Search results for: metabolite fluxes

Authors: Janneth Gonzalez, Andrés Pinzon Velasco, Maria Angarita

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Astrocytes play an important role in various processes in the brain, including pathological conditions such as neurodegenerative diseases. Recent studies have shown that the increase in saturated fatty acids such as palmitic acid (PA) triggers pro-inflammatorypathways in the brain. The use of synthetic neurosteroids such as tibolone has demonstrated neuro-protective mechanisms. However, broad studies with a systemic point of view on the neurodegenerative role of PA and the neuro-protective mechanisms of tibolone are lacking. In this study, we performed the integration of multi-omic data (transcriptome and proteome) into a human astrocyte genomic scale metabolic model to study the astrocytic response during palmitate treatment. We evaluated metabolic fluxes in three scenarios (healthy, induced inflammation by PA, and tibolone treatment under PA inflammation). We also applied a control theory approach to identify those reactions that exert more control in the astrocytic system. Our results suggest that PA generates a modulation of central and secondary metabolism, showing a switch in energy source use through inhibition of folate cycle and fatty acid β‐oxidation and upregulation of ketone bodies formation. We found 25 metabolic switches under PA‐mediated cellular regulation, 9 of which were critical only in the inflammatory scenario but not in the protective tibolone one. Within these reactions, inhibitory, total, and directional coupling profiles were key findings, playing a fundamental role in the (de)regulation of metabolic pathways that may increase neurotoxicity and represent potential treatment targets. Finally, the overall framework of our approach facilitates the understanding of complex metabolic regulation, and it can be used for in silico exploration of the mechanisms of astrocytic cell regulation, directing a more complex future experimental work in neurodegenerative diseases.

Keywords: astrocytes, data integration, palmitic acid, computational model, multi-omics

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Authors: Mohamed A. Etman, Mona G. Abd-Elnasser, Mohamed A. Shams-Eldin, Aly H. Nada

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Orally disintegrating tablets (ODTs) are gaining importance as new drug delivery systems and emerged as one of the popular and widely accepted dosage forms, especially for the pediatric and geriatric patients. Their advantages such as administration without water, anywhere, anytime lead to their suitability to geriatric and pediatric patients. They are also suitable for the mentally ill, the bed-ridden and patients who do not have easy access to water. The benefits, in terms of patient compliance, rapid onset of action, increased bioavailability, and good stability make these tablets popular as a dosage form of choice in the current market. These dosage forms dissolve or disintegrate in the oral cavity within a matter of seconds without the need of water or chewing. Desloratadine is a tricyclic antihistaminic, which has a selective and peripheral H1-antagonist action. It is an antagonist at histamine H1 receptors, and an antagonist at all subtypes of the muscarinic acetylcholine receptor. Desloratadine is the major metabolite of loratadine. Twelve different placebos ODT were prepared (F1-F12) using different functional excipients. They were evaluated for their compressibility, hardness and disintegration time. All formulations were non sticky except four formulations; namely (F8, F9, F10, F11). All formulations were compressible with the exception of (F2). Variable disintegration times were found ranging between 20 and 120 seconds. It was found that (F12) showed the least disintegration time (20 secs) without showing any sticking which could be due to the use of high percentage of superdisintegrants. Desloratadine showed bitter taste when formulated as ODT without any treatment. Therefore, different techniques were tried in order to mask its bitter taste. Using Eudragit EPO resulted in complete masking of the bitter taste of the drug and increased the acceptability to volunteers. The compressible non sticky formulations (F1, F3, F4, F5, F6, F7 and F12) were subjected to further evaluation tests after addition of coated desloratadine, including weight uniformity, wetting time, and friability testing.. Fairly good weight uniformity values were observed in all the tested formulations. F12 exhibiting the shortest wetting time (14.7 seconds) and consequently the lowest (20 seconds) disintegration time. Dissolution profile showed that 100% desloratadine release was attained after only 2.5 minutes from the prepared ODT (F12) with dissolution efficiency of 95%.

Keywords: Desloratadine, orally disintegrating tablets (ODTs), formulations, taste masking

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Authors: Boris Nemzer, Tania Reyes-Izquierdo, Zbigniew Pietrzkowski

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Glucosinolate is a family of glucosides that can be found in a family of brassica vegetables. Upon the damage of the plant, glucosinolate breakdown by an internal enzyme myrosinase (thioglucosidase; EC 3.2.3.1) into isothiocyanates, such as sulforaphane. Sulforaphane is formed by glucoraphanin cleaving the sugar off by myrosinase and rearranged. Sulforaphane nitrile is formed in the same reaction as sulforaphane with the active of epithiospecifier protein (ESP). Most common food processing procedure would break the plant and mix the glucoraphanin and myrosinase together, and the formed sulforaphane would be further degraded. The purpose of this study is to understand the glucoraphanin/sulforaphane and the myrosinase activity of broccoli seeds germinated at a different time and technological processing conditions that keep the activity of the enzyme to form sulforaphane. Broccoli seeds were germinated in the house. Myrosinase activities were tested as the glucose content using glucose assay kit and measured UV-Vis spectrophotometer. Glucosinolates were measured by HPLC/DAD. Sulforaphane was measured using HPLC-DAD and GC/MS. The 6 hr germinated sprouts have a myrosinase activity 32.2 mg glucose/g, which is comparable with 12 and 24 hour germinated seeds and higher than dry seeds. The glucoraphanin content in 6 hour germinated sprouts is 13935 µg/g which is comparable to 24 hour germinated seeds and lower than the dry seeds. GC/MS results show that the amount of sulforaphane is higher than the amount of sulforaphane nitrile in seeds, 6 hour and 24 hour germinated seeds. The ratio of sulforaphane and sulforaphane nitrile is high in 6 hour germinated seeds, which indicates the inactivated ESP in the reaction. After evaluating the results, the short time germinated seeds can be used as the source of glucoraphanin and myrosinase supply to form potential higher sulforaphane content. Broccoli contains glucosinolates, glucoraphanin (4-methylsulfinylbutyl glucosinolate), which is an important metabolite with health-promoting effects. In the pilot clinical study, we observed the effects of a glucosinolates/glucoraphanin-rich extract from short time germinated broccoli seeds on blood adenosine triphosphate (ATP), reactive oxygen species (ROS) and lactate levels. A single dose of 50 mg of broccoli sprouts extract increased blood levels of ATP up to 61% (p=0.0092) during the first 2 hours after the ingestion. Interestingly, this effect was not associated with an increase in blood ROS or lactate. When compared to the placebo group, levels of lactate were reduced by 10% (p=0.006). These results indicate that broccoli germinated seed extract may positively affect the generation of ATP in humans. Due to the preliminary nature of this work and promising results, larger clinical trials are justified.

Keywords: broccoli glucosinolates, glucoraphanin, germinated seeds, myrosinase, adenosine triphosphate

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Authors: Maged El-Sayed Mohamed

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Plant tissue culture practices are presented nowadays as the most promising substitute to a whole plant in the terms of secondary metabolites production. They offer the advantages of high production, tunability and they have less effect on plant ecosystems. Lantana camara is a weed, which is common all over the world as an ornamental plant. Weeds can adapt to any type of soil and climate due to their rich cellular machinery for secondary metabolites’ production. This characteristic is found in Lantana camara as a plant of very rich diversity of secondary metabolites with no dominant class of compounds. Aim: This trait has encouraged the author to develop tissue culture experiments for Lantana camara to be a platform for production and manipulation of secondary metabolites through biotransformation. Methodology: The plant was collected in its flowering stage in September 2014, from which explants were prepared from shoot tip, auxiliary bud and leaf. Different types of culture media were tried as well as four phytohormones and their combinations; NAA, 2,4-D, BAP and kinetin. Explants were grown in dark or in 12 hours dark and light cycles at 25°C. A metabolic profile for the produced callus was made and then compared to the whole plant profile. The metabolic profile was made using GC-MS for volatile constituents (extracted by n-hexane) and by HPLC-MS and capillary electrophoresis-mass spectrometry (CE-MS) for non-volatile constituents (extracted by ethanol and water). Results: The best conditions for the callus induction was achieved using MS media supplied with 30 gm sucrose and NAA/BAP (1:0.2 mg/L). Initiation of callus was favoured by incubation in dark for 20 day. The callus produced under these conditions showed yellow colour, which changed to brownish after 30 days. The rate of callus growth was high, expressed in the callus diameter, which reached to 1.15±0.2 cm in 30 days; however, the induction of callus delayed for 15 days. The metabolic profile for both volatile and non-volatile constituents of callus showed more simple background metabolites than the whole plant with two new (unresolved) peaks in the callus’ nonvolatile constituents’ chromatogram. Conclusion: Lantana camara callus production can be itself a source of new secondary metabolites and could be used for biotransformation studies due to its simple metabolic background, which allow easy identification of newly formed metabolites. The callus production gathered the simple metabolic background with the rich cellular secondary metabolite machinery of the plant, which could be elicited to produce valuable medicinally active products.

