Search results for: proteins separation

Authors: Thi Thao Ninh, Yuri Trusov, José Ramón Botella

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Heterotrimeric G proteins, comprised of three subunits, α, β and γ, are involved in signal transduction pathways that mediate a vast number of processes across the eukaryotic kingdom. 23 Gα subunits are present in humans whereas most plant genomes encode for only one canonical Gα. The disparity observed between Arabidopsis, rice, and maize Gα-deficient mutant phenotypes suggest that Gα functions have diversified between eudicots and monocots during evolution. Alternatively, since the only Gα mutations available in dicots have been produced in Arabidopsis, the possibility exists that this species might be an exception to the rule. In order to test this hypothesis, we studied the G protein α subunit (TGA1) in tomato. Four tga1 knockout lines were generated in tomato cultivar Moneymaker using CRISPR/Cas9. The tga1 mutants exhibit a number of auxin-related phenotypes including changes in leaf shape, reduced plant height, fruit size and number of seeds per fruit. In addition, tga1 mutants have increased sensitivity to abscisic acid during seed germination, reduced sensitivity to exogenous auxin during adventitious root formation from cotyledons and excised hypocotyl explants. Our results suggest that Gα mutant phenotypes in tomato are very similar to those observed in monocots, i.e. rice and maize, and cast doubts about the validity of using Arabidopsis as a model system for plant G protein studies.

Keywords: auxin-related phenotypes, CRISPR/Cas9, G protein α subunit, heterotrimeric G proteins, tomato

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Authors: Gadiri Sabiha

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Introduction: Current allergy diagnostic tools using Multiplex technology have made it possible to increase the efficiency of the search for specific IgE. This opportunity is provided by the newly developed “Alex Biochip”, consisting of a panel of 282 allergens in native and molecular form, a CCD inhibitor, and the potential for detecting cross-reactive allergens. We evaluated the performance of this technology in detecting cross-reactivity in previously explored patients. Material/Method: The sera of 39 patients presenting sensitization and polysensitization profiles were explored. The search for specific IgE is carried out by the Alex ® IgE Biochip, and the results are analyzed by nature and by molecular family of allergens using specific software. Results/Discussion: The analysis gave a particular profile of cross-reactivity allergens: 33% for the Ole e1 family, 31% for NPC2, 26% for storage proteins, 20% for Tropomyosin, 10% for LTPs, 10% for Arginine Kinase and 10% for Uteroglobin CCDs were absent in all patients. The “Ole e1” allergen is responsible for a pollen-pollen cross allergy. The storage proteins found and LTP are not species-specific, causing cross-pollen-food allergy. The nDer p2 of the NPC2 family is responsible for cross-reactivity between mite species. Conclusion: The cross-reactivities responsible for mixed syndromes at diagnosis in our patients were dominated by pollen-pollen and pollen-food syndromes. They allow the identification of severity factors linked to the prognosis and the best-adapted immunotherapy.

Keywords: specific IgE, allergy, cross reactivity, molecular allergens

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Authors: C. Lanzerstorfer

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Liquid sprays of water are frequently used in air pollution control for gas cooling purposes and for gas cleaning. Twin-fluid nozzles with internal mixing are often used for these purposes because of the small size of the drops produced. In these nozzles the liquid is dispersed by compressed air or another pressurized gas. In high efficiency scrubbers for particle separation, several nozzles are operated in parallel because of the size of the cross section. In such scrubbers, the scrubbing water has to be re-circulated. Precipitation of some solid material can occur in the liquid circuit, caused by chemical reactions. When such precipitations are detached from the place of formation, they can partly or totally block the liquid flow to a nozzle. Due to the resulting unbalanced supply of the nozzles with water and gas, the efficiency of separation decreases. Thus, the nozzles have to be cleaned if a certain fraction of blockages is reached. The aim of this study was to provide a tool for continuously monitoring the status of the nozzles of a scrubber based on the available operation data (water flow, air flow, water pressure and air pressure). The difference between the air pressure and the water pressure is not well suited for this purpose, because the difference is quite small and therefore very exact calibration of the pressure measurement would be required. Therefore, an equation for the reference air flow of a nozzle at the actual water flow and operation pressure was derived. This flow can be compared with the actual air flow for assessment of the status of the nozzles.

