875 resultados para Metabolic flux analysis
Resumo:
Bangalore is experiencing unprecedented urbanisation in recent times due to concentrated developmental activities with impetus on IT (Information Technology) and BT (Biotechnology) sectors. The concentrated developmental activities has resulted in the increase in population and consequent pressure on infrastructure, natural resources, ultimately giving rise to a plethora of serious challenges such as urban flooding, climate change, etc. One of the perceived impact at local levels is the increase in sensible heat flux from the land surface to the atmosphere, which is also referred as heat island effect. In this communication, we report the changes in land surface temperature (LST) with respect to land cover changes during 1973 to 2007. A novel technique combining the information from sub-pixel class proportions with information from classified image (using signatures of the respective classes collected from the ground) has been used to achieve more reliable classification. The analysis showed positive correlation with the increase in paved surfaces and LST. 466% increase in paved surfaces (buildings, roads, etc.) has lead to the increase in LST by about 2 ºC during the last 2 decades, confirming urban heat island phenomenon. LSTs’ were relatively lower (~ 4 to 7 ºC) at land uses such as vegetation (parks/forests) and water bodies which act as heat sinks.
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In the present work, we report the growth of wurtzite InN epilayers on GaN/Si (1 1 1) substrate by plasma-assisted molecular beam epitaxy (PAMBE). The growth parameters such as indium flux, substrate temperature and RF power affect the crystallographic and morphological properties of InN layers, which were evaluated using high resolution X-ray diffraction (HRXRD) analysis and atomic force microscopy (AFM). It is found that excess indium (In) concentrations and surface roughness were increased with increase in In flux and growth temperature. The intensity of HRXRD (0 0 0 2) peak, corresponding to c-axis orientation has been increased and full width at half maxima (FWHM) has decreased with increase in RF power. It was found that highly c-axis oriented InN epilayers can be grown at 450 degrees C growth temperature, 450 W RF power and 1.30 x 10(-7) mbar In beam equivalent pressure (BEP). The energy gap of InN layers grown by optimizing growth conditions was determined by photoluminescence and optical absorption measurement. (C) 2011 Elsevier B.V. All rights reserved.
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Plant oils are stored in oleosomes or oil bodies, which are surrounded by a monolayer of phospholipids embedded with oleosin proteins that stabilize the structure. Recently, a structural protein, Oleosin3 (OLE3), was shown to exhibit both monoacylglycerol acyltransferase and phospholipase A(2) activities. The regulation of these distinct dual activities in a single protein is unclear. Here, we report that a serine/threonine/tyrosine protein kinase phosphorylates oleosin. Using bimolecular fluorescence complementation analysis, we demonstrate that this kinase interacts with OLE3 and that the fluorescence was associated with chloroplasts. Oleosin-green fluorescent protein fusion protein was exclusively associated with the chloroplasts. Phosphorylated OLE3 exhibited reduced monoacylglycerol acyltransferase and increased phospholipase A(2) activities. Moreover, phosphatidylcholine and diacylglycerol activated oleosin phosphorylation, whereas lysophosphatidylcholine, oleic acid, and Ca2+ inhibited phosphorylation. In addition, recombinant peanut (Arachis hypogaea) kinase was determined to predominantly phosphorylate serine residues, specifically serine-18 in OLE3. Phosphorylation levels of OLE3 during seed germination were determined to be higher than in developing peanut seeds. These findings provide direct evidence for the in vivo substrate selectivity of the dual-specificity kinase and demonstrate that the bifunctional activities of oleosin are regulated by phosphorylation.
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This letter develops theoretical relationships for equilibrium timescale and temperature scale of a vaporizing droplet in a convective and a radiative environment. The transient temperature normalized by the respective scales exhibits a unified profile for both modes of heating. The analysis allows for the prediction of the required laser flux to show its equivalence in a corresponding heated gas stream. The theoretical equivalence shows good agreement with experiments across a range of droplet sizes. Simple experiments can be conducted in a levitator to extrapolate information in realistic convective environments like combustion and spray drying. (C) 2012 American Institute of Physics. http://dx.doi.org/10.1063/1.4720092]
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Lipopolysaccharide (LPS) is an endotoxin, a potent stimulator of immune response and induction of LPS leads to acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). ARDS is a life-threatening disease worldwide with a high mortality rate. The immunological effect of LPS with spleen and thymus is well documented; however the impact on membrane phospholipid during endotoxemia has not yet been studied. Hence we aimed to investigate the influence of LPS on spleen and thymus phospholipid and fatty acid composition by 32P]orthophosphate labeling in rats. The in vitro labeling was carried out with phosphate-free medium (saline). Time course, LPS concentration-dependent, pre- and post-labeling with LPS and fatty acid analysis of phospholipid were performed. Labeling studies showed that 50 mu g LPS specifically altered the major phospholipids, phosphatidylcholine and phosphatidylglycerol in spleen and phosphatidylcholine in thymus. Fatty acid analysis showed a marked alteration of unsaturated fatty acids/saturated fatty acids in spleen and thymus leading to immune impairment via the fatty acid remodeling pathway. Our present in vitro lipid metabolic labeling study could open up new vistas for exploring LPS-induced immune impairment in spleen and thymus, as well as the underlying mechanism.