Keywords: capillary electrophoresis-mass spectrometry, gas chromatography, metabolic profile, plant tissue culture

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Authors: William Kelly, Sorelle Veigne, Xianhua Li, Zuyi Huang, Shyamsundar Subramanian, Eugene Schaefer

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The ambr15™ bioreactor is a single-use microbioreactor for cell line development and process optimization. The ambr system offers fully automatic liquid handling with the possibility of fed-batch operation and automatic control of pH and oxygen delivery. With operating conditions for large scale biopharmaceutical production properly scaled down, micro bioreactors such as the ambr15™ can potentially be used to predict the effect of process changes such as modified media or different cell lines. In this study, gassing rates and dilution rates were varied for a semi-continuous cell culture system in the ambr15™ bioreactor. The corresponding changes to metabolite production and consumption, as well as cell growth rate and therapeutic protein production were measured. Conditions were identified in the ambr15™ bioreactor that produced metabolic shifts and specific metabolic and protein production rates also seen in the corresponding larger (5 liter) scale perfusion process. A Dynamic Flux Balance model was employed to understand and predict the metabolic changes observed. The DFB model-predicted trends observed experimentally, including lower specific glucose consumption when CO₂ was maintained at higher levels (i.e. 100 mm Hg) in the broth. A Computational Fluid Dynamic (CFD) model of the ambr15™ was also developed, to understand transfer of O₂ and CO₂ to the liquid. This CFD model predicted gas-liquid flow in the bioreactor using the ANSYS software. The two-phase flow equations were solved via an Eulerian method, with population balance equations tracking the size of the gas bubbles resulting from breakage and coalescence. Reasonable results were obtained in that the Carbon Dioxide mass transfer coefficient (kLa) and the air hold up increased with higher gas flow rate. Volume-averaged kLa values at 500 RPM increased as the gas flow rate was doubled and matched experimentally determined values. These results form a solid basis for optimizing the ambr15™, using both CFD and FBA modelling approaches together, for use in microscale simulations of larger scale cell culture processes.

Keywords: cell culture, computational fluid dynamics, dynamic flux balance analysis, microbioreactor

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Authors: Antoine Villa, Moustapha Mohamedou, Florence Pilliere, Catherine Verdun-Esquer, Mathieu Molimard, Mohamed Sidatt Cheikh El Moustaph, Mireille Canal-Raffin

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Context: In Mauritania, the activity of the National Center of Oncology (NCO) has steadily risen leading to an increase in the handling of antineoplastic drugs (AD) by healthcare professionals. In this context, the AD contamination of those professionals is a major concern for occupational physicians. It has been evaluated using biological monitoring of occupational exposure (BMOE). Methods: The intervention took place in 2015, in 2 care units, and evaluated nurses preparing and/or infusing AD and agents in charge of hygiene. Participants provided a single urine sample, at the end of the week, at the end of their shift. Five molecules were sought using specific high sensitivity methods (UHPLC-MS/MS) with very low limits of quantification (LOQ) (cyclophosphamide (CP), Ifosfamide (IF), methotrexate (MTX): 2.5ng/L; doxorubicin (Doxo): 10ng/L; α-fluoro-β-alanine (FBAL, 5-FU metabolite): 20ng/L). A healthcare worker was considered as 'contaminated' when an AD was detected at a urine concentration equal to or greater than the LOQ of the analytical method or at trace concentration. Results: Twelve persons participated (6 nurses, 6 agents in charge of hygiene). Twelve urine samples were collected and analyzed. The percentage of contamination was 66.6% for all participants (n=8/12), 100% for nurses (6/6) and 33% for agents in charge of hygiene (2/6). In 62.5% (n=5/8) of the contaminated workers, two to four of the AD were detected in the urine. CP was found in the urine of all contaminated workers. FBAL was found in four, MTX in three and Doxo in one. Only IF was not detected. Urinary concentrations (all drugs combined) ranged from 3 to 844 ng/L for nurses and from 3 to 44 ng/L for agents in charge of hygiene. The median urinary concentrations were 87 ng/L, 15.1 ng/L and 4.4 ng/L for FBAL, CP and MTX, respectively. The Doxo urinary concentration was found 218ng/L. Discussion: There is no current biological exposure index for the interpretation of AD contamination. The contamination of these healthcare professionals is therefore established by the detection of one or more AD in urine. These urinary contaminations are higher than the LOQ of the analytical methods, which must be as low as possible. Given the danger of AD, the implementation of corrective measures is essential for the staff. Biological monitoring of occupational exposure is the most reliable process to identify groups at risk, tracing insufficiently controlled exposures and as an alarm signal. These results show the necessity to educate professionals about the risks of handling AD and/or to care for treated patients.

Keywords: antineoplastic drugs, Mauritania, biological monitoring of occupational exposure, contamination

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Authors: Sidrah Ahmed

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The river and lakes flows are modeled mathematically by shallow water equations that are depth-averaged Reynolds Averaged Navier-Stokes equations under Boussinesq approximation. The temperature stratification dynamics influence the water quality and mixing characteristics. It is mainly due to the atmospheric conditions including air temperature, wind velocity, and radiative forcing. The experimental observations are commonly taken along vertical scales and are not sufficient to estimate small turbulence effects of temperature variations induced characteristics of shallow flows. Wind shear stress over the water surface influence flow patterns, heat fluxes and thermodynamics of water bodies as well. Hence it is crucial to couple temperature gradients with shallow water model to estimate the atmospheric effects on flow patterns. The Ripa system has been introduced to study ocean currents as a variant of shallow water equations with addition of temperature variations within the flow. Ripa model is a hyperbolic system of partial differential equations because all the eigenvalues of the system’s Jacobian matrix are real and distinct. The time steps of a numerical scheme are estimated with the eigenvalues of the system. The solution to Riemann problem of the Ripa model is composed of shocks, contact and rarefaction waves. Solving Ripa model with Riemann initial data with the central schemes is difficult due to the eigen structure of the system.This works presents the comparison of four different finite difference schemes for the numerical solution of Riemann problem for Ripa model. These schemes include Lax-Friedrichs, Lax-Wendroff, MacCormack scheme and a higher order finite difference scheme with WENO method. The numerical flux functions in both dimensions are approximated according to these methods. The temporal accuracy is achieved by employing TVD Runge Kutta method. The numerical tests are presented to examine the accuracy and robustness of the applied methods. It is revealed that Lax-Freidrichs scheme produces results with oscillations while Lax-Wendroff and higher order difference scheme produce quite better results.