Keywords: condition monitoring, dual flow nozzles, flow equation, operation data

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Authors: Azam Zare

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Separation flow control for performance enhancement over airfoils at high incidence angle has become an increasingly important topic. This work details the characteristics of an efficient feedback control of the turbulent subsonic flow over NACA23012 airfoil using forced reduced‐order model based on the proper orthogonal decomposition/Galerkin projection and perturbation method on the compressible Reynolds Averaged Navier‐Stokes equations. The forced reduced‐order model is used in the optimal control of the turbulent separated flow over a NACA23012 airfoil at Mach number of 0.2, Reynolds number of 5×106, and high incidence angle of 24° using blowing/suction controlling jets. The Spallart-Almaras turbulence model is implemented for high Reynolds number calculations. The main shortcoming of the POD/Galerkin projection on flow equations for controlling purposes is that the blowing/suction controlling jet velocity does not show up explicitly in the resulting reduced order model. Combining perturbation method and POD/Galerkin projection on flow equations introduce a forced reduced‐order model that can predict the time-varying influence of the blowing/suction controlling jet velocity. An optimal control theory based on forced reduced‐order system is used to design a control law for a nonlinear reduced‐order model, which attempts to minimize the vorticity content in the turbulent flow field over NACA23012 airfoil. Numerical simulations were performed to help understand the behavior of the controlled suction jet at 12% to 18% chord from leading edge and a pair of blowing/suction jets at 15% to 18% and 24% to 30% chord from leading edge, respectively. Analysis of streamline profiles indicates that the blowing/suction jets are efficient in removing separation bubbles and increasing the lift coefficient up to 22%, while the perturbation method can predict the flow field in an accurate Manner.

Keywords: flow control, POD, Galerkin projection, separation

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Authors: Pawit Tangviroon, Apichit Svang-Ariyaskul

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Rubber seed oil is an attractive alternative feedstock for biodiesel production because it is not related to food-chain plant. Rubber seed oil contains large amount of free fatty acids, which causes problem in biodiesel production. Free fatty acids can react with alkaline catalyst in biodiesel production. Acid esterification is used as pre-treatment to convert unwanted compound to desirable biodiesel. Phase separation of oil and methanol occurs at low ratio of methanol to oil and causes low reaction rate and conversion. Acid esterification requires large excess of methanol in order to increase the miscibility of methanol in oil and accordingly, it is a more expensive separation process. In this work, the kinetics of esterification of rubber seed oil with methanol is developed from available experimental results. Reactive distillation process was designed by using Aspen Plus program. The effects of operating parameters such as feed ratio, molar reflux ratio, feed temperature, and feed stage are investigated in order to find the optimum conditions. Results show that the reactive distillation process is proved to be better than conventional process. It consumes less feed methanol and less energy while yielding higher product purity than the conventional process. This work can be used as a guideline for further development to industrial scale of biodiesel production using reactive distillation.

Keywords: biodiesel, reactive distillation, rubber seed oil, transesterification

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Authors: Nutcha Thianbut

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In the petroleum drilling industry, besides oil and gas, water is also produced from petroleum production. which will have oil droplets dispersed in the water as an emulsion. Commonly referred to as produced water, most industrial water-based produced water methods use the method of pumping water back into wells or catchment areas. because it cannot be utilized further, but in the compression of water each time, the cost is quite high. And the survey found that the amount of water from the petroleum production process has increased every year. In this research, we would like to study the removal of oil in produced water by the Coalescer device using fibers from agricultural waste as an intermediary. As an alternative to reduce the cost of water management in the petroleum drilling industry. The objectives of this research are 1. To study the fiber pretreatment by chemical process for the efficiency of oil-water separation 2. To study and design the fiber-packed coalescer device to destroy the emulsion of crude oil in water. 3. To study the working conditions of coalescer devices in emulsion destruction. using a fiber medium. In this research, the experiment was divided into two parts. The first part will study the absorbency of fibers. It compares untreated fibers with chemically treated alkaline fibers that change over time as well as adjusting the amount of fiber on the absorbency of the fiber and the second part will study the separation of oil from produced water by Coalescer equipment using fiber as medium to study the optimum condition of coalescer equipment for further development and industrial application.