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Background: The set of indispensable genes that are required by an organism to grow and sustain life are termed as essential genes. There is a strong interest in identification of the set of essential genes, particularly in pathogens, not only for a better understanding of the pathogen biology, but also for identifying drug targets and the minimal gene set for the organism. Essentiality is inherently a systems property and requires consideration of the system as a whole for their identification. The available experimental approaches capture some aspects but each method comes with its own limitations. Moreover, they do not explain the basis for essentiality in most cases. A powerful prediction method to recognize this gene pool including rationalization of the known essential genes in a given organism would be very useful. Here we describe a multi-level multi-scale approach to identify the essential gene pool in a deadly pathogen, Mycobacterium tuberculosis. Results: The multi-level workflow analyses the bacterial cell by studying (a) genome-wide gene expression profiles to identify the set of genes which show consistent and significant levels of expression in multiple samples of the same condition, (b) indispensability for growth by using gene expression integrated flux balance analysis of a genome-scale metabolic model, (c) importance for maintaining the integrity and flow in a protein-protein interaction network and (d) evolutionary conservation in a set of genomes of the same ecological niche. In the gene pool identified, the functional basis for essentiality has been addressed by studying residue level conservation and the sub-structure at the ligand binding pockets, from which essential amino acid residues in that pocket have also been identified. 283 genes were identified as essential genes with high-confidence. An agreement of about 73.5% is observed with that obtained from the experimental transposon mutagenesis technique. A large proportion of the identified genes belong to the class of intermediary metabolism and respiration. Conclusions: The multi-scale, multi-level approach described can be generally applied to other pathogens as well. The essential gene pool identified form a basis for designing experiments to probe their finer functional roles and also serve as a ready shortlist for identifying drug targets.
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Experiments involving heating of liquid droplets which are acoustically levitated, reveal specific modes of oscillations. For a given radiation flux, certain fluid droplets undergo distortion leading to catastrophic bag type breakup. The voltage of the acoustic levitator has been kept constant to operate at a nominal acoustic pressure intensity, throughout the experiments. Thus the droplet shape instabilities are primarily a consequence of droplet heating through vapor pressure, surface tension and viscosity. A novel approach is used by employing Legendre polynomials for the mode shape approximation to describe the thermally induced instabilities. The two dominant Legendre modes essentially reflect (a) the droplet size reduction due to evaporation, and (b) the deformation around the equilibrium shape. Dissipation and inter-coupling of modal energy lead to stable droplet shape while accumulation of the same ultimately results in droplet breakup. (C) 2013 Elsevier B.V. All rights reserved.
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The paper analyses the results of experiments on the propagation rate in a fuel bed under gasification conditions in a co-current reactor configuration. Experiments using wood chips with different values of moisture content have been conducted under gasification conditions. The influence of air mass flux on the propagation rate, peak temperature and gas quality is investigated. It is observed from the experiments that the flame front propagation rate initially increases as the air mass flux increased, reaching a peak propagation rate, and further increase in the air mass flux results in a decrease in the propagation rate. However, the bed movement increases with the increase in air mass flux. The experimental results provide an understanding on influence of the fuel properties on propagation front. The surface area per unit volume of the particles in the packed bed plays an important role in the propagation rate. It has been argued that the flaming pyrolysis contributes towards the flame propagation as opposed to the overall combustion process in a packed bed. The calorific value of the producer gas generated is nearly the same over the entire range of air mass flux for bone-dry and 10% moist wood. (C) 2014 Elsevier B.V. All rights reserved.