Keywords: finite difference schemes, Riemann problem, shallow water equations, temperature gradients

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Authors: Anindita Patra, Prasad K. Bhaskaran

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The atmosphere-ocean is a dynamically linked system that influences the exchange of energy, mass, and gas at the air-sea interface. The exchange of energy takes place in the form of sensible heat, latent heat, and momentum commonly referred to as fluxes along the atmosphere-ocean boundary. The large scale features such as El Nino and Southern Oscillation (ENSO) is a classic example on the interaction mechanism that occurs along the air-sea interface that deals with the inter-annual variability of the Earth’s Climate System. Most importantly the ocean and atmosphere as a coupled system acts in tandem thereby maintaining the energy balance of the climate system, a manifestation of the coupled air-sea interaction process. The present work is an attempt to understand the long-term variability in atmospheric parameters (from surface to upper levels) and investigate their role in influencing the surface ocean variables. More specifically the influence of atmospheric circulation and its variability influencing the mean Sea Level Pressure (SLP) has been explored. The study reports on a critical examination of both ocean-atmosphere parameters during a monsoon season over the head Bay of Bengal region. A trend analysis has been carried out for several atmospheric parameters such as the air temperature, geo-potential height, and omega (vertical velocity) for different vertical levels in the atmosphere (from surface to the troposphere) covering a period from 1992 to 2012. The Reanalysis 2 dataset from the National Centers for Environmental Prediction-Department of Energy (NCEP-DOE) was used in this study. The study signifies that the variability in air temperature and omega corroborates with the variation noticed in geo-potential height. Further, the study advocates that for the lower atmosphere the geo-potential heights depict a typical east-west contrast exhibiting a zonal dipole behavior over the study domain. In addition, the study clearly brings to light that the variations over different levels in the atmosphere plays a pivotal role in supporting the observed dipole pattern as clearly evidenced from the trends in SLP, associated surface wind speed and significant wave height over the study domain.

Keywords: air temperature, geopotential height, head Bay of Bengal, long-term variability, NCEP reanalysis 2, omega, wind-waves

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Authors: Nataša Todorović, Jovana Nikolov, Ivana Stojković, Milan Vraneš, Jovana Panić, Slobodan Gadžurić

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The detection of ²¹⁰Pb levels in aquatic environments evokes interest in various scientific studies. Its precise determination is important not only for the radiological assessment of drinking waters but also ²¹⁰Pb, and ²¹⁰Po distribution in the marine environment are significant for the assessment of the removal rates of particles from the ocean and particle fluxes during transport along the coast, as well as particulate organic carbon export in the upper ocean. Measurement techniques for ²¹⁰Pb determination, gamma spectrometry, alpha spectrometry, or liquid scintillation counting (LSC) are either time-consuming or demand expensive equipment or complicated chemical pre-treatments. However, one other possibility is to measure ²¹⁰Pb on an LS counter if it is in equilibrium with its progeny ²¹⁰Bi - through the Cherenkov counting method. It is unaffected by the chemical quenching and assumes easy sample preparation but has the drawback of lower counting efficiencies than standard LSC methods, typically from 10% up to 20%. The aim of the presented research in this paper is to investigate the possible increment of detection efficiency of Cherenkov counting during ²¹⁰Pb/²¹⁰Bi detection on an LS counter Quantulus 1220. Considering naturally low levels of ²¹⁰Pb in aqueous samples, the addition of ionic liquids to the counting vials with the analysed samples has the benefit of detection limit’s decrement during ²¹⁰Pb quantification. Our results demonstrated that ionic liquid, 1-butyl-3-methylimidazolium salicylate, is more efficient in Cherenkov counting efficiency increment than the previously explored 2-hydroxypropan-1-amminium salicylate. Consequently, the impact of a few other ionic liquids that were synthesized with the same cation group (1-butyl-3-methylimidazolium benzoate, 1-butyl-3-methylimidazolium 3-hydroxybenzoate, and 1-butyl-3-methylimidazolium 4-hydroxybenzoate) was explored in order to test their potential influence on Cherenkov counting efficiency. It was confirmed that, among the explored ones, only ionic liquids in the form of salicylates exhibit a wavelength shifting effect. Namely, the addition of small amounts (around 0.8 g) of 1-butyl-3-methylimidazolium salicylate increases the detection efficiency from 16% to >70%, consequently reducing the detection threshold by more than four times. Moreover, the addition of ionic liquids could find application in the quantification of other radionuclides besides ²¹⁰Pb/²¹⁰Bi via Cherenkov counting method.

Keywords: liquid scintillation counting, ionic liquids, Cherenkov counting, ²¹⁰PB/²¹⁰BI in water

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Authors: Qianqian He, Naian Liu, Xiaodong Xie, Linhe Zhang, Yang Zhang, Weidong Yan

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In the wild, the vegetation layer with exceptionally complex fuel composition and heterogeneous spatial distribution strongly affects the rate of fire spread (ROS) and fire intensity. Clarifying the influence of fuel bed structure on fire spread behavior is of great significance to wildland fire management and prediction. The packing ratio is one of the key physical parameters describing the property of the fuel bed. There is a threshold value of the packing ratio for ROS, but little is known about the controlling mechanism. In this study, to address this deficiency, a series of fire spread experiments were performed across a discrete fuel bed composed of some regularly arranged laser-cut cardboards, with constant wind speed and different packing ratios (0.0125-0.0375). The experiment aims to explore the relative importance of the internal and surface heat transfer with packing ratio. The dependence of the measured ROS on the packing ratio was almost consistent with the previous researches. The data of the radiative and total heat fluxes show that the internal heat transfer and surface heat transfer are both enhanced with increasing packing ratio (referred to as ‘Stage 1’). The trend agrees well with the variation of the flame length. The results extracted from the video show that the flame length markedly increases with increasing packing ratio in Stage 1. Combustion intensity is suggested to be increased, which, in turn, enhances the heat radiation. The heat flux data shows that the surface heat transfer appears to be more important than the internal heat transfer (fuel preheating inside the fuel bed) in Stage 1. On the contrary, the internal heat transfer dominates the fuel preheating mechanism when the packing ratio further increases (referred to as ‘Stage 2’) because the surface heat flux keeps almost stable with the packing ratio in Stage 2. As for the heat convection, the flow velocity was measured using Pitot tubes both inside and on the upper surface of the fuel bed during the fire spread. Based on the gas velocity distribution ahead of the flame front, it is found that the airflow inside the fuel bed is restricted in Stage 2, which can reduce the internal heat convection in theory. However, the analysis indicates not the influence of inside flow on convection and combustion, but the decreased internal radiation of per unit fuel is responsible for the decrease of ROS.

Keywords: discrete fuel bed, fire spread, packing ratio, wildfire

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Authors: Ruchi Pathania, Ahmad Ahmad, Shireesh Srivastava

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Cyanobacteria is a promising microbe that can capture and convert atmospheric CO₂ and light into valuable industrial bio-products like biofuels, biodegradable plastics, etc. Among their most attractive traits are faster autotrophic growth, whole year cultivation using non-arable land, high photosynthetic activity, much greater biomass and productivity and easy for genetic manipulations. Cyanobacteria store carbon in the form of glycogen which can be hydrolyzed to release glucose and fermented to form bioethanol or other valuable products. Marine cyanobacterial species are especially attractive for countries with scarcity of freshwater. We recently identified a marine native cyanobacterium Synechococcus sp. BDU 130192 which has good growth rate and high level of polyglucans accumulation compared to Synechococcus PCC 7002. In this study, firstly we sequenced the whole genome and the sequences were annotated using the RAST server. Genome scale metabolic model (GSMM) was reconstructed through COBRA toolbox. GSMM is a computational representation of the metabolic reactions and metabolites of the target strain. GSMMs construction through the application of Flux Balance Analysis (FBA), which uses external nutrient uptake rates and estimate steady state intracellular and extracellular reaction fluxes, including maximization of cell growth. The model, which we have named isyn942, includes 942 reactions and 913 metabolites having 831 metabolic, 78 transport and 33 exchange reactions. The phylogenetic tree obtained by BLAST search revealed that the strain was a close relative of Synechococcus PCC 7002. The flux balance analysis (FBA) was applied on the model iSyn942 to predict the theoretical yields (mol product produced/mol CO₂ consumed) for native and non-native products like acetone, butanol, etc. under phototrophic condition by applying metabolic engineering strategies. The reported strain can be a viable strain for biotechnological applications, and the model will be helpful to researchers interested in understanding the metabolism as well as to design metabolic engineering strategies for enhanced production of various bioproducts.