Keywords: produced water, fiber, surface modification, coalescer

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Authors: Parminder Kaur Heer, Alexei Lapkin

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Due to global challenges of increased competition and demand for more sustainable products/processes, there is a rising pressure on the industry to develop innovative processes. Through Process Intensification (PI) the existing and new processes may be able to attain higher efficiency. However, very few PI options are generally considered. This is because processes are typically analysed at a unit operation level, thus limiting the search space for potential process options. PI performed at more detailed levels of a process can increase the size of the search space. The different levels at which PI can be achieved is unit operations, functional and phenomena level. Physical/chemical phenomena form the lowest level of aggregation and thus, are expected to give the highest impact because all the intensification options can be described by their enhancement. The objective of the current work is thus, generation of numerous process alternatives based on phenomena, and development of their corresponding computer aided models. The methodology comprises: a) automated generation of process options, and b) automated generation of process models. The process under investigation is disintegrated into functions viz. reaction, separation etc., and these functions are further broken down into the phenomena required to perform them. E.g., separation may be performed via vapour-liquid or liquid-liquid equilibrium. A list of phenomena for the process is formed and new phenomena, which can overcome the difficulties/drawbacks of the current process or can enhance the effectiveness of the process, are added to the list. For instance, catalyst separation issue can be handled by using solid catalysts; the corresponding phenomena are identified and added. The phenomena are then combined to generate all possible combinations. However, not all combinations make sense and, hence, screening is carried out to discard the combinations that are meaningless. For example, phase change phenomena need the co-presence of the energy transfer phenomena. Feasible combinations of phenomena are then assigned to the functions they execute. A combination may accomplish a single or multiple functions, i.e. it might perform reaction or reaction with separation. The combinations are then allotted to the functions needed for the process. This creates a series of options for carrying out each function. Combination of these options for different functions in the process leads to the generation of superstructure of process options. These process options, which are formed by a list of phenomena for each function, are passed to the model generation algorithm in the form of binaries (1, 0). The algorithm gathers the active phenomena and couples them to generate the model. A series of models is generated for the functions, which are combined to get the process model. The most promising process options are then chosen subjected to a performance criterion, for example purity of product, or via a multi-objective Pareto optimisation. The methodology was applied to a two-step process and the best route was determined based on the higher product yield. The current methodology can identify, produce and evaluate process intensification options from which the optimal process can be determined. It can be applied to any chemical/biochemical process because of its generic nature.

Keywords: Phenomena, Process intensification, Process models , Process options

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Authors: Saskya E. Carrera P., Ben Hankamer, Melanie Oey

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The green alga C. reinhardtii provides a platform for the cheap, scalable, and safe production of complex proteins. Despite gene expression in photosynthetic organisms being tightly regulated by light, most expression studies have analysed chloroplast recombinant protein production under constant light. Here the influence of illumination time and intensity on GFP and a GFP-PlyGBS (bacterial-lysin) fusion protein expression was investigated. The expression of both proteins was strongly influenced by the light regime (6-24 hr illumination per day), the light intensity (0-450 E m⁻²s⁻¹) and growth condition (photoautotrophic, mixotrophic and heterotrophic). Heterotrophic conditions resulted in relatively low recombinant protein yields per unit volume, despite high protein yields per cell, due to low growth rates. Mixotrophic conditions exhibited the highest yields at 6 hrs illumination at 200µE m⁻²s⁻¹ and under continuous low light illumination (13-16 mg L⁻¹ GFP and 1.2-1.6 mg L⁻¹ GFP-PlyGBS), as these conditions supported good cell growth and cellular protein yields. A ~23-fold increase in protein accumulation per cell and ~9-fold increase L⁻¹ culture was observed compared to standard constant 24 hr illumination for GFP-PlyGBS. The highest yields under photoautotrophic conditions were obtained under 9 hrs illumination (6 mg L⁻¹ GFP and 2.1 mg L⁻¹ GFP-PlyGBS). This represents a ~4-fold increase in cellular protein accumulation for GFP-PlyGBS. On a volumetric basis the highest yield was at 15 hrs illumination (~2-fold increase L⁻¹ over the constant light for GFP-PlyGBS). Optimising illumination conditions to balance growth and protein expression can thus significantly enhance overall recombinant protein production in C. reinhardtii cultures.