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The present study combines field and satellite observations to investigate how hydrographical transformations influence phytoplankton size structure in the southern Bay of Bengal during the peak Southwest Monsoon/Summer Monsoon (July-August). The intrusion of the Summer Monsoon Current (SMC) into the Bay of Bengal and associated changes in sea surface chemistry, traceable eastward up to 90 degrees E along 8 degrees N, seems to influence biology of the region significantly. Both in situ and satellite (MODIS) data revealed low surface chlorophyll except in the area influenced by the SMC During the study period, two well-developed cydonic eddies (north) and an anti-cyclonic eddy (south), closely linked to the main eastward flow of the SMC, were sampled. Considering the capping effect of the low-saline surface water that is characteristic of the Bay of Bengal, the impact of the cyclonic eddy, estimated in terms of enhanced nutrients and chlorophyll, was mostly restricted to the subsurface waters (below 20 m depth). Conversely, the anti-cyclonic eddy aided by the SMC was characterized by considerably higher nutrient concentration and chlorophyll in the upper water column (upper 60 m), which was contrary to the general characteristic of such eddies. Albeit smaller phytoplankton predominated the southern Bay of Bengal (60-95% of the total chlorophyll), the contribution of large phytoplankton was double in the regions influenced by the SMC and associated eddies. Multivariate analysis revealed the extent to which SMC-associated eddies spatially influence phytoplankton community structure. The study presents the first direct quantification of the size structure of phytoplankton from the southern Bay of Bengal and demonstrates that the SMC-associated hydrographical ramifications significantly increase the phytoplankton biomass contributed by larger phytoplankton and thereby influence the vertical opal and organic carbon flux in the region. (C) 2014 Elsevier B.V. All rights reserved.
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In cells, N-10-formyltetrahydrofolate (N-10-fTHF) is required for formylation of eubacterial/organellar initiator tRNA and purine nucleotide biosynthesis. Biosynthesis of N-10-fTHF is catalyzed by 5,10-methylene-tetrahydrofolate dehydrogenase/cyclohydrolase (FolD) and/or 10-formyltetrahydrofolate synthetase (Fhs). All eubacteria possess FolD, but some possess both FolD and Fhs. However, the reasons for possessing Fhs in addition to FolD have remained unclear. We used Escherichia coli, which naturally lacks fhs, as our model. We show that in E. coli, the essential function of folD could be replaced by Clostridium perfringens fhs when it was provided on a medium-copy-number plasmid or integrated as a single-copy gene in the chromosome. The fhs-supported folD deletion (Delta folD) strains grow well in a complex medium. However, these strains require purines and glycine as supplements for growth in M9 minimal medium. The in vivo levels of N-10-fTHF in the Delta folD strain (supported by plasmid-borne fhs) were limiting despite the high capacity of the available Fhs to synthesize N-10-fTHF in vitro. Auxotrophy for purines could be alleviated by supplementing formate to the medium, and that for glycine was alleviated by engineering THF import into the cells. The Delta folD strain (harboring fhs on the chromosome) showed a high NADP(+)-to-NADPH ratio and hypersensitivity to trimethoprim. The presence of fhs in E. coli was disadvantageous for its aerobic growth. However, under hypoxia, E. coli strains harboring fhs outcompeted those lacking it. The computational analysis revealed a predominant natural occurrence of fhs in anaerobic and facultative anaerobic bacteria.
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The transcriptional regulation of gene expression is orchestrated by complex networks of interacting genes. Increasing evidence indicates that these `transcriptional regulatory networks' (TRNs) in bacteria have an inherently hierarchical architecture, although the design principles and the specific advantages offered by this type of organization have not yet been fully elucidated. In this study, we focussed on the hierarchical structure of the TRN of the gram-positive bacterium Bacillus subtilis and performed a comparative analysis with the TRN of the gram-negative bacterium Escherichia coli. Using a graph-theoretic approach, we organized the transcription factors (TFs) and sigma-factors in the TRNs of B. subtilis and E. coli into three hierarchical levels (Top, Middle and Bottom) and studied several structural and functional properties across them. In addition to many similarities, we found also specific differences, explaining the majority of them with variations in the distribution of s-factors across the hierarchical levels in the two organisms. We then investigated the control of target metabolic genes by transcriptional regulators to characterize the differential regulation of three distinct metabolic subsystems (catabolism, anabolism and central energy metabolism). These results suggest that the hierarchical architecture that we observed in B. subtilis represents an effective organization of its TRN to achieve flexibility in response to a wide range of diverse stimuli.
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Phase-change cooling technique is a suitable method for thermal management of electronic equipment subjected to transient or cyclic heat loads. The thermal performance of a phase-change based heat sink under cyclic heat load depends on several design parameters, namely, applied heat flux, cooling heat transfer coefficient, thermophysical properties of phase-change materials (PCMs), and physical dimensions of phase-change storage system during melting and freezing processes. A one-dimensional conduction heat transfer model is formulated to evaluate the effectiveness of preliminary design of practical PCM-based energy storage units. In this model, the phase-change process of the PCM is divided into melting and solidification subprocesses, for which separate equations are written. The equations are solved sequentially and an explicit closed-form solution is obtained. The efficacy of analytical model is estimated by comparing with a finite-volume-based numerical solution for both transient and cyclic heat loads.