Keywords: cyanobacteria, flux balance analysis, genome scale metabolic model, metabolic engineering

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Authors: Savitha Janakiraman, Shivakumar M. C

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Anidulafungin, an advanced class of antifungal agent used for the treatment of chronic fungal infections, is derived from Echinocandin B nucleus, an intermediate metabolite of Echinocandin B produced by Aspergillus nidulans. The enzyme acylase derived from the fermentation broth of Actinoplanes utahensis (NRRL 12052) plays a key role in the bioconversion of echinocandin B to echinocandin B nucleus. The membrane-bound nature of acylase and low levels of expression contributes to the rate-limiting process of enzymatic deacylation, hence low yields of ECB nucleus and anidulafungin. In the present study, this is addressed through novel genetic engineering approaches of overexpression and heterologous expression studies, immobilization of whole cells of Actinoplanes utahensis (NRRL 12052) and Co-cultivation studies. Overexpression of the acylase gene in Actinoplanes utahensis (NRRL 12052) was done by increasing the gene copy number to increase the echinocandin B nucleus production. Echinocandin B acylase gene, under the control of a PermE* promoter, was cloned in pSET152 vector and introduced into Actinoplanes utahensis (NRRL12052) by a ɸC31-directed site-specific recombination method. The resultant recombinant strain (C2-18) showed a 3-fold increase in acylase expression, which was confirmed by HPLC analysis. Pichia pastoris is one of the most effective and versatile host systems for the production of heterologous proteins. The ECB acylase gene was cloned into pPIC9K vector with AOX1 promoter and was transformed into Pichia pastoris (GS115). The acylase expression was confirmed by protein expression and bioconversion studies. The heterologous expression of acylase in Pichia pastoris, is a milestone in the development of antifungals. Actively growing cells of Actinoplanes utahensis (NRRL 12052) were immobilized and tested for bioconversion ability which showed >90% conversion in each cycle. The stability of immobilized cell beads retained the deacylation ability up to 60 days and reusability was confirmed up to 4 cycles. The significant findings from the study have revealed that immobilization of whole cells of Actinoplanes utahensis (NRRL 12052) could be an alternative option for bioconversion of echinocandin B to echinocandin B nucleus, which has not been reported to date. The concept of co-cultivation of Aspergillus nidulans and Actinoplanes utahensis strains for the production of the echinocandin B nucleus was also carried out in order to produce echinocandin B nucleus. The process completely reduced the ECB purification step and, therefore, could be recommended as an ingenious method to improve the yield of the ECB nucleus.

Keywords: acylase, anidulafungin, antifungals, Aspergillus nidulans

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Authors: Ivana Čavlina Tomašević, Višnjica Vučetić, Maja Telišman Prtenjak, Barbara Malečić

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The Split wildfire on the mid-Adriatic Coast in July 2017 is one of the most severe wildfires in Croatian history, given the size and unexpected fire behavior, and it is used in this research as a case study to run the Weather Research and Forecasting Spread Fire (WRF SFIRE) model. This coupled fire-atmosphere model was successfully run for the first time ever for one Croatian wildfire case. Verification of coupled simulations was possible by using the detailed reconstruction of the Split wildfire. Specifically, precise information on ignition time and location, together with mapped fire progressions and spotting within the first 30 hours of the wildfire, was used for both – to initialize simulations and to evaluate the model’s ability to simulate fire’s propagation and final fire scar. The preliminary simulations were obtained using high-resolution vegetation and topography data for the fire area, additionally interpolated to fire grid spacing at 33.3 m. The results demonstrated that the WRF SFIRE model has the ability to work with real data from Croatia and produce adequate results for forecasting fire spread. As the model in its setup has the ability to include and exclude the energy fluxes between the fire and the atmosphere, this was used to investigate possible fire-atmosphere interactions during the Split wildfire. Finally, successfully coupled simulations provided the first numerical evidence that a wildfire from the Adriatic coast region can modify the dynamical structure of the surrounding atmosphere, which agrees with observations from fire grounds. This study has demonstrated that the WRF SFIRE model has the potential for operational application in Croatia with more accurate fire predictions in the future, which could be accomplished by inserting the higher-resolution input data into the model without interpolation. Possible uses for fire management in Croatia include prediction of fire spread and intensity that may vary under changing weather conditions, available fuels and topography, planning effective and safe deployment of ground and aerial firefighting forces, preventing wildland-urban interface fires, effective planning of evacuation routes etc. In addition, the WRF SFIRE model results from this research demonstrated that the model is important for fire weather research and education purposes in order to better understand this hazardous phenomenon that occurs in Croatia.

Keywords: meteorology, agrometeorology, fire weather, wildfires, couple fire-atmosphere model

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Authors: Geminne Manzano, Porfirio Aliño, Clairecynth Yu, Lilibeth Salvador-Reyes, Viviene Santiago

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Many marine benthic organisms produce secondary metabolites that serve as ecological roles to different biological and environmental factors. The secondary metabolites found in these organisms like algae, sponges, tunicates and worms exhibit variation at different scales. Understanding the chemical variation can be essential in deriving the evolutionary and ecological function of the secondary metabolites that may explain their patterns. Ecological surveys were performed on two collection sites representing from two Philippine marine biogeographic regions – in Oriental Mindoro located on the West Philippine Sea (WPS) and in Zamboanga del Sur located at Celebes Sea (CS), where a total of 39 Xestospongia sp. sponges were collected using SCUBA. The sponge samples were transported to the laboratory for taxonomic identification and chemical analysis. Biological and environmental factors were investigated to determine their relation to the abundance and distribution patterns and its spatial variability of their secondary metabolite production. Extracts were subjected to thin-layer chromatography and anti-proliferative assays to confirm the presence of Renieramycin-M and to test its cytotoxicity. The blue sponges were found to be more abundant on the WPS than in CS. Both the benthic community and the fish community in Oriental Mindoro, WPS and Zamboanga del Sur, CS sites are characterized by high species diversity and abundance and a very high biomass category. Environmental factors like depth and monsoonal exposure were also compared showing that wave exposure and depth are associated with the abundance and distribution of the sponges. Renieramycin-M presence using the TLC profiles between the sponge extracts from WPS and from CS showed differences in the Reniermycin-M presence and the presence of other functional groups were observed between the two sites. In terms of bioactivity, different responses were also exhibited by the sponge extracts coming from the different region. Different responses were also noted on its bioactivity depending on the cell lines tested. Exploring the influence of ecological parameters on the chemical variation can provide deeper chemical ecological insights in the knowledge and their potential varied applications at different scales. The results of this study provide further impetus in pursuing studies into patterns and processes of the chemical diversity of the Philippine blue sponge, Xestospongia sp. and the chemical ecological significance of the coral triangle.

Keywords: chemical ecology, porifera, renieramycin-m, spatial variability, Xestospongia sp.

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Authors: Thomas Rowan, Mohammed Seaid

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A fast finite volume solver for multi-layered shallow water flows with mass exchange and an erodible bed is developed. This enables the user to solve a number of complex sediment-based problems including (but not limited to), dam-break over an erodible bed, recirculation currents and bed evolution as well as levy and dyke failure. This research develops methodologies crucial to the under-standing of multi-sediment fluvial mechanics and waterway design. In this model mass exchange between the layers is allowed and, in contrast to previous models, sediment and fluid are able to transfer between layers. In the current study we use a two-step finite volume method to avoid the solution of the Riemann problem. Entrainment and deposition rates are calculated for the first time in a model of this nature. In the first step the governing equations are rewritten in a non-conservative form and the intermediate solutions are calculated using the method of characteristics. In the second stage, the numerical fluxes are reconstructed in conservative form and are used to calculate a solution that satisfies the conservation property. This method is found to be considerably faster than other comparative finite volume methods, it also exhibits good shock capturing. For most entrainment and deposition equations a bed level concentration factor is used. This leads to inaccuracies in both near bed level concentration and total scour. To account for diffusion, as no vertical velocities are calculated, a capacity limited diffusion coefficient is used. The additional advantage of this multilayer approach is that there is a variation (from single layer models) in bottom layer fluid velocity: this dramatically reduces erosion, which is often overestimated in simulations of this nature using single layer flows. The model is used to simulate a standard dam break. In the dam break simulation, as expected, the number of fluid layers utilised creates variation in the resultant bed profile, with more layers offering a higher deviation in fluid velocity . These results showed a marked variation in erosion profiles from standard models. The overall the model provides new insight into the problems presented at minimal computational cost.