Keywords: chlamydomonas reinhardtii, light, mixotrophic, recombinant protein

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Authors: Elisa Y. M. Ang, Teng Yong Ng, Jingjie Yeo, Rongming Lin, Zishun Liu, K. R. Geethalakshmi

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By stacking carbon nanotubes (CNT) one on top of another, single layer CNT arrays can perform water-salt separation with ultra-high permeability and selectivity. Such outer-wall CNT slit membrane is named as the transverse flow CNT membrane. By adjusting the slit size between neighboring CNTs, the membrane can be configured to sieve out different solutes, right down to the separation of monovalent salt ions from water. Molecular dynamics (MD) simulation results show that the permeability of transverse flow CNT membrane is more than two times that of conventional axial-flow CNT membranes, and orders of magnitude higher than current reverse osmosis membrane. In addition, by carrying out MD simulations with different CNT size, it was observed that the variance in desalination performance with CNT size is small. This insensitivity of the transverse flow CNT membrane’s performance to CNT size is a distinct advantage over axial flow CNT membrane designs. Not only does the membrane operate well under constant pressure desalination operation, but MD simulations further indicate that oscillatory operation can further enhance the membrane’s desalination performance, making it suitable for operation such as electrodialysis reversal. While there are still challenges that need to be overcome, particularly on the physical fabrication of such membrane, it is hope that this versatile membrane design can bring the idea of using low dimensional structures for desalination closer to reality.

Keywords: carbon nanotubes, membrane desalination, transverse flow carbon nanotube membrane, molecular dynamics

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Authors: Rahul Madathiparambil Visalakshan, Alex Cavallaro, John Hayball, Krasimir Vasilev

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The role of biomaterial surface nanotopograhy on fibrinogen adsorption and unfolding, and the subsequent immune response were studied. Inconsistent topography and varying chemical functionalities along with a lack of reproducibility pose a challenge in determining the specific effects of nanotopography or chemistry on proteins and cells. It is important to have a well-defined nanotopography with a homogeneous chemistry to study the real effect of nanotopography on biological systems. Therefore, we developed a technique that can produce well-defined and highly reproducible topography to identify the role of specific roughness, size, height and density with the presence of homogeneous chemical functionality. Using plasma polymerisation of oxazoline monomers and immobilized gold nanoparticles we created surfaces with an equal number density of nanoparticles of different sizes. This surface was used to study the role of surface nanotopography and the interplay of surface chemistry on proteins and immune cells. The effect of nanotopography on fibrinogen adsorption was investigated using Quartz Cristal Microbalance with Dissipation and micro BCA. The mass of fibrinogen adsorbed on the surface increased with increasing size of nano-topography. Protein structural changes up on adsorption to the nano rough surface was studied using circular dichroism spectroscopy. Fibrinogen unfolding varied depending on the specific nanotopography of the surfaces. It was revealed that the in vitro immune response to the nanotopography surfaces changed due to this protein unfolding.

Keywords: biomaterial inflammation, protein and cell responses, protein unfolding, surface nanotopography

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Authors: Serges Djoyou Kamga, Gerard Emmanuel Kamdem Kamga

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Any serious constitutionalism entails a system of government characterised by the separation of powers between the executive, the legislature, and the judiciary. The latter is generally in charge of upholding the rule of law and the respect for human rights which are vital for the functioning of any democracy. Therefore, for the judiciary to play its role as a watchdog, it should be independent from other branches of government. The aim of this paper is to examine the independence of the judiciary in South Africa after 20 of democracy. Defining judicial independence as the courts’ ability ‘to decide cases on the basis of established law and the merits of the case, without interference from other political or governmental agents’, the paper examines the extent to which the South African judiciary is independent after twenty years of democracy. As part of assessing the independence of the judiciary, the paper begins by looking at the situation during apartheid, then proceeds with an examination of the post-apartheid legal order. It also examines the institutional independence of the judiciary by looking into its day to day activities which revolve around its self-governance, or administrative and financial independence. In addition, the paper assesses the judges’ individual independence by examining whether judicial appointment, security of tenure, judges’ remuneration and disciplinary actions and the removal of judges from office do not contain loopholes that can hinder judicial independence. Ultimately, the chapter argues that although the South African model of judicial independence is yet to be perfect, it is a good practice that can be emulated by other African countries.

