based on the Helmholtz decomposition of deformations parameterized by stationary velocity fields. We use the scalar pressure map associated to the irrotational component of the deformation to discover the critical regions of volume change. These regions are used to consistently quantify the associated measure of volume change by the probabilistic integration of the flux of the longitudinal deformations across the boundaries. The presented framework unifies voxel-based and regional approaches, and robustly describes the volume changes at both group-wise and subject-specific level as a spatial process governed by consistently defined regions. Our experiments on the large cohorts of the ADNI dataset show that the regional flux analysis is a powerful and flexible instrument for the study of Alzheimer's disease in a wide range of scenarios: cross-sectional deformation based morphometry, longitudinal discovery and quantification of group-wise volume changes, and statistically powered and robust quantification of hippocampal and ventricular atrophy.

  15. Sensitivity Analysis of the Land Surface Model NOAH-MP for Different Model Fluxes

    NASA Astrophysics Data System (ADS)

    Mai, Juliane; Thober, Stephan; Samaniego, Luis; Branch, Oliver; Wulfmeyer, Volker; Clark, Martyn; Attinger, Sabine; Kumar, Rohini; Cuntz, Matthias

    2015-04-01

    Land Surface Models (LSMs) use a plenitude of process descriptions to represent the carbon, energy and water cycles. They are highly complex and computationally expensive. Practitioners, however, are often only interested in specific outputs of the model such as latent heat or surface runoff. In model applications like parameter estimation, the most important parameters are then chosen by experience or expert knowledge. Hydrologists interested in surface runoff therefore chose mostly soil parameters while biogeochemists interested in carbon fluxes focus on vegetation parameters. However, this might lead to the omission of parameters that are important, for example, through strong interactions with the parameters chosen. It also happens during model development that some process descriptions contain fixed values, which are supposedly unimportant parameters. However, these hidden parameters remain normally undetected although they might be highly relevant during model calibration. Sensitivity analyses are used to identify informative model parameters for a specific model output. Standard methods for sensitivity analysis such as Sobol indexes require large amounts of model evaluations, specifically in case of many model parameters. We hence propose to first use a recently developed inexpensive sequential screening method based on Elementary Effects that has proven to identify the relevant informative parameters. This reduces the number parameters and therefore model evaluations for subsequent analyses such as sensitivity analysis or model calibration. In this study, we quantify parametric sensitivities of the land surface model NOAH-MP that is a state-of-the-art LSM and used at regional scale as the land surface scheme of the atmospheric Weather Research and Forecasting Model (WRF). NOAH-MP contains multiple process parameterizations yielding a considerable amount of parameters (˜ 100). Sensitivities for the three model outputs (a) surface runoff, (b) soil drainage

  16. Potential emission flux to aerosol pollutants over Bengal Gangetic plain through combined trajectory clustering and aerosol source fields analysis

    NASA Astrophysics Data System (ADS)

    Kumar, D. Bharath; Verma, S.

    2016-09-01

    A hybrid source-receptor analysis was carried out to evaluate the potential emission flux to winter monsoon (WinMon) aerosols over Bengal Gangetic plain urban (Kolkata, Kol) and semi-urban atmospheres (Kharagpur, Kgp). This was done through application of fuzzy c-mean clustering to back-trajectory data combined with emission flux and residence time weighted aerosols analysis. WinMon mean aerosol optical depth (AOD) and angstrom exponent (AE) at Kol (AOD: 0.77; AE: 1.17) were respectively slightly higher than and nearly equal to that at Kgp (AOD: 0.71; AE: 1.18). Out of six source region clusters over Indian subcontinent and two over Indian oceanic region, the cluster mean AOD was the highest when associated with the mean path of air mass originating from the Bay of Bengal and the Arabian sea clusters at Kol and that from the Indo-Gangetic plain (IGP) cluster at Kgp. Spatial distribution of weighted AOD fields showed the highest potential source of aerosols over the IGP, primarily over upper IGP (e.g. Punjab, Haryana), lower IGP (e.g. Uttarpradesh) and eastern region (e.g. west Bengal, Bihar, northeast India) clusters. The emission flux contribution potential (EFCP) of fossil fuel (FF) emissions at surface (SL) of Kol/Kgp, elevated layer (EL) of Kol, and of biomass burning (BB) emissions at SL of Kol were primarily from upper, lower, upper/lower IGP clusters respectively. The EFCP of FF/BB emissions at Kgp-EL/SL, and that of BB at EL of Kol/Kgp were mainly from eastern region and Africa (AFR) clusters respectively. Though the AFR cluster was constituted of significantly high emission flux source potential of dust emissions, the EFCP of dust from northwest India (NWI) was comparable to that from AFR at Kol SL/EL.

  17. Analysis of the sensible heat flux from the exterior surface of buildings using time sequential thermography

    NASA Astrophysics Data System (ADS)

    Hoyano, Akira; Asano, Kohichi; Kanamaru, Takehisa

    In this study, the distribution of surface temperature on the surface of two buildings having different characteristics was measured using a thermal infrared camera. Measurements were made in the summer, the period in Japan during which heat flux from buildings is of major interest, and again in the winter for comparison purposes. The heat characteristics of each building were obtained throughout the day by time-sequential thermography (TST), and the surface temperature of each physical element was classified according to temperature, shape, material and position. When the temperature of a surface could not be determined by an infrared camera mounted on the top of a building or a pole, temperature measurements were made using a hand-held IR camera. In addition, the sensible heat flux from each surface was calculated using TST and the surface area of each element as calculated from blueprints of the buildings.

  18. A new sensitivity analysis and solution method for scintillometer measurements of area-average turbulent fluxes

    NASA Astrophysics Data System (ADS)

    Gruber, Matthew

    Scintillometer measurements of the turbulence inner-scale length lo and refractive index structure function C2n allow for the retrieval of large-scale area-averaged turbulent fluxes in the atmospheric surface layer. This retrieval involves the solution of the non-linear set of equations defined by the Monin-Obukhov similarity hypothesis. A new method that uses an analytic solution to the set of equations is presented, which leads to a stable and efficient numerical method of computation that has the potential of eliminating computational error. Mathematical expressions are derived that map out the sensitivity of the turbulent flux measurements to uncertainties in source measurements such as lo. These sensitivity functions differ from results in the previous literature; the reasons for the differences are explored.

  19. A New Sensitivity Analysis and Solution Method for Scintillometer Measurements of Area-Averaged Turbulent Fluxes

    NASA Astrophysics Data System (ADS)

    Gruber, Matthew; Fochesatto, Gilberto J.

    2013-07-01

    Scintillometer measurements of the turbulence inner-scale length l_o and refractive index structure function C_n^2 allow for the retrieval of large-scale area-averaged turbulent fluxes in the atmospheric surface layer. This retrieval involves the solution of the non-linear set of equations defined by the Monin-Obukhov similarity hypothesis. A new method that uses an analytic solution to the set of equations is presented, which leads to a stable and efficient numerical method of computation that has the potential of eliminating computational error. Mathematical expressions are derived that map out the sensitivity of the turbulent flux measurements to uncertainties in source measurements such as l_o. These sensitivity functions differ from results in the previous literature; the reasons for the differences are explored.

  20. FluxCTTX: A LIMS-based tool for management and analysis of cytotoxicity assays data

    PubMed Central

    2015-01-01

    Background Cytotoxicity assays have been used by researchers to screen for cytotoxicity in compound libraries. Researchers can either look for cytotoxic compounds or screen "hits" from initial high-throughput drug screens for unwanted cytotoxic effects before investing in their development as a pharmaceutical. These assays may be used as an alternative to animal experimentation and are becoming increasingly important in modern laboratories. However, the execution of these assays in large scale and different laboratories requires, among other things, the management of protocols, reagents, cell lines used as well as the data produced, which can be a challenge. The management of all this information is greatly improved by the utilization of computational tools to save time and guarantee quality. However, a tool that performs this task designed specifically for cytotoxicity assays is not yet available. Results In this work, we have used a workflow based LIMS -- the Flux system -- and the Together Workflow Editor as a framework to develop FluxCTTX, a tool for management of data from cytotoxicity assays performed at different laboratories. The main work is the development of a workflow, which represents all stages of the assay and has been developed and uploaded in Flux. This workflow models the activities of cytotoxicity assays performed as described in the OECD 129 Guidance Document. Conclusions FluxCTTX presents a solution for the management of the data produced by cytotoxicity assays performed at Interlaboratory comparisons. Its adoption will contribute to guarantee the quality of activities in the process of cytotoxicity tests and enforce the use of Good Laboratory Practices (GLP). Furthermore, the workflow developed is complete and can be adapted to other contexts and different tests for management of other types of data. PMID:26696462

  1. Heat Flux Analysis of a Reacting Thermite Spray Impingent on a Substrate

    SciTech Connect

    Eric S. Collins; Michelle L. Pantoya; Michael A. Daniels; Daniel J. Prentice; Eric D. Steffler; Steven P. D'Arche

    2012-03-01

    Spray combustion from a thermite reaction is a new area of research relevant to localized energy generation applications, such as welding or cutting. In this study, we characterized the heat flux of combustion spray impinging on a target from a nozzle for three thermite mixtures. The reactions studied include aluminum (Al) with iron oxide (Fe2O3), Al with copper oxide (CuO), and Al with molybdenum oxide (MoO3). Several standoff distances (i.e., distance from the nozzle exit to the target) were analyzed. A fast response heat flux sensor was engineered for this purpose and is discussed in detail. Results correlated substrate damage to a threshold heat flux of 4550 W/cm2 for a fixed-nozzle configuration. Also, higher gas-generating thermites were shown to produce a widely dispersed spray and be less effective at imparting kinetic energy damage to a target. These results provide an understanding of the role of thermal and physical properties (i.e., such as heat of combustion, gas generation, and particle size) on thermite spray combustion performance measured by damaging a target substrate.

  2. An analysis of the decadal variability of Carbon fluxes in three evergreen European forests through modelling

    NASA Astrophysics Data System (ADS)

    Delpierre, N.; Dufrêne, E.

    2009-04-01

    With several sites measuring mass and energy turbulent fluxes for more than ten years, the CarboEurope database appears as a valuable resource for addressing the question of the determinism of the interannual variability of carbon (C) and water balances in forests ecosystems. Apart from major climate-driven anomalies during the anomalous 2003 summer and 2007 spring, little is known about the factors driving interannual variability (IAV) of the C balance in forest ecosystems. We used the CASTANEA process-based model to simulate the C and W fluxes and balances of three European evergreen forests for the 2000-2007 period (FRPue Quercus ilex, 44°N; DETha Picea abies, 51°N; FIHyy Pinus sylvestris, 62°N). The model fairly reproduced the day-to-day variability of measured fluxes, accounting for 70-81%, 77-91% and 59-90% of the daily variance of measured NEP, GPP and TER, respectively. However, the model was challenged in representing the IAV of fluxes integrated on an annual time scale. It reproduced ca. 80% of the interannual variance of measured GPP, but no significant relationship could be established between annual measured and modelled NEP or TER. Accordingly, CASTANEA appeared as a suitable tool for disentangling the influence of climate and biological processes on GPP at mutiple time scales. We show that climate and biological processes relative influences on the modelled GPP vary from year to year in European evergreen forests. Water-stress related and phenological processes (i.e. release of the winter thermal constraint on photosynthesis in evergreens) appear as primary drivers for the particular 2003 and 2007 years, respectively, but the relative influence of other climatic factors widely varies for less remarkable years at all sites. We discuss shortcomings of the method, as related to the influence of compensating errors in the simulated fluxes, and assess the causes of the model poor ability to represent the IAV of the annual sums of NEP and TER.

  3. A Sensitivity Analysis of the Impact of Rain on Regional and Global Sea-Air Fluxes of CO2

    PubMed Central

    Shutler, J. D.; Land, P. E.; Woolf, D. K.; Quartly, G. D.

    2016-01-01

    The global oceans are considered a major sink of atmospheric carbon dioxide (CO2). Rain is known to alter the physical and chemical conditions at the sea surface, and thus influence the transfer of CO2 between the ocean and atmosphere. It can influence gas exchange through enhanced gas transfer velocity, the direct export of carbon from the atmosphere to the ocean, by altering the sea skin temperature, and through surface layer dilution. However, to date, very few studies quantifying these effects on global net sea-air fluxes exist. Here, we include terms for the enhanced gas transfer velocity and the direct export of carbon in calculations of the global net sea-air fluxes, using a 7-year time series of monthly global climate quality satellite remote sensing observations, model and in-situ data. The use of a non-linear relationship between the effects of rain and wind significantly reduces the estimated impact of rain-induced surface turbulence on the rate of sea-air gas transfer, when compared to a linear relationship. Nevertheless, globally, the rain enhanced gas transfer and rain induced direct export increase the estimated annual oceanic integrated net sink of CO2 by up to 6%. Regionally, the variations can be larger, with rain increasing the estimated annual net sink in the Pacific Ocean by up to 15% and altering monthly net flux by > ± 50%. Based on these analyses, the impacts of rain should be included in the uncertainty analysis of studies that estimate net sea-air fluxes of CO2 as the rain can have a considerable impact, dependent upon the region and timescale. PMID:27673683

  4. A Sensitivity Analysis of the Impact of Rain on Regional and Global Sea-Air Fluxes of CO2.

    PubMed

    Ashton, I G; Shutler, J D; Land, P E; Woolf, D K; Quartly, G D

    The global oceans are considered a major sink of atmospheric carbon dioxide (CO2). Rain is known to alter the physical and chemical conditions at the sea surface, and thus influence the transfer of CO2 between the ocean and atmosphere. It can influence gas exchange through enhanced gas transfer velocity, the direct export of carbon from the atmosphere to the ocean, by altering the sea skin temperature, and through surface layer dilution. However, to date, very few studies quantifying these effects on global net sea-air fluxes exist. Here, we include terms for the enhanced gas transfer velocity and the direct export of carbon in calculations of the global net sea-air fluxes, using a 7-year time series of monthly global climate quality satellite remote sensing observations, model and in-situ data. The use of a non-linear relationship between the effects of rain and wind significantly reduces the estimated impact of rain-induced surface turbulence on the rate of sea-air gas transfer, when compared to a linear relationship. Nevertheless, globally, the rain enhanced gas transfer and rain induced direct export increase the estimated annual oceanic integrated net sink of CO2 by up to 6%. Regionally, the variations can be larger, with rain increasing the estimated annual net sink in the Pacific Ocean by up to 15% and altering monthly net flux by > ± 50%. Based on these analyses, the impacts of rain should be included in the uncertainty analysis of studies that estimate net sea-air fluxes of CO2 as the rain can have a considerable impact, dependent upon the region and timescale.

  5. Analysis of the Effects of Vitiates on Surface Heat Flux in Ground Tests of Hypersonic Vehicles

    NASA Technical Reports Server (NTRS)

    Cuda, Vincent; Gaffney, Richard L

    2008-01-01

    To achieve the high enthalpy conditions associated with hypersonic flight, many ground test facilities burn fuel in the air upstream of the test chamber. Unfortunately, the products of combustion contaminate the test gas and alter gas properties and the heat fluxes associated with aerodynamic heating. The difference in the heating rates between clean air and a vitiated test medium needs to be understood so that the thermal management system for hypersonic vehicles can be properly designed. This is particularly important for advanced hypersonic vehicle concepts powered by air-breathing propulsion systems that couple cooling requirements, fuel flow rates, and combustor performance by flowing fuel through sub-surface cooling passages to cool engine components and preheat the fuel prior to combustion. An analytical investigation was performed comparing clean air to a gas vitiated with methane/oxygen combustion products to determine if variations in gas properties contributed to changes in predicted heat flux. This investigation started with simple relationships, evolved into writing an engineering-level code, and ended with running a series of CFD cases. It was noted that it is not possible to simultaneously match all of the gas properties between clean and vitiated test gases. A study was then conducted selecting various combinations of freestream properties for a vitiated test gas that matched clean air values to determine which combination of parameters affected the computed heat transfer the least. The best combination of properties to match was the free-stream total sensible enthalpy, dynamic pressure, and either the velocity or Mach number. This combination yielded only a 2% difference in heating. Other combinations showed departures of up to 10% in the heat flux estimate.

  6. Leakage effects in car underhood aerothermal management: temperature and heat flux analysis

    NASA Astrophysics Data System (ADS)

    Khaled, Mahmoud; Habchi, Charbel; Harambat, Fabien; Elmarakbi, Ahmed; Peerhossaini, Hassan

    2014-10-01

    Air leakage from the engine compartment of a vehicle comes mainly from the junctions of the vehicle hood and the front end grill, the vehicle wings, the optical and the windshield. The present paper studies the thermal impact of these air leakage zones on the components of the vehicle engine compartment through temperature and heat-flux measurements. The front wheels of the test vehicle are positioned on a dynamometer and driven by the vehicle engine. The engine compartment is instrumented with almost 100 surface and air thermocouples and 20 fluxmeters of normal gradients. Measurements were made for three different thermal operating points. Five leak-sealing configurations are studied.

  7. Metabolic flux analysis in complex isotopolog space. Recycling of glucose in tobacco plants.

    PubMed

    Ettenhuber, Christian; Radykewicz, Tanja; Kofer, Waltraud; Koop, Hans-Ulrich; Bacher, Adelbert; Eisenreich, Wolfgang

    2005-02-01

    Tobacco plants grown in vitro were supplied with a mixture of [U-13C6]glucose and unlabelled sucrose via the root system. After 20 days, leaves were harvested and extracted with water. Glucose was isolated from the extract and was analysed by 13C NMR spectroscopy. All 13C signals appeared as complex multiplets due to 13C-13C coupling. The abundance of 21 isotopologous glucose species was determined from the 13C NMR signal integrals by numerical deconvolution using a genetic algorithm. The relative fractions of specific isotopologs in the overall excess of 13C-labelled specimens establish flux contributions via glycolysis/glucogenesis, pentose phosphate pathway, citric acid cycle and Calvin cycle including 13CO2 refixation. The fluxes were modelled and reconstructed in silico by a novel rule-based approach yielding the contributions of circular pathways and the degree of multiple cycling events. The data indicate that the vast majority of the proffered [U-13C6]glucose molecules had been modified by catabolism and subsequent glucogenesis from catabolic fragments, predominantly via passage through the citric acid cycle and the pentose phosphate pathway.

  8. Flux analysis of central metabolic pathways in Geobactermetallireducens during reduction of solubleFe(III)-NTA

    SciTech Connect

    Tang, Yinjie J.; Chakraborty, Romy; Garcia-Martin, Hector; Chu,Jeannie; Hazen, Terry C.; Keasling, Jay D.

    2007-01-01

    We analyzed the carbon fluxes in the central metabolism ofGeobacter metallireducens strain GS-15 using 13C isotopomer modeling.Acetate labeled in the 1st or 2nd position was the sole carbon source,and Fe-NTA was the sole terminal electron acceptor. The measured labeledacetate uptake rate was 21 mmol/gdw/h in the exponential growth phase.The resulting isotope labeling pattern of amino acids allowed an accuratedetermination of the in vivo global metabolic reaction rates (fluxes)through the central metabolic pathways using a computational isotopomermodel. The tracer experiments showed that G. metallireducens containedcomplete biosynthesis pathways for essential metabolism, and this strainmight also have an unusual isoleucine biosynthesis route (usingacetyl-CoA and pyruvate as the precursors). The model indicated that over90 percent of the acetate was completely oxidized to CO2 via a completetricarboxylic acid (TCA) cycle while reducing iron. Pyruvate carboxylaseand phosphoenolpyruvate carboxykinase were present under theseconditions, but enzymes in the glyoxylate shunt and malic enzyme wereabsent. Gluconeogenesis and the pentose phosphate pathway were mainlyemployed for biosynthesis and accounted for less than 3 percent of totalcarbon consumption. The model also indicated surprisingly highreversibility in the reaction between oxoglutarate and succinate. Thisstep operates close to the thermodynamic equilibrium possibly becausesuccinate is synthesized via a transferase reaction, and the conversionof oxoglutarate to succinate is a rate limiting step for carbonmetabolism. These findings enable a better understanding of therelationship between genome annotation and extant metabolic pathways inG. metallireducens.

  9. Validation of the MCNP computational model for neutron flux distribution with the neutron activation analysis measurement

    NASA Astrophysics Data System (ADS)

    Tiyapun, K.; Chimtin, M.; Munsorn, S.; Somchit, S.

    2015-05-01

    The objective of this work is to demonstrate the method for validating the predication of the calculation methods for neutron flux distribution in the irradiation tubes of TRIGA research reactor (TRR-1/M1) using the MCNP computer code model. The reaction rate using in the experiment includes 27Al(n, α)24Na and 197Au(n, γ)198Au reactions. Aluminium (99.9 wt%) and gold (0.1 wt%) foils and the gold foils covered with cadmium were irradiated in 9 locations in the core referred to as CT, C8, C12, F3, F12, F22, F29, G5, and G33. The experimental results were compared to the calculations performed using MCNP which consisted of the detailed geometrical model of the reactor core. The results from the experimental and calculated normalized reaction rates in the reactor core are in good agreement for both reactions showing that the material and geometrical properties of the reactor core are modelled very well. The results indicated that the difference between the experimental measurements and the calculation of the reactor core using the MCNP geometrical model was below 10%. In conclusion the MCNP computational model which was used to calculate the neutron flux and reaction rate distribution in the reactor core can be used for others reactor core parameters including neutron spectra calculation, dose rate calculation, power peaking factors calculation and optimization of research reactor utilization in the future with the confidence in the accuracy and reliability of the calculation.

  10. Analysis of cell flux in the parallel plate flow chamber: implications for cell capture studies.

    PubMed

    Munn, L L; Melder, R J; Jain, R K

    1994-08-01

    The parallel plate flow chamber provides a controlled environment for determinations of the shear stress at which cells in suspension can bind to endothelial cell monolayers. By decreasing the flow rate of cell-containing media over the monolayer and assessing the number of cells bound at each wall shear stress, the relationship between shear force and binding efficiency can be determined. The rate of binding should depend on the delivery of cells to the surface as well as the intrinsic cell-surface interactions; thus, only if the cell flux to the surface is known can the resulting binding curves be interpreted correctly. We present the development and validation of a mathematical model based on the sedimentation rate and velocity profile in the chamber for the delivery of cells from a flowing suspension to the chamber surface. Our results show that the flux depends on the bulk cell concentration, the distance from the entrance point, and the flow rate of the cell-containing medium. The model was then used in a normalization procedure for experiments in which T cells attach to TNF-alpha-stimulated HUVEC monolayers, showing that a threshold for adhesion occurs at a shear stress of about 3 dyn/cm2.

  11. Analysis of non-destructive current simulators of flux compression generators

    NASA Astrophysics Data System (ADS)

    O'Connor, K. A.; Curry, R. D.

    2014-06-01

    Development and evaluation of power conditioning systems and high power microwave components often used with flux compression generators (FCGs) requires repeated testing and characterization. In an effort to minimize the cost and time required for testing with explosive generators, non-destructive simulators of an FCG's output current have been developed. Flux compression generators and simulators of FCGs are unique pulsed power sources in that the current waveform exhibits a quasi-exponential increasing rate at which the current rises. Accurately reproducing the quasi-exponential current waveform of a FCG can be important in designing electroexplosive opening switches and other power conditioning components that are dependent on the integral of current action and the rate of energy dissipation. Three versions of FCG simulators have been developed that include an inductive network with decreasing impedance in time. A primary difference between these simulators is the voltage source driving them. It is shown that a capacitor-inductor-capacitor network driving a constant or decreasing inductive load can produce the desired high-order derivatives of the load current to replicate a quasi-exponential waveform. The operation of the FCG simulators is reviewed and described mathematically for the first time to aid in the design of new simulators. Experimental and calculated results of two recent simulators are reported with recommendations for future designs.

  12. Flux analysis of the human proximal colon using anaerobic digestion model 1.

    PubMed

    Motelica-Wagenaar, Anne Marieke; Nauta, Arjen; van den Heuvel, Ellen G H M; Kleerebezem, Robbert

    2014-08-01

    The colon can be regarded as an anaerobic digestive compartment within the gastro intestinal tract (GIT). An in silico model simulating the fluxes in the human proximal colon was developed on basis of the anaerobic digestion model 1 (ADM1), which is traditionally used to model waste conversion to biogas. Model calibration was conducted using data from in vitro fermentation of the proximal colon (TIM-2), and, amongst others, supplemented with the bio kinetics of prebiotic galactooligosaccharides (GOS) fermentation. The impact of water and solutes absorption by the host was also included. Hydrolysis constants of carbohydrates and proteins were estimated based on total short chain fatty acids (SCFA) and ammonia production in vitro. Model validation was established using an independent dataset of a different in vitro model: an in vitro three-stage continuous culture system. The in silico model was shown to provide quantitative insight in the microbial community structure in terms of functional groups, and the substrate and product fluxes between these groups as well as the host, as a function of the substrate composition, pH and the solids residence time (SRT). The model confirms the experimental observation that methanogens are washed out at low pH or low SRT-values. The in silico model is proposed as useful tool in the design of experimental setups for in vitro experiments by giving insight in fermentation processes in the proximal human colon.

  13. Heat-flux measurements for the rotor of a full-stage turbine. II - Description of analysis technique and typical time-resolved measurements

    NASA Technical Reports Server (NTRS)

    Dunn, M. G.; George, W. K.; Rae, W. J.; Woodward, S. H.; Moller, J. C.

    1986-01-01

    An analytical technique for obtaining the time-resolved heat flux of a turbine blade is applied to the case of a TFE 731-2 hp full-stage rotating turbine. In order to obtain the heat flux values from the thin film gage temperature histories, a finite difference procedure is used to solve the heat equation with variable thermal properties. After setting out the data acquisition and analysis procedures, their application is illustrated for three midspan locations on the blade and operation at the design flow function. Results demonstrate that the magnitude of the heat flux fluctuation due to vane-balde interaction is large by comparison to the time-averaged heat flux at all investigated locations; FFT of a portion of the heat flux record illustrates that the dominant frequencies occur at the wake-cutting frequency and its harmonics.

  14. NASA's Carbon Monitoring System Flux-Pilot Project: A Multi-Component Analysis System for Carbon-Cycle Research and Monitoring

    NASA Technical Reports Server (NTRS)

    Pawson, S.; Gunson, M.; Potter, C.; Jucks, K.