Keywords: erosion, finite volume method, sediment transport, shallow water equations

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Authors: Maria Sofia Palagonia, Doriano Brogioli, Fabio La Mantia

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In the last decade, lithium has become an important raw material in various sectors, in particular for rechargeable batteries. Its production is expected to grow more and more in the future, especially for mobile energy storage and electromobility. Until now it is mostly produced by the evaporation of water from salt lakes, which led to a huge water consumption, a large amount of waste produced and a strong environmental impact. A new, clean and faster electrochemical technique to recover lithium has been recently proposed: electrochemical ion pumping. It consists in capturing lithium ions from a feed solution by intercalation in a lithium-selective material, followed by releasing them into a recovery solution; both steps are driven by the passage of a current. In this work, a new configuration of the electrochemical cell is presented, used to study and optimize the process of the intercalation of lithium ions through the hydrodynamic condition. Lithium Manganese Oxide (LiMn₂O₄) was used as a cathode to intercalate lithium ions selectively during the reduction, while Nickel Hexacyano Ferrate (NiHCF), used as an anode, releases positive ion. The effect of hydrodynamics on the process has been studied by conducting the experiments at various fluxes of the electrolyte through the electrodes, in terms of charge circulated through the cell, captured lithium per unit mass of material and overvoltage. The result shows that flowing the electrolyte inside the cell improves the lithium capture, in particular at low lithium concentration. Indeed, in Atacama feed solution, at 40 mM of lithium, the amount of lithium captured does not increase considerably with the flux of the electrolyte. Instead, when the concentration of the lithium ions is 5 mM, the amount of captured lithium in a single capture cycle increases by increasing the flux, thus leading to the conclusion that the slowest step in the process is the transport of the lithium ion in the liquid phase. Furthermore, an influence of the concentration of other cations in solution on the process performance was observed. In particular, the capturing of the lithium using a different concentration of NaCl together with 5 mM of LiCl was performed, and the results show that the presence of NaCl limits the amount of the captured lithium. Further studies can be performed in order to understand why the full capacity of the material is not reached at the highest flow rate. This is probably due to the porous structure of the material since the liquid phase is likely not affected by the convection flow inside the pores. This work proves that electrochemical ion pumping, with a suitable hydrodynamic design, enables the recovery of lithium from feed solutions at the lower concentration than the sources that are currently exploited, down to 1 mM.

Keywords: desalination battery, electrochemical ion pumping, hydrodynamic, lithium

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Authors: Elif Gencturk, Ekin Yurdakul, Ahmet Y. Celik, Senol Mutlu, Kutlu O. Ulgen

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Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Keywords: COP, HMF, ribosome biogenesis, thermoplastic microbioreactor, yeast

Procedia PDF Downloads 171

Authors: E. K. K. Agyeman, P. Mousseau, A. Sarda, D. Edelin

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During the cooling of hot metals by the circulation of water in canals formed by boring holes in the metal, the rapid phase change of the water due to the high initial temperature of the metal leads to a non homogenous distribution of the phases within the canals. The liquid phase dominates towards the entrance of the canal while the gaseous phase dominates towards the exit. As a result of the different thermal properties of both phases, the metal is not uniformly cooled. This poses a problem during the cooling of moulds, where a uniform temperature distribution is needed in order to ensure the integrity of the part being formed. In this study, the simultaneous use of multiple water jets and an airflow for the uniform and controlled cooling of a steel block is investigated. A circular hole is bored at the centre of the steel block along its length and a perforated steel pipe is inserted along the central axis of the hole. Water jets that impact the internal surface of the steel block are generated from the perforations in the steel pipe when the water within it is put under pressure. These jets are oriented in the opposite direction to that of gravity. An intermittent airflow is imposed in the annular space between the steel pipe and the surface of hole bored in the steel block. The evolution of the temperature with respect to time of the external surface of the block is measured with the help of thermocouples and an infrared camera. Due to the high initial temperature of the steel block (350 °C), the water changes phase when it impacts the internal surface of the block. This leads to high heat fluxes. The strategy used to control the cooling speed of the block is the intermittent impingement of its internal surface by the jets. The intervals of impingement and of non impingement are varied in order to achieve the desired result. An airflow is used during the non impingement periods as an additional regulator of the cooling speed and to improve the temperature homogeneity of the impinged surface. After testing different jet positions, jet speeds and impingement intervals, it’s observed that the external surface of the steel block has a uniform temperature distribution along its length. However, the temperature distribution along its width isn’t uniform with the maximum temperature difference being between the centre of the block and its edge. Changing the positions of the jets has no significant effect on the temperature distribution on the external surface of the steel block. It’s also observed that reducing the jet impingement interval and increasing the non impingement interval slows down the cooling of the block and improves upon the temperature homogeneity of its external surface while increasing the duration of jet impingement speeds up the cooling process.

Keywords: cooling speed, homogenous cooling, jet impingement, phase change

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Authors: Hammad Majeed, Hanna Knuutila, Magne Hilestad, Hallvard Svendsen

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Formation of aerosols can cause serious complications in industrial exhaust gas CO2 capture processes. SO3 present in the flue gas can cause aerosol formation in an absorption based capture process. Small mist droplets and fog formed can normally not be removed in conventional demisting equipment because their submicron size allows the particles or droplets to follow the gas flow. As a consequence of this aerosol based emissions in the order of grams per Nm3 have been identified from PCCC plants. In absorption processes aerosols are generated by spontaneous condensation or desublimation processes in supersaturated gas phases. Undesired aerosol development may lead to amine emissions many times larger than what would be encountered in a mist free gas phase in PCCC development. It is thus of crucial importance to understand the formation and build-up of these aerosols in order to mitigate the problem.Rigorous modelling of aerosol dynamics leads to a system of partial differential equations. In order to understand mechanics of a particle entering an absorber an implementation of the model is created in Matlab. The model predicts the droplet size, the droplet internal variable profiles and the mass transfer fluxes as function of position in the absorber. The Matlab model is based on a subclass method of weighted residuals for boundary value problems named, orthogonal collocation method. The model comprises a set of mass transfer equations for transferring components and the essential diffusion reaction equations to describe the droplet internal profiles for all relevant constituents. Also included is heat transfer across the interface and inside the droplet. This paper presents results describing the basic simulation tool for the characterization of aerosols formed in CO2 absorption columns and gives examples as to how various entering droplets grow or shrink through an absorber and how their composition changes with respect to time. Below are given some preliminary simulation results for an aerosol droplet composition and temperature profiles. Results: As an example a droplet of initial size of 3 microns, initially containing a 5M MEA, solution is exposed to an atmosphere free of MEA. Composition of the gas phase and temperature is changing with respect to time throughout the absorber.

Keywords: amine solvents, emissions, global climate change, simulation and modelling, aerosol generation

Procedia PDF Downloads 265

Authors: K. Thiel, P. Patrikainen, C. Nagy, D. Fitzpatrick, E.-M. Aro, P. Kallio

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Photosynthetic cyanobacteria have been recognized as potential future biotechnological hosts for the direct conversion of CO₂ into chemicals of interest using sunlight as the solar energy source. However, in order to develop commercially viable systems, the flux of electrons from the photosynthetic light reactions towards specified target chemicals must be significantly improved. The objective of the study was to investigate whether the autotrophic production efficiency of specified end-metabolites can be improved in engineered cyanobacterial cells by rescuing excited electrons that are normally lost to molecular oxygen due to the cyanobacterial flavodiiron protein Flv1/3. Natively Flv1/3 dissipates excess electrons in the photosynthetic electron transfer chain by directing them to molecular oxygen in Mehler-like reaction to protect photosystem I. To evaluate the effect of flavodiiron inactivation on autotrophic production efficiency in the cyanobacterial host Synechocystis sp. PCC 6803 (Synechocystis), sucrose was selected as the quantitative reporter and a representative of a potential end-product of interest. The concept is based on the native property of Synechocystis to produce sucrose as an intracellular osmoprotectant when exposed to high external ion concentrations, in combination with the introduction of a heterologous sucrose permease (CscB from Escherichia coli), which transports the sucrose out from the cell. In addition, cell growth, photosynthetic gas fluxes using membrane inlet mass spectrometry and endogenous storage compounds were analysed to illustrate the consequent effects of flv deletion on pathway flux distributions. The results indicate that a significant proportion of the electrons can be lost to molecular oxygen via Flv1/3 even when the cells are grown under high CO₂ and that the inactivation of flavodiiron activity can enhance the photosynthetic electron flux towards optionally available sinks. The flux distribution is dependent on the light conditions and the genetic context of the Δflv mutants, and favors the production of either sucrose or one of the two storage compounds, glycogen or polyhydroxybutyrate. As a conclusion, elimination of the native Flv1/3 reaction and concomitant introduction of an engineered product pathway as an alternative sink for excited electrons could enhance the photosynthetic electron flux towards the target endproduct without compromising the fitness of the host.