Keywords: judical independence, South Africa, democracy, separation of powers

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Authors: Silvia Gastaldello, Maria Grillo, Luca Tassoni, Claudio Maran, Stefano Balbo

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Antioxidants are nowadays attractive not only for the countless benefits to the human and animal health, but also for the perspective of use as food preservative instead of synthetic chemical molecules. In this study, the radical scavenging activity of six protein extracts from pulse and oleaginous seeds was evaluated. The selected matrices are Pisum sativum (yellow pea from two different origins), Carthamus tinctorius (safflower), Helianthus annuus (sunflower), Lupinus luteus cv Mister (lupin) and Glycine max (soybean), since they are economically interesting for both human and animal nutrition. The seeds were grinded and proteins extracted from 20mg powder with a specific vegetal-extraction kit. Proteins have been quantified through Bradford protocol and scavenging activity was revealed using DPPH assay, based on radical DPPH (2,2-diphenyl-1-picrylhydrazyl) absorbance decrease in the presence of antioxidants molecules. Different concentrations of the protein extract (1, 5, 10, 50, 100, 500 µg/ml) were mixed with DPPH solution (DPPH 0,004% in ethanol 70% v/v). Ascorbic acid was used as a scavenging activity standard reference, at the same six concentrations of protein extracts, while DPPH solution was used as control. Samples and standard were prepared in triplicate and incubated for 30 minutes in dark at room temperature, the absorbance was read at 517nm (ABS30). Average and standard deviation of absorbance values were calculated for each concentration of samples and standard. Statistical analysis using t-students and p-value were performed to assess the statistical significance of the scavenging activity difference between the samples (or standard) and control (ABSctrl). The percentage of antioxidant activity has been calculated using the formula [(ABSctrl-ABS30)/ABSctrl]*100. The obtained results demonstrate that all matrices showed antioxidant activity. Ascorbic acid, used as standard, exhibits a 96% scavenging activity at the concentration of 500 µg/ml. At the same conditions, sunflower, safflower and yellow peas revealed the highest antioxidant performance among the matrices analyzed, with an activity of 74%, 68% and 70% respectively (p < 0.005). Although lupin and soybean exhibit a lower antioxidant activity compared to the other matrices, they showed a percentage of 46 and 36 respectively. All these data suggest the possibility to use undervalued edible matrices as antioxidants source. However, further studies are necessary to investigate a possible synergic effect of several matrices as well as the impact of industrial processes for a large-scale approach.

Keywords: antioxidants, DPPH assay, natural matrices, vegetal proteins

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Authors: M. Latha, Achintya K. Dolui, P. Vijayaraj

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Vegetable oils mainly triacylglycerol (TAG) are an essential nutrient in the human diet as well as one of the major global commodity. There is a pressing need to enhance the yield of oil production to meet the world’s growing demand. Oil content is controlled by the balance between synthesis and breakdown in the cells. Several studies have established to increase the oil content by the overexpression of oil biosynthetic enzymes. Interestingly the significant oil accumulation was observed with impaired TAG hydrolysis. Unfortunately, the structural, as well as the biochemical properties of the lipase enzymes, is widely unknown, and so far, no candidate gene was identified in seeds except sugar-dependent1 (SDP1). Evidence has shown that SDP1directly responsible for initiation of oil breakdown in the seeds during germination. The present study is the identification of seed-specific acyl-hydrolases by activity based proteome profiling (ABPP) using Arabidopsis thaliana as a model system. The ABPP reveals that around 8 to 10 proteins having the serine hydrolase domain and are expressed during germination of Arabidopsis seed. The N-term sequencing, as well as LC-MS/MS analysis, was performed for the differentially expressed protein during germination. The coding region of the identified proteins was cloned, and lipases activity was assessed with purified recombinant protein. The enzyme assay was performed against various lipid substrates, and we have observed the acylhydrolase activity towards lysophosphatidylcholine and monoacylglycerol. Further, the functional characteristic of the identified protein will reveal the physiological significance the enzyme in oil accumulation.