    2012-01-01

    The importance of greenhouse gas increases for climate motivates NASA s observing strategy for CO2 from space, including the forthcoming Orbiting Carbon Observatory (OCO-2) mission. Carbon cycle monitoring, including attribution of atmospheric concentrations to regional emissions and uptake, requires a robust modeling and analysis infrastructure to optimally extract information from the observations. NASA's Carbon-Monitoring System Flux-Pilot Project (FPP) is a prototype for such analysis, combining a set of unique tools to facilitate analysis of atmospheric CO2 along with fluxes between the atmosphere and the terrestrial biosphere or ocean. NASA's analysis system is unique, in that it combines information and expertise from the land, oceanic, and atmospheric branches of the carbon cycle and includes some estimates of uncertainty. Numerous existing space-based missions provide information of relevance to the carbon cycle. This study describes the components of the FPP framework, assessing the realism of computed fluxes, thus providing the basis for research and monitoring applications. Fluxes are computed using data-constrained terrestrial biosphere models and physical ocean models, driven by atmospheric observations and assimilating ocean-color information. Use of two estimates provides a measure of uncertainty in the fluxes. Along with inventories of other emissions, these data-derived fluxes are used in transport models to assess their consistency with atmospheric CO2 observations. Closure is achieved by using a four-dimensional data assimilation (inverse) approach that adjusts the terrestrial biosphere fluxes to make them consistent with the atmospheric CO2 observations. Results will be shown, illustrating the year-to-year variations in land biospheric and oceanic fluxes computed in the FPP. The signals of these surface-flux variations on atmospheric CO2 will be isolated using forward modeling tools, which also incorporate estimates of transport error. The

  15. A Combined Maximum-likelihood Analysis of the High-energy Astrophysical Neutrino Flux Measured with IceCube

    NASA Astrophysics Data System (ADS)

    Aartsen, M. G.; Abraham, K.; Ackermann, M.; Adams, J.; Aguilar, J. A.; Ahlers, M.; Ahrens, M.; Altmann, D.; Anderson, T.; Archinger, M.; Arguelles, C.; Arlen, T. C.; Auffenberg, J.; Bai, X.; Barwick, S. W.; Baum, V.; Bay, R.; Beatty, J. J.; Becker Tjus, J.; Becker, K.-H.; Beiser, E.; BenZvi, S.; Berghaus, P.; Berley, D.; Bernardini, E.; Bernhard, A.; Besson, D. Z.; Binder, G.; Bindig, D.; Bissok, M.; Blaufuss, E.; Blumenthal, J.; Boersma, D. J.; Bohm, C.; Börner, M.; Bos, F.; Bose, D.; Böser, S.; Botner, O.; Braun, J.; Brayeur, L.; Bretz, H.-P.; Brown, A. M.; Buzinsky, N.; Casey, J.; Casier, M.; Cheung, E.; Chirkin, D.; Christov, A.; Christy, B.; Clark, K.; Classen, L.; Coenders, S.; Cowen, D. F.; Cruz Silva, A. H.; Daughhetee, J.; Davis, J. C.; Day, M.; de André, J. P. A. M.; De Clercq, C.; Dembinski, H.; De Ridder, S.; Desiati, P.; de Vries, K. D.; de Wasseige, G.; de With, M.; DeYoung, T.; Díaz-Vélez, J. C.; Dumm, J. P.; Dunkman, M.; Eagan, R.; Eberhardt, B.; Ehrhardt, T.; Eichmann, B.; Euler, S.; Evenson, P. A.; Fadiran, O.; Fahey, S.; Fazely, A. R.; Fedynitch, A.; Feintzeig, J.; Felde, J.; Filimonov, K.; Finley, C.; Fischer-Wasels, T.; Flis, S.; Fuchs, T.; Gaisser, T. K.; Gaior, R.; Gallagher, J.; Gerhardt, L.; Ghorbani, K.; Gier, D.; Gladstone, L.; Glagla, M.; Glüsenkamp, T.; Goldschmidt, A.; Golup, G.; Gonzalez, J. G.; Goodman, J. A.; Góra, D.; Grant, D.; Gretskov, P.; Groh, J. C.; Gross, A.; Ha, C.; Haack, C.; Haj Ismail, A.; Hallgren, A.; Halzen, F.; Hansmann, B.; Hanson, K.; Hebecker, D.; Heereman, D.; Helbing, K.; Hellauer, R.; Hellwig, D.; Hickford, S.; Hignight, J.; Hill, G. C.; Hoffman, K. D.; Hoffmann, R.; Holzapfel, K.; Homeier, A.; Hoshina, K.; Huang, F.; Huber, M.; Huelsnitz, W.; Hulth, P. O.; Hultqvist, K.; In, S.; Ishihara, A.; Jacobi, E.; Japaridze, G. S.; Jero, K.; Jurkovic, M.; Kaminsky, B.; Kappes, A.; Karg, T.; Karle, A.; Kauer, M.; Keivani, A.; Kelley, J. L.; Kemp, J.; Kheirandish, A.; Kiryluk, J.; Kläs, J.; Klein, S. R.; Kohnen, G.; Kolanoski, H.; Konietz, R.; Koob, A.; Köpke, L.; Kopper, C.; Kopper, S.; Koskinen, D. J.; Kowalski, M.; Krings, K.; Kroll, G.; Kroll, M.; Kunnen, J.; Kurahashi, N.; Kuwabara, T.; Labare, M.; Lanfranchi, J. L.; Larson, M. J.; Lesiak-Bzdak, M.; Leuermann, M.; Leuner, J.; Lünemann, J.; Madsen, J.; Maggi, G.; Mahn, K. B. M.; Maruyama, R.; Mase, K.; Matis, H. S.; Maunu, R.; McNally, F.; Meagher, K.; Medici, M.; Meli, A.; Menne, T.; Merino, G.; Meures, T.; Miarecki, S.; Middell, E.; Middlemas, E.; Miller, J.; Mohrmann, L.; Montaruli, T.; Morse, R.; Nahnhauer, R.; Naumann, U.; Niederhausen, H.; Nowicki, S. C.; Nygren, D. R.; Obertacke, A.; Olivas, A.; Omairat, A.; O'Murchadha, A.; Palczewski, T.; Paul, L.; Pepper, J. A.; Pérez de los Heros, C.; Pfendner, C.; Pieloth, D.; Pinat, E.; Posselt, J.; Price, P. B.; Przybylski, G. T.; Pütz, J.; Quinnan, M.; Rädel, L.; Rameez, M.; Rawlins, K.; Redl, P.; Reimann, R.; Relich, M.; Resconi, E.; Rhode, W.; Richman, M.; Richter, S.; Riedel, B.; Robertson, S.; Rongen, M.; Rott, C.; Ruhe, T.; Ruzybayev, B.; Ryckbosch, D.; Saba, S. M.; Sabbatini, L.; Sander, H.-G.; Sandrock, A.; Sandroos, J.; Sarkar, S.; Schatto, K.; Scheriau, F.; Schimp, M.; Schmidt, T.; Schmitz, M.; Schoenen, S.; Schöneberg, S.; Schönwald, A.; Schukraft, A.; Schulte, L.; Seckel, D.; Seunarine, S.; Shanidze, R.; Smith, M. W. E.; Soldin, D.; Spiczak, G. M.; Spiering, C.; Stahlberg, M.; Stamatikos, M.; Stanev, T.; Stanisha, N. A.; Stasik, A.; Stezelberger, T.; Stokstad, R. G.; Stössl, A.; Strahler, E. A.; Ström, R.; Strotjohann, N. L.; Sullivan, G. W.; Sutherland, M.; Taavola, H.; Taboada, I.; Ter-Antonyan, S.; Terliuk, A.; Tešić, G.; Tilav, S.; Toale, P. A.; Tobin, M. N.; Tosi, D.; Tselengidou, M.; Unger, E.; Usner, M.; Vallecorsa, S.; Vandenbroucke, J.; van Eijndhoven, N.; Vanheule, S.; van Santen, J.; Veenkamp, J.; Vehring, M.; Voge, M.; Vraeghe, M.; Walck, C.; Wallace, A.; Wallraff, M.; Wandkowsky, N.; Weaver, Ch.; Wendt, C.; Westerhoff, S.; Whelan, B. J.; Whitehorn, N.; Wichary, C.; Wiebe, K.; Wiebusch, C. H.; Wille, L.; Williams, D. R.; Wissing, H.; Wolf, M.; Wood, T. R.; Woschnagg, K.; Xu, D. L.; Xu, X. W.; Xu, Y.; Yanez, J. P.; Yodh, G.; Yoshida, S.; Zarzhitsky, P.; Zoll, M.; IceCube Collaboration

    2015-08-01

    Evidence for an extraterrestrial flux of high-energy neutrinos has now been found in multiple searches with the IceCube detector. The first solid evidence was provided by a search for neutrino events with deposited energies ≳ 30 TeV and interaction vertices inside the instrumented volume. Recent analyses suggest that the extraterrestrial flux extends to lower energies and is also visible with throughgoing, νμ-induced tracks from the Northern Hemisphere. Here, we combine the results from six different IceCube searches for astrophysical neutrinos in a maximum-likelihood analysis. The combined event sample features high-statistics samples of shower-like and track-like events. The data are fit in up to three observables: energy, zenith angle, and event topology. Assuming the astrophysical neutrino flux to be isotropic and to consist of equal flavors at Earth, the all-flavor spectrum with neutrino energies between 25 TeV and 2.8 PeV is well described by an unbroken power law with best-fit spectral index -2.50 ± 0.09 and a flux at 100 TeV of ({6.7}-1.2+1.1)× {10}-18 {{GeV}}-1 {{{s}}}-1 {{sr}}-1 {{cm}}-2. Under the same assumptions, an unbroken power law with index -2 is disfavored with a significance of 3.8σ (p = 0.0066%) with respect to the best fit. This significance is reduced to 2.1σ (p = 1.7%) if instead we compare the best fit to a spectrum with index -2 that has an exponential cut-off at high energies. Allowing the electron-neutrino flux to deviate from the other two flavors, we find a νe fraction of 0.18 ± 0.11 at Earth. The sole production of electron neutrinos, which would be characteristic of neutron-decay-dominated sources, is rejected with a significance of 3.6σ (p = 0.014%).

  16. Contrasting ecosystem CO2 fluxes of inland and coastal wetlands: a meta-analysis of eddy covariance data.

    PubMed

    Lu, Weizhi; Xiao, Jingfeng; Liu, Fang; Zhang, Yue; Liu, Chang'an; Lin, Guanghui

    2017-03-01

    Wetlands play an important role in regulating the atmospheric carbon dioxide (CO2 ) concentrations and thus affecting the climate. However, there is still lack of quantitative evaluation of such a role across different wetland types, especially at the global scale. Here, we conducted a meta-analysis to compare ecosystem CO2 fluxes among various types of wetlands using a global database compiled from the literature. This database consists of 143 site-years of eddy covariance data from 22 inland wetland and 21 coastal wetland sites across the globe. Coastal wetlands had higher annual gross primary productivity (GPP), ecosystem respiration (Re ), and net ecosystem productivity (NEP) than inland wetlands. On a per unit area basis, coastal wetlands provided large CO2 sinks, while inland wetlands provided small CO2 sinks or were nearly CO2 neutral. The annual CO2 sink strength was 93.15 and 208.37 g C m(-2) for inland and coastal wetlands, respectively. Annual CO2 fluxes were mainly regulated by mean annual temperature (MAT) and mean annual precipitation (MAP). For coastal and inland wetlands combined, MAT and MAP explained 71%, 54%, and 57% of the variations in GPP, Re , and NEP, respectively. The CO2 fluxes of wetlands were also related to leaf area index (LAI). The CO2 fluxes also varied with water table depth (WTD), although the effects of WTD were not statistically significant. NEP was jointly determined by GPP and Re for both inland and coastal wetlands. However, the NEP/Re and NEP/GPP ratios exhibited little variability for inland wetlands and decreased for coastal wetlands with increasing latitude. The contrasting of CO2 fluxes between inland and coastal wetlands globally can improve our understanding of the roles of wetlands in the global C cycle. Our results also have implications for informing wetland management and climate change policymaking, for example, the efforts being made by international organizations and enterprises to restore coastal wetlands for

  17. Natural isotopic signatures of variations in body nitrogen fluxes: a compartmental model analysis.

    PubMed

    Poupin, Nathalie; Mariotti, François; Huneau, Jean-François; Hermier, Dominique; Fouillet, Hélène

    2014-10-01

    Body tissues are generally 15N-enriched over the diet, with a discrimination factor (Δ15N) that varies among tissues and individuals as a function of their nutritional and physiopathological condition. However, both 15N bioaccumulation and intra- and inter-individual Δ15N variations are still poorly understood, so that theoretical models are required to understand their underlying mechanisms. Using experimental Δ15N measurements in rats, we developed a multi-compartmental model that provides the first detailed representation of the complex functioning of the body's Δ15N system, by explicitly linking the sizes and Δ15N values of 21 nitrogen pools to the rates and isotope effects of 49 nitrogen metabolic fluxes. We have shown that (i) besides urea production, several metabolic pathways (e.g., protein synthesis, amino acid intracellular metabolism, urea recycling and intestinal absorption or secretion) are most probably associated with isotope fractionation and together contribute to 15N accumulation in tissues, (ii) the Δ15N of a tissue at steady-state is not affected by variations of its P turnover rate, but can vary according to the relative orientation of tissue free amino acids towards oxidation vs. protein synthesis, (iii) at the whole-body level, Δ15N variations result from variations in the body partitioning of nitrogen fluxes (e.g., urea production, urea recycling and amino acid exchanges), with or without changes in nitrogen balance, (iv) any deviation from the optimal amino acid intake, in terms of both quality and quantity, causes a global rise in tissue Δ15N, and (v) Δ15N variations differ between tissues depending on the metabolic changes involved, which can therefore be identified using simultaneous multi-tissue Δ15N measurements. This work provides proof of concept that Δ15N measurements constitute a new promising tool to investigate how metabolic fluxes are nutritionally or physiopathologically reorganized or altered. The existence of such

  18. Natural Isotopic Signatures of Variations in Body Nitrogen Fluxes: A Compartmental Model Analysis

    PubMed Central

    Poupin, Nathalie; Mariotti, François; Huneau, Jean-François; Hermier, Dominique; Fouillet, Hélène

    2014-01-01

    Body tissues are generally 15N-enriched over the diet, with a discrimination factor (Δ15N) that varies among tissues and individuals as a function of their nutritional and physiopathological condition. However, both 15N bioaccumulation and intra- and inter-individual Δ15N variations are still poorly understood, so that theoretical models are required to understand their underlying mechanisms. Using experimental Δ15N measurements in rats, we developed a multi-compartmental model that provides the first detailed representation of the complex functioning of the body's Δ15N system, by explicitly linking the sizes and Δ15N values of 21 nitrogen pools to the rates and isotope effects of 49 nitrogen metabolic fluxes. We have shown that (i) besides urea production, several metabolic pathways (e.g., protein synthesis, amino acid intracellular metabolism, urea recycling and intestinal absorption or secretion) are most probably associated with isotope fractionation and together contribute to 15N accumulation in tissues, (ii) the Δ15N of a tissue at steady-state is not affected by variations of its P turnover rate, but can vary according to the relative orientation of tissue free amino acids towards oxidation vs. protein synthesis, (iii) at the whole-body level, Δ15N variations result from variations in the body partitioning of nitrogen fluxes (e.g., urea production, urea recycling and amino acid exchanges), with or without changes in nitrogen balance, (iv) any deviation from the optimal amino acid intake, in terms of both quality and quantity, causes a global rise in tissue Δ15N, and (v) Δ15N variations differ between tissues depending on the metabolic changes involved, which can therefore be identified using simultaneous multi-tissue Δ15N measurements. This work provides proof of concept that Δ15N measurements constitute a new promising tool to investigate how metabolic fluxes are nutritionally or physiopathologically reorganized or altered. The existence of such

  19. Seahorse Xfe 24 Extracellular Flux Analyzer-Based Analysis of Cellular Respiration in Caenorhabditis elegans.

    PubMed

    Luz, Anthony L; Smith, Latasha L; Rooney, John P; Meyer, Joel N

    2015-11-02

    Mitochondria are critical for their role in ATP production as well as multiple nonenergetic functions, and mitochondrial dysfunction is causal in myriad human diseases. Less well appreciated is the fact that mitochondria integrate environmental and intercellular as well as intracellular signals to modulate function. Because mitochondria function in an organismal milieu, there is need for assays capable of rapidly assessing mitochondrial health in vivo. Here, using the Seahorse XF(e) 24 Extracellular Flux Analyzer and the pharmacological inhibitors dicyclohexylcarbodiimide (DCCD, ATP synthase inhibitor), carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP, mitochondrial uncoupler), and sodium azide (cytochrome c oxidase inhibitor), we describe how to obtain in vivo measurements of the fundamental parameters [basal oxygen consumption rate (OCR), ATP-linked respiration, maximal OCR, spare respiratory capacity, and proton leak] of the mitochondrial respiratory chain in the model organism Caenorhabditis elegans.

  20. Seahorse Xfe24 Extracellular Flux Analyzer-based analysis of cellular respiration in Caenorhabditis elegans

    PubMed Central

    Luz, Anthony L.; Smith, Latasha L.; Rooney, John P.

    2015-01-01

    Mitochondria are critical for their role in ATP production as well as multiple nonenergetic functions, and mitochondrial dysfunction is causal in myriad human diseases. Less well appreciated is the fact that mitochondria integrate environmental and inter- as well as intracellular signals to modulate function. Because mitochondria function in an organismal milieu, there is need for assays capable of rapidly assessing mitochondrial health in vivo. Here, using the Seahorse XFe24 Extracellular Flux Analyzer and the pharmacological inhibitors dicyclohexylcarbodiimide (DCCD, ATP synthase inhibitor), carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP, mitochondrial uncoupler) and sodium azide (cytochrome c oxidase inhibitor), we describe how to obtain in vivo measurements of the fundamental parameters (basal oxygen consumption rate (OCR), ATP-linked respiration, maximal OCR, spare respiratory capacity and proton leak) of the mitochondrial respiratory chain in the model organism Caenorhabditis elegans. PMID:26523474

  1. High Flux Isotope Reactor Core Analysis-Challenges and Recent Enhancements in Modeling and Simulation

    SciTech Connect

    Ilas, Germina

    2016-01-01

    A concerted effort over the past few years has focused on enhancing the core depletion models for the High Flux Isotope Reactor (HFIR) as part of a comprehensive study for designing a HFIR core that would use low-enriched uranium (LEU) fuel. A HFIR core depletion model that is based on current state-of-the-art methods and nuclear data was needed for use as a reference for the design of an LEU fuel for HFIR and to improve the basis for analyses that support HFIR s current operation with high-enriched uranium (HEU) fuel. This paper summarizes the recent improvements in modeling and simulation for HFIR core analyses, with a focus on core depletion models.

  2. Inference of physical phenomena from FFTF (Fast Flux Test Facility) noise analysis

    SciTech Connect

    Thie, J.A.; Damiano, B.; Campbell, L.R.

    1989-01-01

    The source of features observed in noise spectra collected by an automated data collection system operated by the Oak Ridge National Laboratory at the Fast Flux Test Facility (FFTF) can be identified using a methodology based on careful data observation and intuition. When a large collection of data is available, as in this case, automatic pattern recognition and parameter storage and retrieval using a data base can be used to extract useful information. However, results can be limited to empirical signature comparison monitoring unless an effort is made to determine the noise sources. This paper describes the identification of several FFTF noise data phenomena and suggests how this understanding may lead to new or enhanced monitoring. 13 refs., 4 figs.

  3. Analysis and Simulation on the Effect of Rotor Interturn Short Circuit on Magnetic Flux Density of Turbo-Generator

    NASA Astrophysics Data System (ADS)

    He, Yu-Ling; Ke, Meng-Qiang; Tang, Gui-Ji; Jiang, Hong-Chun; Yuan, Xing-Hua

    2016-09-01

    The intent of this paper is to investigate the effect of the interturn short circuit fault (ISCF) in rotor on the magnetic flux density (MFD) of turbo-generator. Different from other studies, this work not only pays attention to the influence of the faulty degrees on the general magnetic field, but also investigates the effect of the short circuit positions on the harmonic components of MFD. The theoretical analysis and the digital simulation through the FEM software Ansoft are performed for a QSFN-600-2YHG turbo-generator. Several significant formulas and conclusions drawn from the analysis and the simulation results are obtained to indicate the relation between the harmonic amplitude of the MFD and the faulty degree (via nm, the number of the short circuit turns), and the relation between the MFD harmonic amplitude and the faulty position (via αr, the angle of the two slots in which the interturn short circuit occurs). Also, the developing tendency of the general magnetic field intensity, the distribution of the magnetic flux lines, and the peak-to-peak value of MFD are presented.

  4. Hydroacoustic and spatial analysis of sediment fluxes and accumulation rates in two Virginia reservoirs, USA.

    PubMed

    Clark, E V; Odhiambo, B K; Yoon, S; Pilati, L

    2015-06-01

    Watershed sediment fluxes and reservoir sediment accumulation rates were analyzed in two contrasting reservoir systems in central and western Virginia. Lake Pelham, located in the Piedmont geologic province, is a human-impacted reservoir with a watershed dominated by agricultural, residential and industrial land uses. Conversely, Lake Moomaw has a largely undeveloped watershed characterized by very steep slopes and forested land use located in the Valley and Ridge province. The Revised Universal Soil Loss Equation (RUSLE) and sediment delivery ratios (SDRs) were used to estimate soil losses in the two watersheds. Bathymetric and sediment accumulation surveys of the two reservoirs were also conducted using a multi-frequency hydroacoustic surveying system. The RUSLE/SDR erosion model estimates 2150 kg ha(-1) year(-1) for Lake Pelham and 2720 kg ha(-1) year(-1) for Lake Moomaw, a 410 and 13 % increase from assumed pristine (100 % forested) land use for the respective basins. Mean sediment accumulation rates of 1.51 and 0.60 cm year(-1) were estimated from the hydroacoustic survey of Lake Pelham and Lake Moomaw, respectively. Overall, Lake Moomaw has relatively low sediment accumulation rates; however, the reservoir is vulnerable to increases in sediment fluxes with further human development due to the steep slopes and highly erodible colluvial soils that characterize the basin. Higher erosion and sediment accumulation rates in Lake Pelham are most likely reflecting the impact of human development on sedimentation processes, where the loss of vegetal buffers and increase in impervious surfaces exacerbates both the surficial soil losses as well as intrinsic stream sediment production leading to the current annual reservoir capacity loss of 0.4 %.

  5. Intensive flux measurements and analysis of greenhouse gases from an upland cabbage field at Kunsan, Korea

    NASA Astrophysics Data System (ADS)

    Kim, D.; Na, U.

    2010-12-01

    It has been recognized that intensively managed agricultural soil is a dominant source of atmospheric N2O through increase in use of nitrogen fertilizer and soil microbial processes, contributing to about 57% (9Tg y-1) of total N2O annual global emission. Organic carbons in soil and wetland sediment including tidal flat affect the CO2 and CH4 emission in such environments depending on their physicochemical conditions. From October 2009 to June 2010, CO2, CH4, and N2O (GHG) soil emission measurements were conducted from upland cabbage field at Kunsan (35o56’23’’N, 126o43’14’’E), Korea by using closed static chamber method. During the experimental period, hourly GHG emissions were conducted mostly from 1000 to 1800LST in each field measurement day (total 28 days). After placing each chamber over soil surface of two neighboring plots, 50 ml of air sample inside the chambers was taken for every 15 min over a 30 min period by using plastic syringes (total of three samples). GHG concentrations were simultaneously analyzed in the laboratory by using a GC equipped with a methanizer, FID and ECD (Varian CP3800). The GHG fluxes were calculated from a linear regression of the changes in the concentrations. Negative values indicate GHG uptake by the soil surface, and positive values indicate GHG emission to the atmosphere. In addition, soil parameters (e.g. soil moisture, temperature, pH, organic C, soil N) were measured at the sampling plot. The average soil pH and soil moisture during the experimental period was ~pH5.4±0.4 and 70.0±19.7 %WFPS, respectively. The average fluxes and ranges of GHG during the experimental period were -0.004±0.032 mg-m-2 hr-1 (-0.087 ~ 0.045 mg-m-2 hr-1) for CH4, 5.32±57.63 mg-m-2 hr-1 (-92.96 ~ 139.38 mg-m-2 hr-1) for CO2, and 1.119±1.918 mg-m-2 hr-1 (0.077 ~ 8.409 mg-m-2 hr-1) for N2O, respectively. Monthly base flux measurement results revealed that monthly means of CO2 and CH4 flux during October (fall) was positive and

  6. Development of a benthic-flux chamber for measurement of ground-water seepage and water sampling for mercury analysis at the sediment-water interface

    USGS Publications Warehouse

    Menheer, M.A.

    2004-01-01

    A benthic-flux chamber was constructed to collect data to determine the relation between ground- and surface-water interaction and mercury concentrations in water at the sediment- water interface. The benthic-flux chamber was successfully used to measure the rate of ground water seeping to surface water or surface water seeping to ground water, and to collect water samples for mercury analysis from the sedimentwater interface in a lake setting. The benthic-flux chamber was designed to be deployed in relatively calm fresh water lakes, in areas of water less than 2 meters deep. The groundwater seepage rate data were comparable to data from an in-line flow meter in a calibration tank and with data from two 55-gallon drum seepage meters concurrently deployed in two different lakes. The benthic-flux chamber was used to collect possible water samples for analysis of total mercury and methylmercury concentrations.

  7. Wavelet and Fractal Analysis of Remotely Sensed Surface Temperature with Applications to Estimation of Surface Sensible Heat Flux Density

    NASA Technical Reports Server (NTRS)

    Schieldge, John

    2000-01-01

    Wavelet and fractal analyses have been used successfully to analyze one-dimensional data sets such as time series of financial, physical, and biological parameters. These techniques have been applied to two-dimensional problems in some instances, including the analysis of remote sensing imagery. In this respect, these techniques have not been widely used by the remote sensing community, and their overall capabilities as analytical tools for use on satellite and aircraft data sets is not well known. Wavelet and fractal analyses have the potential to provide fresh insight into the characterization of surface properties such as temperature and emissivity distributions, and surface processes such as the heat and water vapor exchange between the surface and the lower atmosphere. In particular, the variation of sensible heat flux density as a function of the change In scale of surface properties Is difficult to estimate, but - in general - wavelets and fractals have proved useful in determining the way a parameter varies with changes in scale. We present the results of a limited study on the relationship between spatial variations in surface temperature distribution and sensible heat flux distribution as determined by separate wavelet and fractal analyses. We analyzed aircraft imagery obtained in the thermal infrared (IR) bands from the multispectral TIMS and hyperspectral MASTER airborne sensors. The thermal IR data allows us to estimate the surface kinetic temperature distribution for a number of sites in the Midwestern and Southwestern United States (viz., San Pedro River Basin, Arizona; El Reno, Oklahoma; Jornada, New Mexico). The ground spatial resolution of the aircraft data varied from 5 to 15 meters. All sites were instrumented with meteorological and hydrological equipment including surface layer flux measuring stations such as Bowen Ratio systems and sonic anemometers. The ground and aircraft data sets provided the inputs for the wavelet and fractal analyses

  8. Altered swelling and ion fluxes in articular cartilage as a biomarker in osteoarthritis and joint immobilization: a computational analysis

    PubMed Central

    Manzano, Sara; Manzano, Raquel; Doblaré, Manuel; Doweidar, Mohamed Hamdy

    2015-01-01

    In healthy cartilage, mechano-electrochemical phenomena act together to maintain tissue homeostasis. Osteoarthritis (OA) and degenerative diseases disrupt this biological equilibrium by causing structural deterioration and subsequent dysfunction of the tissue. Swelling and ion flux alteration as well as abnormal ion distribution are proposed as primary indicators of tissue degradation. In this paper, we present an extension of a previous three-dimensional computational model of the cartilage behaviour developed by the authors to simulate the contribution of the main tissue components in its behaviour. The model considers the mechano-electrochemical events as concurrent phenomena in a three-dimensional environment. This model has been extended here to include the effect of repulsion of negative charges attached to proteoglycans. Moreover, we have studied the fluctuation of these charges owning to proteoglycan variations in healthy and pathological articular cartilage. In this sense, standard patterns of healthy and degraded tissue behaviour can be obtained which could be a helpful diagnostic tool. By introducing measured properties of unhealthy cartilage into the computational model, the severity of tissue degeneration can be predicted avoiding complex tissue extraction and subsequent in vitro analysis. In this work, the model has been applied to monitor and analyse cartilage behaviour at different stages of OA and in both short (four, six and eight weeks) and long-term (11 weeks) fully immobilized joints. Simulation results showed marked differences in the corresponding swelling phenomena, in outgoing cation fluxes and in cation distributions. Furthermore, long-term immobilized patients display similar swelling as well as fluxes and distribution of cations to patients in the early stages of OA, thus, preventive treatments are highly recommended to avoid tissue deterioration. PMID:25392400

  9. Assessing the spatial representativeness of eddy-covariance measurements of AmeriFlux network based on remote sensing and footprint analysis

    NASA Astrophysics Data System (ADS)

    Fu, D.; Zhang, L.; Chen, B.

    2015-12-01

    The eddy-covariance towers of AmeriFlux network are important for the analysis of terrestrial ecosystem-atmosphere interactions, and they have been used to improve our understanding of the mechanism behind terrestrial carbon cycle and upscaling from site to landscape and regional scales. However, the spatial representativeness of AmeriFlux network has not been assessed, especially accounting for the effects of land cover change on it using high spatial resolution data. Here we demonstrated an approach for evaluating the spatial representativeness of flux tower measurements based on footprint climatology analyses, land cover change data and remotely sensed vegetation indices. This method was applied to 79 flux towers of AmeriFlux network located in the continental United States, covering evergreen forest, deciduous forest, mixed forest, grass, cropland, shrub, and wetland biomes. For each site, monthly and annual footprint climatologies (i.e. monthly or annual accumulative footprints) were calculated using the Simple Analytical Footprint model on Eulerian coordinates (SAFE-f). The footprint climatologies were then overlaid on the images of Normalized Difference Vegetation Index (NDVI) and National Land Cover Database (NLCD) for the years (2001, 2006 and 2011), which were used as surrogates of land surface fluxes to assess the spatial representativeness. For most sites of AmeriFlux network, the results show that (i) the percentages of the target vegetation functional type (dominant land cover) observed by the AmeriFlux towers were higher than 60%; (ii) to some extent, most of the AmeriFlux sites presented anisotropically distributed patterns of NDVI within the 90% annual footprint climatology area; (iii) the land surface heterogeneity within the flux footprint area differed among sites; and (iv) the land cover types had changed higher than 10% within 6 km*6 km area centered at the flux tower for 5 AmeriFlux sites. We conclude that the footprint modeling based on high

  10. Genealogy profiling through strain improvement by using metabolic network analysis: metabolic flux genealogy of several generations of lysine-producing corynebacteria.

    PubMed

    Wittmann, Christoph; Heinzle, Elmar

    2002-12-01

    A comprehensive approach of metabolite balancing, (13)C tracer studies, gas chromatography-mass spectrometry, matrix-assisted laser desorption ionization-time of flight mass spectrometry, and isotopomer modeling was applied for comparative metabolic network analysis of a genealogy of five successive generations of lysine-producing Corynebacterium glutamicum. The five strains examined (C. glutamicum ATCC 13032, 13287, 21253, 21526, and 21543) were previously obtained by random mutagenesis and selection. Throughout the genealogy, the lysine yield in batch cultures increased markedly from 1.2 to 24.9% relative to the glucose uptake flux. Strain optimization was accompanied by significant changes in intracellular flux distributions. The relative pentose phosphate pathway (PPP) flux successively increased, clearly corresponding to the product yield. Moreover, the anaplerotic net flux increased almost twofold as a consequence of concerted regulation of C(3) carboxylation and C(4) decarboxylation fluxes to cover the increased demand for lysine formation; thus, the overall increase was a consequence of concerted regulation of C(3) carboxylation and C(4) decarboxylation fluxes. The relative flux through isocitrate dehydrogenase dropped from 82.7% in the wild type to 59.9% in the lysine-producing mutants. In contrast to the NADPH demand, which increased from 109 to 172% due to the increasing lysine yield, the overall NADPH supply remained constant between 185 and 196%, resulting in a decrease in the apparent NADPH excess through strain optimization. Extrapolated to industrial lysine producers, the NADPH supply might become a limiting factor. The relative contributions of PPP and the tricarboxylic acid cycle to NADPH generation changed markedly, indicating that C. glutamicum is able to maintain a constant supply of NADPH under completely different flux conditions. Statistical analysis by a Monte Carlo approach revealed high precision for the estimated fluxes, underlining the

  11. Isotopically nonstationary 13C flux analysis of changes in Arabidopsis thaliana leaf metabolism due to high light acclimation

    SciTech Connect

    Ma, Fangfang; Jazmin, Lara J.; Young, Jamey D.; Allen, Doug K.

    2014-11-03

    Improving plant productivity is an important aim for metabolic engineering. There are few comprehensive methods that quantitatively describe leaf metabolism, although such information would be valuable for increasing photosynthetic capacity, enhancing biomass production, and rerouting carbon flux toward desirable end products. Isotopically nonstationary metabolic flux analysis (INST-MFA) has been previously applied to map carbon fluxes in photoautotrophic bacteria, which involves model-based regression of transient 13C-labeling patterns of intracellular metabolites. However, experimental and computational difficulties have hindered its application to terrestrial plant systems. Here, we performed in vivo isotopic labeling of Arabidopsis thaliana rosettes with 13CO2 and estimated fluxes throughout leaf photosynthetic metabolism by INST-MFA. Plants grown at 200 µmol m$-$2s$-$1 light were compared with plants acclimated for 9 d at an irradiance of 500 µmol∙m$-$2∙s$-$1. Approximately 1,400 independent mass isotopomer measurements obtained from analysis of 37 metabolite fragment ions were regressed to estimate 136 total fluxes (54 free fluxes) under each condition. The results provide a comprehensive description of changes in carbon partitioning and overall photosynthetic flux after long-term developmental acclimation of leaves to high light. Despite a doubling in the carboxylation rate, the photorespiratory flux increased from 17 to 28% of net CO2 assimilation with high-light acclimation (Vc/Vo: 3.5:1 vs. 2.3:1, respectively). In conclusion, this study highlights the potential of 13C INST-MFA to describe emergent flux phenotypes that respond to environmental conditions or plant physiology and cannot be obtained by other complementary approaches.