Keywords: cyanobacterial engineering, flavodiiron proteins, redirecting electron flux, sucrose

Procedia PDF Downloads 125

Authors: Sayantan Saha, Sambit Supriya Dash, Vinayak Malhotra

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Efficient propulsive systems development is an area of major interest and concern in aerospace industry. Combustion forms the most reliable and basic form of propulsion for ground and space applications. The generation of large amount of energy from a small volume relates mostly to the flaming combustion. This study deals with instabilities associated with flaming combustion. Combustion is always accompanied by acoustics be it external or internal. Chemical propulsion oriented rockets and space systems are well known to encounter acoustic instabilities. Acoustic brings in changes in inter-energy conversion and alter the reaction rates. The modified heat fluxes, owing to wall temperature, reaction rates, and non-linear heat transfer are observed. The thermoacoustic instabilities significantly result in reduced combustion efficiency leading to uncontrolled liquid rocket engine performance, serious hazards to systems, assisted testing facilities, enormous loss of resources and every year a substantial amount of money is spent to prevent them. Present work attempts to fundamentally understand the mechanisms governing the thermoacoustic combustion in liquid rocket engine using a simplified experimental setup comprising a butane cylinder and an impinging acoustic source. Rocket engine produces sound pressure level in excess of 153 Db. The RL-10 engine generates noise of 180 Db at its base. Systematic studies are carried out for varying fuel flow rates, acoustic levels and observations are made on the flames. The work is expected to yield a good physical insight into the development of acoustic devices that when coupled with the present propulsive devices could effectively enhance combustion efficiency leading to better and safer missions. The results would be utilized to develop impinging acoustic devices that impinge sound on the combustion chambers leading to stable combustion thus, improving specific fuel consumption, specific impulse, reducing emissions, enhanced performance and fire safety. The results can be effectively applied to terrestrial and space application.

Keywords: combustion instability, fire safety, improved performance, liquid rocket engines, thermoacoustics

Procedia PDF Downloads 146

Authors: V. W. Kamgang, A. Van Der Goot, N. C. Bennett, A. Ganswindt

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The roan antelope (Hippotragus equinus) is the second largest antelope species in Africa. These past decades, populations of roan antelope are declining drastically throughout Africa. This situation resulted in the development of intensive breeding programmes for this species in Southern African, where they are popular game ranching herbivores in with increasing numbers in captivity. Nowadays, avoidance of stress is important when managing wildlife to ensure animal welfare. In this regard, a non-invasive approach to monitor the adrenocortical function as a measure of stress would be preferable, since animals are not disturbed during sample collection. However, to date, a non-invasive method has not been established for the roan antelope. In this study, we validated a non-invasive technique to monitor the adrenocortical function in this species. Herein, we performed an adrenocorticotropic hormone (ACTH) stimulation test at Lapalala reserve Wilderness, South Africa, using adult captive roan antelopes to determine the stress-related physiological responses. Two individually housed roan antelope (a male and a female) received an intramuscular injection with Synacthen depot (Norvatis) loaded into a 3ml syringe (Pneu-Dart) at an estimated dose of 1 IU/kg. A total number of 86 faecal samples (male: 46, female: 40) were collected 5 days before and 3 days post-injection. All samples were then lyophilised, pulverized and extracted with 80% ethanol (0,1g/3ml) and the resulting faecal extracts were analysed for immunoreactive faecal glucocorticoid metabolite (fGCM) concentrations using five enzyme immunoassays (EIAs); (i) 11-oxoaetiocholanolone I (detecting 11,17 dioxoandrostanes), (ii) 11-oxoaetiocholanolone II (detecting fGCM with a 5α-pregnane-3α-ol-11one structure), (iii) a 5α-pregnane-3β-11β,21-triol-20-one (measuring 3β,11β-diol CM), (iv) a cortisol and (v) a corticosterone. In both animals, all EIAs detected an increase in fGCM concentration 100% post-ACTH administration. However, the 11-oxoaetiocholanolone I EIA performed best, with a 20-fold increase in the male (baseline: 0.384 µg/g, DW; peak: 8,585 µg/g DW) and a 17-fold in the female (baseline: 0.323 µg/g DW, peak: 7,276 µg/g DW), measured 17 hours and 12 hours post-administration respectively. These results are important as the ability to assess adrenocortical function non-invasively in roan can now be used as an essential prerequisite to evaluate the effects of stressful circumstances; such as variation of environmental conditions or reproduction in other to improve management strategies for the conservation of this iconic antelope species.

Keywords: adrenocorticotropic hormone challenge, adrenocortical function, captive breeding, non-invasive method, roan antelope

Procedia PDF Downloads 145

Authors: Kaushal Kishore, Suman Mukhopadhyay, Susovan Das, Manashi Adhikary, Sandip Bhattacharyya

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In integrated steel plants, conveyor belts are widely used for transferring raw materials from one location to another. An output pin bush coupling attached with a conveyor transferring iron ore fines and fluxes failed after two years of service life. This led to an operational delay of approximately 15 hours. This study is focused on failure analysis of the coupling and recommending counter-measures to prevent any such failures in the future. Investigation consisted of careful visual observation, checking of operating parameters, stress calculation and analysis, macro and micro-fractography, material characterizations like chemical and metallurgical analysis and tensile and impact testings. The fracture occurred from an unusually sharp double step. There were multiple corrosion pits near the step that aggravated the situation. Inner contact surface of the coupling revealed differential abrasion that created a macroscopic difference in the height of the component. This pointed towards misalignment of the coupling beyond a threshold limit. In addition to these design and installation issues, material of the coupling did not meet the quality standards. These were made up of grey cast iron having graphite morphology intermediate between random distribution (Type A) and rosette pattern (Type B). This manifested as a marked reduction in impact toughness and tensile strength of the component. These findings corroborated well with the brittle mode of fracture that might have occurred during minor impact loading while loading of conveyor belt with raw materials from height. Simulated study was conducted to examine the effect of corrosion pits on tensile and impact toughness of grey cast iron. It was observed that pitting marginally reduced tensile strength and ductility. However, there was marked (up to 45%) reduction in impact toughness due to pitting. Thus, it became evident that failure of the coupling occurred due to combination of factors like inferior material, misalignment, poor step design and corrosion pitting. Recommendation for life enhancement of coupling included the use of tougher SG 500/7 grade, incorporation of proper fillet radius for the step, correction of alignment and application of corrosion resistant organic coating to prevent pitting.