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Hydrogen, either in pure form or as a gaseous fuel mixture specie enhances the fuel conversion efficiency and reduce emissions in an internal combustion engine. This is due to the reduction in combustion duration attributed to higher laminar flame speeds. Hydrogen is also expected to increase the engine convective heat flux, attributed (directly or indirectly) to parameters like higher adiabatic flame temperature, laminar flame speed, thermal conductivity and diffusivity and lower flame quenching distance. These factors (adversely) affect the thermo-kinematic response and offset some of the benefits. The current work addresses the influence of mixture hydrogen fraction in syngas on the engine energy balance and the thermo-kinematic response for close to stoichiometric operating conditions. Four different bio-derived syngas compositions with fuel calorific value varying from 3.14 MJ/kg to 7.55 MJ/kg and air fuel mixture hydrogen fraction varying from 7.1% to 14.2% by volume are used. The analysis comprises of (a) use of chemical kinetics simulation package CHEMKIN for quantifying the thermo-physical properties (b) 0-D model for engine in-cylinder analysis and (c) in-cylinder investigations on a two-cylinder engine in open loop cooling mode for quantifying the thermo-kinematic response and engine energy balance. With lower adiabatic flame temperature for Syngas, the in-cylinder heat transfer analysis suggests that temperature has little effect in terms of increasing the heat flux. For typical engine like conditions (700 K and 25 bar at CR of 10), the laminar flame speed for syngas exceeds that of methane (55.5 cm/s) beyond mixture hydrogen fraction of 11% and is attributed to the increase in H based radicals. This leads to a reduction in the effective Lewis number and laminar flame thickness, potentially inducing flame instability and cellularity. Use of a thermodynamic model to assess the isolated influence of thermal conductivity and diffusivity on heat flux suggests an increase in the peak heat flux between 2% and 15% for the lowest (0.420 MW/m(2)) and highest (0.480 MW/m(2)) hydrogen containing syngas over methane (0.415 MW/m(2)) fueled operation. Experimental investigations indicate the engine cooling load for syngas fueled engine is higher by about 7% and 12% as compared to methane fueled operation; the losses are seen to increase with increasing mixture hydrogen fraction. Increase in the gas to electricity efficiency is observed from 18% to 24% as the mixture hydrogen fraction increases from 7.1% to 9.5%. Further increase in mixture hydrogen fraction to 14.2% results in the reduction of efficiency to 23%; argued due to the changes in the initial and terminal stages of combustion. On doubling of mixture hydrogen fraction, the flame kernel development and fast burn phase duration decrease by about 7% and 10% respectively and the terminal combustion duration, corresponding to 90%-98% mass burn, increases by about 23%. This increase in combustion duration arises from the cooling of the near wall mixture in the boundary layer attributed to the presence of hydrogen. The enhancement in engine cooling load and subsequent reduction in the brake thermal efficiency with increasing hydrogen fraction is evident from the engine energy balance along with the cumulative heat release profiles. Copyright (C) 2015, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.
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We report the diffusion characteristics of water vapor through two different porous media, viz., membrane electrode assembly (MEA) and gas diffusion layer (GDL) in a nonoperational fuel cell. Tunable diode laser absorption spectroscopy (TDLAS) was employed for measuring water vapor concentration in the test channel. Effects of the membrane pore size and the inlet humidity on the water vapor transport are quantified through mass flux and diffusion coefficient. Water vapor transport rate is found to be higher for GDL than for MEA. The flexibility and wide range of application of TDLAS in a fuel cell setup is demonstrated through experiments with a stagnant flow field on the dry side.
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Identifying cellular processes in terms of metabolic pathways is one of the avowed goals of metabolomics studies. Currently, this is done after relevant metabolites are identified to allow their mapping onto specific pathways. This task is daunting due to the complex nature of cellular processes and the difficulty in establishing the identity of individual metabolites. We propose here a new method: ChemSMP (Chemical Shifts to Metabolic Pathways), which facilitates rapid analysis by identifying the active metabolic pathways directly from chemical shifts obtained from a single two-dimensional (2D) C-13-H-1] correlation NMR spectrum without the need for identification and assignment of individual metabolites. ChemSMP uses a novel indexing and scoring system comprised of a ``uniqueness score'' and a ``coverage score''. Our method is demonstrated on metabolic pathways data from the Small Molecule Pathway Database (SMPDB) and chemical shifts from the Human Metabolome Database (HMDB). Benchmarks show that ChemSMP has a positive prediction rate of >90% in the presence of deduttered data and can sustain the same at 60-70% even in the presence of noise, such as deletions of peaks and chemical shift deviations. The method tested on NMR data acquired for a mixture of 20 amino acids shows a success rate of 93% in correct recovery of pathways. When used on data obtained from the cell lysate of an unexplored oncogenic cell line, it revealed active metabolic pathways responsible for regulating energy homeostasis of cancer cells. Our unique tool is thus expected to significantly enhance analysis of NMIR-based metabolomics data by reducing existing impediments.