  12. ANALYSIS OF THE FLUX OF AN ENDOCRINE DISRUPTING DICARBOXIMIDE AND ITS DEGRADATION PRODUCTS FROM THE SOIL TO THE LOWER TROPOSPHERE

    EPA Science Inventory

    A method for measuring the atmospheric flux of the antiandrogenic dicarboxirnide, vinclozolin, and its degradation products was investigated. A nitric oxide laboratory chamber was modified to measure the flux of semi-volatile compounds. Pesticide application systems and soil in...

  13. System identification algorithm analysis of acupuncture effect on mean blood flux of contralateral hegu acupoint.

    PubMed

    Wang, Guangjun; Han, Jianguo; Litscher, Gerhard; Zhang, Weibo

    2012-01-01

    Background. Acupoints (belonging to 12 meridians) which have the same names are symmetrically distributed on the body. It has been proved that acupoints have certain biological specificities different from the normal parts of the body. However, there is little evidence that acupoints which have the same name and are located bilaterally and symmetrically have lateralized specificity. Thus, researching the lateralized specificity and the relationship between left-side and right-side acupuncture is of special importance. Methodology and Principal Findings. The mean blood flux (MBF) in both Hegu acupoints was measured by Moor full-field laser perfusion imager. With the method of system identification algorithm, the output distribution in different groups was acquired, based on different acupoint stimulation and standard signal input. It is demonstrated that after stimulation of the right Hegu acupoint by needle, the output value of MBF in contralateral Hegu acupoint was strongly amplified, while after acupuncturing the left Hegu acupoint, the output value of MBF in either side Hegu acupoint was amplified moderately. Conclusions and Significance. This paper indicates that the Hegu acupoint has lateralized specificity. After stimulating the ipsilateral Hegu acupoint, symmetry breaking will be produced in contrast to contralateral Hegu acupoint stimulation.

  14. A Data-Centered Collaboration Portal to Support Global Carbon-Flux Analysis

    SciTech Connect

    Agarwal, Deborah A.; Humphrey, Marty; Beekwilder, Norm; Jackson, Keith; Goode, Monte; van Ingen, Catharine

    2009-04-07

    Carbon-climate, like other environmental sciences, has been changing. Large-scalesynthesis studies are becoming more common. These synthesis studies are often conducted by science teams that are geographically distributed and on datasets that are global in scale. A broad array of collaboration and data analytics tools are now available that could support these science teams. However, building tools that scientists actually use is hard. Also, moving scientists from an informal collaboration structure to one mediated by technology often exposes inconsistencies in the understanding of the rules of engagement between collaborators. We have developed a scientific collaboration portal, called fluxdata.org, which serves the community of scientists providing and analyzing the global FLUXNET carbon-flux synthesis dataset. Key things we learned or re-learned during our portal development include: minimize the barrier to entry, provide features on a just-in-time basis, development of requirements is an on-going process, provide incentives to change leaders and leverage the opportunity they represent, automate as much as possible, and you can only learn how to make it better if people depend on it enough to give you feedback. In addition, we also learned that splitting the portal roles between scientists and computer scientists improved user adoption and trust. The fluxdata.org portal has now been in operation for ~;;1.5 years and has become central to the FLUXNET synthesis efforts.

  15. Flux Balance Analysis Inspired Bioprocess Upgrading for Lycopene Production by a Metabolically Engineered Strain of Yarrowia lipolytica

    PubMed Central

    Nambou, Komi; Jian, Xingxing; Zhang, Xinkai; Wei, Liujing; Lou, Jiajia; Madzak, Catherine; Hua, Qiang

    2015-01-01

    Genome-scale metabolic models embody a significant advantage of systems biology since their applications as metabolic flux simulation models enable predictions for the production of industrially-interesting metabolites. The biotechnological production of lycopene from Yarrowia lipolytica is an emerging scope that has not been fully scrutinized, especially for what concerns cultivation conditions of newly generated engineered strains. In this study, by combining flux balance analysis (FBA) and Plackett-Burman design, we screened chemicals for lycopene production from a metabolically engineered strain of Y. lipolytica. Lycopene concentrations of 126 and 242 mg/L were achieved correspondingly from the FBA-independent and the FBA-assisted designed media in fed-batch cultivation mode. Transcriptional studies revealed upregulations of heterologous genes in media designed according to FBA, thus implying the efficiency of model predictions. Our study will potentially support upgraded lycopene and other terpenoids production from existing or prospect bioengineered strains of Y. lipolytica and/or closely related yeast species. PMID:26703753

  16. Estimating regional greenhouse gas fluxes: An uncertainty analysis of planetary boundary layer techniques and bottom-up inventories

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Quantification of regional greenhouse gas (GHG) fluxes is essential for establishing mitigation strategies and evaluating their effectiveness. Here, we used multiple top-down approaches and multiple trace gas observations at a tall tower to estimate GHG regional fluxes and evaluate the GHG fluxes de...

  17. CeCaFDB: a curated database for the documentation, visualization and comparative analysis of central carbon metabolic flux distributions explored by 13C-fluxomics.

    PubMed

    Zhang, Zhengdong; Shen, Tie; Rui, Bin; Zhou, Wenwei; Zhou, Xiangfei; Shang, Chuanyu; Xin, Chenwei; Liu, Xiaoguang; Li, Gang; Jiang, Jiansi; Li, Chao; Li, Ruiyuan; Han, Mengshu; You, Shanping; Yu, Guojun; Yi, Yin; Wen, Han; Liu, Zhijie; Xie, Xiaoyao

    2015-01-01

    The Central Carbon Metabolic Flux Database (CeCaFDB, available at http://www.cecafdb.org) is a manually curated, multipurpose and open-access database for the documentation, visualization and comparative analysis of the quantitative flux results of central carbon metabolism among microbes and animal cells. It encompasses records for more than 500 flux distributions among 36 organisms and includes information regarding the genotype, culture medium, growth conditions and other specific information gathered from hundreds of journal articles. In addition to its comprehensive literature-derived data, the CeCaFDB supports a common text search function among the data and interactive visualization of the curated flux distributions with compartmentation information based on the Cytoscape Web API, which facilitates data interpretation. The CeCaFDB offers four modules to calculate a similarity score or to perform an alignment between the flux distributions. One of the modules was built using an inter programming algorithm for flux distribution alignment that was specifically designed for this study. Based on these modules, the CeCaFDB also supports an extensive flux distribution comparison function among the curated data. The CeCaFDB is strenuously designed to address the broad demands of biochemists, metabolic engineers, systems biologists and members of the -omics community.

  18. Neonatal infrared thermography imaging: Analysis of heat flux during different clinical scenarios

    NASA Astrophysics Data System (ADS)

    Abbas, Abbas K.; Heimann, Konrad; Blazek, Vladimir; Orlikowsky, Thorsten; Leonhardt, Steffen

    2012-11-01

    IntroductionAn accurate skin temperature measurement of Neonatal Infrared Thermography (NIRT) imaging requires an appropriate calibration process for compensation of external effects (e.g. variation of environmental temperature, variable air velocity or humidity). Although modern infrared cameras can perform such calibration, an additional compensation is required for highly accurate thermography. This compensation which corrects any temperature drift should occur during the NIRT imaging process. We introduce a compensation technique which is based on modeling the physical interactions within the measurement scene and derived the detected temperature signal of the object. Materials and methodsIn this work such compensation was performed for different NIRT imaging application in neonatology (e.g. convective incubators, kangaroo mother care (KMC), and an open radiant warmer). The spatially distributed temperatures of 12 preterm infants (average gestation age 31 weeks) were measured under these different infant care arrangements (i.e. closed care system like a convective incubator, and open care system like kangaroo mother care, and open radiant warmer). ResultsAs errors in measurement of temperature were anticipated, a novel compensation method derived from infrared thermography of the neonate's skin was developed. Moreover, the differences in temperature recording for the 12 preterm infants varied from subject to subject. This variation could be arising from individual experimental setting applied to the same region of interest over the neonate's body. The experimental results for the model-based corrections is verified over the selected patient group. ConclusionThe proposed technique relies on applying model-based correction to the measured temperature and reducing extraneous errors during NIRT. This application specific method is based on different heat flux compartments present in neonatal thermography scene. Furthermore, these results are considered to be

  19. Effects of experimental nitrogen deposition on peatland carbon pools and fluxes: a modelling analysis

    NASA Astrophysics Data System (ADS)

    Wu, Y.; Blodau, C.; Moore, T. R.; Bubier, J.; Juutinen, S.; Larmola, T.

    2015-01-01

    Nitrogen (N) pollution of peatlands alters their carbon (C) balances, yet long-term effects and controls are poorly understood. We applied the model PEATBOG to explore impacts of long-term nitrogen (N) fertilization on C cycling in an ombrotrophic bog. Simulations of summer gross ecosystem production (GEP), ecosystem respiration (ER) and net ecosystem exchange (NEE) were evaluated against 8 years of observations and extrapolated for 80 years to identify potential effects of N fertilization and factors influencing model behaviour. The model successfully simulated moss decline and raised GEP, ER and NEE on fertilized plots. GEP was systematically overestimated in the model compared to the field data due to factors that can be related to differences in vegetation distribution (e.g. shrubs vs. graminoid vegetation) and to high tolerance of vascular plants to N deposition in the model. Model performance regarding the 8-year response of GEP and NEE to N input was improved by introducing an N content threshold shifting the response of photosynthetic capacity (GEPmax) to N content in shrubs and graminoids from positive to negative at high N contents. Such changes also eliminated the competitive advantages of vascular species and led to resilience of mosses in the long-term. Regardless of the large changes of C fluxes over the short-term, the simulated GEP, ER and NEE after 80 years depended on whether a graminoid- or shrub-dominated system evolved. When the peatland remained shrub-Sphagnum-dominated, it shifted to a C source after only 10 years of fertilization at 6.4 g N m-2 yr-1, whereas this was not the case when it became graminoid-dominated. The modelling results thus highlight the importance of ecosystem adaptation and reaction of plant functional types to N deposition, when predicting the future C balance of N-polluted cool temperate bogs.

  20. Effects of experimental nitrogen deposition on peatland carbon pools and fluxes: a modeling analysis

    NASA Astrophysics Data System (ADS)

    Wu, Y.; Blodau, C.; Moore, T. R.; Bubier, J. L.; Juutinen, S.; Larmola, T.

    2014-07-01

    Nitrogen (N) pollution of peatlands alters their carbon (C) balances, yet long-term effects and controls are poorly understood. We applied the model PEATBOG to analyze impacts of long-term nitrogen (N) fertilization on C cycling in an ombrotrophic bog. Simulations of summer gross ecosystem production (GEP), ecosystem respiration (ER) and net ecosystem exchange (NEE) were evaluated against 8 years of observations and extrapolated for 80 years to identify potential effects of N fertilization and factors influencing model behavior. The model successfully simulated moss decline and raised GEP, ER and NEE on fertilized plots. GEP was systematically overestimated in the model compared to the field data due to high tolerance of Sphagnum to N deposition in the model. Model performance regarding the 8 year response of GEP and NEE to N was improved by introducing an N content threshold shifting the response of photosynthesis capacity to N content in shrubs and graminoids from positive to negative at high N contents. Such changes also eliminated the competitive advantages of vascular species and led to resilience of mosses in the long-term. Regardless of the large changes of C fluxes over the short-term, the simulated GEP, ER and NEE after 80 years depended on whether a graminoid- or shrub-dominated system evolved. When the peatland remained shrub-Sphagnum dominated, it shifted to a C source after only 10 years of fertilization at 6.4 g N m-2 yr-1, whereas this was not the case when it became graminoid-dominated. The modeling results thus highlight the importance of ecosystem adaptation and reaction of plant functional types to N deposition, when predicting the future C balance of N-polluted cool temperate bogs.

  1. Statistical analysis of the flux of micrometeoroids at Mercury from both cometary and asteroidal components

    NASA Astrophysics Data System (ADS)

    Borin, P.; Cremonese, G.; Marzari, F.

    2016-01-01

    Context. Meteoroid impacts are an important source of neutral atoms for the exosphere of Mercury. We previously estimated the contribution of meteoroids originating in the asteroid belt for vapor release. In this paper, we concentrate on the cometary component of particles impacting the planet. Comets and asteroids are considered to be the two major sources of interplanetary dust particles in the solar system. The debate about which source contributes most to dust populating the solar system is still ongoing. Aims: In this work, we compute the orbital evolution of dust particles produced by Jupiter-family comets (JFC) via N-body numerical integrations. From our numerical simulations, we compute the fraction of particles hitting Earth and Mercury's surface and the corresponding distribution of impact velocities. According to some authors more than 80% of all the incoming mass of meteoroids entering the Earth's atmosphere is concentrated in the mass range 10-7-10-3 g. In our model, we considered a slightly different range, 10-9 to 10-6 g, to include possible uncertainty. Methods: The orbital evolution of dust particles of different sizes is computed with a numerical integration code, which includes the effects of Poynting-Robertson drag, solar wind drag, and planetary perturbations. Results: By comparing the impact frequency of grains evolving either from main belt asteroids or JFC we find that the cometary component is significantly less efficient in releasing dust particles on Mercury than on the Earth. The opposite occurs in the case of dust coming from the main belt with a flux higher at Mercury than on the Earth. This is mostly due to the different dynamical histories of the grains from their release until impact. This may have important implications for the vapor production rate on Mercury. We compare our results with previous estimates given by different authors.

  2. Reactor physics input to the safety analysis report for the High Flux Isotope Reactor

    SciTech Connect

    Primm, R.T. III.

    1992-03-01

    HFIR specific, few group neutron and coupled neutron-gamma libraries have been prepared. These are based on data from ENDF/B-V and beginning-of-life (BOL) conditions. The neutron library includes actinide data for curium target rods. Six critical experiments, collectively designated HFIR critical experiment 4, were analyzed. Calculated k-effective was 2% high at BOL-typical conditions but was 1.0 at end-of-life-typical conditions. The local power density distributions were calculated for each of the critical experiments. The axially averaged values at a given radius were frequently within experimental error. However at individual points, the calculated local power densities were significantly different from the experimentally derived values (several times greater than experimental uncertainty). A reassessment of the foil activation data with transport theory techniques seems desirable. Using the results of the critical experiments study, a model of current HFIR configuration was prepared. As with the critical experiments, BOL k-effective was high (3%). However, end-of-life k-effective was high (2%). The end-of-life concentrations of fission products were compared to those generated using the ORIGEN code. Agreement was generally good through differences in the inventories of some important nuclides, Xe and I, need to be understood. End-of-cycle curium target isotopics based on measured, discharged target rods were compared to calculated values and agreement was good. Axial flux plots at various irradiation positions were generated. Time-dependent power distributions based on two-dimensional calculations were provided.

  3. GRACE Mass Flux Measurements of Inland and Marginal Seas from Mascons: Analysis and Validation

    NASA Astrophysics Data System (ADS)

    Loomis, B.; Luthcke, S. B.; Sabaka, T. J.

    2015-12-01

    The latest GRACE time-variable gravity mascon solution from the NASA Goddard Space Flight Center (GSFC) applies an optimized set of models and constraints towards the direct measurement of 1-arc-degree global mass flux parameters each month. Separate mascon spatial constraint regions have been defined for the largest inland and marginal seas: Mediterranean Sea, Black Sea, Caspian Sea, Red Sea, and Hudson Bay. The mascon estimation approach, when applied with well-designed constraints, minimizes signal leakage across regional boundaries and eliminates the need for post-processing strategies. These post-processing techniques (e.g. smoothed averaging kernels) are necessary for computing regional mass change from the unconstrained spherical harmonics provided by the GRACE project to reduce the effect of noisy high degree and order terms, but introduce signal leakage into and out of the considered region. These mass signals are also difficult to obtain from altimetry measurements due to the comparatively sparse temperature and salinity data in these regions, which is needed to compute and remove the steric component of sea level variations. We provide new GSFC mascon measurements of these inland and marginal seas and compare to results obtained from kernel-averaged spherical harmonic solutions and steric-corrected altimetry measurements. The relative accuracy of the various solutions is determined by incorporating their output into the set of forward models applied in our processing of the GRACE Level-1B data and analyzing the effect on the inter-satellite range-rate residuals, where a reduction in residuals is a direct validation of improved solution quality.

  4. A method for accounting for maintenance costs in flux balance analysis improves the prediction of plant cell metabolic phenotypes under stress conditions.

    PubMed

    Cheung, C Y Maurice; Williams, Thomas C R; Poolman, Mark G; Fell, David A; Ratcliffe, R George; Sweetlove, Lee J

    2013-09-01

    Flux balance models of metabolism generally utilize synthesis of biomass as the main determinant of intracellular fluxes. However, the biomass constraint alone is not sufficient to predict realistic fluxes in central heterotrophic metabolism of plant cells because of the major demand on the energy budget due to transport costs and cell maintenance. This major limitation can be addressed by incorporating transport steps into the metabolic model and by implementing a procedure that uses Pareto optimality analysis to explore the trade-off between ATP and NADPH production for maintenance. This leads to a method for predicting cell maintenance costs on the basis of the measured flux ratio between the oxidative steps of the oxidative pentose phosphate pathway and glycolysis. We show that accounting for transport and maintenance costs substantially improves the accuracy of fluxes predicted from a flux balance model of heterotrophic Arabidopsis cells in culture, irrespective of the objective function used in the analysis. Moreover, when the new method was applied to cells under control, elevated temperature and hyper-osmotic conditions, only elevated temperature led to a substantial increase in cell maintenance costs. It is concluded that the hyper-osmotic conditions tested did not impose a metabolic stress, in as much as the metabolic network is not forced to devote more resources to cell maintenance.

  5. Adjoint-based sensitivity analysis for high-energy density radiative transfer using flux-limited diffusion

    NASA Astrophysics Data System (ADS)

    Humbird, Kelli D.; McClarren, Ryan G.

    2017-03-01

    Uncertainty quantification and sensitivity analyses are a vital component for predictive modeling in the sciences and engineering. The adjoint approach to sensitivity analysis requires solving a primary system of equations and a mathematically related set of adjoint equations. The information contained in the equations can be combined to produce sensitivity information in a computationally efficient manner. In this work, sensitivity analyses are performed on systems described by flux-limited radiative diffusion using the adjoint approach. The sensitivities computed are shown to agree with standard perturbation theory and require significantly less computational time. The adjoint approach saves the computational cost of one forward solve per sensitivity, making the method attractive when multiple sensitivities are of interest.

  6. Characteristic and magnetic field analysis of a high temperature superconductor axial-flux coreless induction maglev motor

    NASA Astrophysics Data System (ADS)

    Wei, Qin; Yu, Fan; Jin, Fang; Shuo, Li; Guoguo, Li; Gang, Lv

    2012-04-01

    A new high temperature superconductor axial-flux coreless maglev motor (HTS AFIM) is proposed, of which the primary windings are made of HTS tapes and the secondary is a non-magnetic conductor. The main works of this paper are the magnetic-field computation and characteristics analysis of HTS AFIM. For the first one, the reduction of magnetic fields near outer and inner radius of the HTS AFIM is solved by introducing the sub-loop electro-magnetic model along the radial position. For the second one, the AC losses of HTS coils are calculated. The relationships between the device's characteristics and device parameters are presented, and the results indicate that under certain frequency and current levitation device can output enough lift force. The conclusions are verified by finite element calculations.

  7. Fermentation characterization and flux analysis of recombinant strains of Clostridium acetobutylicum with an inactivated solR gene.

    PubMed

    Harris, L M; Blank, L; Desai, R P; Welker, N E; Papoutsakis, E T

    2001-11-01

    The effect of solR inactivation on the metabolism of Clostridium acetobutylicum was examined using fermentation characterization and metabolic flux analysis. The solR-inactivated strain (SolRH) of this study had a higher rate of glucose utilization and produced higher solvent concentrations (by 25%, 14%, and 81%, respectively, for butanol, acetone, and ethanol) compared to the wild type. Strain SolRH(pTAAD), carrying a plasmid-encoded copy of the bifunctional alcohol/aldehyde dehydrogenase gene (aad) used in butanol production, produced even higher concentrations of solvents (by 21%, 45%, and 62%, respectively, for butanol, acetone, and ethanol) than strain SolRH. Clarithromycin used for strain SolRH maintenance during SolRH(pTAAD) fermentations did not alter product formation; however, tetracycline used for pTAAD maintenance resulted in 90% lower solvent production.

  8. Analysis of the cumulative neutrino flux from Fermi LAT blazar populations using 3 years of IceCube data

    NASA Astrophysics Data System (ADS)

    Glüsenkamp, Thorsten

    2016-07-01

    The recent discovery of a diffuse neutrino flux up to PeV energies raises the question of which populations of astrophysical sources contribute to this diffuse signal. One extragalactic candidate source population to produce high-energy neutrinos are Blazars. We present results from a likelihood analysis searching for cumulative neutrino emission from Blazar populations selected with the 2nd Fermi LAT AGN catalogue (2LAC) using an IceCube data set that has been optimized for the detection of individual sources. In contrast to previous searches with IceCube, the investigated populations contain up to hundreds of sources, the biggest one being the entire Blazar sample measured by the Fermi-LAT. No significant neutrino signal was found from any of these populations. Some implications of this non-observation for the origin of the observed PeV diffuse signal will be discussed.

  9. Understanding alternative fluxes/effluxes through comparative metabolic pathway analysis of phylum actinobacteria using a simplified approach.

    PubMed

    Verma, Mansi; Lal, Devi; Saxena, Anjali; Anand, Shailly; Kaur, Jasvinder; Kaur, Jaspreet; Lal, Rup

    2013-12-01

    Actinobacteria are known for their diverse metabolism and physiology. Some are dreadful human pathogens whereas some constitute the natural flora for human gut. Therefore, the understanding of metabolic pathways is a key feature for targeting the pathogenic bacteria without disturbing the symbiotic ones. A big challenge faced today is multiple drug resistance by Mycobacterium and other pathogens that utilize alternative fluxes/effluxes. With the availability of genome sequence, it is now feasible to conduct the comparative in silico analysis. Here we present a simplified approach to compare metabolic pathways so that the species specific enzyme may be traced and engineered for future therapeutics. The analyses of four key carbohydrate metabolic pathways, i.e., glycolysis, pyruvate metabolism, tri carboxylic acid cycle and pentose phosphate pathway suggest the presence of alternative fluxes. It was found that the upper pathway of glycolysis was highly variable in the actinobacterial genomes whereas lower glycolytic pathway was highly conserved. Likewise, pentose phosphate pathway was well conserved in contradiction to TCA cycle, which was found to be incomplete in majority of actinobacteria. The clustering based on presence and absence of genes of these metabolic pathways clearly revealed that members of different genera shared identical pathways and, therefore, provided an easy method to identify the metabolic similarities/differences between pathogenic and symbiotic organisms. The analyses could identify isoenzymes and some key enzymes that were found to be missing in some pathogenic actinobacteria. The present work defines a simple approach to explore the effluxes in four metabolic pathways within the phylum actinobacteria. The analysis clearly reflects that actinobacteria exhibit diverse routes for metabolizing substrates. The pathway comparison can help in finding the enzymes that can be used as drug targets for pathogens without effecting symbiotic organisms

  10. A 25-Year Retrospective Analysis of River Nitrogen Fluxes in the Atchafalaya

    NASA Astrophysics Data System (ADS)

    Xu, Y.

    2005-05-01

    Nitrogen enrichment from the upper Mississippi River Basin has been attributed to be the major cause for the hypoxia in the Northern Gulf of Mexico. The hypoxia threatens not only the aquatic ecosystem health but Louisiana's fishery industry directly among other problems. Although fresh water diversion from the lower Mississippi River into the region's wetlands has been considered an alternative means for reducing nitrogen loading, it is largely uncertain how much nitrogen can actually be retained from the overflowing waters in these natural wetlands. Generally, there is a knowledge gap in what tools are available for accurate assessment of nitrogen inflow, outflow and removal potential for the complex and diverse coastal floodplain systems. This study is to seek answers to three critical questions: (1) Does the Atchafalaya River Swamp remove a significant amount of nitrogen from the overflowing water or release more nitrogen into the Gulf than removing it? (2) How seasonally and annually do the nitrogen removal or release rates fluctuate? (3) What are the relationships between the nitrogen removal capacity and the basin's hydrologic conditions such as river stage and discharge? By utilizing river's long-term discharge and water quality data (1978-2002), monthly and annual nitrogen fluxes were quantified, and their relationships with the basin's hydrologic conditions were investigated. A total Kjeldahl nitrogen (TKN) mass input-output balance between the upstream (Simmesport) and downstream (Morgan City and Wax Lake Outlet) locations was established to examine the organic nitrogen removal potential for this largest freshwater swamp basin in North America. The results showed that on average, TKN input into the Atchafalaya was 200,323 Mg yr-1 and TKN output leaving the basin was 145,917 Mg yr-1, resulting in a 27% removal rate of nitrogen. Monthly nitrogen input and output in the basin were highest from March to June (input vs. output: 25,000 vs. 18,000 Mg mon-1) and

  11. A Mass Balance Analysis of Total Mercury Flux Through a Large, Managed Floodplain

    NASA Astrophysics Data System (ADS)

    Springborn, M.; Singer, M. B.; Dunne, T.

    2005-05-01

    The fate and transport of mercury are of critical concern in lowland floodplains worldwide. Increasing attention has been paid to the uncertainty of mercury sources and sinks in the Sacramento Valley, which is still recovering from decades of gold mining that used mercury for gold separation. Active in floods, Yolo Bypass is the largest flood-control bypass (or conveyance floodway) on the Sacramento River and is a key conduit for flow (up to 15,000 m(3)s(-1)) and the transport of fine sediment and adsorbed mercury to the San Francisco-Sacramento Bay-Delta. The 24,000 hectare bypass located in the lower Sacramento Valley has been recently implicated as a likely storage site for mercury with a high risk for methylation and transmission into the food chain at the primary wintering stop on the Pacific Flyway. In order to assess contaminant risk in the bypass, quantitative relationships between (1) total mercury concentration and suspended sediment concentration and (2) suspended sediment concentration and flow were developed for each of its major inputs and outputs using event-based sample data from various sources. These relationships were improved by incorporating dynamics of seasonal exhaustion and intraflood exhaustion (hysteresis) of sediment and mercury. From this characterization of how the flow-sediment-mercury transport system functions we were able to characterize the relative contributions of the various inputs. While the main inflow to the bypass is via flood weirs along the Sacramento River, two major creeks and an agricultural runoff canal made significant contributions to flow, sediment and mercury loads. Using the continuous record of flow to estimate sediment transport and sediment transport to estimate mercury flux we computed the net transfer of mercury through the bypass over a five-year period. Based on the volume and source of the expected change in flow and sediment, we were able to evaluate how mercury loading might change in the future due to

  12. Single-step enantioselective amino acid flux analysis by capillary electrophoresis using on-line sample preconcentration with chemical derivatization.

    PubMed

    Ptolemy, Adam S; Tran, Lara; Britz-McKibbin, Philip

    2006-07-15

    Capillary electrophoresis (CE) represents a versatile platform for integrating sample pretreatment with chemical analysis because of its ability to tune analyte electromigration and band dispersion properties in discontinuous electrolyte systems. In this article, a single-step method that combines on-line sample preconcentration with in-capillary chemical derivatization is developed for rapid, sensitive, and enantioselective analysis of micromolar levels of amino acids that lack intrinsic chromophores by CE with UV detection. Time-resolved electrophoretic studies revealed two distinct stages of amino acid band narrowing within the original long sample injection plug occurring both prior to and after in-capillary labeling via zone passing by ortho-phthalaldehyde/N-acetyl l-cysteine (OPA/NAC). This technique enabled direct analysis of d-amino acids in a 95% enantiomeric excess mixture with sub-micromolar detection limits and minimal sample handling, where the capillary functions as a preconcentrator, microreactor, and chiral selector. On-line sample preconcentration with chemical derivatization CE (SPCD-CE) was applied to study the enantioselective amino acid flux in Escherichia coli bacteria cultures, which demonstrated a unique l-Ala efflux into the extracellular medium. New strategies for high-throughput analyses of low-abundance metabolites are important for understanding fundamental physiological processes in bacteria required for screening the efficacy of new classes of antibiotics as well as altered metabolism in genetically modified mutant strains.

  13. Efficient energy transfer in light-harvesting systems: quantum-classical comparison, flux network, and robustness analysis.

    PubMed

    Wu, Jianlan; Liu, Fan; Ma, Jian; Silbey, Robert J; Cao, Jianshu

    2012-11-07

    Following the calculation of optimal energy transfer in thermal environment in our first paper [J. L. Wu, F. Liu, Y. Shen, J. S. Cao, and R. J. Silbey, New J. Phys. 12, 105012 (2010)], full quantum dynamics and leading-order "classical" hopping kinetics are compared in the seven-site Fenna-Matthews-Olson (FMO) protein complex. The difference between these two dynamic descriptions is due to higher-order quantum corrections. Two thermal bath models, classical white noise (the Haken-Strobl-Reineker (HSR) model) and quantum Debye model, are considered. In the seven-site FMO model, we observe that higher-order corrections lead to negligible changes in the trapping time or in energy transfer efficiency around the optimal and physiological conditions (2% in the HSR model and 0.1% in the quantum Debye model for the initial site at BChl 1). However, using the concept of integrated flux, we can identify significant differences in branching probabilities of the energy transfer network between hopping kinetics and quantum dynamics (26% in the HSR model and 32% in the quantum Debye model for the initial site at BChl 1). This observation indicates that the quantum coherence can significantly change the distribution of energy transfer pathways in the flux network with the efficiency nearly the same. The quantum-classical comparison of the average trapping time with the removal of the bottleneck site, BChl 4, demonstrates the robustness of the efficient energy transfer by the mechanism of multi-site quantum coherence. To reconcile with the latest eight-site FMO model which is also investigated in the third paper [J. Moix, J. L. Wu, P. F. Huo, D. F. Coker, and J. S. Cao, J. Phys. Chem. Lett. 2, 3045 (2011)], the quantum-classical comparison with the flux network analysis is summarized in Appendix C. The eight-site FMO model yields similar trapping time and network structure as the seven-site FMO model but leads to a more disperse distribution of energy transfer pathways.

  14. Advanced digital methods for blood flow flux analysis using µPIV approach

    NASA Astrophysics Data System (ADS)

    Kurochkin, Maxim A.; Timoshina, Polina A.; Fedosov, Ivan V.; Tuchin, Valery V.