Keywords: lipase, lipids, vegetable oil, triacylglycerol

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Authors: Kanchan Yadav, Ai-Chuan Chou, Rajesh Kumar Ulaganathan, Hua-De Gao, Hsien-Ming Lee, Chien-Yuan Pan, Yit-Tsong Chen

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Optogenetics is an innovative technology now widely adopted by researchers in different fields of the biological sciences. However, due to the weak tissue penetration capability of the short wavelengths used to activate light-sensitive proteins, an invasive light guide has been used in animal studies for photoexcitation of target tissues. Upconverting nanoparticles (UCNPs), which transform near-infrared (NIR) light to short-wavelength emissions, can help address this issue. To improve optogenetic performance, we enhance the target selectivity for optogenetic controls by specifically conjugating the UCNPs with light-sensitive proteins at a molecular level, which shortens the distance as well as enhances the efficiency of energy transfer. We tagged V5 and Lumio epitopes to the extracellular N-terminal of channelrhodopsin-2 with an mCherry conjugated at the intracellular C-terminal (VL-ChR2m) and then bound NeutrAvidin-functionalized UCNPs (NAv-UCNPs) to the VL-ChR2m via a biotinylated antibody against V5 (bV5-Ab). We observed an apparent energy transfer from the excited UCNP (donor) to the bound VL-ChR2m (receptor) by measuring emission-intensity changes at the donor-receptor complex. The successful patch-clamp electrophysiological test and an intracellular Ca2+ elevation observed in the designed UCNP-ChR2 system under optogenetic manipulation confirmed the practical employment of UCNP-assisted NIR-optogenetic functionality. This work represents a significant step toward improving therapeutic optogenetics.

Keywords: Channelrhodopsin-2, near infrared, optogenetics, upconverting nanoparticles

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Authors: Orsolya Horvath, Laszlo Deres, Krisztian Eros, Katalin Ordog, Tamas Habon, Balazs Sumegi, Kalman Toth, Robert Halmosi

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Introduction: Oxidative stress induces an imbalance in mitochondrial fusion and fission processes, finally leading to cell death. The two antioxidant molecules, BGP-15 and L2286 have beneficial effects on mitochondrial functions and on cellular oxidative stress response. In this work, we studied the effects of these compounds on the processes of mitochondrial quality control. Methods: We used H9c2 cardiomyoblast and isolated neonatal rat cardiomyocytes (NRCM) for the experiments. The concentration of stressors and antioxidants was beforehand determined with MTT test. We applied 1-Methyl-3-nitro-1-nitrosoguanidine (MNNG) in 125 µM, 400 µM and 800 µM concentrations for 4 and 8 hours on H9c2 cells. H₂O₂ was applied in 150 µM and 300 µM concentration for 0.5 and 4 hours on both models. L2286 was administered in 10 µM, while BGP-15 in 50 µM doses. Cellular levels of the key proteins playing role in mitochondrial dynamics were measured in Western blot samples. For the analysis of mitochondrial network dynamics, we applied electron microscopy and immunocytochemistry. Results: Due to MNNG treatment the level of fusion proteins (OPA1, MFN2) decreased, while the level of fission protein DRP1 elevated markedly. The levels of fusion proteins OPA1 and MNF2 increased in the L2286 and BGP-15 treated groups. During the 8 hour treatment period, the level of DRP1 also increased in the treated cells (p < 0.05). In the H₂O₂ stressed cells, administration of L2286 increased the level of OPA1 in both H9c2 and NRCM models. MFN2 levels in isolated neonatal rat cardiomyocytes raised considerably due to BGP-15 treatment (p < 0.05). L2286 administration decreased the DRP1 level in H9c2 cells (p < 0.05). We observed that the H₂O₂-induced mitochondrial fragmentation could be decreased by L2286 treatment. Conclusion: Our results indicated that the PARP-inhibitor L2286 has beneficial effect on mitochondrial dynamics during oxidative stress scenario, and also in the case of directly induced DNA damage. We could make the similar conclusions in case of BGP-15 administration, which, via reducing ROS accumulation, propagates fusion processes, this way aids preserving cellular viability. Funding: GINOP-2.3.2-15-2016-00049; GINOP-2.3.2-15-2016-00048; GINOP-2.3.3-15-2016-00025; EFOP-3.6.1-16-2016-00004; ÚNKP-17-4-I-PTE-209

Keywords: H9c2, mitochondrial dynamics, neonatal rat cardiomyocytes, oxidative stress

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Authors: S. Teimouri, S. Abbasi