Keywords: brittle fracture, cast iron, coupling, double step, pitting, simulated impact tests

Procedia PDF Downloads 133

Authors: Angelo Garofalo, Laura Donato, Maria Concetta Carnevale, Enrico Drioli, Omar Alharbi, Saad Aljlil, Alessandra Criscuoli, Catia Algieri

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Nowadays, water scarcity may be considered one of the most important and serious questions concerning our community: in fact, there is a remarkable mismatch between water supply and water demand. Exploitation of natural fresh water resources combined with higher water demand has led to an increased requirement for alternative water resources. In this context, desalination provides such an alternative source, offering water otherwise not accessible for irrigational, industrial and municipal use. Considering the various drawbacks of the polymeric membranes, zeolite membranes represent a potential device for water desalination owing to their high thermal and chemical stability. In this area wide attention was focused on the MFI (silicalite, ZSM-5) membranes, having a pore size lower (about 5.5 Å) than the major kinetic diameters of hydrated ions. In the present work, a scale-up for the preparation of supported silicalite membranes was performed. Therefore, tubular membranes 30 cm long were synthesized by using the secondary growth method coupled with the cross flow seeding procedure. The secondary growth presents two steps: seeding and growth of zeolite crystals on the support. This process, decoupling zeolite nucleation from crystals growth, permits to control the conditions of each step separately. The seeding procedure consists of a cross-flow filtration through a porous support coupled with the support rotation and tilting. The combination of these three different aspects allows a homogeneous and uniform coverage of the support with the zeolite seeds. After characterization by scanning electron microscope (SEM), X-ray diffractometry (XRD) and Energy-dispersive X-ray (EDX) analysis, the prepared membranes were tested by means of single gas permeation and then by Vacuum Membrane Distillation (VMD) using both deionized water and NaCl solutions. The experimental results evidenced the possibility to perform the scale up for the preparation of almost defect free silicalite membranes. VMD tests indicated the possibility to prepare membranes that exhibit interesting performance in terms of fluxes and salt rejections for concentrations from 0.2 M to 0.9 M. Furthermore, it was possible to restore the original performance of the membrane after an identified cleaning procedure. Acknowledgements: The authors gratefully acknowledge the support of the King Abdulaziz City for Science and Technology (KACST) for funding the research Project 895/33 entitled ‘Preparation and Characterization of Zeolite Membranes for Water Treatment’.

Keywords: desalination, MFI membranes, secondary growth, vacuum membrane distillation

Procedia PDF Downloads 256

Authors: Wesley A. Saltarelli, Nicolas R. Finkler, Adriana C. P. Miwa, Maria C. Calijuri, Davi G. F. Cunha

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The metabolism of the streams is an indicator of ecosystem disturbance due to the influences of the catchment on the structure of the water bodies. The study of the respiration and photosynthesis allows the estimation of energy fluxes through the food webs and the analysis of the autotrophic and heterotrophic processes. We aimed at evaluating the metabolism in streams located in the Brazilian savannah, Cerrado (Sao Carlos, SP), by determining and modeling the daily changes of dissolved oxygen (DO) in the water during one year. Three water bodies with minimal anthropogenic interference in their surroundings were selected, Espraiado (ES), Broa (BR) and Canchim (CA). Every two months, water temperature, pH and conductivity are measured with a multiparameter probe. Nitrogen and phosphorus forms are determined according to standard methods. Also, canopy cover percentages are estimated in situ with a spherical densitometer. Stream flows are quantified through the conservative tracer (NaCl) method. For the metabolism study, DO (PME-MiniDOT) and light (Odyssey Photosynthetic Active Radiation) sensors log data for at least three consecutive days every ten minutes. The reaeration coefficient (k2) is estimated through the method of the tracer gas (SF6). Finally, we model the variations in DO concentrations and calculate the rates of gross and net primary production (GPP and NPP) and respiration based on the one station method described in the literature. Three sampling were carried out in October and December 2015 and February 2016 (the next will be in April, June and August 2016). The results from the first two periods are already available. The mean water temperatures in the streams were 20.0 +/- 0.8C (Oct) and 20.7 +/- 0.5C (Dec). In general, electrical conductivity values were low (ES: 20.5 +/- 3.5uS/cm; BR 5.5 +/- 0.7uS/cm; CA 33 +/- 1.4 uS/cm). The mean pH values were 5.0 (BR), 5.7 (ES) and 6.4 (CA). The mean concentrations of total phosphorus were 8.0ug/L (BR), 66.6ug/L (ES) and 51.5ug/L (CA), whereas soluble reactive phosphorus concentrations were always below 21.0ug/L. The BR stream had the lowest concentration of total nitrogen (0.55mg/L) as compared to CA (0.77mg/L) and ES (1.57mg/L). The average discharges were 8.8 +/- 6L/s (ES), 11.4 +/- 3L/s and CA 2.4 +/- 0.5L/s. The average percentages of canopy cover were 72% (ES), 75% (BR) and 79% (CA). Significant daily changes were observed in the DO concentrations, reflecting predominantly heterotrophic conditions (respiration exceeded the gross primary production, with negative net primary production). The GPP varied from 0-0.4g/m2.d (in Oct and Dec) and the R varied from 0.9-22.7g/m2.d (Oct) and from 0.9-7g/m2.d (Dec). The predominance of heterotrophic conditions suggests increased vulnerability of the ecosystems to artificial inputs of organic matter that would demand oxygen. The investigation of the metabolism in the pristine streams can help defining natural reference conditions of trophic state.

Keywords: low-order streams, metabolism, net primary production, trophic state

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Authors: S. Mehrab Amiri, Nasser Talebbeydokhti

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Modeling sediment transport processes by means of numerical approach often poses severe challenges. In this way, a number of techniques have been suggested to solve flow and sediment equations in decoupled, semi-coupled or fully coupled forms. Furthermore, in order to capture flow discontinuities, a number of techniques, like artificial viscosity and shock fitting, have been proposed for solving these equations which are mostly required careful calibration processes. In this research, a numerical scheme for solving shallow water and Exner equations in fully coupled form is presented. First-Order Centered scheme is applied for producing required numerical fluxes and the reconstruction process is carried out toward using Monotonic Upstream Scheme for Conservation Laws to achieve a high order scheme.  In order to satisfy C-property of the scheme in presence of bed topography, Surface Gradient Method is proposed. Combining the presented scheme with fourth order Runge-Kutta algorithm for time integration yields a competent numerical scheme. In addition, to handle non-prismatic channels problems, Cartesian Cut Cell Method is employed. A trained Multi-Layer Perceptron Artificial Neural Network which is of Feed Forward Back Propagation (FFBP) type estimates sediment flow discharge in the model rather than usual empirical formulas. Hydrodynamic part of the model is tested for showing its capability in simulation of flow discontinuities, transcritical flows, wetting/drying conditions and non-prismatic channel flows. In this end, dam-break flow onto a locally non-prismatic converging-diverging channel with initially dry bed conditions is modeled. The morphodynamic part of the model is verified simulating dam break on a dry movable bed and bed level variations in an alluvial junction. The results show that the model is capable in capturing the flow discontinuities, solving wetting/drying problems even in non-prismatic channels and presenting proper results for movable bed situations. It can also be deducted that applying Artificial Neural Network, instead of common empirical formulas for estimating sediment flow discharge, leads to more accurate results.

Keywords: artificial neural network, morphodynamic model, sediment continuity equation, shallow water equations

Procedia PDF Downloads 188

Authors: Man Fung Ho, Lap So, Jiaqi Zhang, Yuheng Zhao, Huiyang Lu, Tat Shing Choi, K. Y. Michael Wong

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Nowadays, a tremendous amount of data is available in the transportation system, enabling the development of various machine learning approaches to make short-term latency predictions. A natural question is then the choice of relevant information to enable accurate predictions. Using traffic data collected from the Taiwan Freeway System, we consider the prediction of short-term latency of a freeway segment with a length of 17 km covering 5 measurement points, each collecting vehicle-by-vehicle data through the electronic toll collection system. The processed data include the past latencies of the freeway segment with different time lags, the traffic conditions of the individual segments (the accumulations, the traffic fluxes, the entrance and exit rates), the total accumulations, and the weekday latency profiles obtained by Gaussian process regression of past data. We arrive at several important conclusions about how data should be refined to obtain accurate predictions, which have implications for future system-wide latency predictions. (1) We find that the prediction of median latency is much more accurate and meaningful than the prediction of average latency, as the latter is plagued by outliers. This is verified by machine-learning prediction using XGBoost that yields a 35% improvement in the mean square error of the 5-minute averaged latencies. (2) We find that the median latency of the segment 15 minutes ago is a very good baseline for performance comparison, and we have evidence that further improvement is achieved by machine learning approaches such as XGBoost and Long Short-Term Memory (LSTM). (3) By analyzing the feature importance score in XGBoost and calculating the mutual information between the inputs and the latencies to be predicted, we identify a sequence of inputs ranked in importance. It confirms that the past latencies are most informative of the predicted latencies, followed by the total accumulation, whereas inputs such as the entrance and exit rates are uninformative. It also confirms that the inputs are much less informative of the average latencies than the median latencies. (4) For predicting the latencies of segments composed of two or three sub-segments, summing up the predicted latencies of each sub-segment is more accurate than the one-step prediction of the whole segment, especially with the latency prediction of the downstream sub-segments trained to anticipate latencies several minutes ahead. The duration of the anticipation time is an increasing function of the traveling time of the upstream segment. The above findings have important implications to predicting the full set of latencies among the various locations in the freeway system.