    2015-03-01

    A digital optical system focused on work with laboratory animals for intravital capillaroscopy has been developed. It implements the particle image velocimetry (PIV) based approach for measurements of red blood cells velocity in laboratory rat stomach capillaries. We propose a method of involuntary displacement compensation of the capillary network images. Image stabilization algorithm is based on correlation of feature tracking. The efficiency of designed image stabilization algorithm was experimentally demonstrated. The results of capillary blood flow analysis are demonstrated.

  15. Genome-Based Metabolic Mapping and 13C Flux Analysis Reveal Systematic Properties of an Oleaginous Microalga Chlorella protothecoides

    SciTech Connect

    Wu, Chao; Xiong, Wei; Dai, Junbiao; Wu, Qingyu

    2014-12-15

    We report that integrated and genome-based flux balance analysis, metabolomics, and 13C-label profiling of phototrophic and heterotrophic metabolism in Chlorella protothecoides, an oleaginous green alga for biofuel. The green alga Chlorella protothecoides, capable of autotrophic and heterotrophic growth with rapid lipid synthesis, is a promising candidate for biofuel production. Based on the newly available genome knowledge of the alga, we reconstructed the compartmentalized metabolic network consisting of 272 metabolic reactions, 270 enzymes, and 461 encoding genes and simulated the growth in different cultivation conditions with flux balance analysis. Phenotype-phase plane analysis shows conditions achieving theoretical maximum of the biomass and corresponding fatty acid-producing rate for phototrophic cells (the ratio of photon uptake rate to CO2 uptake rate equals 8.4) and heterotrophic ones (the glucose uptake rate to O2 consumption rate reaches 2.4), respectively. Isotope-assisted liquid chromatography-mass spectrometry/mass spectrometry reveals higher metabolite concentrations in the glycolytic pathway and the tricarboxylic acid cycle in heterotrophic cells compared with autotrophic cells. We also observed enhanced levels of ATP, nicotinamide adenine dinucleotide (phosphate), reduced, acetyl-Coenzyme A, and malonyl-Coenzyme A in heterotrophic cells consistently, consistent with a strong activity of lipid synthesis. To profile the flux map in experimental conditions, we applied nonstationary 13C metabolic flux analysis as a complementing strategy to flux balance analysis. We found that the result reveals negligible photorespiratory fluxes and a metabolically low active tricarboxylic acid cycle in phototrophic C. protothecoides. In comparison, high throughput of amphibolic reactions and the tricarboxylic acid cycle with no glyoxylate shunt activities were measured for heterotrophic cells. Lastly, taken together, the

  16. Hybrid metabolic flux analysis and recombinant protein prediction in Pichia pastoris X-33 cultures expressing a single-chain antibody fragment.

    PubMed

    Isidro, Inês A; Portela, Rui M; Clemente, João J; Cunha, António E; Oliveira, Rui

    2016-09-01

    Despite the growing importance of the Pichia pastoris expression system as industrial workhorse, the literature is almost absent in systematic studies on how culture medium composition affects central carbon fluxes and heterologous protein expression. In this study we investigate how 26 variations of the BSM+PTM1 medium impact central carbon fluxes and protein expression in a P. pastoris X-33 strain expressing a single-chain antibody fragment. To achieve this goal, we adopted a hybrid metabolic flux analysis (MFA) methodology, which is a modification of standard MFA to predict the rate of synthesis of recombinant proteins. Hybrid MFA combines the traditional parametric estimation of central carbon fluxes with non-parametric statistical modeling of product-related quantitative or qualitative measurements as a function of central carbon fluxes. It was observed that protein yield variability was 53.6 % (relative standard deviation) among the different experiments. Protein yield is much more sensitive to medium composition than biomass growth, which is mainly determined by the carbon source availability and main salts. Hybrid MFA was able to describe accurately the protein yield with normalized RMSE of 6.3 % over 5 independent experiments. The metabolic state that promotes high protein yields is characterized by high overall metabolic rates through main central carbon pathways concomitantly with a relative shift of carbon flux from biosynthetic towards energy generating pathways.

  17. Systematic Analysis of the Effect of Small Scale Permeability Heterogeneity on Hyporheic Exchange Flux and Residence Times

    NASA Astrophysics Data System (ADS)

    Laube, G.; Schmidt, C.; Fleckenstein, J. H.

    2014-12-01

    The hyporheic zone (HZ) contributes significantly to whole stream biogeochemical cycling. Biogeochemical reactions within the HZ are often transport limited, thus, understanding these reactions requires knowledge about the magnitude of hyporheic fluxes (HF) and the residence time (RT) of these fluxes within the HZ. While the hydraulics of HF are relatively well understood, studies addressing the influence of permeability heterogeneity lack systematic analysis and have even produced contradictory results (e.g. [1] vs. [2]). In order to close this gap, this study uses a statistical numerical approach to elucidate the influence of permeability heterogeneity on HF and RT. We simulated and evaluated 3750 2D-scenarios of sediment heterogeneity by means of Gaussian random fields with focus on total HF and RT distribution. The scenarios were based on ten realizations of each of all possible combinations of 15 different correlation lengths, 5 dipping angles and 5 permeability variances. Roughly 500 hyporheic stream traces were analyzed per simulation, for a total of almost two million stream traces analyzed for correlations between permeability heterogeneity, HF, and RT. Total HF and the RT variance positively correlated with permeability variance while the mean RT negatively correlated with permeability variance. In contrast, changes in correlation lengths and dipping angles had little effect on the examined properties RT and HF. These results provide a possible explanation of the seemingly contradictory conclusions of recent studies, given that the permeability variances in these studies differ by several orders of magnitude. [1] Bardini, L., Boano, F., Cardenas, M.B, Sawyer, A.H, Revelli, R. and Ridolfi, L. "Small-Scale Permeability Heterogeneity Has Negligible Effects on Nutrient Cycling in Streambeds." Geophysical Research Letters, 2013. doi:10.1002/grl.50224. [2] Zhou, Y., Ritzi, R. W., Soltanian, M. R. and Dominic, D. F. "The Influence of Streambed Heterogeneity on

  18. A detailed analysis of inviscid flux splitting algorithms for real gases with equilibrium or finite-rate chemistry

    NASA Technical Reports Server (NTRS)

    Shuen, Jian-Shun; Liou, Meng-Sing; Van Leer, Bram

    1989-01-01

    The extension of the known flux-vector and flux-difference splittings to real gases via rigorous mathematical procedures is demonstrated. Formulations of both equilibrium and finite-rate chemistry for real-gas flows are described, with emphasis on derivations of finite-rate chemistry. Split-flux formulas from other authors are examined. A second-order upwind-based TVD scheme is adopted to eliminate oscillations and to obtain a sharp representation of discontinuities.

  19. Analysis of a link of embrittlement mechanisms and neutron flux effect as applied to reactor pressure vessel materials of WWER

    NASA Astrophysics Data System (ADS)

    Margolin, B. Z.; Yurchenko, E. V.; Morozov, A. M.; Pirogova, N. E.; Brumovsky, M.

    2013-03-01

    The effect of neutron flux on embrittlement of WWER RPV materials is analyzed for cases when different radiation defects prevail. Data bases on the ductile-brittle transition temperature shifts obtained in the surveillance specimens programs and the research programs are used. The material embrittlement mechanisms for which the flux effect is practically absent and for which the flux effect is remarkable are determined. For case when the phosphorus segregation mechanism dominates the theoretical justification of the absence of the flux effect is performed on the basis of the theory of radiation-enhanced diffusion.

  20. Overcoming the metabolic burden of protein secretion in Schizosaccharomyces pombe--a quantitative approach using 13C-based metabolic flux analysis.

    PubMed

    Klein, Tobias; Lange, Sabrina; Wilhelm, Nadine; Bureik, Matthias; Yang, Tae-Hoon; Heinzle, Elmar; Schneider, Konstantin

    2014-01-01

    Protein secretion in yeast is generally associated with a burden to cellular metabolism. To investigate this metabolic burden in Schizosaccharomyces pombe, we constructed a set of strains secreting the model protein maltase in different amounts. We quantified the influence of protein secretion on the metabolism applying (13)C-based metabolic flux analysis in chemostat cultures. Analysis of the macromolecular biomass composition revealed an increase in cellular lipid content at elevated levels of protein secretion and we observed altered metabolic fluxes in the pentose phosphate pathway, the TCA cycle, and around the pyruvate node including mitochondrial NADPH supply. Supplementing acetate to glucose or glycerol minimal media was found to improve protein secretion, accompanied by an increased cellular lipid content and carbon flux through the TCA cycle as well as increased mitochondrial NADPH production. Thus, systematic metabolic analyses can assist in identifying factors limiting protein secretion and in deriving strategies to overcome these limitations.

  1. Bioremediation in marine ecosystems: a computational study combining ecological modeling and flux balance analysis.

    PubMed

    Taffi, Marianna; Paoletti, Nicola; Angione, Claudio; Pucciarelli, Sandra; Marini, Mauro; Liò, Pietro

    2014-01-01

    The pressure to search effective bioremediation methodologies for contaminated ecosystems has led to the large-scale identification of microbial species and metabolic degradation pathways. However, minor attention has been paid to the study of bioremediation in marine food webs and to the definition of integrated strategies for reducing bioaccumulation in species. We propose a novel computational framework for analysing the multiscale effects of bioremediation at the ecosystem level, based on coupling food web bioaccumulation models and metabolic models of degrading bacteria. The combination of techniques from synthetic biology and ecological network analysis allows the specification of arbitrary scenarios of contaminant removal and the evaluation of strategies based on natural or synthetic microbial strains. In this study, we derive a bioaccumulation model of polychlorinated biphenyls (PCBs) in the Adriatic food web, and we extend a metabolic reconstruction of Pseudomonas putida KT2440 (iJN746) with the aerobic pathway of PCBs degradation. We assess the effectiveness of different bioremediation scenarios in reducing PCBs concentration in species and we study indices of species centrality to measure their importance in the contaminant diffusion via feeding links. The analysis of the Adriatic sea case study suggests that our framework could represent a practical tool in the design of effective remediation strategies, providing at the same time insights into the ecological role of microbial communities within food webs.

  2. Can the whole be less than the sum of its parts? Pathway analysis in genome-scale metabolic networks using elementary flux patterns

    PubMed Central

    Kaleta, Christoph; de Figueiredo, Luís Filipe; Schuster, Stefan

    2009-01-01

    Elementary modes represent a valuable concept in the analysis of metabolic reaction networks. However, they can only be computed in medium-size systems, preventing application to genome-scale metabolic models. In consequence, the analysis is usually constrained to a specific part of the known metabolism, and the remaining system is modeled using abstractions like exchange fluxes and external species. As we show by the analysis of a model of the central metabolism of Escherichia coli that has been previously analyzed using elementary modes, the choice of these abstractions heavily impacts the pathways that are detected, and the results are biased by the knowledge of the metabolic capabilities of the network by the user. In order to circumvent these problems, we introduce the concept of elementary flux patterns, which explicitly takes into account possible steady-state fluxes through a genome-scale metabolic network when analyzing pathways through a subsystem. By being similar to elementary mode analysis, our concept now allows for the application of many elementary-mode-based tools to genome-scale metabolic networks. We present an algorithm to compute elementary flux patterns and analyze a model of the tricarboxylic acid cycle and adjacent reactions in E. coli. Thus, we detect several pathways that can be used as alternative routes to some central metabolic pathways. Finally, we give an outlook on further applications like the computation of minimal media, the development of knockout strategies, and the analysis of combined genome-scale networks. PMID:19541909

  3. Development and analysis of prognostic equations for mesoscale kinetic energy and mesoscale (subgrid scale) fluxes for large-scale atmospheric models

    NASA Technical Reports Server (NTRS)

    Avissar, Roni; Chen, Fei

    1993-01-01

    Generated by landscape discontinuities (e.g., sea breezes) mesoscale circulation processes are not represented in large-scale atmospheric models (e.g., general circulation models), which have an inappropiate grid-scale resolution. With the assumption that atmospheric variables can be separated into large scale, mesoscale, and turbulent scale, a set of prognostic equations applicable in large-scale atmospheric models for momentum, temperature, moisture, and any other gaseous or aerosol material, which includes both mesoscale and turbulent fluxes is developed. Prognostic equations are also developed for these mesoscale fluxes, which indicate a closure problem and, therefore, require a parameterization. For this purpose, the mean mesoscale kinetic energy (MKE) per unit of mass is used, defined as E-tilde = 0.5 (the mean value of u'(sub i exp 2), where u'(sub i) represents the three Cartesian components of a mesoscale circulation (the angle bracket symbol is the grid-scale, horizontal averaging operator in the large-scale model, and a tilde indicates a corresponding large-scale mean value). A prognostic equation is developed for E-tilde, and an analysis of the different terms of this equation indicates that the mesoscale vertical heat flux, the mesoscale pressure correlation, and the interaction between turbulence and mesoscale perturbations are the major terms that affect the time tendency of E-tilde. A-state-of-the-art mesoscale atmospheric model is used to investigate the relationship between MKE, landscape discontinuities (as characterized by the spatial distribution of heat fluxes at the earth's surface), and mesoscale sensible and latent heat fluxes in the atmosphere. MKE is compared with turbulence kinetic energy to illustrate the importance of mesoscale processes as compared to turbulent processes. This analysis emphasizes the potential use of MKE to bridge between landscape discontinuities and mesoscale fluxes and, therefore, to parameterize mesoscale fluxes

  4. Orientation-free and differentially pumped addition of a low-flux reactive gas beam to a surface analysis system

    NASA Astrophysics Data System (ADS)

    Harthcock, Colin; Jahanbekam, Abdolreza; Eskelsen, Jeremy R.; Lee, David Y.

    2016-11-01

    We describe an example of a piecewise gas chamber that can be customized to incorporate a low flux of gas-phase radicals with an existing surface analysis chamber for in situ and stepwise gas-surface interaction experiments without any constraint in orientation. The piecewise nature of this gas chamber provides complete angular freedom and easy alignment and does not require any modification of the existing surface analysis chamber. In addition, the entire gas-surface system is readily differentially pumped with the surface chamber kept under ultra-high-vacuum during the gas-surface measurements. This new design also allows not only straightforward reconstruction to accommodate the orientation of different surface chambers but also for the addition of other desired features, such as an additional pump to the current configuration. Stepwise interaction between atomic oxygen and a highly ordered pyrolytic graphite surface was chosen to test the effectiveness of this design, and the site-dependent O-atom chemisorption and clustering on the graphite surface were resolved by a scanning tunneling microscope in the nm-scale. X-ray photoelectron spectroscopy was used to further confirm the identity of the chemisorbed species on the graphite surface as oxygen.

  5. Forward flux sampling-type schemes for simulating rare events: efficiency analysis.

    PubMed

    Allen, Rosalind J; Frenkel, Daan; ten Wolde, Pieter Rein

    2006-05-21

    We analyze the efficiency of several simulation methods which we have recently proposed for calculating rate constants for rare events in stochastic dynamical systems in or out of equilibrium. We derive analytical expressions for the computational cost of using these methods and for the statistical error in the final estimate of the rate constant for a given computational cost. These expressions can be used to determine which method to use for a given problem, to optimize the choice of parameters, and to evaluate the significance of the results obtained. We apply the expressions to the two-dimensional nonequilibrium rare event problem proposed by Maier and Stein [Phys. Rev. E 48, 931 (1993)]. For this problem, our analysis gives accurate quantitative predictions for the computational efficiency of the three methods.

  6. Analysis of shape of porous cooled medium for an imposed surface heat flux and temperature

    NASA Technical Reports Server (NTRS)

    Siegel, R.

    1973-01-01

    The surface of a porous cooled medium is to be maintained at a specified design temperature while being subjected to uniform heating by an external source. An analytical method is given for determining the shape of the medium surface that will satisfy these boundary conditions. The analysis accounts for temperature dependent variations of fluid density and viscosity and for temperature dependent matrix thermal conductivity. The energy equation is combined with Darcy's law in such a way that a potential can be defined that satisfies Laplace's equation. All of the heat-transfer and flow quantities are expressed in terms of this potential. The determination of the shape of the porous cooled region is thereby reduced to a free-boundary problem such as in inviscid free jet theory. Two illustrative examples are carried out: a porous leading edge with coolant supplied through a slot and a porous cooled duct with a rectangular outer boundary.

  7. Metabolic Flux Analysis of Lipid Biosynthesis in the Yeast Yarrowia lipolytica Using 13C-Labled Glucose and Gas Chromatography-Mass Spectrometry.

    PubMed

    Zhang, Huaiyuan; Wu, Chao; Wu, Qingyu; Dai, Junbiao; Song, Yuanda

    2016-01-01

    The oleaginous yeast Yarrowia lipolytica has considerable potential for producing single cell oil, which can be converted to biodiesel, a sustainable alternative to fossil fuels. However, extensive fundamental and engineering efforts must be carried out before commercialized production become cost-effective. Therefore, in this study, metabolic flux analysis of Y. lipolytica was performed using 13C-labeled glucose as a sole carbon source in nitrogen sufficient and insufficient media. The nitrogen limited medium inhibited cell growth while promoting lipid accumulation (from 8.7% of their biomass to 14.3%). Metabolic flux analysis showed that flux through the pentose phosphate pathway was not significantly regulated by nitrogen concentration, suggesting that NADPH generation is not the limiting factor for lipid accumulation in Y. lipolytica. Furthermore, metabolic flux through malic enzyme was undetectable, confirming its non-regulatory role in lipid accumulation in this yeast. Nitrogen limitation significantly increased flux through ATP:citrate lyase (ACL), implying that ACL plays a key role in providing acetyl-CoA for lipid accumulation in Y. lipolytica.

  8. Metabolic Flux Analysis of Lipid Biosynthesis in the Yeast Yarrowia lipolytica Using 13C-Labled Glucose and Gas Chromatography-Mass Spectrometry

    PubMed Central

    Zhang, Huaiyuan; Wu, Chao; Wu, Qingyu; Dai, Junbiao; Song, Yuanda

    2016-01-01

    The oleaginous yeast Yarrowia lipolytica has considerable potential for producing single cell oil, which can be converted to biodiesel, a sustainable alternative to fossil fuels. However, extensive fundamental and engineering efforts must be carried out before commercialized production become cost-effective. Therefore, in this study, metabolic flux analysis of Y. lipolytica was performed using 13C-labeled glucose as a sole carbon source in nitrogen sufficient and insufficient media. The nitrogen limited medium inhibited cell growth while promoting lipid accumulation (from 8.7% of their biomass to 14.3%). Metabolic flux analysis showed that flux through the pentose phosphate pathway was not significantly regulated by nitrogen concentration, suggesting that NADPH generation is not the limiting factor for lipid accumulation in Y. lipolytica. Furthermore, metabolic flux through malic enzyme was undetectable, confirming its non-regulatory role in lipid accumulation in this yeast. Nitrogen limitation significantly increased flux through ATP:citrate lyase (ACL), implying that ACL plays a key role in providing acetyl-CoA for lipid accumulation in Y. lipolytica. PMID:27454589

  9. Analysis on proton fluxes during several solar events with the PAMELA experiment

    NASA Astrophysics Data System (ADS)

    Martucci, Matteo

    2015-04-01

    The charged particle production during solar events have been widely modelized in the past decades. The satellite-borne PAMELA experiment has been continuously collecting data since 2006. This apparatus is designed to study charged particles in the cosmic radiation. The combination of permanent magnet, silicon strip spectrometer and silicon-tungsten imaging calorimeter, with the redundancy of instrumentation allows very precise studies on the physics of cosmic rays in a wide energy range and with high statistics. This makes PAMELA a very suitable instrument for Solar Energetic Particle (SEP) observations. Not only does it span the energy range between the ground-based neutron monitor data and the observations of SEPs from space, but also PAMELA carries out the first direct measurements of the composition for the highest energy SEP events. PAMELA has registered many SEP events in solar cycle 24, offering unique opportunities to address the question of high-energy SEP origin. A preliminar analysis on proton spectra during several events of the 24th solar cycle is presented.

  10. Uncertainty Analysis on an Operational Simplified Surface Energy Balance algorithm for Estimation of Evapotranspiration at Multiple Flux Tower Sites

    NASA Astrophysics Data System (ADS)

    Chen, M.; Senay, G. B.; Verdin, J. P.; Rowland, J.

    2014-12-01

    Current regional to global and daily to annual Evapotranspiration ( ET) estimation mainly relies on surface energy balance (SEB) ET models or statistical empirical methods driven by remote sensing data and various meteorology databases. However, these ET models face challenging issues—large uncertainties from inevitable input errors, poorly defined parameters, and inadequate model structures. The eddy covariance measurements on water, energy, and carbon fluxes at globally available FLUXNET tower sites provide a feasible opportunity to assess the ET modelling uncertainties. In this study, we focused on uncertainty analysis on an operational simplified surface energy balance (SSEBop) algorithm for ET estimation at multiple Ameriflux tower sites with diverse land cover characteristics and climatic conditions. The input land surface temperature (LST) data of the algorithm were adopted from the 8-day composite1-km Moderate Resolution Imaging Spectroradiometer (MODIS) land surface temperature. The other input data were taken from the Ameriflux database. Results of statistical analysis indicated that uncertainties or random errors from input variables and parameters of SSEBop led to daily and seasonal ET estimates with relative errors around 20% across multiple flux tower sites distributed across different biomes. This uncertainty of SSEBop lies in the error range of 20-30% of similar SEB-based ET algorithms, such as, Surface Energy Balance System and Surface Energy Balance Algorithm for Land. The R2 between daily and seasonal ET estimates by SSEBop and ET eddy covariance measurements at multiple Ameriflux tower sites exceed 0.7, and even up to 0.9 for croplands, grasslands, and forests, suggesting systematic error or bias of the SSEBop is acceptable. In summary, the uncertainty assessment verifies that the SSEBop is a reliable method for wide-area ET calculation and especially useful for detecting drought years and relative drought severity for agricultural production

  11. High-frequency analysis of the complex linkage between soil CO(2) fluxes, photosynthesis and environmental variables.

    PubMed

    Martin, Jonathan G; Phillips, Claire L; Schmidt, Andres; Irvine, James; Law, Beverly E

    2012-01-01

    High-frequency soil CO(2) flux data are valuable for providing new insights into the processes of soil CO(2) production. A record of hourly soil CO(2) fluxes from a semi-arid ponderosa pine stand was spatially and temporally deconstructed in attempts to determine if variation could be explained by logical drivers using (i) CO(2) production depths, (ii) relationships and lags between fluxes and soil temperatures, or (iii) the role of canopy assimilation in soil CO(2) flux variation. Relationships between temperature and soil fluxes were difficult to establish at the hourly scale because diel cycles of soil fluxes varied seasonally, with the peak of flux rates occurring later in the day as soil water content decreased. Using a simple heat transport/gas diffusion model to estimate the time and depth of CO(2) flux production, we determined that the variation in diel soil CO(2) flux patterns could not be explained by changes in diffusion rates or production from deeper soil profiles. We tested for the effect of gross ecosystem productivity (GEP) by minimizing soil flux covariance with temperature and moisture using only data from discrete bins of environmental conditions (±1 °C soil temperature at multiple depths, precipitation-free periods and stable soil moisture). Gross ecosystem productivity was identified as a possible driver of variability at the hourly scale during the growing season, with multiple lags between ~5, 15 and 23 days. Additionally, the chamber-specific lags between GEP and soil CO(2) fluxes appeared to relate to combined path length for carbon flow (top of tree to chamber center). In this sparse and heterogeneous forested system, the potential link between CO(2) assimilation and soil CO(2) flux may be quite variable both temporally and spatially. For model applications, it is important to note that soil CO(2) fluxes are influenced by many biophysical factors, which may confound or obscure relationships with logical environmental drivers and act at

  12. Statistical analysis of error rate of large-scale single flux quantum logic circuit by considering fluctuation of timing parameters

    NASA Astrophysics Data System (ADS)

    Yamanashi, Yuki; Masubuchi, Kota; Yoshikawa, Nobuyuki

    2016-11-01

    The relationship between the timing margin and the error rate of the large-scale single flux quantum logic circuits is quantitatively investigated to establish a timing design guideline. We observed that the fluctuation in the set-up/hold time of single flux quantum logic gates caused by thermal noises is the most probable origin of the logical error of the large-scale single flux quantum circuit. The appropriate timing margin for stable operation of the large-scale logic circuit is discussed by taking the fluctuation of setup/hold time and the timing jitter in the single flux quantum circuits. As a case study, the dependence of the error rate of the 1-million-bit single flux quantum shift register on the timing margin is statistically analyzed. The result indicates that adjustment of timing margin and the bias voltage is important for stable operation of a large-scale SFQ logic circuit.

  13. Tropospheric ozone reduces carbon assimilation in trees: estimates from analysis of continuous flux measurements.

    PubMed

    Fares, Silvano; Vargas, Rodrigo; Detto, Matteo; Goldstein, Allen H; Karlik, John; Paoletti, Elena; Vitale, Marcello

    2013-08-01

    High ground-level ozone concentrations are typical of Mediterranean climates. Plant exposure to this oxidant is known to reduce carbon assimilation. Ozone damage has been traditionally measured through manipulative experiments that do not consider long-term exposure and propagate large uncertainty by up-scaling leaf-level observations to ecosystem-level interpretations. We analyzed long-term continuous measurements (>9 site-years at 30 min resolution) of environmental and eco-physiological parameters at three Mediterranean ecosystems: (i) forest site dominated by Pinus ponderosa in the Sierra Mountains in California, USA; (ii) forest site composed of a mixture of Quercus spp. and P. pinea in the Tyrrhenian sea coast near Rome, Italy; and (iii) orchard site of Citrus sinensis cultivated in the California Central Valley, USA. We hypothesized that higher levels of ozone concentration in the atmosphere result in a decrease in carbon assimilation by trees under field conditions. This hypothesis was tested using time series analysis such as wavelet coherence and spectral Granger causality, and complemented with multivariate linear and nonlinear statistical analyses. We found that reduction in carbon assimilation was more related to stomatal ozone deposition than to ozone concentration. The negative effects of ozone occurred within a day of exposure/uptake. Decoupling between carbon assimilation and stomatal aperture increased with the amount of ozone pollution. Up to 12-19% of the carbon assimilation reduction in P. ponderosa and in the Citrus plantation was explained by higher stomatal ozone deposition. In contrast, the Italian site did not show reductions in gross primary productivity either by ozone concentration or stomatal ozone deposition, mainly due to the lower ozone concentrations in the periurban site over the shorter period of investigation. These results highlight the importance of plant adaptation/sensitivity under field conditions, and the importance of

  14. Transient-state analysis of metabolic fluxes in crabtree-positive and crabtree-negative yeasts.

    PubMed

    Van Urk, H; Voll, W S; Scheffers, W A; Van Dijken, J P

    1990-01-01

    In bakers' yeast, an immediate alcoholic fermentation begins when a glucose pulse is added to glucose-limited, aerobically grown cells. The mechanism of this short-term Crabtree effect was investigated via a comparative enzymic analysis of eight yeast species. It was established that the fermentation rate of the organisms upon transition from glucose limitation to glucose excess is positively correlated with the level of pyruvate decarboxylase (EC 4.1.1.1). In the Crabtree-negative yeasts, the pyruvate decarboxylase activity was low and did not increase when excess glucose was added. In contrast, in the Crabtree-positive yeasts, the activity of this enzyme was on the average sixfold higher and increased after exposure to glucose excess. In Crabtree-negative species, relatively high activities of acetaldehyde dehydrogenases (EC 1.2.1.4 and EC 1.2.1.5) and acetyl coenzyme A synthetase (EC 6.2.1.1), in addition to low pyruvate decarboxylase activities, were present. Thus, in these yeasts, acetaldehyde can be effectively oxidized via a bypass that circumvents the reduction of acetaldehyde to ethanol. Growth rates of most Crabtree-positive yeasts did not increase upon transition from glucose limitation to glucose excess. In contrast, the Crabtree-negative yeasts exhibited enhanced rates of biomass production which in most cases could be ascribed to the intracellular accumulation of reserve carbohydrates. Generally, the glucose consumption rate after a glucose pulse was higher in the Crabtree-positive yeasts than in the Crabtree-negative yeasts. However, the respiratory capacities of steady-state cultures of Crabtree-positive yeasts were not significantly different from those of Crabtree-negative yeasts. Thus, a limited respiratory capacity is not the primary cause of the Crabtree effect in yeasts. Instead, the difference between Crabtree-positive and Crabtree-negative yeasts is attributed to differences in the kinetics of glucose uptake, synthesis of reserve

  15. A Method of Accounting for Enzyme Costs in Flux Balance Analysis Reveals Alternative Pathways and Metabolite Stores in an Illuminated Arabidopsis Leaf.

    PubMed

    Cheung, C Y Maurice; Ratcliffe, R George; Sweetlove, Lee J

    2015-11-01

    Flux balance analysis of plant metabolism is an established method for predicting metabolic flux phenotypes and for exploring the way in which the plant metabolic network delivers specific outcomes in different cell types, tissues, and temporal phases. A recurring theme is the need to explore the flexibility of the network in meeting its objectives and, in particular, to establish the extent to which alternative pathways can contribute to achieving specific outcomes. Unfortunately, predictions from conventional flux balance analysis minimize the simultaneous operation of alternative pathways, but by introducing flux-weighting factors to allow for the variable intrinsic cost of supporting each flux, it is possible to activate different pathways in individual simulations and, thus, to explore alternative pathways by averaging thousands of simulations. This new method has been applied to a diel genome-scale model of Arabidopsis (Arabidopsis thaliana) leaf metabolism to explore the flexibility of the network in meeting the metabolic requirements of the leaf in the light. This identified alternative flux modes in the Calvin-Benson cycle revealed the potential for alternative transitory carbon stores in leaves and led to predictions about the light-dependent contribution of alternative electron flow pathways and futile cycles in energy rebalancing. Notable features of the analysis include the light-dependent tradeoff between the use of carbohydrates and four-carbon organic acids as transitory storage forms and the way in which multiple pathways for the consumption of ATP and NADPH can contribute to the balancing of the requirements of photosynthetic metabolism with the energy available from photon capture.

  16. A Method of Accounting for Enzyme Costs in Flux Balance Analysis Reveals Alternative Pathways and Metabolite Stores in an Illuminated Arabidopsis Leaf1[OPEN

    PubMed Central

    Cheung, C.Y. Maurice; Ratcliffe, R. George; Sweetlove, Lee J.