Keywords: data refinement, machine learning, mutual information, short-term latency prediction

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Authors: L. C. Sánchez, L. C. Corrales, A. G. Lancheros, E. Castañeda, Y. Ariza, L. S. Fuentes, L. Sierra, J. L. Cuervo

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The importance of environment friendly alternatives in agricultural processes is the reason why the research group Ceparium, the Colegio Mayor de Cundinamarca University, Colombia, investigated the genus Bacillus and its applicability for improving crops of economic importance in Colombia. In this investigation, we presented a study in which the genus Bacillus plays a leading role as beneficial microorganism. The objective was to identify the biochemical potential of three indigenous species of Bacillus, which were able to carry out actions for biological control against pathogens and pests or promoted growth to improve productivity of crops in Colombia. The procedures were performed in three phases: first, the production of biomass of an indigenous strain and a reference strain starting from culture media for production of spores and toxins were made. Spore count was done in a Neubauer chamber, concentrations of spores of Bacillus sphaericus were prepared and a bioassay was done at the Laboratory of Entomology at the University Jorge Tadeo Lozano of Plutella xylostella larvae, insect pest of crucifers in several Colombian regions. The second phase included the extraction in the liquid state fermentation, a secondary metabolite that has antibiosis action against fungi, call iturin B, and was obtained from strains of Bacillus subtilis. The molecule was identified using High Resolution Chromatography (HPLC) and its biocontrol effect on Fusarium sp fungus causes vascular wilt in economically important plant varieties, was confirmed using testing of antagonism in Petri dish. In the third phase, an initial procedure in that let recover and identify microorganisms of the genus Bacillus from the rhizosphere in two aromatic herbs, Rosmarinus officinalis and Thymus vulgaris L. was used. Subsequently, testing of antagonism against Fusarium sp were made and an assay was done under greenhouse conditions to observe biocontrol and growth promoting action by comparing growth in length and dry weight. In the first experiment, native Bacillus sphaericus was lethal to 92% Plutella xylostella larvae in 10 DDA. In the second experiment, iturin B was identified and biological control of Fusarium sp was demonstrated. In the third study, all strains demonstrated biological control and the B14 strain identified as Bacillus megaterium increased root length and productivity of the two plants in terms of weight. It was concluded that the native microorganisms of the genus Bacillus has a great biochemical potential that provides a beneficial interactions with plants, improve their growth and development and therefore a greater impact on production.

Keywords: genus bacillus, biological control, PGPRs, biochemical potential

Procedia PDF Downloads 436

Authors: Cinzia Federico, Gaetano Giudice, Salvatore Inguaggiato, Marco Liuzzo, Maria Pedone, Fabio Vita, Christoph Kern, Leonardo La Pica, Giovannella Pecoraino, Lorenzo Calderone, Vincenzo Francofonte

Abstract:

The periods of increased fumarolic activity at La Fossa volcano have been characterized, since early 80's, by changes in the gas chemistry and in the output rate of fumaroles. Excepting the direct measurements of the steam output from fumaroles performed from 1983 to 1995, the mass output of the single gas species has been recently measured, with various methods, only sporadically or for short periods. Since 2008, a scanning DOAS system is operating in the Palizzi area for the remote measurement of the in-plume SO₂ flux. On these grounds, the need of a cross-comparison of different methods for the in situ measurement of the output rate of different gas species is envisaged. In 2015, two field campaigns have been carried out, aimed at: 1. The mapping of the concentration of CO₂, H₂S and SO₂ in the fumarolic plume at 1 m from the surface, by using specific open-path diode tunable lasers (GasFinder Boreal Europe Ltd.) and an Active DOAS for SO₂, respectively; these measurements, coupled to simultaneous ultrasonic wind speed and meteorological data, have been elaborated to obtain the dispersion map and the output rate of single species in the overall fumarolic field; 2. The mapping of the concentrations of CO₂, H₂S, SO₂, H₂O in the fumarolic plume at 0.5 m from the soil, by using an integrated system, including IR spectrometers and specific electrochemical sensors; this has provided the concentration ratios of the analysed gas species and their distribution in the fumarolic field; 3. The in-fumarole sampling of vapour and measurement of the steam output, to validate the remote measurements. The dispersion map of CO₂, obtained from the tunable laser measurements, shows a maximum CO₂ concentration at 1m from the soil of 1000 ppmv along the rim, and 1800 ppmv in the inner slopes. As observed, the largest contribution derives from a wide fumarole of the inner-slope, despite its present outlet temperature of 230°C, almost 200°C lower than those measured at the rim fumaroles. Actually, fumaroles in the inner slopes are among those emitting the largest amount of magmatic vapour and, during the 1989-1991 crisis, reached the temperature of 690°C. The estimated CO₂ and H₂S fluxes are 400 t/d and 4.4 t/d, respectively. The coeval SO₂ flux, measured by the scanning DOAS system, is 9±1 t/d. The steam output, recomputed from CO₂ flux measurements, is about 2000 t/d. The various direct and remote methods (as described at points 1-3) have produced coherent results, which encourage to the use of daily and automatic DOAS SO₂ data, coupled with periodic in-plume measurements of different acidic gases, to obtain the total mass rates.

Keywords: DOAS, fumaroles, plume, tunable laser

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Authors: Remy Mpulumba Badiambile, Paul Musa Obadia, Malick Useni Mutayo, Jeef Numbi Mukanya, Patient Nkulu Banza, Tony Kayembe Kitenge, Erik Smolders, Jean-François Picron, Vincent Haufroid, Célestin Banza Lubaba Nkulu, Benoit Nemery

Abstract:

Background: In July 2021, the Tshikapa river became heavily polluted by mining wastes from a diamond mine in neighboring Angola, leading to massive killing of fish, as well as disease and even deaths among residents living along the Tshikapa and Kasai rivers, a major contributory of the Congo river. The exact nature of the pollutants was unknown. Methods: In a cross-sectional study conducted in the city of Tshikapa in August 2021, we enrolled by opportunistic sampling 65 residents (11 children < 16y) living alongside the polluted rivers and 65 control residents (5 children) living alongside a non-affected portion of the Kasai river (upstream from the Tshikapa-Kasai confluence). We administered a questionnaire and obtained spot urine samples for measurements of thiocyanate (a metabolite of cyanide) and 26 trace metals (by ICP-MS). Metals (and pH) were also measured in samples of river water. Results: Participants from both groups consumed river water. In the area affected by the pollution, most participants had eaten dead fish. Prevalences of reported health symptoms were higher in the exposed group than among controls: skin rashes (52% vs 0%), diarrhea (40% vs 8%), abdominal pain (8% vs 3%), nausea (3% vs 0%). In polluted water, concentrations [median (range)] were only higher for nickel [(2.2(1.4–3.5)µg/L] and uranium [78(71–91)ng/L] than in non-polluted water [0.8(0.6–1.9)µg/L; 9(7–19)ng/L]. In urine, concentrations [µg/g creatinine, median(IQR)] were significantly higher in the exposed group than in controls for lithium [19.5(12.4–27.3) vs 6.9(5.9–12.1)], thallium [0.41(0.31–0.57) vs 0.19(0.16–0.39)], and uranium [0.026(0.013–0.037)] vs 0.012(0.006–0.024)]. Other elements did not differ between the groups, but levels were higher than reference values for several metals (including manganese, cobalt, nickel, and lead). Urinary thiocyanate concentrations did not differ. Conclusion: This study, after an ecological disaster in the DRC, has documented contamination of river water by nickel and uranium and high urinary levels of some trace metals among affected riverine populations. However, the exact cause of the massive fish kill and disease among residents remains elusive. The capacity to rapidly investigate toxic pollution events must be increased in the area.

Keywords: metal contaminants, river water and human urine, pollution by mining wastes, DR Congo

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