    2015-01-01

    Flux balance analysis of plant metabolism is an established method for predicting metabolic flux phenotypes and for exploring the way in which the plant metabolic network delivers specific outcomes in different cell types, tissues, and temporal phases. A recurring theme is the need to explore the flexibility of the network in meeting its objectives and, in particular, to establish the extent to which alternative pathways can contribute to achieving specific outcomes. Unfortunately, predictions from conventional flux balance analysis minimize the simultaneous operation of alternative pathways, but by introducing flux-weighting factors to allow for the variable intrinsic cost of supporting each flux, it is possible to activate different pathways in individual simulations and, thus, to explore alternative pathways by averaging thousands of simulations. This new method has been applied to a diel genome-scale model of Arabidopsis (Arabidopsis thaliana) leaf metabolism to explore the flexibility of the network in meeting the metabolic requirements of the leaf in the light. This identified alternative flux modes in the Calvin-Benson cycle revealed the potential for alternative transitory carbon stores in leaves and led to predictions about the light-dependent contribution of alternative electron flow pathways and futile cycles in energy rebalancing. Notable features of the analysis include the light-dependent tradeoff between the use of carbohydrates and four-carbon organic acids as transitory storage forms and the way in which multiple pathways for the consumption of ATP and NADPH can contribute to the balancing of the requirements of photosynthetic metabolism with the energy available from photon capture. PMID:26265776

  17. Ecosystem function and particle flux dynamics across the Mackenzie Shelf (Beaufort Sea, Arctic Ocean): an integrative analysis of spatial variability and biophysical forcings

    NASA Astrophysics Data System (ADS)

    Forest, A.; Babin, M.; Stemmann, L.; Picheral, M.; Sampei, M.; Fortier, L.; Gratton, Y.; Bélanger, S.; Devred, E.; Sahlin, J.; Doxaran, D.; Joux, F.; Ortega-Retuerta, E.; Jeffrey, W. H.; Martín, J.; Gasser, B.; Miquel, J. C.

    2012-08-01

    A better understanding of how environmental changes affect organic matter fluxes in Arctic marine ecosystems is sorely needed. Here, we combine mooring times-series, ship-based measurements and remote-sensing to assess the variability and forcing factors of vertical fluxes of particulate organic carbon (POC) across the Mackenzie Shelf in 2009. We developed a geospatial model of these fluxes to proceed to an integrative analysis of their biophysical determinants in summer. Flux data were obtained with sediment traps and via a regional empirical algorithm applied to particle size-distributions (17 classes from 0.08-4.2 mm) measured by an Underwater Vision Profiler 5. Redundancy analyses and forward selection of abiotic/biotic parameters, linear trends, and spatial structures (i.e. principal coordinates of neighbor matrices, PCNM), were conducted to partition the variation of POC flux size-classes. Flux variability was explained at 69.5 % by the addition of a linear temporal trend, 7 significant PCNM and 9 biophysical variables. The interaction of all these factors explained 27.8 % of the variability. The first PCNM canonical axis (44.4 % of spatial variance) reflected a shelf-basin gradient controlled by bottom depth and ice concentration (p < 0.01), but a complex assemblage of fine-to-broad scale patterns was also identified. Among biophysical parameters, bacterial production and northeasterly wind (upwelling-favorable) were the two strongest explanatory variables (r2 cum. = 0.37), suggesting that bacteria were associated with sinking material, which was itself partly linked to upwelling-induced productivity. The second most important spatial structure corresponded actually to the two areas where shelf break upwelling is known to occur under easterlies. Copepod biomass was negatively correlated (p < 0.05) with vertical POC fluxes, implying that metazoans played a significant role in the regulation of export fluxes. The low fractal dimension of settling particles (1

  18. Metabolic flux analysis model for optimizing xylose conversion into ethanol by the natural C5-fermenting yeast Candida shehatae.

    PubMed

    Bideaux, Carine; Montheard, Julie; Cameleyre, Xavier; Molina-Jouve, Carole; Alfenore, Sandrine

    2016-02-01

    A metabolic flux analysis (MFA) model was developed to optimize the xylose conversion into ethanol using Candida shehatae strain. This metabolic model was compartmented and constructed with xylose as carbon substrate integrating the enzymatic duality of the first step of xylose degradation via an algebraic coefficient. The model included the pentose phosphate pathway, glycolysis, synthesis of major metabolites like ethanol, acetic acid and glycerol, the tricarboxylic acid cycle as well as the respiratory chain, the cofactor balance, and the maintenance. The biomass composition and thus production were integrated considering the major biochemical synthesis reactions from monomers to each constitutive macromolecule (i.e., proteins, lipids, polysaccharides, nucleic acids). The construction of the model resulted into a 122-linear equation system to be resolved. A first experiment allowed was to verify the accuracy of the model by comparing calculated and experimental data. The metabolic model was utilized to determine the theoretical yield taking into account oxido-reductive balance and to optimize ethanol production. The maximal theoretical yield was calculated at 0.62 Cmolethanol/Cmolxylose for an oxygen requirement of 0.33 moloxygen/molxylose linked to the cofactors of the xylose reductase. Cultivations in chemostat mode allowed the fine tuning of both xylose and oxygen uptakes and showed that lower was the oxygen/xylose ratio, higher was the ethanol production yield. The best experimental ethanol production yield (0.51 Cmolethanol/Cmolxylose) was obtained for an oxygen supply of 0.47 moloxygen/molxylose.

  19. Semi-automated Curation of Metabolic Models via Flux Balance Analysis: A Case Study with Mycoplasma gallisepticum

    PubMed Central

    Szczepanek, Steven M.; Johnson, Erik L.; Tulman, Edan R.; Ching, Wei-Mei; Geary, Steven J.; Srivastava, Ranjan

    2013-01-01

    Primarily used for metabolic engineering and synthetic biology, genome-scale metabolic modeling shows tremendous potential as a tool for fundamental research and curation of metabolism. Through a novel integration of flux balance analysis and genetic algorithms, a strategy to curate metabolic networks and facilitate identification of metabolic pathways that may not be directly inferable solely from genome annotation was developed. Specifically, metabolites involved in unknown reactions can be determined, and potentially erroneous pathways can be identified. The procedure developed allows for new fundamental insight into metabolism, as well as acting as a semi-automated curation methodology for genome-scale metabolic modeling. To validate the methodology, a genome-scale metabolic model for the bacterium Mycoplasma gallisepticum was created. Several reactions not predicted by the genome annotation were postulated and validated via the literature. The model predicted an average growth rate of 0.358±0.12, closely matching the experimentally determined growth rate of M. gallisepticum of 0.244±0.03. This work presents a powerful algorithm for facilitating the identification and curation of previously known and new metabolic pathways, as well as presenting the first genome-scale reconstruction of M. gallisepticum. PMID:24039564

  20. Synchrotron emission from the blazar PG 1553+113. An analysis of its flux and polarization variability

    NASA Astrophysics Data System (ADS)

    Raiteri, C. M.; Nicastro, F.; Stamerra, A.; Villata, M.; Larionov, V. M.; Blinov, D.; Acosta-Pulido, J. A.; Arévalo, M. J.; Arkharov, A. A.; Bachev, R.; Borman, G. A.; Carnerero, M. I.; Carosati, D.; Cecconi, M.; Chen, W.-P.; Damljanovic, G.; Di Paola, A.; Ehgamberdiev, Sh. A.; Frasca, A.; Giroletti, M.; González-Morales, P. A.; Griñon-Marín, A. B.; Grishina, T. S.; Huang, P.-C.; Ibryamov, S.; Klimanov, S. A.; Kopatskaya, E. N.; Kurtanidze, O. M.; Kurtanidze, S. O.; Lähteenmäki, A.; Larionova, E. G.; Larionova, L. V.; Lázaro, C.; Leto, G.; Liodakis, I.; Martínez-Lombilla, C.; Mihov, B.; Mirzaqulov, D. O.; Mokrushina, A. A.; Moody, J. W.; Morozova, D. A.; Nazarov, S. V.; Nikolashvili, M. G.; Ohlert, J. M.; Panopoulou, G. V.; Pastor Yabar, A.; Pinna, F.; Protasio, C.; Rizzi, N.; Sadun, A. C.; Savchenko, S. S.; Semkov, E.; Sigua, L. A.; Slavcheva-Mihova, L.; Strigachev, A.; Tornikoski, M.; Troitskaya, Yu. V.; Troitsky, I. S.; Vasilyev, A. A.; Vera, R. J. C.; Vince, O.; Zanmar Sanchez, R.

    2017-04-01

    In 2015 July 29-September 1, the satellite XMM-Newton pointed at the BL Lac object PG 1553+133 six times, collecting data for 218 h. During one of these epochs, simultaneous observations by the Swift satellite were requested to compare the results of the X-ray and optical-UV instruments. Optical, near-infrared and radio monitoring was carried out by the Whole Earth Blazar Telescope (WEBT) collaboration for the whole observing season. We here present the results of the analysis of all these data, together with an investigation of the source photometric and polarimetric behaviour over the last 3 yr. The 2015 EPIC spectra show slight curvature and the corresponding light curves display fast X-ray variability with a time-scale of the order of 1 h. In contrast to previous results, during the brightest X-ray states detected in 2015 the simple log-parabolic model that best fits the XMM-Newton data also reproduces reasonably well the whole synchrotron bump, suggesting a peak in the near-UV band. We found evidence of a wide rotation of the polarization angle in 2014, when the polarization degree was variable, but the flux remained almost constant. This is difficult to interpret with deterministic jet emission models, while it can be easily reproduced by assuming some turbulence of the magnetic field.

  1. MID Max: LC-MS/MS Method for Measuring the Precursor and Product Mass Isotopomer Distributions of Metabolic Intermediates and Cofactors for Metabolic Flux Analysis Applications.

    PubMed

    McCloskey, Douglas; Young, Jamey D; Xu, Sibei; Palsson, Bernhard O; Feist, Adam M

    2016-01-19

    The analytical challenges to acquire accurate isotopic data of intracellular metabolic intermediates for stationary, nonstationary, and dynamic metabolic flux analysis (MFA) are numerous. This work presents MID Max, a novel LC-MS/MS workflow, acquisition, and isotopomer deconvolution method for MFA that takes advantage of additional scan types that maximizes the number of mass isotopomer distributions (MIDs) that can be acquired in a given experiment. The analytical method was found to measure the MIDs of 97 metabolites, corresponding to 74 unique metabolite-fragment pairs (32 precursor spectra and 42 product spectra) with accuracy and precision. The compounds measured included metabolic intermediates in central carbohydrate metabolism and cofactors of peripheral metabolism (e.g., ATP). Using only a subset of the acquired MIDs, the method was found to improve the precision of flux estimations and number of resolved exchange fluxes for wild-type E. coli compared to traditional methods and previously published data sets.

  2. Dynamic flux balance analysis of the metabolism of Saccharomyces cerevisiae during the shift from fully respirative or respirofermentative metabolic states to anaerobiosis.

    PubMed

    Jouhten, Paula; Wiebe, Marilyn; Penttilä, Merja

    2012-09-01

    Dynamic flux balance analysis was utilized to simulate the metabolic behaviour of initially fully respirative and respirofermentative steady-state cultures of Saccharomyces cerevisiae during sudden oxygen depletion. The hybrid model for the dynamic flux balance analysis included a stoichiometric genome-scale metabolic model as a static part and dynamic equations for the uptake of glucose and the cessation of respirative metabolism. The yeast consensus genome-scale metabolic model [Herrgård MJ et al. (2008) Nat Biotechnol 26, 1155-1160; Dobson PD et al. (2010) BMC Syst Biol 4, 145] was refined with respect to oxygen-dependent energy metabolism and further modified to reflect S. cerevisiae anabolism in the absence of oxygen. Dynamic flux balance analysis captured well the essential features of the dynamic metabolic behaviour of S. cerevisiae during adaptation to anaerobiosis. Modelling and simulation enabled the identification of short time-scale flux distribution dynamics under the transition to anaerobic metabolism, during which the specific growth rate was reduced, as well as longer time-scale process dynamics when the specific growth rate recovered. Expression of the metabolic genes was set into the context of the identified dynamics. Metabolic gene expression responses associated with the specific growth rate and with the cessation of respirative metabolism were distinguished.

  3. Metabolic Flux Analysis of the Synechocystis sp. PCC 6803 ΔnrtABCD Mutant Reveals a Mechanism for Metabolic Adaptation to Nitrogen-Limited Conditions.

    PubMed

    Nakajima, Tsubasa; Yoshikawa, Katsunori; Toya, Yoshihiro; Matsuda, Fumio; Shimizu, Hiroshi

    2017-03-01

    Metabolic flux redirection during nitrogen-limited growth was investigated in the Synechocystis sp. PCC 6803 glucose-tolerant (GT) strain under photoautotrophic conditions by isotopically non-stationary metabolic flux analysis (INST-MFA). A ΔnrtABCD mutant of Synechocystis sp. PCC 6803 was constructed to reproduce phenotypes arising during nitrogen starvation. The ΔnrtABCD mutant and the wild-type GT strain were cultured under photoautotrophic conditions by a photobioreactor. Intracellular metabolites were labeled over a time course using NaH13CO3 as a carbon source. Based on these data, the metabolic flux distributions in the wild-type and ΔnrtABCD cells were estimated by INST-MFA. The wild-type GT and ΔnrtABCD strains displayed similar distribution patterns, although the absolute levels of metabolic flux were lower in ΔnrtABCD. Furthermore, the relative flux levels for glycogen metabolism, anaplerotic reactions and the oxidative pentose phosphate pathway were increased in ΔnrtABCD. This was probably due to the increased expression of enzyme genes that respond to nitrogen depletion. Additionally, we found that the ratio of ATP/NADPH demand increased slightly in the ΔnrtABCD mutant. These results indicated that futile ATP consumption increases under nitrogen-limited conditions because the Calvin-Benson cycle and the oxidative pentose phosphate pathway form a metabolic futile cycle that consumes ATP without CO2 fixation and NADPH regeneration.

  4. Integration of a constraint-based metabolic model of Brassica napus developing seeds with 13C-metabolic flux analysis

    SciTech Connect

    Hay, Jordan O.; Shi, Hai; Heinzel, Nicolas; Hebbelmann, Inga; Rolletschek, Hardy; Schwender, Jorg

    2014-12-19

    The use of large-scale or genome-scale metabolic reconstructions for modeling and simulation of plant metabolism and integration of those models with large-scale omics and experimental flux data is becoming increasingly important in plant metabolic research. Here we report an updated version of bna572, a bottom-up reconstruction of oilseed rape (Brassica napus L.; Brassicaceae) developing seeds with emphasis on representation of biomass-component biosynthesis. New features include additional seed-relevant pathways for isoprenoid, sterol, phenylpropanoid, flavonoid, and choline biosynthesis. Being now based on standardized data formats and procedures for model reconstruction, bna572+ is available as a COBRA-compliant Systems Biology Markup Language (SBML) model and conforms to the Minimum Information Requested in the Annotation of Biochemical Models (MIRIAM) standards for annotation of external data resources. Bna572+ contains 966 genes, 671 reactions, and 666 metabolites distributed among 11 subcellular compartments. It is referenced to the Arabidopsis thaliana genome, with gene-protein-reaction (GPR) associations resolving subcellular localization. Detailed mass and charge balancing and confidence scoring were applied to all reactions. Using B. napus seed specific transcriptome data, expression was verified for 78% of bna572+ genes and 97% of reactions. Alongside bna572+ we also present a revised carbon centric model for 13C-Metabolic Flux Analysis (13C-MFA) with all its reactions being referenced to bna572+ based on linear projections. By integration of flux ratio constraints obtained from 13C-MFA and by elimination of infinite flux bounds around thermodynamically infeasible loops based on COBRA loopless methods, we demonstrate improvements in predictive power of Flux Variability Analysis (FVA). In conclusion, using this combined approach we characterize the difference in metabolic flux of developing seeds of two B. napus

  5. Fuel-Coolant-Interaction modeling and analysis work for the High Flux Isotope Reactor Safety Analysis Report

    SciTech Connect

    Taleyarkhan, R.P.; Georgevich, V.; Nestor, C.W.; Chang, S.J.; Freels, J.; Gat, U.; Lepard, B.L.; Gwaltney, R.C.; Luttrell, C.; Kirkpatrick, J.

    1993-07-01

    A brief historical background and a description of short- and long-term task plan development for effective closure of this important safety issue for the HFIR are given. Short-term aspects deal with Fuel-Coolant-Interaction (FCI) issues experimentation, modeling, and analysis for the flow-blockage-induced steam explosion events in direct support of the SAR. Long-term aspects deal with addressing FCI issues resulting from other accidents in conjunction with issues dealing with aluminum ignition, which can result in an order of magnitude increase in overall energetics. Problem formulation, modeling, and computer code simulation for the various phases of steam explosions are described. The evaluation of core melt initiation propagation, and melt superheat are described. Core melt initiation and propagation have been studied using simple conservative models as well as from modeling and analysis using RELAP5. Core debris coolability, heatup, and melting/freezing aspects have been studied by use of the two-dimensional melting/freezing analysis code 2DKO, which was also benchmarked with MELCOR code predictions. Descriptions are provided for the HM, BH, FCIMOD, and CTH computer codes that have been implemented for studying steam explosion energetics from the standpoint of evaluating bounding loads by thermodynamic models or best-estimate loads from one- and two-dimensional simulations of steam explosion energetics. Vessel failure modeling and analysis was conducted using the principles of probabilistic fracture mechanics in conjunction with ADINA code calculations. Top head bolts failure modeling has also been conducted where the failure criterion was based upon stresses in the bolts exceeding the material yield stress for a given time duration. Missile transport modeling and analysis was conducted by setting up a one-dimensional mathematical model that accounts for viscous dissipation, virtual mass effects, and material inertia.

  6. Metabolic flux responses to genetic modification for shikimic acid production by Bacillus subtilis strains

    PubMed Central

    2014-01-01

    Background Shikimic acid (SA) is a key chiral starting molecule for the synthesis of the neuramidase inhibitor GS4104 against viral influenza. Microbial production of SA has been extensively investigated in Escherichia coli, and to a less extent in Bacillus subtilis. However, metabolic flux of the high SA-producing strains has not been explored. In this study, we constructed with genetic manipulation and further determined metabolic flux with 13C-labeling test of high SA-producing B. subtilis strains. Results B. subtilis 1A474 had a mutation in SA kinase gene (aroI) and accumulated 1.5 g/L of SA. Overexpression of plasmid-encoded aroA, aroB, aroC or aroD in B. subtilis revealed that aroD had the most significantly positive effects on SA production. Simultaneous overexpression of genes for 3-deoxy-D-arabinoheptulosonate-7-phosphate synthase (aroA) and SA dehydrogenase (aroD) in B. subtilis BSSA/pSAAroA/pDGSAAroD resulted in SA production of 3.2 g/L. 13C-Metabolic flux assay (MFA) on the two strains BSSA/pHCMC04/pDG148-stu and BSSA/pSAAroA/pDGSAAroD indicated the carbon flux from glucose to SA increased to 4.6% in BSSA/pSAAroA/pDGSAAroD from 1.9% in strain BSSA/pHCMC04/pDG148-stu. The carbon flux through tricarboxylic acid cycle significantly reduced, while responses of the pentose phosphate pathway and the glycolysis to high SA production were rather weak, in the strain BSSA/pSAAroA/pDGSAAroD. Based on the results from MFA, two potential targets for further optimization of SA production were identified. Experiments on genetic deletion of phosphoenoylpyruvate kinase gene confirmed its positive influence on SA production, while the overexpression of the transketolase gene did not lead to increase in SA production. Conclusion Of the genes involved in shikimate pathway in B. subtilis, aroD exerted most significant influence on SA accumulation. Overexpression of plasmid-encoded aroA and aroD doubled SA production than its parent strain. MFA revealed metabolic flux

  7. Optimal fluxes and Reynolds stresses

    NASA Astrophysics Data System (ADS)

    Jiménez, Javier

    2016-12-01

    It is remarked that fluxes in conservation laws, such as the Reynolds stresses in the momentum equation of turbulent shear flows, or the spectral energy flux in isotropic turbulence, are only defined up to an arbitrary solenoidal field. While this is not usually significant for long-time averages, it becomes important when fluxes are modelled locally in large-eddy simulations, or in the analysis of intermittency and cascades. As an example, a numerical procedure is introduced to compute fluxes in scalar conservation equations in such a way that their total integrated magnitude is minimised. The result is an irrotational vector field that derives from a potential, thus minimising sterile flux `circuits'. The algorithm is generalised to tensor fluxes and applied to the transfer of momentum in a turbulent channel. The resulting instantaneous Reynolds stresses are compared with their traditional expressions, and found to be substantially different.

  8. Levels, fluxes and time trends of persistent organic pollutants in Lake Thun, Switzerland: combining trace analysis and multimedia modeling.

    PubMed

    Bogdal, Christian; Scheringer, Martin; Schmid, Peter; Bläuenstein, Markus; Kohler, Martin; Hungerbühler, Konrad

    2010-08-01

    Levels, mass fluxes, and time trends of polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) in Lake Thun, a peri-Alpine lake, are investigated. We present measurements of PBDEs and PCBs in air, lake water, lake sediment, and tributary water. These measurements are combined with a multimedia fate model, based on site-specific environmental parameters from the lake catchment. Measured loadings of PBDEs and PCBs in air and tributaries were used to drive the model. The model satisfactorily reproduces PBDE and PCB congener patterns in water and sediment, but it tends to yield concentrations in water below the measurements and concentrations in sediment exceeding the measurements. A sensitivity analysis reveals that partitioning of PBDEs and PCBs between the aqueous dissolved phase and suspended particulate matter in the water column strongly affects the model results, in particular the concentrations in water and sediment. For lower-brominated PBDEs, approximately 70% and 30% of input into the lake stems from atmospheric deposition and from tributaries, respectively. For heavier PBDEs and all PCBs, rivers appear to deliver the major load (64-92%). Waste water effluents are of minor importance. 50-90% of the total input is buried in the permanent sediment. Sediment burial makes PBDEs and PCBs less available for recycling in the environment, and reduces concentrations in the outflowing river. If use of deca-BDE increases in the future, levels in Lake Thun will follow the same trend. If the use and resulting environmental emissions decrease, concentrations in water will rapidly decline, according to our calculations, while sediment levels will decrease at a considerably slower rate.

  9. A Global Synthesis Inversion Analysis of Recent Variability in Natural CO2 Fluxes Using GOSAT and In Situ Observations

    NASA Astrophysics Data System (ADS)

    Wang, J. S.; Kawa, S. R.; Collatz, G. J.

    2013-12-01

    Around one-half of the CO2 emissions from fossil fuel combustion and deforestation accumulates in the atmosphere, where it contributes to global warming. The rest is taken up by vegetation and the ocean. The precise contribution of the two, and the location and year-to-year variability of the CO2 sinks are not well understood though. We use a batch Bayesian inversion approach to deduce the global spatiotemporal distributions of CO2 fluxes during 2009-2010. For prior constraints, we utilize fluxes from the CASA-GFED v.3 model of the terrestrial biosphere and biomass burning driven by satellite observations and interannually varying meteorology. We also use measurement-based ocean flux estimates from Takahashi et al. [2009], and fixed fossil CO2 emissions from the CDIAC dataset. Here, we present preliminary results from our inversions that incorporate column CO2 measurements from the GOSAT satellite, ground-based observations (individual flask and afternoon-average continuous observations), and aircraft observations to estimate fluxes in 108 regions over 8-day intervals. Relationships between fluxes and atmospheric concentrations are derived using the PCTM atmospheric transport model run at 2° x 2.5° (latitude/longitude) resolution driven by meteorology from the MERRA reanalysis. We obtain spatiotemporal distributions of fluxes resembling those from other inversions, including NOAA's CarbonTracker. We compare the a posteriori fluxes obtained with and without the addition of GOSAT observations to the in situ network, and discuss possible impacts of biases in the GOSAT data.

  10. A Global Synthesis Inversion Analysis of Recent Variability in Natural CO2 Fluxes Using Gosat and in Situ Observations

    NASA Astrophysics Data System (ADS)

    Wang, J. S.; Kawa, S. R.; Collatz, G. J.

    2014-12-01

    About one-half of the CO2 emissions from fossil fuel combustion and deforestation accumulates in the atmosphere, where it contributes to global warming. The rest is taken up by vegetation and the ocean. The precise contribution of the two, and the location and year-to-year variability of the CO2 sinks are, however, not well understood. We use a batch Bayesian inversion approach to deduce the global spatiotemporal distributions of CO2 fluxes during 2009-2010. For prior constraints, we utilize fluxes from the CASA-GFED model of the terrestrial biosphere and biomass burning driven by satellite observations and interannually varying meteorology. We also use measurement-based ocean flux estimates, and fixed fossil CO2 emissions. Here, we present results from our inversions that incorporate column CO2 measurements from the GOSAT satellite (ACOS retrieval, filtered and bias-corrected) and in situ observations (individual flask and afternoon-average continuous observations) to estimate fluxes in 108 regions over 8-day intervals. Relationships between fluxes and atmospheric concentrations are derived using the PCTM atmospheric transport model run at 2° x 2.5° (latitude/longitude) resolution driven by meteorology from the MERRA reanalysis. We evaluate the posterior CO2 concentrations using independent aircraft and other data sets. The optimized fluxes generally resemble those from other inversion systems using different techniques, for example indicating a net terrestrial biospheric CO2 sink, and a shift in the sink from tropics to northern high latitudes when going from an in-situ-only inversion to a GOSAT inversion. We show that in this inversion framework, GOSAT provides better flux estimates in most regions with its greater spatial coverage, but we also discuss impacts of possible remaining biases in the data.

  11. SAMOS Surface Fluxes

    NASA Astrophysics Data System (ADS)

    Smith, Shawn; Bourassa, Mark

    2014-05-01

    observations and the choices of constants that are used. Analysis of the preliminary SAMOS flux products will be presented, including spatial and temporal coverage for each derived parameter. The unique quality and sampling locations of research vessel observations and their independence from many models and products makes them ideal for validation studies. The strengths and limitations of research observations for flux validation studies will be discussed. The authors welcome a discussion with the flux community regarding expansion of the SAMOS program to include additional international vessels, thus facilitating and expansion of this research vessel-based flux product.

  12. An Inversion Analysis of Recent Variability in Natural CO2 Fluxes Using GOSAT and In Situ Observations

    NASA Technical Reports Server (NTRS)

    Wang, James S.; Kawa, S. Randolph; Collatz, G. James; Baker, David F.; Ott, Lesley

    2015-01-01

    About one-half of the global CO2 emissions from fossil fuel combustion and deforestation accumulates in the atmosphere, where it contributes to global warming. The rest is taken up by vegetation and the ocean. The precise contribution of the two sinks, and their location and year-to-year variability are, however, not well understood. We use two different approaches, batch Bayesian synthesis inversion and variational data assimilation, to deduce the global spatiotemporal distributions of CO2 fluxes during 2009-2010. One of our objectives is to assess different sources of uncertainties in inferred fluxes, including uncertainties in prior flux estimates and observations, and differences in inversion techniques. For prior constraints, we utilize fluxes and uncertainties from the CASA-GFED model of the terrestrial biosphere and biomass burning driven by satellite observations and interannually varying meteorology. We also use measurement-based ocean flux estimates and two sets of fixed fossil CO2 emissions. Here, our inversions incorporate column CO2 measurements from the GOSAT satellite (ACOS retrieval, filtered and bias-corrected) and in situ observations (individual flask and afternoon-average continuous observations) to estimate fluxes in 108 regions over 8-day intervals for the batch inversion and at 3 x 3.75 weekly for the variational system. Relationships between fluxes and atmospheric concentrations are derived consistently for the two inversion systems using the PCTM atmospheric transport model driven by meteorology from the MERRA reanalysis. We compare the posterior fluxes and uncertainties derived using different data sets and the two inversion approaches, and evaluate the posterior atmospheric concentrations against independent data including aircraft measurements. The optimized fluxes generally resemble those from other studies. For example, the results indicate that the terrestrial biosphere is a net CO2 sink, and a GOSAT-only inversion suggests a shift in

  13. An Inversion Analysis of Recent Variability in Natural CO2 Fluxes Using GOSAT and In Situ Observations

    NASA Astrophysics Data System (ADS)

    Wang, J. S.; Kawa, S. R.; Baker, D. F.; Collatz, G. J.; Ott, L. E.

    2015-12-01

    About one-half of the global CO2 emissions from fossil fuel combustion and deforestation accumulates in the atmosphere, where it contributes to global warming. The rest is taken up by vegetation and the ocean. The precise contribution of the two sinks, and their location and year-to-year variability are, however, not well understood. We use two different approaches, batch Bayesian synthesis inversion and variational data assimilation, to deduce the global spatiotemporal distributions of CO2 fluxes during 2009-2010. One of our objectives is to assess different sources of uncertainties in inferred fluxes, including uncertainties in prior flux estimates and observations, and differences in inversion techniques. For prior constraints, we utilize fluxes and uncertainties from the CASA-GFED model of the terrestrial biosphere and biomass burning driven by satellite observations and interannually varying meteorology. We also use measurement-based ocean flux estimates and two sets of fixed fossil CO2 emissions. Here, our inversions incorporate column CO2 measurements from the GOSAT satellite (ACOS retrieval, filtered and bias-corrected) and in situ observations (individual flask and afternoon-average continuous observations) to estimate fluxes in 108 regions over 8-day intervals for the batch inversion and at 3° x 3.75° weekly for the variational system. Relationships between fluxes and atmospheric concentrations are derived consistently for the two inversion systems using the PCTM atmospheric transport model driven by meteorology from the MERRA reanalysis. We compare the posterior fluxes and uncertainties derived using different data sets and the two inversion approaches, and evaluate the posterior atmospheric concentrations against independent data including aircraft measurements. The optimized fluxes generally resemble those from other studies. For example, the results indicate that the terrestrial biosphere is a net CO2 sink, and a GOSAT-only inversion suggests a

  14. Introduction of J-OFURO version 2 surface heat flux data set and its analysis over the North Pacific

    NASA Astrophysics Data System (ADS)

    Tomita, H.; Jubota, M.; Iwasaki, S.; Hihara, T.; Kawatsura, A.

    2007-05-01

    Japanese Ocean Flux Data Sets with Use of Remote Sensing Observations (J-OFURO) includes global ocean surface heat flux data derived from satellite data and are used in many studies related to air-sea interaction. Recently new surface heat flux data was constructed in J-OFURO as the version 2. In the version 2 many points are improved compared with the version 1. Since we used wind speed and specific humidity data derived from one DMSP/SSMI sensor in the version 1, we obtained two data at most one day. Therefore, there may be large sampling errors for the daily-mean value. In order to escape this problem, multi-satellite data (DMSP/SSMI F08- 15, Aqua/AMSR-E, TRMM/TMI, ERS/AMI and QuikScat/SeaWinds) are used in the version 2. As a result we could improve accuracy and temporal resolution from 3-days mean value in version 1 to daily-mean value in version 2. Also we used an Optimum Interpolation method to estimate specific humidity data instead of a simple mean method. We basically need sea surface temperature (SST), specific humidity and wind speed data for estimation of latent heat flux. In version 1 we used NCEP data (Reynolds and Smith, 1994) as SST data. However, the temporal resolution of the data is based on weekly and considerably low. Recently there are many kinds of global SST data because we can obtain SST data using a microwave radiometer sensor such as TRMM/MI and Aqua/AMSR-E. Therefore, we compared many SST products and determined to use Merged satellite and in situ data Global Daily (MGD) SST provided by Japan Meteorological Agency. A bulk algorithm used for estimation of turbulent heat flux is changed from Kondo (1975) to COASRE 3.0(Fairall et al., 2003). Shortwave and longwave radiation data are based on the ISCCP product and some modifications are carried out for longwave radiation. Finally surface latent and sensible flux data and shortwave and longwave radiation data are extended to1989- 2004. In this presentation we will introduce surface heat flux

  15. Metabolic flux ratio analysis and multi-objective optimization revealed a globally conserved and coordinated metabolic response of E. coli to paraquat-induced oxidative stress.

    PubMed

    Shen, Tie; Rui, Bin; Zhou, Hong; Zhang, Ximing; Yi, Yin; Wen, Han; Zheng, Haoran; Wu, Jihui; Shi, Yunyu

    2013-01-27

    The ability of a microorganism to adapt to changes in the environment, such as in nutrient or oxygen availability, is essential for its competitive fitness and survival. The cellular objective and the strategy of the metabolic response to an extreme environment are therefore of tremendous interest and, thus, have been increasingly explored. However, the cellular objective of the complex regulatory structure of the metabolic changes has not yet been fully elucidated and more details regarding the quantitative behaviour of the metabolic flux redistribution are required to understand the systems-wide biological significance of this response. In this study, the intracellular metabolic flux ratios involved in the central carbon metabolism were determined by fractional (13)C-labeling and metabolic flux ratio analysis (MetaFoR) of the wild-type E. coli strain JM101 at an oxidative environment in a chemostat. We observed a significant increase in the flux through phosphoenolpyruvate carboxykinase (PEPCK), phosphoenolpyruvate carboxylase (PEPC), malic enzyme (MEZ) and serine hydroxymethyltransferase (SHMT). We applied an ε-constraint based multi-objective optimization to investigate the trade-off relationships between the biomass yield and the generation of reductive power using the in silico iJR904 genome-scale model of E. coli K-12. The theoretical metabolic redistribution supports that the trans-hydrogenase pathway should not play a direct role in the defence mounted by E. coli against oxidative stress. The agreement between the measured ratio and the theoretical redistribution established the significance of NADPH synthesis as the goal of the metabolic reprogramming that occurs in response to oxidative stress. Our work presents a framework that combines metabolic flux ratio analysis and multi-objective optimization to investigate the metabolic trade-offs that occur under varied environmental conditions. Our results led to the proposal that the metabolic response of E

  16. Metabolic flux analysis of Arthrobacter sp. CGMCC 3584 for cAMP production based on 13C tracer experiments and gas chromatography-mass spectrometry.

    PubMed

    Niu, Huanqing; Chen, Yong; Yao, Shiwei; Liu, Lixia; Yang, Chen; Li, Bingbing; Liu, Dong; Xie, Jingjing; Chen, Xiaochun; Wu, Jinglan; Ying, Hanjie

    2013-12-01

    Arthrobacter sp. CGMCC 3584 are able to produce cAMP from glucose by the purine synthesis pathway via de novo or salvage biosynthesis. In order to gain an improved understanding of its metabolism, (13)C-labeling experiment and gas chromatography-mass spectrometry (GC-MS) analysis were employed to determine the metabolic network structure and estimate the intracellular fluxes. GC-MS analysis helps to reflect the activity of the intracellular pathways and reactions. The metabolic network mainly contains glycolytic and pentose phosphate pathways, the tricarboxylic acid cycle, and the inactive glyoxylate shunt. Hypoxanthine as a precursor of cAMP and sodium fluoride as an inhibitor of glycolysis were found to increase the cAMP production, as well as the flux through the PP pathway. The effects of adding hypoxanthine and sodium fluoride are discussed based on the enzyme assays and metabolic flux analysis. In conclusion, our results provide quantitative insights into how cells manipulate the metabolic network under different culture conditions and this may be of value in metabolic regulation for desirable production.

  17. Metabolic changes in Klebsiella oxytoca in response to low oxidoreduction potential, as revealed by comparative proteomic profiling integrated with flux balance analysis.

    PubMed

    Zhu, Yan; Li, Dan; Bao, Guanhui; Wang, Shaohua; Mao, Shaoming; Song, Jiangning; Li, Yin; Zhang, Yanping

    2014-05-01

    Oxidoreduction potential (ORP) is an important physiological parameter for biochemical production in anaerobic or microaerobic processes. However, the effect of ORP on cellular physiology remains largely unknown, which hampers the design of engineering strategies targeting proteins associated with ORP response. Here we characterized the effect of altering ORP in a 1,3-propanediol producer, Klebsiella oxytoca, by comparative proteomic profiling combined with flux balance analysis. Decreasing the extracellular ORP from -150 to -240 mV retarded cell growth and enhanced 1,3-propanediol production. Comparative proteomic analysis identified 61 differentially expressed proteins, mainly involved in carbohydrate catabolism, cellular constituent biosynthesis, and reductive stress response. A hypothetical oxidoreductase (HOR) that catalyzes 1,3-propanediol production was markedly upregulated, while proteins involved in biomass precursor synthesis were downregulated. As revealed by subsequent flux balance analysis, low ORP induced a metabolic shift from glycerol oxidation to reduction and rebalancing of redox and energy metabolism. From the integrated protein expression profiles and flux distributions, we can construct a rational analytic framework that elucidates how (facultative) anaerobes respond to extracellular ORP changes.

  18. Ecosystem function and particle flux dynamics across the Mackenzie Shelf (Beaufort Sea, Arctic Ocean): an integrative analysis of spatial variability and biophysical forcings

    NASA Astrophysics Data System (ADS)

    Forest, A.; Babin, M.; Stemmann, L.; Picheral, M.; Sampei, M.; Fortier, L.; Gratton, Y.; Bélanger, S.; Devred, E.; Sahlin, J.; Doxaran, D.; Joux, F.; Ortega-Retuerta, E.; Martín, J.; Jeffrey, W. H.; Gasser, B.; Miquel, J. Carlos

    2013-05-01

    A better understanding of how environmental changes affect organic matter fluxes in Arctic marine ecosystems is sorely needed. Here we combine mooring times series, ship-based measurements and remote sensing to assess the variability and forcing factors of vertical fluxes of particulate organic carbon (POC) across the Mackenzie Shelf in 2009. We developed a geospatial model of these fluxes to proceed to an integrative analysis of their determinants in summer. Flux data were obtained with sediment traps moored around 125 m and via a regional empirical algorithm applied to particle size distributions (17 classes from 0.08-4.2 mm) measured by an Underwater Vision Profiler 5. The low fractal dimension (i.e., porous, fluffy particles) derived from the algorithm (1.26 ± 0.34) and the dominance (~ 77%) of rapidly sinking small aggregates (< 0.5 mm) in total fluxes suggested that settling material was the product of recent aggregation processes between marine detritus, gel-like substances, and ballast minerals. Modeled settling velocity of small and large aggregates was, respectively, higher and lower than in previous studies within which a high fractal dimension (i.e., more compact particles) was consequential of deep-trap collection (~400-1300 m). Redundancy analyses and forward selection of abiotic/biotic parameters, linear trends, and spatial structures (i.e., principal coordinates of neighbor matrices, PCNM) were conducted to partition the variation of the 17 POC flux size classes. Flux variability was explained at 69.5% by the addition of a temporal trend, 7 significant PCNM, and 9 biophysical variables. The first PCNM canonical axis (44.5% of spatial variance) reflected the total magnitude of POC fluxes through a shelf-basin gradient controlled by bottom depth and sea ice concentration (p < 0.01). The second most important spatial structure (5.0%) corresponded to areas where shelf break upwelling is known to occur under easterlies and where phytoplankton was

  19. Evaluation of the effect of a cover layer on radon exhalation from uranium mill tailings: transient radon flux analysis.

    PubMed

    Ferry, Cécile; Richon, Patrick; Beneito, Alain; Robé, Marie-Christine

    2002-01-01

    An experimental study concerning the transport of 222Rn in uranium mill tailings (UMTs) and in the cover layer was launched in 1997 with the participation of the French uranium mining company (COGEMA). Evaluation of the cover layer's effectiveness in reducing 222Rn flux emanating from UMTs was one of its objectives. In the first phase, the 222Rn flux densities were measured regularly on a UMT layer. In the second phase, the UMT was covered with a one-meter layer of compacted material consisting of crushed waste rock derived from mining activities. Radon-222 flux was then measured at the surface of this cover layer. Observations were compared with radon flux calculated using TRACI, a model for vertical water and gas flow and radon transport. The results show that the calculations bear a fair resemblance to the observations in both cases. They also show that the effectiveness of the cover layer calculated with TRACI, using the thickness and textural properties of the cover material, is very close to the measured effectiveness.

  20. Analysis on current limiting characteristics of a flux-lock type SFCL with two triggering current levels

    NASA Astrophysics Data System (ADS)

    Lim, S. H.

    2011-11-01

    In this paper, the flux-lock type superconducting fault current limiter (SFCL) with two triggering current levels was suggested and its current limiting characteristics were analyzed. The structure of the suggested flux-lock type SFCL with two triggering current levels largely consists of two parts. One is the flux-lock type SFCL with two magnetically coupled coils with series or parallel connection and one high-TC superconducting (HTSC) element, which is connected in series or parallel with one of two coils. The other is constructed in the third coil with another HTSC element. This suggested flux-lock type SFCL can limit the fault current by generating its limiting impedance with two different amplitudes, which are dependent on the initial amplitude of the fault current in case of the fault occurrence. To confirm the usefulness of the proposed SFCL, the limiting tests of the SFCL according to the fault angle, which is one of the effective fault conditions to affect the amplitude of the initial fault current, were carried out and its effective limiting operations were discussed.

  1. Neural network analysis on the effect of heat fluxes on greenhouse gas emissions from anaerobic swine waste treatment lagoon

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this study, we examined the various meteorological factors (i.e., air temperatures, solar radiation, and heat fluxes) that potentially affect greenhouse gas (GHG) emissions from swine waste lagoon. GHG concentrations (methane, carbon dioxide, and nitrous oxide) were monitored using a photoacous...

  2. Analysis of neutron flux distribution for the validation of computational methods for the optimization of research reactor utilization.

    PubMed

    Snoj, L; Trkov, A; Jaćimović, R; Rogan, P; Zerovnik, G; Ravnik, M

    2011-01-01

    In order to verify and validate the computational methods for neutron flux calculation in TRIGA research reactor calculations, a series of experiments has been performed. The neutron activation method was used to verify the calculated neutron flux distribution in the TRIGA reactor. Aluminium (99.9 wt%)-Gold (0.1 wt%) foils (disks of 5mm diameter and 0.2mm thick) were irradiated in 33 locations; 6 in the core and 27 in the carrousel facility in the reflector. The experimental results were compared to the calculations performed with Monte Carlo code MCNP using detailed geometrical model of the reactor. The calculated and experimental normalized reaction rates in the core are in very good agreement for both isotopes indicating that the material and geometrical properties of the reactor core are modelled well. In conclusion one can state that our computational model describes very well the neutron flux and reaction rate distribution in the reactor core. In the reflector however, the accuracy of the epithermal and thermal neutron flux distribution and attenuation is lower, mainly due to lack of information about the material properties of the graphite reflector surrounding the core, but the differences between measurements and calculations are within 10%. Since our computational model properly describes the reactor core it can be used for calculations of reactor core parameters and for optimization of research reactor utilization.

  3. Soil heat flux calculation for sunlit and shaded surfaces under row crops: 1 - Model Development and sensitivity analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Soil heat flux at the surface (G0) is strongly influenced by whether the soil is shaded or sunlit, and therefore can have large spatial variability for incomplete vegetation cover, such as across the interrows of row crops. Most practical soil-plant-atmosphere energy balance models calculate G0 as a...

  4. Quantitative analysis of flux along the gluconeogenic, glycolytic and pentose phosphate pathways under reducing conditions in hepatocytes isolated from fed rats.

    PubMed Central

    Crawford, J M; Blum, J J

    1983-01-01

    Hepatocytes were isolated from the livers of fed rats and incubated with a mixture of glucose (10 mM), ribose (1 mM), mannose (4 mM), glycerol (3 mM), acetate (1.25 mM), and ethanol (5 mM) with one substrate labelled with 14C in any given incubation. Incorporation of label into CO2, glucose, glycogen, lipid glycerol and fatty acids, acetate and C-1 of glucose was measured at 20 and 40 min after the start of the incubation. The data (about 48 measurements for each interval) were used in conjunction with a single-compartment model of the reactions of the gluconeogenic, glycolytic and pentose phosphate pathways and a simplified model of the relevant mitochondrial reactions. An improved method of computer analysis of the equations describing the flow of label through each carbon atom of each metabolite under steady-state conditions was used to compute values for the 34 independent flux parameters in this model. A good fit to the data was obtained, thereby permitting good estimates of most of the fluxes in the pathways under consideration. The data show that: net flux above the level of the triose phosphates is gluconeogenic; label in the hexose phosphates is fully equilibrated by the second 20 min interval; the triose phosphate isomerase step does not equilibrate label between the triose phosphates; substrate cycles are operating at the glucose-glucose 6-phosphate, fructose 6-phosphate-fructose 1,6-bisphosphate and phosphoenolpyruvate-pyruvate-oxaloacetate cycles; and, although net flux through the enzymes catalysing the non-oxidative steps of the pentose phosphate pathway is small, bidirectional fluxes are large. PMID:6411069

  5. An investigation of the neutron flux in bone-fluorine phantoms comparing accelerator based in vivo neutron activation analysis and FLUKA simulation data

    NASA Astrophysics Data System (ADS)

    Mostafaei, F.; McNeill, F. E.; Chettle, D. R.; Matysiak, W.; Bhatia, C.; Prestwich, W. V.

    2015-01-01

    We have tested the Monte Carlo code FLUKA for its ability to assist in the development of a better system for the in vivo measurement of fluorine. We used it to create a neutron flux map of the inside of the in vivo neutron activation analysis irradiation cavity at the McMaster Accelerator Laboratory. The cavity is used in a system that has been developed for assessment of fluorine levels in the human hand. This study was undertaken to (i) assess the FLUKA code, (ii) find the optimal hand position inside the cavity and assess the effects on precision of a hand being in a non-optimal position and (iii) to determine the best location for our γ-ray detection system within the accelerator beam hall. Simulation estimates were performed using FLUKA. Experimental measurements of the neutron flux were performed using Mn wires. The activation of the wires was measured inside (1) an empty bottle, (2) a bottle containing water, (3) a bottle covered with cadmium and (4) a dry powder-based fluorine phantom. FLUKA was used to simulate the irradiation cavity, and used to estimate the neutron flux in different positions both inside, and external to, the cavity. The experimental results were found to be consistent with the Monte Carlo simulated neutron flux. Both experiment and simulation showed that there is an optimal position in the cavity, but that the effect on the thermal flux of a hand being in a non-optimal position is less than 20%, which will result in a less than 10% effect on the measurement precision. FLUKA appears to be a code that can be useful for modeling of this type of experimental system.

  6. Transition of an Anaerobic Escherichia coli Culture to Aerobiosis: Balancing mRNA and Protein Levels in a Demand-Directed Dynamic Flux Balance Analysis

    PubMed Central

    von Wulffen, Joachim; Sawodny, Oliver; Feuer, Ronny

    2016-01-01

    The facultative anaerobic bacterium Escherichia coli is frequently forced to adapt to changing environmental conditions. One important determinant for metabolism is the availability of oxygen allowing a more efficient metabolism. Especially in large scale bioreactors, the distribution of oxygen is inhomogeneous and individual cells encounter frequent changes. This might contribute to observed yield losses during process upscaling. Short-term gene expression data exist of an anaerobic E. coli batch culture shifting to aerobic conditions. The data reveal temporary upregulation of genes that are less efficient in terms of energy conservation than the genes predicted by conventional flux balance analyses. In this study, we provide evidence for a positive correlation between metabolic fluxes and gene expression. We then hypothesize that the more efficient enzymes are limited by their low expression, restricting flux through their reactions. We define a demand that triggers expression of the demanded enzymes that we explicitly include in our model. With these features we propose a method, demand-directed dynamic flux balance analysis, dddFBA, bringing together elements of several previously published methods. The introduction of additional flux constraints proportional to gene expression provoke a temporary demand for less efficient enzymes, which is in agreement with the transient upregulation of these genes observed in the data. In the proposed approach, the applied objective function of growth rate maximization together with the introduced constraints triggers expression of metabolically less efficient genes. This finding is one possible explanation for the yield losses observed in large scale bacterial cultivations where steady oxygen supply cannot be warranted. PMID:27384956

  7. Control analysis of lipid biosynthesis in tissue cultures from oil crops shows that flux control is shared between fatty acid synthesis and lipid assembly.

    PubMed Central

    Ramli, Umi S; Baker, Darren S; Quant, Patti A; Harwood, John L

    2002-01-01

    Top-Down (Metabolic) Control Analysis (TDCA) was used to examine, quantitatively, lipid biosynthesis in tissue cultures from two commercially important oil crops, olive (Olea europaea L.) and oil palm (Elaeis guineensis Jacq.). A conceptually simplified system was defined comprising two blocks of reactions: fatty acid synthesis (Block A) and lipid assembly (Block B), which produced and consumed, respectively, a common and unique system intermediate, cytosolic acyl-CoA. We manipulated the steady-state levels of the system intermediate by adding exogenous oleic acid and, using two independent assays, measured the effect of the addition on the system fluxes (J(A) and J(B)). These were the rate of incorporation of radioactivity: (i) through Block A from [1-(14)C]acetate into fatty acids and (ii) via Block B from [U-(14)C]glycerol into complex lipids respectively. The data showed that fatty acid formation (Block A) exerted higher control than lipid assembly (Block B) in both tissues with the following group flux control coefficients (C):(i) Oil palm: *C(J(TL))(BlkA)=0.64+/-0.05 and *C(J(TL))(BlkB)=0.36+/-0.05(ii) Olive: *C(J(TL))(BlkA)=0.57+/-0.10 and *C(J(TL))(BlkB)=0.43+/-0.10where *C indicates the group flux control coefficient over the lipid biosynthesis flux (J(TL)) and the subscripts BlkA and BlkB refer to defined blocks of the system, Block A and Block B. Nevertheless, because both parts of the lipid biosynthetic pathway exert significant flux control, we suggest strongly that manipulation of single enzyme steps will not affect product yield appreciably. The present study represents the first use of TDCA to examine the overall lipid biosynthetic pathway in any tissue, and its findings are of immediate academic and economic relevance to the yield and nutritional quality of oil crops. PMID:12023882

  8. Control analysis of lipid biosynthesis in tissue cultures from oil crops shows that flux control is shared between fatty acid synthesis and lipid assembly.

    PubMed

    Ramli, Umi S; Baker, Darren S; Quant, Patti A; Harwood, John L

    2002-06-01

    Top-Down (Metabolic) Control Analysis (TDCA) was used to examine, quantitatively, lipid biosynthesis in tissue cultures from two commercially important oil crops, olive (Olea europaea L.) and oil palm (Elaeis guineensis Jacq.). A conceptually simplified system was defined comprising two blocks of reactions: fatty acid synthesis (Block A) and lipid assembly (Block B), which produced and consumed, respectively, a common and unique system intermediate, cytosolic acyl-CoA. We manipulated the steady-state levels of the system intermediate by adding exogenous oleic acid and, using two independent assays, measured the effect of the addition on the system fluxes (J(A) and J(B)). These were the rate of incorporation of radioactivity: (i) through Block A from [1-(14)C]acetate into fatty acids and (ii) via Block B from [U-(14)C]glycerol into complex lipids respectively. The data showed that fatty acid formation (Block A) exerted higher control than lipid assembly (Block B) in both tissues with the following group flux control coefficients (C):(i) Oil palm: *C(J(TL))(BlkA)=0.64+/-0.05 and *C(J(TL))(BlkB)=0.36+/-0.05(ii) Olive: *C(J(TL))(BlkA)=0.57+/-0.10 and *C(J(TL))(BlkB)=0.43+/-0.10where *C indicates the group flux control coefficient over the lipid biosynthesis flux (J(TL)) and the subscripts BlkA and BlkB refer to defined blocks of the system, Block A and Block B. Nevertheless, because both parts of the lipid biosynthetic pathway exert significant flux control, we suggest strongly that manipulation of single enzyme steps will not affect product yield appreciably. The present study represents the first use of TDCA to examine the overall lipid biosynthetic pathway in any tissue, and its findings are of immediate academic and economic relevance to the yield and nutritional quality of oil crops.

  9. A priori analysis of subgrid scale pressure and heat flux in high pressure mixing and reacting shear layers

    NASA Astrophysics Data System (ADS)

    Ma, Zhiyuan; Korucu, Ayse; Miller, Richard Steven

    2015-11-01

    Direct Numerical Simulation (DNS) data on high pressure H2/O2 and H2/air flames using the compressible flow formulation, detailed kinetics, a real fluid equation of state, and generalised diffusion are analysed. The DNS is filtered over a range of filter widths to provide exact terms in the Large Eddy Simulation (LES) governing equations, including unclosed terms. The filtered pressure and the filtered heat flux vector are extensively compared with the pressure and the heat flux vector calculated as a function of the filtered primitive variables (i.e. the exact LES term is compared with its form available within an actual LES). The difference between these forms defines the subgrid pressure and the subgrid heat flux vector. The analyses are done both globally across the entire flame, as well as by conditionally averaging over specific regions of the flame; including regions of large subgrid kinetic energy, subgrid scalar dissipation, subgrid temperature variance, flame temperature, etc. In this work, although negligible for purely mixing cases, the gradient of the subgrid pressure is shown to be of the same order as, and larger than, the corresponding divergence of the turbulent subgrid stresses for reacting cases. This is despite the fact that all species behave essentially as ideal gases for this flame and holds true even when the ideal gas law is used to calculate the pressure. The ratio of the subgrid pressure gradient to the subgrid stress tensor divergence is shown to increase with increasing Reynolds number. Both the subgrid heat flux vector and its divergence are found to be substantially larger in reacting flows in comparison with mixing due to the associated larger temperature gradients. However, the divergence of the subgrid heat flux vector tends to be significantly smaller than other unclosed terms in the energy equation with decreasing significance with increasing Reynolds number.

  10. Establishing ozone flux-response relationships for winter wheat: Analysis of uncertainties based on data for UK and Polish genotypes

    NASA Astrophysics Data System (ADS)

    Gonzalez–Fernandez, Ignacio; Kaminska, Agnieszka; Dodmani, Mahmadali; Goumenaki, Eleni; Quarrie, Steve; Barnes, Jeremy D.

    2010-02-01

    The work outlined in this paper had three objectives. The first was to explore the effects of ozone pollution on grain yield and quality of commercially-grown winter wheat cultivars. The second was to derive a stomatal ozone flux model for winter wheat and compare with those already developed for spring wheat. The third was to evaluate exposure- versus flux-response approaches from a risk assessment perspective, and explore the implications of genetic variation in modelled ozone flux. Fifteen winter wheat cultivars were grown in open-top chambers where they were exposed to four levels of ozone. During fumigation, stomatal conductance measurements were made over the lifespan of the flag leaf across a range of environmental conditions. Although significant intra-specific variation in 'ozone sensitivity' (in terms of impacts on yield) was identified, yield was inversely related ( R2 = 0.63, P < 0.001) to the accumulated hourly averaged ozone exposure above 40 ppb during daylight hours (AOT40) across the dataset. The adverse effect of ozone on yield was principally due to a decline in seed weight. Algorithms defining the influence of environmental variables on stomatal uptake were subtly different from those currently in use, based on data for spring wheat, to map ozone impacts on pan-European cereal yield. Considerable intra-specific variation in phenological effects was identified. This meant that an 'average behaviour' had to be derived which reduced the predictive capability of the derived stomatal flux model ( R2 = 0.49, P < 0.001, 15 cultivars included). Indeed, given the intra-specific variability encountered, the flux model that was derived from the full dataset was no better in predicting O 3 impacts on wheat yield than was the AOT40 index. The study highlights the need to use ozone risk assessment tools appropriate to specific vegetation types when modelling and mapping ozone impacts at the regional level.

  11. Analysis of the Variation of Energetic Electron Flux with Respect to Longitude and Distance Normal to the Magnetic Equatorial Plane for Galileo Energetic Particle Detector Data

    NASA Technical Reports Server (NTRS)

    Swimm, Randall; Garrett, Henry B.; Jun, Insoo; Evans, Robin W.

    2004-01-01

    In this study we examine ten-minute omni-directional averages of energetic electron data measured by the Galileo spacecraft Energetic Particle Detector (EPD). Count rates from electron channels B1, DC2, and DC3 are evaluated using a power law model to yield estimates of the differential electron fluxes from 1 MeV to 11 MeV at distances between 8 and 51 Jupiter radii. Whereas the orbit of the Galileo spacecraft remained close to the rotational equatorial plane of Jupiter, the approximately 11 degree tilt of the magnetic axis of Jupiter relative to its rotational axis allowed the EPD instrument to sample high energy electrons at limited distances normal to the magnetic equatorial plane. We present a Fourier analysis of the semi-diurnal variation of electron fluxes with longitude.

  12. Dynamic Characteristics of the Main Indexes of Space Weather and Their Application to the Analysis Monitoring Observations Flux Densities of Power Radio Sources on RT

    NASA Astrophysics Data System (ADS)

    Guglya, L.; Ryabov, M.; Panishko, S.; Suharev, A.

    On radio telescope "URAN-4" of the Odessa observatory of Radio-astronomical Institute during twenty five years (since 1987 till present) monitoring of power galactic and extragalactic radio sources on frequencies 25 and 20MHz has been carried out. Data of the observation was spent in a current of the 22-23th cycles of solar activity and in the beginning of the 24th cycle. Long-term variations density fluxes of radio sources connection with change of a condition of ionosphere in a cycle of solar activity are considered. Means Fourier and Wavelet analysis determine dynamics of changes of the main indexes of space weather and the basic periods of activity are revealed. The obtained data will be used for interpretation of the observation changes flux of radio sources for during all investigated cycle of activity and periods of extreme developments of space weather.

  13. Assessment of energetic costs of AhR activation by β-naphthoflavone in rainbow trout (Oncorhynchus mykiss) hepatocytes using metabolic flux analysis

    SciTech Connect

    Nault, Rance; Abdul-Fattah, Hiba; Mironov, Gleb G.; Berezovski, Maxim V.; Moon, Thomas W.

    2013-08-15

    Exposure to environmental contaminants such as activators of the aryl hydrocarbon receptor (AhR) leads to the induction of defense and detoxification mechanisms. While these mechanisms allow organisms to metabolize and excrete at least some of these environmental contaminants, it has been proposed that these mechanisms lead to significant energetic challenges. This study tests the hypothesis that activation of the AhR by the model agonist β-naphthoflavone (βNF) results in increased energetic costs in rainbow trout (Oncorhynchus mykiss) hepatocytes. To address this hypothesis, we employed traditional biochemical approaches to examine energy allocation and metabolism including the adenylate energy charge (AEC), protein synthesis rates, Na{sup +}/K{sup +}-ATPase activity, and enzyme activities. Moreover, we have used for the first time in a fish cell preparation, metabolic flux analysis (MFA) an in silico approach for the estimation of intracellular metabolic fluxes. Exposure of trout hepatocytes to 1 μM βNF for 48 h did not alter hepatocyte AEC, protein synthesis, or Na{sup +}/K{sup +}-ATPase activity but did lead to sparing of glycogen reserves and changes in activities of alanine aminotransferase and citrate synthase suggesting altered metabolism. Conversely, MFA did not identify altered metabolic fluxes, although we do show that the dynamic metabolism of isolated trout hepatocytes poses a significant challenge for this type of approach which should be considered in future studies. - Highlights: • Energetic costs of AhR activation by βNF was examined in rainbow trout hepatocytes. • Metabolic flux analysis was performed on a fish cell preparation for the first time. • Exposure to βNF led to sparing of glycogen reserves and altered enzyme activities. • Adenylate energy charge was maintained despite temporal changes in metabolism.

  14. Combined trajectory clustering and aerosol fields analysis to evaluate the potential emission flux to aerosol pollutants in an urban and semi-urban atmospheres in eastern India

    NASA Astrophysics Data System (ADS)

    Kumar, B. D.; Verma, S.

    2015-12-01

    A hybrid source-receptor analysis was carried out to evaluate the potential emission flux to winter monsoon (WinMon) aerosols over eastern India urban (Kolkata, Kol) and semi-urban atmospheres (Kharagpur, Kgp). This was done through application of fuzzy c-mean clustering to back-trajectory data combined with emission flux and residence time weighted aerosols analysis. WinMon mean aerosol optical depth (AOD) and angstrom exponent (AE) at Kol were respectively slightly higher than and nearly equal to that at Kgp. Out of six source region clusters over Indian subcontinent and two over Indian oceanic region, the cluster mean AOD was the highest when associated with the mean path of air mass originating from the Bay of Bengal (BoB) and the Arabian sea (AS) clusters at Kol and that from the Indo-Gangetic plain (IGP)cluster at Kgp. Spatial distribution of weighted AOD fields showed the highest potential source of aerosols over the IGP, majorly over upper IGP (IGP-U), lower IGP (IGP-L) and eastern region (ER) clusters. The emission flux contribution potential (EFCP) of fossil fuel (FF) emissions at surface (SL) of Kol/Kgp , elevated layer (EL) of Kol, and of biomass burning (BB) emissions at SL of Kol were majorly from IGP-U, IGP-L and IGP-U/L clusters respectively. The EFCP of FF/BB emissions at Kgp-EL/SL, and that of BB at EL of Kol/Kgp were mainly from ER and Africa (AFR) clusters respectively. Though the AFR cluster was constituted of significantly high emission flux source potential of dust emissions, the EFCP of dust from NWI was comparable to that from AFR at Kol SL/EL.

  15. Unusual flux-distance relationship for pulsars suggested by analysis of the Australia national telescopy facility pulsar catalogue

    SciTech Connect

    Singleton, John; Perez, M R; Singleton, J; Ardavan, H; Ardavan, A

    2009-01-01

    We analyze pulsar fluxes at 1400 MHz (S(1400)) and distances d taken from the Australia National Telescope Facility (ATNF) Pulsar Catalogue. Under the assumption that pulsar populations in different parts of the Galaxy are similar, we find that either (a) pulsar fluxes diminish with distance according to a non-standard power law (we suggest S(1400){proportional_to} 1/d rather than {proportional_to} 1/d{sup 2}) or (b) that there are very significant (i.e. order of magnitude) errors in the distance estimates quoted in the ATNF Catalogue. The former conclusion (a) supports a recent model for pulsar emission that has also successfully explained the frequency spectrum of the Crab pulsar over 16 orders of magnitude of frequency, whilst alternative (b) would necessitate a radical re-evaluation of both the dispersion method for estimating pulsar distances and current ideas about the distribution of pulsars within our Galaxy.

  16. Numerical Analysis of Nucleate Boiling on High Heat-Flux and High Subcooling Condition for Reactivity Initiation Accident

    SciTech Connect

    Heo, S.; Koshizuka, S.; Oka, Y.

    2002-07-01

    This paper shows the numerical simulation study on the growth of the bubble in the transient pool boiling using MPS-MAFL method. The growth process of a bubble with the different initial radii is calculated in a high heat-flux and high subcooling condition expected in nuclear reactor core during RIA. The smaller initial radius is, the earlier the growth starts. The initial bubble radius has little effect on the growth initiation time and the bubble departure radius. (authors)

  17. A Time Series Analysis of Energetic Electron Fluxes (1.2 - 16 MeV) at Geosynchronous Altitude.

    DTIC Science & Technology

    1986-12-01

    charging of spacecraft surfaces and resultant discharges" (Grajek and McPherson , 1979:769). They went on to observe that though most of these anomalies...difficult problems for our spacecraft deployed in geosynchronous earth orbits inside the magnetosphere . High levels of flux (defined as the measured...or cancer) (Spjeldvik and Rothwell, 1983:121). Therefore, given that man has found it necessary to operate spacecraft in the magnetosphere for a host

  18. STABILITY AND DYNAMICS OF A FLUX ROPE FORMED VIA FLUX EMERGENCE INTO THE SOLAR ATMOSPHERE

    SciTech Connect

    An, J. M.; Magara, T. E-mail: magara@khu.ac.kr

    2013-08-10

    We study the stability and dynamics of a flux rope formed through the emergence of a twisted magnetic flux tube into the solar atmosphere. A three-dimensional magnetohydrodynamic simulation has been performed to investigate several key factors affecting the dynamics of the flux rope. The stability of the flux rope is examined by deriving the decay index of the coronal magnetic field surrounding the flux rope. We investigate a transition between the quasi-static and dynamic states of the flux rope through an analysis of the curvature and scale height of emerging magnetic field. A practical application of this analysis for global eruptions is also considered.

  19. Interannual Variability in Soil Trace Gas (CO2, N2O, NO) Fluxes and Analysis of Controllers

    NASA Technical Reports Server (NTRS)

    Potter, C.; Klooster, S.; Peterson, David L. (Technical Monitor)

    1997-01-01

    Interannual variability in flux rates of biogenic trace gases must be quantified in order to understand the differences between short-term trends and actual long-term change in biosphere-atmosphere interactions. We simulated interannual patterns (1983-1988) of global trace gas fluxes from soils using the NASA Ames model version of CASA (Carnegie-Ames-Stanford Approach) in a transient simulation mode. This ecosystem model has been recalibrated for simulations driven by satellite vegetation index data from the NOAA Advanced Very High Resolution Radiometer (AVHRR) over the mid-1980s. The predicted interannual pattern of soil heterotropic CO2 emissions indicates that relatively large increases in global carbon flux from soils occurred about three years following the strong El Nino Southern Oscillation (ENSO) event of 1983. Results for the years 1986 and 1987 showed an annual increment of +1 Pg (1015 g) C-CO2 emitted from soils, which tended to dampen the estimated global increase in net ecosystem production with about a two year lag period relative to plant carbon fixation. Zonal discrimination of model results implies that 80-90 percent of the yearly positive increments in soil CO2 emission during 1986-87 were attributable to soil organic matter decomposition in the low-latitudes (between 30 N and 30 S). Soils of the northern middle-latitude zone (between 30 N and 60 N) accounted for the residual of these annual increments. Total annual emissions of nitrogen trace gases (N2O and NO) from soils were estimated to vary from 2-4 percent over the time period modeled, a level of variability which is consistent with predicted interannual fluctuations in global soil CO2 fluxes. Interannual variability of precipitation in tropical and subtropical zones (30 N to 20 S appeared to drive the dynamic inverse relationship between higher annual emissions of NO versus emissions of N2O. Global mean emission rates from natural (heterotrophic) soil sources over the period modeled (1983

  20. Numerical prediction of heat-flux to massive calorimeters engulfed in regulatory fires with the cask analysis fire environment (CAFE) model

    SciTech Connect

    KOSKI,JORMAN A.; SUO-ANTITLA,AHTI; KRAMER,M. ALEX; GREINER,MILES

    2000-05-11

    Recent observations show that the thermal boundary conditions within large-scale fires are significantly affected by the presence of thermally massive objects. These objects cool the soot and gas near their surfaces, and these effects reduce the incoming radiant heat-flux to values lower than the levels expected from simple {sigma}T{sub fire}{sup 4} models. They also affect the flow and temperature fields in the fire far from their surfaces. The Cask Analysis Fire Environment (CAFE) code has been developed at Sandia National Laboratories to provide an enhanced fire boundary condition for the design of radioactive material packages. CAFE is a set of computer subroutines that use computational fluid mechanics methods to predict convective heat transfer and mixing. It also includes models for fuel and oxygen transport, chemical reaction, and participating-media radiation heat transfer. This code uses two-dimensional computational models so that it has reasonably short turnaround times on standard workstations and is well suited for design and risk studies. In this paper, CAFE is coupled with a commercial finite-element program to model a large cylindrical calorimeter fully engulfed in a pool fire. The time-dependent heat-flux to the calorimeter and the calorimeter surface temperature are determined for several locations around the calorimeter circumference. The variation of heat-flux with location is determined for calorimeters with different diameters and wall thickness, and the observed effects discussed.

  1. Fast flux locked loop

    DOEpatents

    Ganther, Jr., Kenneth R.; Snapp, Lowell D.

    2002-09-10

    A flux locked loop for providing an electrical feedback signal, the flux locked loop employing radio-frequency components and technology to extend the flux modulation frequency and tracking loop bandwidth. The flux locked loop of the present invention has particularly useful application in read-out electronics for DC SQUID magnetic measurement systems, in which case the electrical signal output by the flux locked loop represents an unknown magnetic flux applied to the DC SQUID.

  2. Application of the principles of systems biology and Wiener's cybernetics for analysis of regulation of energy fluxes in muscle cells in vivo.

    PubMed

    Guzun, Rita; Saks, Valdur

    2010-03-08

    The mechanisms of regulation of respiration and energy fluxes in the cells are analyzed based on the concepts of systems biology, non-equilibrium steady state kinetics and applications of Wiener's cybernetic principles of feedback regulation. Under physiological conditions cardiac function is governed by the Frank-Starling law and the main metabolic characteristic of cardiac muscle cells is metabolic homeostasis, when both workload and respiration rate can be changed manifold at constant intracellular level of phosphocreatine and ATP in the cells. This is not observed in skeletal muscles. Controversies in theoretical explanations of these observations are analyzed. Experimental studies of permeabilized fibers from human skeletal muscle vastus lateralis and adult rat cardiomyocytes showed that the respiration rate is always an apparent hyperbolic but not a sigmoid function of ADP concentration. It is our conclusion that realistic explanations of regulation of energy fluxes in muscle cells require systemic approaches including application of the feedback theory of Wiener's cybernetics in combination with detailed experimental research. Such an analysis reveals the importance of limited permeability of mitochondrial outer membrane for ADP due to interactions of mitochondria with cytoskeleton resulting in quasi-linear dependence of respiration rate on amplitude of cyclic changes in cytoplasmic ADP concentrations. The system of compartmentalized creatine kinase (CK) isoenzymes functionally coupled to ANT and ATPases, and mitochondrial-cytoskeletal interactions separate energy fluxes (mass and energy transfer) from signalling (information transfer) within dissipative metabolic structures - intracellular energetic units (ICEU). Due to the non-equilibrium state of CK reactions, intracellular ATP utilization and mitochondrial ATP regeneration are interconnected by the PCr flux from mitochondria. The feedback regulation of respiration occurring via cyclic fluctuations of

  3. Application of the Principles of Systems Biology and Wiener’s Cybernetics for Analysis of Regulation of Energy Fluxes in Muscle Cells in Vivo

    PubMed Central

    Guzun, Rita; Saks, Valdur

    2010-01-01

    The mechanisms of regulation of respiration and energy fluxes in the cells are analyzed based on the concepts of systems biology, non-equilibrium steady state kinetics and applications of Wiener’s cybernetic principles of feedback regulation. Under physiological conditions cardiac function is governed by the Frank-Starling law and the main metabolic characteristic of cardiac muscle cells is metabolic homeostasis, when both workload and respiration rate can be changed manifold at constant intracellular level of phosphocreatine and ATP in the cells. This is not observed in skeletal muscles. Controversies in theoretical explanations of these observations are analyzed. Experimental studies of permeabilized fibers from human skeletal muscle vastus lateralis and adult rat cardiomyocytes showed that the respiration rate is always an apparent hyperbolic but not a sigmoid function of ADP concentration. It is our conclusion that realistic explanations of regulation of energy fluxes in muscle cells require systemic approaches including application of the feedback theory of Wiener’s cybernetics in combination with detailed experimental research. Such an analysis reveals the importance of limited permeability of mitochondrial outer membrane for ADP due to interactions of mitochondria with cytoskeleton resulting in quasi-linear dependence of respiration rate on amplitude of cyclic changes in cytoplasmic ADP concentrations. The system of compartmentalized creatine kinase (CK) isoenzymes functionally coupled to ANT and ATPases, and mitochondrial-cytoskeletal interactions separate energy fluxes (mass and energy transfer) from signalling (information transfer) within dissipative metabolic structures – intracellular energetic units (ICEU). Due to the non-equilibrium state of CK reactions, intracellular ATP utilization and mitochondrial ATP regeneration are interconnected by the PCr flux from mitochondria. The feedback regulation of respiration occurring via cyclic fluctuations

  4. The Virtual Observatory Service TheoSSA: Establishing a Database of Synthetic Stellar Flux Standards II. NLTE Spectral Analysis of the OB-Type Subdwarf Feige 110

    NASA Technical Reports Server (NTRS)

    Rauch, T.; Rudkowski, A.; Kampka, D.; Werner, K.; Kruk, J. W.; Moehler, S.

    2014-01-01

    Context. In the framework of the Virtual Observatory (VO), the German Astrophysical VO (GAVO) developed the registered service TheoSSA (Theoretical Stellar Spectra Access). It provides easy access to stellar spectral energy distributions (SEDs) and is intended to ingest SEDs calculated by any model-atmosphere code, generally for all effective temperatures, surface gravities, and elemental compositions. We will establish a database of SEDs of flux standards that are easily accessible via TheoSSA's web interface. Aims. The OB-type subdwarf Feige 110 is a standard star for flux calibration. State-of-the-art non-local thermodynamic equilibrium stellar-atmosphere models that consider opacities of species up to trans-iron elements will be used to provide a reliable synthetic spectrum to compare with observations. Methods. In case of Feige 110, we demonstrate that the model reproduces not only its overall continuum shape from the far-ultraviolet (FUV) to the optical wavelength range but also the numerous metal lines exhibited in its FUV spectrum. Results. We present a state-of-the-art spectral analysis of Feige 110. We determined Teff =47 250 +/- 2000 K, log g=6.00 +/- 0.20, and the abundances of He, N, P, S, Ti, V, Cr, Mn, Fe, Co, Ni, Zn, and Ge. Ti, V, Mn, Co, Zn, and Ge were identified for the first time in this star. Upper abundance limits were derived for C, O, Si, Ca, and Sc. Conclusions. The TheoSSA database of theoretical SEDs of stellar flux standards guarantees that the flux calibration of astronomical data and cross-calibration between different instruments can be based on models and SEDs calculated with state-of-the-art model atmosphere codes.

  5. Metabolic flux analysis gives an insight on verapamil induced changes in central metabolism of HL-1 cells.

    PubMed

    Strigun, Alexander; Noor, Fozia; Pironti, Alejandro; Niklas, Jens; Yang, Tae Hoon; Heinzle, Elmar

    2011-09-20

    Verapamil has been shown to inhibit glucose transport in several cell types. However, the consequences of this inhibition on central metabolism are not well known. In this study we focused on verapamil induced changes in metabolic fluxes in a murine atrial cell line (HL-1 cells). These cells were adapted to serum free conditions and incubated with 4 μM verapamil and [U-¹³C₅] glutamine. Specific extracellular metabolite uptake/production rates together with mass isotopomer fractions in alanine and glutamate were implemented into a metabolic network model to calculate metabolic flux distributions in the central metabolism. Verapamil decreased specific glucose consumption rate and glycolytic activity by 60%. Although the HL-1 cells show Warburg effect with high lactate production, verapamil treated cells completely stopped lactate production after 24 h while maintaining growth comparable to the untreated cells. Calculated fluxes in TCA cycle reactions as well as NADH/FADH₂ production rates were similar in both treated and untreated cells. This was confirmed by measurement of cell respiration. Reduction of lactate production seems to be the consequence of decreased glucose uptake due to verapamil. In case of tumors, this may have two fold effects; firstly depriving cancer cells of substrate for anaerobic glycolysis on which their growth is dependent; secondly changing pH of the tumor environment, as lactate secretion keeps the pH acidic and facilitates tumor growth. The results shown in this study may partly explain recent observations in which verapamil has been proposed to be a potential anticancer agent. Moreover, in biotechnological production using cell lines, verapamil may be used to reduce glucose uptake and lactate secretion thereby increasing protein production without introduction of genetic modifications and application of more complicated fed-batch processes.

  6. Determination of aluminum and phosphorus in biological materials by reactor activation analysis using germanium as integral flux monitor and comparator.

    PubMed

    Furnari, J C; Cohen, I M

    1994-01-01

    A method for determination of aluminum and phosphorus in biological materials, based on activation in a nuclear reactor and measurement of 28Al, produced by the 27Al(n, gamma)28Al and 31P(n, alpha)28Al reactions, is described. Irradiations in the undisturbed and epicadmium spectra provide a two-equation system in order to determine the contributions of aluminum and phosphorus to the total activities. Germanium is used as an integral flux monitor and comparator, through the reactions: 74Ge(n, gamma)75Ge, 76Ge(n, gamma)77Ge, and 72Ge(n,p)72Ga.

  7. An improved spectral analysis of the stray flux component for the detection of air-gap irregularities in squirrel cage motors.

    PubMed

    Salah, Mohamed; Bacha, Khmais; Chaari, Abdelkader

    2014-05-01

    For machines׳ monitoring purpose, the classical motor current signature analysis has shown its weakness in distinguishing the eccentricity occurrence in presence of others mechanical faults. Although Park׳s vector approach can cover this drawback, the high cost due to the requirement to use three current sensors associated with an advanced processing technique, makes it less desired by industrialists. In this paper, we suggest an alternative diagnosis method based on a suitable processing of the stray flux data. The experimental results have revealed the potential of a simple search coil for the detection and the distinction of the accurate eccentricity nature even in presence of similar mechanical faults.

  8. Use of magnetic Barkhausen noise and magnetic flux leakage signals for analysis of defects in pipeline steel

    SciTech Connect

    Mandal, K.; Dufour, D.; Atherton, D.L.

    1999-05-01

    Oil and gas pipelines are normally buried. They are inspected, while they are in service, by pumping tools through the lines. Inspection tools based on magnetic flux leakage (MFL) measurements are the most economical method used for detecting metal loss, such as corrosion pits. Here the authors report on studies of the effects of hoop and axial tensile stresses on magnetic flux leakage (MFL) signals from 50% penetration electrochemically milled pits eroded in line pipe steel. It is observed that stress can change the MFL signal by more than 50%, depending on the magnetization of the pipe wall. The studies were performed on pits created in both the absence and presence of 330-MPa hoop or axial tensile stress. The MFL results obtained in the two cases show detectable differences if the applied stress is high enough to create plastic deformation in the regions of stress concentration near the pits. The effect of stress applied during pit erosion is less than that when the same stress is applied during the subsequent measurements. Magnetic Barkhausen noise (MBN) measurements have been used to study the stress concentrations around electrochemically milled and mechanically drilled defects and have shown that significant additional stress and plastic deformation can be introduced during mechanical drilling. The MBN results are used to assist interpretation of the stress-dependent MFL results.

  9. Flux analysis of central metabolic pathways in the Fe(III)-reducing organism Geobacter metallireducens via 13C isotopiclabeling

    SciTech Connect

    Tang, Yinjie J.; Chakraborty, Romy; Martin, Hector Garcia; Chu,Jeannie; Hazen, Terry C.; Keasling, Jay D.

    2007-08-13

    We analyzed the carbon fluxes in the central metabolism ofGeobacter metallireducens strain GS-15 using 13C isotopomer modeling.Acetate labeled in the 1st or 2nd position was the sole carbon source,and Fe-NTA was the sole terminal electron acceptor. The measured labeledacetate uptake rate was 21 mmol/gdw/h in the exponential growth phase.The resulting isotope labeling pattern of amino acids allowed an accuratedetermination of the in vivo global metabolic reaction rates (fluxes)through the central metabolic pathways using a computational isotopomermodel. The model indicated that over 90 percent of the acetate wascompletely oxidized to CO2 via a complete tricarboxylic acid (TCA) cyclewhile reducing iron. Pyruvate carboxylase and phosphoenolpyruvatecarboxykinase were present under these conditions, but enzymes in theglyoxylate shunt and malic enzyme were absent. Gluconeogenesis and thepentose phosphate pathway were mainly employed for biosynthesis andaccounted for less than 3 percent of total carbon consumption. The modelalso indicated surprisingly high reversibility in the reaction betweenoxoglutarate and succinate. This step operates close to the thermodynamicequilibrium possibly because succinate is synthesized via a transferasereaction, and its product, acetyl-CoA, inhibits the conversion ofoxoglutarate to succinate. These findings enable a better understandingof the relationship between genome annotation and extant metabolicpathways in G. metallireducens.

  10. Coupling river hydrochemical information with catchment properties for multi-scale-analysis of lateral matter fluxes in the Earth system

    NASA Astrophysics Data System (ADS)

    Hartmann, Jens; Lauerwald, Ronny; Moosdorf, Nils

    2016-04-01

    Over the last decade the number of regional to global scale studies of river chemical fluxes and their steering factors increased rapidly, entailing a growing demand for appropriate databases to calculate mass budgets, to calibrate models, or to test hypotheses [1, 2]. Research applying compilations of hydrochemical data are related to questions targeting different time and spatial scales, as for example the annual to centennial scale. In focus are often the alteration of land-ocean matter fluxes due anthropogenic disturbance, the climate sensitivity of chemical weathering fluxes [3], or nutrient fluxes and their evolution [2, 4]. We present an overview of the GLObal RIver CHemistry database GLORICH, which combines an assemblage of hydrochemical data from varying sources with catchment characteristics of the sampling locations [1]. The information provided include e.g. catchment size, lithology, soil, climate, land cover, net primary production, population density and average slope gradient. The data base comprises 1.27 million samples distributed over 17,000 sampling locations [1]. It will be shown how large assemblages of data are useful to target some major questions about the alteration of land ocean element fluxes due to climate or land use change while coupling hydrochemical data with catchment properties in a homogenized database. An extension by isotopic data will be in the focus of future work [c.f. 5]. Further, applications in climate change studies for understanding feedbacks in the Earth system will be discussed [6]. References: [1] Hartmann, J., Lauerwald, R., & Moosdorf, N. (2014). A brief overview of the GLObal RIver CHemistry Database, GLORICH. Procedia Earth and Planetary Science, 10, 23-27. [2] Hartmann, J., Levy, J., & Kempe, S. (2011). Increasing dissolved silica trends in the Rhine River: an effect of recovery from high P loads?. Limnology, 12(1), 63-73. [3] Hartmann, J., Moosdorf, N., Lauerwald, R., Hinderer, M., & West, A. J. (2014). Global

  11. Metabolic profiling and flux analysis of MEL-2 human embryonic stem cells during exponential growth at physiological and atmospheric oxygen concentrations.

    PubMed

    Turner, Jennifer; Quek, Lake-Ee; Titmarsh, Drew; Krömer, Jens O; Kao, Li-Pin; Nielsen, Lars; Wolvetang, Ernst; Cooper-White, Justin

    2014-01-01

    As human embryonic stem cells (hESCs) steadily progress towards regenerative medicine applications there is an increasing emphasis on the development of bioreactor platforms that enable expansion of these cells to clinically relevant numbers. Surprisingly little is known about the metabolic requirements of hESCs, precluding the rational design and optimisation of such platforms. In this study, we undertook an in-depth characterisation of MEL-2 hESC metabolic behaviour during the exponential growth phase, combining metabolic profiling and flux analysis tools at physiological (hypoxic) and atmospheric (normoxic) oxygen concentrations. To overcome variability in growth profiles and the problem of closing mass balances in a complex environment, we developed protocols to accurately measure uptake and production rates of metabolites, cell density, growth rate and biomass composition, and designed a metabolic flux analysis model for estimating internal rates. hESCs are commonly considered to be highly glycolytic with inactive or immature mitochondria, however, whilst the results of this study confirmed that glycolysis is indeed highly active, we show that at least in MEL-2 hESC, it is supported by the use of oxidative phosphorylation within the mitochondria utilising carbon sources, such as glutamine to maximise ATP production. Under both conditions, glycolysis was disconnected from the mitochondria with all of the glucose being converted to lactate. No difference in the growth rates of cells cultured under physiological or atmospheric oxygen concentrations was observed nor did this cause differences in fluxes through the majority of the internal metabolic pathways associated with biogenesis. These results suggest that hESCs display the conventional Warburg effect, with high aerobic activity despite high lactate production, challenging the idea of an anaerobic metabolism with low mitochondrial activity. The results of this study provide new insight that can be used in

  12. Magnetic-flux pump

    NASA Technical Reports Server (NTRS)

    Hildebrandt, A. F.; Elleman, D. D.; Whitmore, F. C. (Inventor)

    1966-01-01

    A magnetic flux pump is described for increasing the intensity of a magnetic field by transferring flux from one location to the magnetic field. The device includes a pair of communicating cavities formed in a block of superconducting material, and a piston for displacing the trapped magnetic flux into the secondary cavity producing a field having an intense flux density.

  13. Quantitative analysis of some mechanisms affecting the yield of oxidative phosphorylation: dependence upon both fluxes and forces.

    PubMed

    Rigoulet, M; Leverve, X; Fontaine, E; Ouhabi, R; Guérin, B

    1998-07-01

    The purpose of this work was to show how the quantitative definition of the different parameters involved in mitochondrial oxidative phosphorylation makes it possible to characterize the mechanisms by which the yield of ATP synthesis is affected. Three different factors have to be considered: (i) the size of the different forces involved (free energy of redox reactions and ATP synthesis, proton electrochemical difference); (ii) the physical properties of the inner mitochondrial membrane in terms of leaks (H+ and cations); and finally (iii) the properties of the different proton pumps involved in this system (kinetic properties, regulation, modification of intrinsic stoichiometry). The data presented different situations where one or more of these parameters are affected, leading to a different yield of oxidative phosphorylation. (1) By manipulating the actual flux through each of the respiratory chain units at constant protonmotive force in yeast mitochondria, we show that the ATP/O ratio decreases when the flux increases. Moreover, the highest efficiency was obtained when the respiratory rate was low and almost entirely controlled by the electron supply. (2) By using almitrine in different kinds of mitochondria, we show that this drug leads to a decrease in ATP synthesis efficiency by increasing the H+/ATP stoichiometry ofATP synthase (Rigoulet M et al. Biochim Biophys Acta 1018: 91-97, 1990). Since this enzyme is reversible, it was possible to test the effect of this drug on the reverse reaction of the enzyme i.e. extrusion of protons catalyzed by ATP hydrolysis. Hence, we are able to prove that, in this case, the decrease in efficiency of oxidative phosphorylation is due to a change in the mechanistic stoichiometry of this proton pump. To our knowledge, this is the first example of a modification in oxidative phosphorylation yield by a change in mechanistic stoichiometry of one of the proton pumps involved. (3) In a model of polyunsaturated fatty acid deficiency

  14. Analysis of single pulse radio flux measurements of PSR B1133+16 at 4.85 and 8.35 GHz

    NASA Astrophysics Data System (ADS)

    Krzeszowski, K.; Maron, O.; Słowikowska, A.; Dyks, J.; Jessner, A.

    2014-05-01

    We show the results of microsecond resolution radio data analysis focused on flux measurements of single pulses of PSR B1133+16. The data were recorded at 4.85 and 8.35 GHz with 0.5- and 1.1-GHz bandwidth, respectively, using Radio Telescope Effelsberg (Max-Planck-Institut für Radioastronomie). The most important conclusion of the analysis is that the strongest single pulse emission at 4.85 and 8.35 GHz contributes almost exclusively to the trailing part of the leading component of the pulsar mean profile, whereas studies at lower frequencies report that the contribution is spread almost uniformly, covering all phases of the pulsar mean profile. We also estimate the radio emission heights to be around 1-2 per cent of the light-cylinder radius, which is in agreement with previous studies. Additionally, these observations allowed us to add two more measurements of the flux density to the PSR B1133+16 broad-band radio spectrum, covering frequencies from 16.7 MHz up to 32 GHz. We fit two different models to the spectrum: a broken power law and a spectrum based on the flicker-noise model, which represents the spectrum in a simpler, but similarly accurate, way.

  15. Fine-Scale Community Structure Analysis of ANME in Nyegga Sediments with High and Low Methane Flux

    PubMed Central

    Roalkvam, Irene; Dahle, Håkon; Chen, Yifeng; Jørgensen, Steffen Leth; Haflidason, Haflidi; Steen, Ida Helene

    2012-01-01

    To obtain knowledge on how regional variations in methane seepage rates influence the stratification, abundance, and diversity of anaerobic methanotrophs (ANME), we analyzed the vertical microbial stratification in a gravity core from a methane micro-seeping area at Nyegga by using 454-pyrosequencing of 16S rRNA gene tagged amplicons and quantitative PCR. These data were compared with previously obtained data from the more active G11 pockmark, characterized by higher methane flux. A down core stratification and high relative abundance of ANME were observed in both cores, with transition from an ANME-2a/b dominated community in low-sulfide and low methane horizons to ANME-1 dominance in horizons near the sulfate-methane transition zone. The stratification was over a wider spatial region and at greater depth in the core with lower methane flux, and the total 16S rRNA copy numbers were two orders of magnitude lower than in the sediments at G11 pockmark. A fine-scale view into the ANME communities at each location was achieved through operational taxonomical units (OTU) clustering of ANME-affiliated sequences. The majority of ANME-1 sequences from both sampling sites clustered within one OTU, while ANME-2a/b sequences were represented in unique OTUs. We suggest that free-living ANME-1 is the most abundant taxon in Nyegga cold seeps, and also the main consumer of methane. The observation of specific ANME-2a/b OTUs at each location could reflect that organisms within this clade are adapted to different geochemical settings, perhaps due to differences in methane affinity. Given that the ANME-2a/b population could be sustained in less active seepage areas, this subgroup could be potential seed populations in newly developed methane-enriched environments. PMID:22715336

  16. Comparison and analysis of empirical equations for soil heat flux for different cropping systems and irrigation methods

    USGS Publications Warehouse

    Irmak, A.; Singh, R.K.; Walter-Shea, Elizabeth; Verma, S.B.; Suyker, A.E.

    2011-01-01

    We evaluated the performance of four models for estimating soil heat flux density (G) in maize (Zea mays L.) and soybean (Glycine max L.) fields under different irrigation methods (center-pivot irrigated fields at Mead, Nebraska, and subsurface drip irrigated field at Clay Center, Nebraska) and rainfed conditions at Mead. The model estimates were compared against measurements made during growing seasons of 2003, 2004, and 2005 at Mead and during 2005, 2006, and 2007 at Clay Center. We observed a strong relationship between the G and net radiation (Rn) ratio (G/Rn) and the normalized difference vegetation index (NDVI). When a significant portion of the ground was bare soil, G/Rn ranged from 0.15 to 0.30 and decreased with increasing NDVI. In contrast to the NDVI progression, the G/Rn ratio decreased with crop growth and development. The G/Rn ratio for subsurface drip irrigated crops was smaller than for the center-pivot irrigated crops. The seasonal average G was 13.1%, 15.2%, 10.9%, and 12.8% of Rn for irrigated maize, rainfed maize, irrigated soybean, and rainfed soybean, respectively. Statistical analyses of the performance of the four models showed a wide range of variation in G estimation. The root mean square error (RMSE) of predictions ranged from 15 to 81.3 W m-2. Based on the wide range of RMSE, it is recommended that local calibration of the models should be carried out for remote estimation of soil heat flux.

  17. Pulse flux measuring device

    DOEpatents

    Riggan, William C.

    1985-01-01

    A device for measuring particle flux comprises first and second photodiode detectors for receiving flux from a source and first and second outputs for producing first and second signals representing the flux incident to the detectors. The device is capable of reducing the first output signal by a portion of the second output signal, thereby enhancing the accuracy of the device. Devices in accordance with the invention may measure distinct components of flux from a single source or fluxes from several sources.

  18. Analysis of riverine suspended particulate matter fluxes (Gulf of Lion, Mediterranean Sea) using a synergy of ocean color observations with a 3-D hydrodynamic sediment transport model

    NASA Astrophysics Data System (ADS)

    Le Fouest, Vincent; Chami, Malik; Verney, Romaric

    2015-02-01

    The export of riverine suspended particulate matter (SPM) in the coastal ocean has major implications for the biogeochemical cycles. In the Mediterranean Sea (France), the Rhone River inputs of SPM into the Gulf of Lion (GoL) are highly variable in time, which severely impedes the assessment of SPM fluxes. The objectives of this study are (i) to investigate the prediction of the land-to-ocean flux of SPM using the complementarity (i.e., synergy) between a hydrodynamic sediment transport model and satellite observations, and (ii) to analyze the spatial distribution of the SPM export. An original approach that combines the MARS-3D model with satellite ocean color data is proposed. Satellite-derived SPM and light penetration depth are used to initialize MARS-3D and to validate its predictions. A sensitivity analysis is performed to quantify the impact of riverine SPM size composition and settling rate on the horizontal export of SPM. The best agreement between the model and the satellite in terms of SPM spatial distribution and export is obtained for two conditions: (i) when the relative proportion of "heavy and fast" settling particles significantly increases relative to the "light and slow" ones, and (ii) when the settling rate of heavy and light SPM increases by fivefold. The synergy between MARS-3D and the satellite data improved the SPM flux predictions by 48% near the Rhone River mouth. Our results corroborate the importance of implementing satellite observations within initialization procedures of ocean models since data assimilation techniques may fail for river floods showing strong seasonal variability.

  19. A high continental weathering flux into Paleoarchean seawater revealed by strontium isotope analysis of 3.26 Ga barite

    NASA Astrophysics Data System (ADS)

    Satkoski, Aaron M.; Lowe, Donald R.; Beard, Brian L.; Coleman, Max L.; Johnson, Clark M.

    2016-11-01

    Controls on Archean seawater chemistry remain controversial. Many studies have suggested that it was largely controlled by oceanic hydrothermal fluid circulation. Recent work, however, from clastic sequences, Hf-O isotope data from detrital zircons, and models for the Rb/Sr evolution of the continental crust suggest that intense continental weathering and low-temperature surface alteration were more important than previously thought during the early Archean. This is consistent with biogeochemical studies that suggest the Archean had a diverse microbial ecology, which would, in part, need to be sustained by nutrients (e.g., phosphorus) that were derived from continental weathering. To further quantify continental weathering during the early Archean, we analyzed 3.26 Ga barite from the Fig Tree Group, South Africa for strontium, oxygen, and sulfur isotope compositions. We propose that the seawater component of the barite is characterized by 87Sr/86Sr ratios >0.701, which is significantly more radiogenic than contemporaneous mantle (∼0.7007-0.7008). The radiogenic nature of seawater at this time suggests that the continental weathering flux at 3.26 Ga had a large impact on ocean chemistry 400 million years earlier than previously suggested.

  20. Statistical analysis of short-term water stress conditions at Riggs Creek OzFlux tower site

    NASA Astrophysics Data System (ADS)

    Azmi, Mohammad; Rüdiger, Christoph; Walker, Jeffrey P.

    2016-08-01

    A large range of indices and proxies are available to describe the water stress conditions of an area subject to different applications, which have varying capabilities and limitations depending on the prevailing local climatic conditions and land cover. The present study uses a range of spatio-temporally high-resolution (daily and within daily) data sources to evaluate a number of drought indices (DIs) for the Riggs Creek OzFlux tower site in southeastern Australia. Therefore, the main aim of this study is to evaluate the statistical characteristics of individual DIs subject to short-term water stress conditions. In order to derive a more general and therefore representative DI, a new criterion is required to specify the statistical similarity between each pair of indices to allow determining the dominant drought types along with their representative DIs. The results show that the monitoring of water stress at this case study area can be achieved by evaluating the individual behaviour of three clusters of (i) vegetation conditions, (ii) water availability and (iii) water consumptions. This indicates that it is not necessary to assess all individual DIs one by one to derive a comprehensive and informative data set about the water stress of an area; instead, this can be achieved by analysing one of the DIs from each cluster or deriving a new combinatory index for each cluster, based on established combination methods.

  1. Is reduced benthic flux related to the Diporeia decline? Analysis of spring blooms and whiting events in Lake Ontario

    USGS Publications Warehouse

    Watkins, James M.; Rudstam, Lars G.; Crabtree, Darran L.; Walsh, Maureen

    2013-01-01

    Benthic monitoring by USGS off the southern shore of Lake Ontario from October 1993 to October 1995 provides a detailed view of the early stages of the decline of the native amphipod Diporeia. A loss of the 1994 and 1995 year classes of Diporeia preceded the disappearance of the native amphipod at sites near Oswego and Rochester at depths from 55 to 130 m. In succeeding years, Diporeia populations continued to decline in Lake Ontario and were nearly extirpated by 2008. Explanations for Diporeia 's decline in the Great Lakes include several hypotheses often linked to the introduction and expansion of exotic zebra and quagga mussels (Dreissena sp.). We compare the timeline of the Diporeia decline in Lake Ontario with trends in two sources of organic matter to the sediments — spring diatom blooms and late summer whiting events. The 1994–95 decline of Diporeia coincided with localized dreissenid effects on phytoplankton in the nearshore and a year (April 1994 to May 1995) of decreased flux of organic carbon recorded by sediment traps moored offshore of Oswego. Later declines of profundal (> 90 m) Diporeia populations in 2003 were poorly associated with trends in spring algal blooms and late summer whiting events. Lake Ontario/Diporeia/Dreissena/remote sensing.

  2. Metabolomic and flux-balance analysis of age-related decline of hypoxia tolerance in Drosophila muscle tissue

    PubMed Central

    Coquin, Laurence; Feala, Jacob D; McCulloch, Andrew D; Paternostro, Giovanni

    2008-01-01

    The fruitfly Drosophila melanogaster is increasingly used as a model organism for studying acute hypoxia tolerance and for studying aging, but the interactions between these two factors are not well known. Here we show that hypoxia tolerance degrades with age in post-hypoxic recovery of whole-body movement, heart rate and ATP content. We previously used 1H NMR metabolomics and a constraint-based model of ATP-generating metabolism to discover the end products of hypoxic metabolism in flies and generate hypotheses for the biological mechanisms. We expand the reactions in the model using tissue- and age-specific microarray data from the literature, and then examine metabolomic profiles of thoraxes after 4 h at 0.5% O2 and after 5 min of recovery in 40- versus 3-day-old flies. Model simulations were constrained to fluxes calculated from these data. Simulations suggest that the decreased ATP production during reoxygenation seen in aging flies can be attributed to reduced recovery of mitochondrial respiration pathways and concomitant overdependence on the acetate production pathway as an energy source. PMID:19096360

  3. The virtual observatory service TheoSSA: Establishing a database of synthetic stellar flux standards . II. NLTE spectral analysis of the OB-type subdwarf Feige 110

    NASA Astrophysics Data System (ADS)

    Rauch, T.; Rudkowski, A.; Kampka, D.; Werner, K.; Kruk, J. W.; Moehler, S.

    2014-06-01

    Context. In the framework of the Virtual Observatory (VO), the German Astrophysical VO (GAVO) developed the registered service TheoSSA (Theoretical Stellar Spectra Access). It provides easy access to stellar spectral energy distributions (SEDs) and is intended to ingest SEDs calculated by any model-atmosphere code, generally for all effective temperatures, surface gravities, and elemental compositions. We will establish a database of SEDs of flux standards that are easily accessible via TheoSSA's web interface. Aims: The OB-type subdwarf Feige 110 is a standard star for flux calibration. State-of-the-art non-local thermodynamic equilibrium stellar-atmosphere models that consider opacities of species up to trans-iron elements will be used to provide a reliable synthetic spectrum to compare with observations. Methods: In case of Feige 110, we demonstrate that the model reproduces not only its overall continuum shape from the far-ultraviolet (FUV) to the optical wavelength range but also the numerous metal lines exhibited in its FUV spectrum. Results: We present a state-of-the-art spectral analysis of Feige 110. We determined , log g = 6.00 ± 0.20, and the abundances of He, N, P, S, Ti, V, Cr, Mn, Fe, Co, Ni, Zn, and Ge. Ti, V, Mn, Co, Zn, and Ge were identified for the first time in this star. Upper abundance limits were derived for C, O, Si, Ca, and Sc. Conclusions: The TheoSSA database of theoretical SEDs of stellar flux standards guarantees that the flux calibration of astronomical data and cross-calibration between different instruments can be based on models and SEDs calculated with state-of-the-art model-atmosphere codes. Based on observations with the NASA/ESA Hubble Space Telescope, obtained at the Space Telescope Science Institute, which is operated by the Association of Universities for Research in Astronomy, Inc., under NASA contract NAS5-26666. Based on observations made with the NASA-CNES-CSA Far Ultraviolet Spectroscopic Explorer. Table 2, Figs. 3 and

  4. Numerical analysis of the heat flux characteristics in the helical scrape-off layer of the limiter start up configuration at Wendelstein 7-X

    NASA Astrophysics Data System (ADS)

    Effenberg, Florian; Feng, Y.; Schmitz, O.; Frerichs, H.; Barbui, T.; Geiger, J.; Jakubowski, M.; Koenig, R.; Krychowiak, M.; Niemann, H.; Pedersen, T. S.; Stephey, L.; Wurden, G.; W7-X Team

    2016-10-01

    A crucial topic for the stellarator W7-X is the power dissipation by impurities for future island divertor scenarios. The investigation of the related heat flux distribution and profiles including the radial power fall-off length λq in the 3D stellarator SOL is less straight forward as in toroidally symmetric tokamaks. Studies with the 3D plasma edge transport code EMC3-Eirene predicted a modulation of plasma parameters with LC and correlated heterogenous heat and particle loads onto the limiters during start-up operation. The relative simple start up geometry at W7-X allows for a detailed analysis of the heat fluxes in separate helical transport channels featuring different ∥ to ⊥ transport ratios. It is shown that the SOL has two characteristic fall off domains - a near SOL and a far SOL domain which both have different power decay lengths. An increase of λq with LC in the order of 1-1.5cm in the near SOL and 1.8-2.8cm in the far SOL for a power scan in the range of P = 0.5-2MW at nLCFS = 2 × 1018m-3 has been found. First comparisons with IR camera data will be discussed. This work was supported by the U.S. Department of Energy (DOE) under Grant DE-SC0014210 and by EUROfusion under Grant No 633053.

  5. Correlating two-photon excited fluorescence imaging of breast cancer cellular redox state with seahorse flux analysis of normalized cellular oxygen consumption

    NASA Astrophysics Data System (ADS)

    Hou, Jue; Wright, Heather J.; Chan, Nicole; Tran, Richard; Razorenova, Olga V.; Potma, Eric O.; Tromberg, Bruce J.

    2016-06-01

    Two-photon excited fluorescence (TPEF) imaging of the cellular cofactors nicotinamide adenine dinucleotide and oxidized flavin adenine dinucleotide is widely used to measure cellular metabolism, both in normal and pathological cells and tissues. When dual-wavelength excitation is used, ratiometric TPEF imaging of the intrinsic cofactor fluorescence provides a metabolic index of cells-the "optical redox ratio" (ORR). With increased interest in understanding and controlling cellular metabolism in cancer, there is a need to evaluate the performance of ORR in malignant cells. We compare TPEF metabolic imaging with seahorse flux analysis of cellular oxygen consumption in two different breast cancer cell lines (MCF-7 and MDA-MB-231). We monitor metabolic index in living cells under both normal culture conditions and, for MCF-7, in response to cell respiration inhibitors and uncouplers. We observe a significant correlation between the TPEF-derived ORR and the flux analyzer measurements (R=0.7901, p<0.001). Our results confirm that the ORR is a valid dynamic index of cell metabolism under a range of oxygen consumption conditions relevant for cancer imaging.

  6. Evaluation of neutron flux parameters in irradiation sites of research reactor using the Westcott-formalism for the k0 neutron activation analysis method

    NASA Astrophysics Data System (ADS)

    Kasban, H.; Hamid, Ashraf

    2015-12-01

    Instrumental Neutron Activation Analysis using k0 (k0-INAA) method has been used to determine a number of elements in sediment samples collected from El-Manzala Lake in Egypt. k0-INAA according to Westcott's formalism has been implemented using the complete irradiation kit of the fast pneumatic rabbit and some selected manually loaded irradiation sites for short and long irradiation at Egypt Second Research Reactor (ETRR-2). Zr-Au and Co sets as neutron flux monitors are used to determine the neutron flux parameters (f and α) in each irradiation sites. Two reference materials IAEA Soil-7 samples have been inserted and implemented for data validation and an internal monostandard multi monitor used (k0 based IM-NAA). It was given a good agreement between the experimental analyzed values and that obtained of the certified values. The major and trace elements in the sediment samples have been evaluated with the use of Co as an internal and Au as an external monostandard comparators. The concentrations of the elements (Cr, Mn and Zn) in the sediment samples of the present work are discussed regarding to those obtained from other sites.

  7. The Mimas ghost revisited - An analysis of the electron flux and electron microsignatures observed in the vicinity of Mimas at Saturn

    NASA Technical Reports Server (NTRS)

    Chenette, D. L.; Stone, E. C.

    1983-01-01

    An analysis of the electron-absorption signature observed by the cosmic-ray system on Voyager 2 near the orbit of Mimas is presented. It is found that these observations cannot be explained as the absorption signature of Mimas. By combining Pioneer 11 and Voyager 2 measurements of the electron flux at Mimas's orbit (L = 3.1), an electron spectrum is found in which most of the flux above about 100 keV is concentrated near 1 to 3 MeV. This spectral form is qualitatively consistent with the bandpass filter model of Van Allen et al. (1980). The expected Mimas absorption signature is calculated from this spectrum neglecting radial diffusion. Since no Mimas absorption signature was observed in the inbound Voyager 2 data, a lower limit on the diffusion coefficient for MeV electrons at L = 3.1 of D greater than 10 to the -8th sq Saturn radii/sec is obtained. With a diffusion coefficient this large, both the Voyager 2 and the Pioneer 11 small-scale electron-absorption-signature observations in Mimas's orbit are enigmatic. Thus the mechanism for producing these signatures is referred to as the Mimas ghost. A cloud of material in orbit with Mimas may account for the observed electron signature if the cloud is at least 1-percent opaque to electrons across a region extending over a few hundred kilometers.

  8. The Mimas ghost revisited: An analysis of the electron flux and electron microsignatures observed in the vicinity of Mimas at Saturn

    NASA Technical Reports Server (NTRS)

    Chenette, D. L.; Stone, E. C.

    1983-01-01

    An analysis of the electron absorption signature observed by the Cosmic Ray System (CRS) on Voyage 2 near the orbit of Mimas is presented. We find that these observations cannot be explained as the absorption signature of Mimas. Combing Pioneer 11 and Voyager 2 measurements of the electron flux at Mimas's orbit (L=3.1), we find an electron spectrum where most of the flux above approx 100 keV is concentrated near 1 to 3 MeV. The expected Mimas absorption signature is calculated from this spectrum neglecting radial diffusion. A lower limit on the diffusion coefficient for MeV electrons is obtained. With a diffusion coefficient this large, both the Voyager 2 and the Pioneer 11 small-scale electron absorption signature observations in Mimas's orbit are enigmatic. Thus we refer to the mechanism for producing these signatures as the Mimas ghost. A cloud of material in orbit with Mimas may account for the observed electron signature if the cloud is at least 1% opaque to electrons across a region extending over a few hundred kilometers.

  9. Metabolic control analysis of the penicillin biosynthetic pathway: the influence of the LLD-ACV:bisACV ratio on the flux control.

    PubMed

    Theilgaard, H A; Nielsen, J

    1999-01-01

    An extended kinetic model for the first two steps of the penicillin biosynthetic pathway in Penicillium chrysogenum is set up. It includes the formation and reduction of the dimer bis-delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (bisACV) from the first pathway intermediate LLD-ACV and their parallel inhibition of the enzyme ACV synthetase (ACVS). The kinetic model is based on Michaelis-Menten type kinetics, with non-competitive inhibition of the ACVS by both LLD-ACV and bisACV, and competitive inhibition of the isopenicillin N synthetase (IPNS) by glutathione. The inhibition constant of LLD-ACV, KACV is determined to be 0.54 mm. With the kinetic model metabolic control analysis is performed to identify the distribution of rate-control in the pathway at all ratios of LLD-ACV:bisACV. It is concluded that the flux control totally resides at the IPNS. This is a result of the regulation of the ACVS by both the LLD-ACV and bisACV demanding a higher flux through the IPNS enzyme to alleviate their inhibition. The measurement of an intracellular ratio of LLD-ACV:bisACV to be in the range of 1-2 moles per moles emphasises the importance of a fast conversion of LLD-ACV to IPN, and accumulation of LLD-ACV above the K(m)-value of the IPNS should therefore be avoided.

  10. Soil erosion and sediment fluxes analysis: a watershed study of the Ni Reservoir, Spotsylvania County, VA, USA.

    PubMed

    Pope, Ian C; Odhiambo, Ben K

    2014-03-01

    Anthropogenic forces that alter the physical landscape are known to cause significant soil erosion, which has negative impact on surface water bodies, such as rivers, lakes/reservoirs, and coastal zones, and thus sediment control has become one of the central aspects of catchment management planning. The revised universal soil loss equation empirical model, erosion pins, and isotopic sediment core analyses were used to evaluate watershed erosion, stream bank erosion, and reservoir sediment accumulation rates for Ni Reservoir, in central Virginia. Land-use and land cover seems to be dominant control in watershed soil erosion, with barren land and human-disturbed areas contributing the most sediment, and forest and herbaceous areas contributing the least. Results show a 7 % increase in human development from 2001 (14 %) to 2009 (21.6 %), corresponding to an increase in soil loss of 0.82 Mg ha(-1) year(-1) in the same time period. (210)Pb-based sediment accumulation rates at three locations in Ni Reservoir were 1.020, 0.364, and 0.543 g cm(-2) year(-1) respectively, indicating that sediment accumulation and distribution in the reservoir is influenced by reservoir configuration and significant contributions from bedload. All three locations indicate an increase in modern sediment accumulation rates. Erosion pin results show variability in stream bank erosion with values ranging from 4.7 to 11.3 cm year(-1). These results indicate that urban growth and the decline in vegetative cover has increased sediment fluxes from the watershed and poses a significant threat to the long-term sustainability of the Ni Reservoir as urbanization continues to increase.

  11. Fast flux module detection using matroid theory.

    PubMed

    Reimers, Arne C; Bruggeman, Frank J; Olivier, Brett G; Stougie, Leen

    2015-05-01

    Flux balance analysis (FBA) is one of the most often applied methods on genome-scale metabolic networks. Although FBA uniquely determines the optimal yield, the pathway that achieves this is usually not unique. The analysis of the optimal-yield flux space has been an open challenge. Flux variability analysis is only capturing some properties of the flux space, while elementary mode analysis is intractable due to the enormous number of elementary modes. However, it has been found by Kelk et al. (2012) that the space of optimal-yield fluxes decomposes into flux modules. These decompositions allow a much easier but still comprehensive analysis of the optimal-yield flux space. Using the mathematical definition of module introduced by Müller and Bockmayr (2013b), we discovered useful connections to matroid theory, through which efficient algorithms enable us to compute the decomposition into modules in a few seconds for genome-scale networks. Using that every module can be represented by one reaction that represents its function, in this article, we also present a method that uses this decomposition to visualize the interplay of modules. We expect the new method to replace flux variability analysis in the pipelines for metabolic networks.

  12. IMFIT Integrated Modeling Applications Supporting Experimental Analysis: Multiple Time-Slice Kinetic EFIT Reconstructions, MHD Stability Limits, and Energy and Momentum Flux Analyses

    NASA Astrophysics Data System (ADS)

    Collier, A.; Lao, L. L.; Abla, G.; Chu, M. S.; Prater, R.; Smith, S. P.; St. John, H. E.; Guo, W.; Li, G.; Pan, C.; Ren, Q.; Park, J. M.; Bisai, N.; Srinivasan, R.; Sun, A. P.; Liu, Y.; Worrall, M.

    2010-11-01

    This presentation summarizes several useful applications provided by the IMFIT integrated modeling framework to support DIII-D and EAST research. IMFIT is based on Python and utilizes modular task-flow architecture with a central manager and extensive GUI support to coordinate tasks among component modules. The kinetic-EFIT application allows multiple time-slice reconstructions by fetching pressure profile data directly from MDS+ or from ONETWO or PTRANSP. The stability application analyzes a given reference equilibrium for stability limits by performing parameter perturbation studies with MHD codes such as DCON, GATO, ELITE, or PEST3. The transport task includes construction of experimental energy and momentum fluxes from profile analysis and comparison against theoretical models such as MMM95, GLF23, or TGLF.

  13. Numerical analysis for radial MHD and mixed convection effects in peristalsis of non-Newtonian nanomaterial with zero mass flux conditions

    NASA Astrophysics Data System (ADS)

    Hayat, T.; Farooq, S.; Alsaedi, A.; Ahmad, B.

    Here peristalsis of Carreau nanofluid in presence of mixed convection and radial magnetohydrodynamics is addressed. Flow in a curved channel is addressed. Heat and mass transfer analysis includes Brownian motion and thermophoresis impacts. Zero mass flux at the channel walls is taken. The relevant equations are first modelled and then simplified through lubrication technique. The system of non-linear equations is solved numerically. Plots for velocity, temperature and concentration are studied through numerical solution. Heat and mass transfer rates at the upper wall of the curved channel is also discussed. Enhancement in concentration is observed. Temperature of the nanoliquid enhances for larger radial magnetic parameter when velocity of the nanoliquids reduces. Significant enhancement in mass transfer rate is seen for larger theromophoretic parameter.

  14. Analysis of energy fluxes and vegetation-atmosphere parameters in irrigated and natural ecosystems of semi-arid Brazil

    NASA Astrophysics Data System (ADS)

    Teixeira, A. H. de Castro; Bastiaanssen, W. G. M.; Ahmad, M. D.; Moura, M. S. B.; Bos, M. G.

    2008-11-01

    SummaryKnowledge on evapotranspiration is essential in quantifying water use depletion and to allocate scarce water resources to competing uses. Despite that an extensive literature describes the theoretical mechanisms of turbulent water vapour transport above and within crop canopies fewer studies have examined land surface parameters within composite landscapes of irrigated crops and semi-arid natural vegetation. Aiming to improve parameterizations of the radiation and energy balance in irrigated crops and natural vegetation, micro-climatic measurements were carried out on irrigated land (vineyards and mango orchard) and natural vegetation (caatinga) in the semi-arid zone of the São Francisco River basin (Brazil) from 2002 to 2005. The fractions of 24 h incident solar radiation available for net radiation were 46%, 55%, 51% and 53%, for wine grape, table grape, mango orchard and caatinga, respectively. Daily evaporative fractions of the net available energy used as latent heat flux ( λE) were 0.80, 0.88, 0.75 and 0.33 respectively. The daylight values of bulk surface resistances ( rs) averaged 128 s m -1, 73 s m -1, 133 s m -1 and 1940 s m -1 for wine grape, table grape, mango orchard and caatinga, respectively. Simplified parameterizations on roughness and evaporation resistances were performed. It could be concluded that net radiation can be estimated by means of a linear expression with incident global solar radiation depending on the type of vegetation. The variability of aerodynamic resistance ( ra) could be mainly explained by the friction velocity ( u ∗) which on turn depends on the surface roughness length for momentum transport ( z 0m). The experimental data showed that for sparse canopies z 0m being 9% of the mean vegetation height is a doable operational rule for the semi-arid region of São Francisco River basin. The seasonal values of rs for irrigated crops were highly correlated with water vapour pressure deficit. The availability of analytical

  15. Effect of amino acid supplementation on titer and glycosylation distribution in hybridoma cell cultures-Systems biology-based interpretation using genome-scale metabolic flux balance model and multivariate data analysis.

    PubMed

    Reimonn, Thomas M; Park, Seo-Young; Agarabi, Cyrus D; Brorson, Kurt A; Yoon, Seongkyu

    2016-09-01

    Genome-scale flux balance analysis (FBA) is a powerful systems biology tool to characterize intracellular reaction fluxes during cell cultures. FBA estimates intracellular reaction rates by optimizing an objective function, subject to the constraints of a metabolic model and media uptake/excretion rates. A dynamic extension to FBA, dynamic flux balance analysis (DFBA), can calculate intracellular reaction fluxes as they change during cell cultures. In a previous study by Read et al. (2013), a series of informed amino acid supplementation experiments were performed on twelve parallel murine hybridoma cell cultures, and this data was leveraged for further analysis (Read et al., Biotechnol Prog. 2013;29:745-753). In order to understand the effects of media changes on the model murine hybridoma cell line, a systems biology approach is applied in the current study. Dynamic flux balance analysis was performed using a genome-scale mouse metabolic model, and multivariate data analysis was used for interpretation. The calculated reaction fluxes were examined using partial least squares and partial least squares discriminant analysis. The results indicate media supplementation increases product yield because it raises nutrient levels extending the growth phase, and the increased cell density allows for greater culture performance. At the same time, the directed supplementation does not change the overall metabolism of the cells. This supports the conclusion that product quality, as measured by glycoform assays, remains unchanged because the metabolism remains in a similar state. Additionally, the DFBA shows that metabolic state varies more at the beginning of the culture but less by the middle of the growth phase, possibly due to stress on the cells during inoculation. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1163-1173, 2016.

  16. FLUXES FOR MECHANIZED ELECTRIC WELDING,

    DTIC Science & Technology

    WELDING FLUXES, WELDING ), (* WELDING , WELDING FLUXES), ARC WELDING , WELDS, STABILITY, POROSITY, WELDING RODS, STEEL, CERAMIC MATERIALS, FLUXES(FUSION), TITANIUM ALLOYS, ALUMINUM ALLOYS, COPPER ALLOYS, ELECTRODEPOSITION

  17. Comparative Analysis of Genomics and Proteomics in the New Isolated Bacillus thuringiensis X022 Revealed the Metabolic Regulation Mechanism of Carbon Flux Following Cu(2+) Treatment.

    PubMed

    Quan, Meifang; Xie, Junyan; Liu, Xuemei; Li, Yang; Rang, Jie; Zhang, Tong; Zhou, Fengjuan; Xia, Liqiu; Hu, Shengbiao; Sun, Yunjun; Ding, Xuezhi

    2016-01-01

    Bacillus thuringiensis (Bt) X022 is a novel strain isolated from soil in China, and showed strong insecticidal activity against several Lepidopteran pests. In this work, we performed whole genome sequencing of this Bt strain using the next-generation sequencing technology, and further conducted a comparative analysis with the proteomics data of the specific spore-release period based on LC-MS/MS approach. The Bt X022 genome consisted of one circular chromosomal DNA and seven plasmids, which were further functionally annotated using the RAST server. Comparative analysis of insecticidal substances showed that X022 contained genes coding for three Cry proteins (Cry1Ac, Cry1Ia and Cry2Ab) and a vegetative insecticidal protein (Vip3A). However, three insecticidal crystal proteins (ICPs) (Cry1Ca, Cry1Ac and Cry1Da) were detected by proteomics in the spore-release period. Moreover, a putative biosynthetic gene cluster and the metabolic pathway for poly-β-hydroxybutyrate in Bt X022 were deduced based on the comparative analysis of genomic and proteomic data, which revealed the metabolic regulation mechanism of carbon flux correlated with increased production of ICPs caused by Cu(2+.) Hence, these results provided a deeper understanding of the genetic background and protein expression profile of Bt X022. This study established a foundation for directed genetic modification and further application of this new isolated Bt strain.

  18. Comparative Analysis of Genomics and Proteomics in the New Isolated Bacillus thuringiensis X022 Revealed the Metabolic Regulation Mechanism of Carbon Flux Following Cu2+ Treatment

    PubMed Central

    Quan, Meifang; Xie, Junyan; Liu, Xuemei; Li, Yang; Rang, Jie; Zhang, Tong; Zhou, Fengjuan; Xia, Liqiu; Hu, Shengbiao; Sun, Yunjun; Ding, Xuezhi

    2016-01-01

    Bacillus thuringiensis (Bt) X022 is a novel strain isolated from soil in China, and showed strong insecticidal activity against several Lepidopteran pests. In this work, we performed whole genome sequencing of this Bt strain using the next-generation sequencing technology, and further conducted a comparative analysis with the proteomics data of the specific spore-release period based on LC-MS/MS approach. The Bt X022 genome consisted of one circular chromosomal DNA and seven plasmids, which were further functionally annotated using the RAST server. Comparative analysis of insecticidal substances showed that X022 contained genes coding for three Cry proteins (Cry1Ac, Cry1Ia and Cry2Ab) and a vegetative insecticidal protein (Vip3A). However, three insecticidal crystal proteins (ICPs) (Cry1Ca, Cry1Ac and Cry1Da) were detected by proteomics in the spore-release period. Moreover, a putative biosynthetic gene cluster and the metabolic pathway for poly-β-hydroxybutyrate in Bt X022 were deduced based on the comparative analysis of genomic and proteomic data, which revealed the metabolic regulation mechanism of carbon flux correlated with increased production of ICPs caused by Cu2+. Hence, these results provided a deeper understanding of the genetic background and protein expression profile of Bt X022. This study established a foundation for directed genetic modification and further application of this new isolated Bt strain. PMID:27303381

  19. Concentration, flux, and the analysis of trends of total and dissolved phosphorus, total nitrogen, and chloride in 18 tributaries to Lake Champlain, Vermont and New York, 1990–2011

    USGS Publications Warehouse

    Medalie, Laura

    2013-01-01

    Annual concentration, flux, and yield for total phosphorus, dissolved phosphorus, total nitrogen, and chloride for 18 tributaries to Lake Champlain were estimated for 1990 through 2011 using a weighted regression method based on time, tributary streamflows (discharges), and seasonal factors. The weighted regression method generated two series of daily estimates of flux and concentration during the period of record: one based on observed discharges and a second based on a flow-normalization procedure that removes random variation due to year-to-year climate-driven effects. The flownormalized estimate for a given date is similar to an average estimate of concentration or flux that would be made if all of the observed discharges for that date were equally likely to have occurred. The flux bias statistic showed that 68 of the 72 flux regression models were minimally biased. Temporal trends in the concentrations and fluxes were determined by calculating percent changes in flow-normalized annual fluxes for the full period of analysis (1990 through 2010) and for the decades 1990–2000 and 2000–2010. Basinwide, flow-normalized total phosphorus