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Milk-fruit juice mixture is a type of soft drinks, which can be produced by mixing milk with pieces of fruits, fruit juices, or fruit juices concentrates. The major problem of these products, mainly the acidic ones, is phase separation which occurs during formulation and storage due to the aggregation of caseins at low pH Short-chain inulin (CLR), long-chain inulin (TEX), native inulin (IQ) and Long-chain inulin (TEX) and short-chain inulin (CLR) combined in different proportions (2o:80, 50:50, and 80:20) were added (2-10 %) to sourcherry juice-milk mixture and their stabilization mechanisms were studied with using rheological and microstructural observations. Stevioside as a bio-sweetener and sugar-replacer was added at last step. Finally, sensory analyses were taken place on stabilized samples. According to the findings, TEX stabilized the mixture at concentration of 8%. MIX and IQ reduced phase separation at high concentration but had not complete effect on stabilization. CLR did not effect on stabilization. Rheological changes and inulin aggregates formation were not observed in CLR samples during the one month storage period. However TEX, MIX and IQ samples formed inulin aggregates and became more thixotropic, elastic and increased the viscosity of mixture. The rate of the inulin aggregates formation and viscosity increasing was in the following order TEX > MIX > IQ. Consequently the mixture which stabilized with inulin and sweetened with stevioside had the prebiotic properties which may suggest to diabetic patients and children.

Keywords: prebiotic, inulin, casein, stabilization, stevioside

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Authors: Zoe Goldberger, Priscilla S. Briquez, Jeffrey A. Hubbell

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Tumor cells have mutations resulting from genetic instability that the immune system can actively recognize. Immune checkpoint immunotherapy (ICI) is commonly used in the clinic to re-activate immune reactions against mutated proteins, called neoantigens, resulting in tumor remission in cancer patients. However, only around 20% of patients show durable response to ICI. While tumor mutational burden (TMB) has been approved by the Food and Drug Administration (FDA) as a criterion for ICI therapy, the relevance of the subcellular localizations of the mutated proteins within the tumor cell has not been investigated. Here, we hypothesized that localization of mutations impacts the effect of immune responsiveness to ICI. We analyzed publicly available tumor mutation sequencing data of ICI treated patients from 3 independent datasets. We extracted the subcellular localization from the UniProtKB/Swiss-Prot database and quantified the proportion of membrane, cytoplasmic, nuclear, or secreted mutations per patient. We analyzed this information in relation to response to ICI treatment and overall survival of patients showing with 1722 ICI-treated patients that high mutational burden localized at the membrane (mTMB), correlate with ICI responsiveness, and improved overall survival in multiple cancer types. We anticipate that our results will ameliorate predictability of cancer patient response to ICI with potential implications in clinical guidelines to tailor ICI treatment. This would not only increase patient survival for those receiving ICI, but also patients’ quality of life by reducing the number of patients enduring non-effective ICI treatments.

Keywords: cancer, immunotherapy, membrane neoantigens, efficacy prediction, biomarkers

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Authors: Mohammed Sabbah, Prospero Di Pierro, Raffaele Porta

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Protein-based edible films and coatings have attracted an increasing interest in recent years since they might be used to protect pharmaceuticals or improve the shelf life of different food products. Among them, several plant proteins represent an abundant, inexpensive and renewable raw source. These natural biopolymers are used as film forming agents, being able to form intermolecular linkages by various interactions. However, without the addition of a plasticizing agent, many biomaterials are brittle and, consequently, very difficult to be manipulated. Plasticizers are generally small and non-volatile organic additives used to increase film extensibility and reduce its crystallinity, brittleness and water vapor permeability. Plasticizers normally act by decreasing the intermolecular forces along the polymer chains, thus reducing the relative number of polymer-polymer contacts, producing a decrease in cohesion and tensile strength and thereby increasing film flexibility allowing its deformation without rupture. The most commonly studied plasticizers are polyols, like glycerol (GLY) and some mono or oligosaccharides. In particular, GLY not only increases film extensibility but also migrates inside the film network often causing the loss of desirable mechanical properties of the material. Therefore, replacing GLY with a different plasticizer might help to improve film characteristics allowing potential industrial applications. To improve film properties, it seemed of interest to test as plasticizers some cationic small molecules like polyamines (PAs). Putrescine, spermidine (SPD), and spermine are PAs widely distributed in nature and of particular interest for their biological activities that may have some beneficial health effects. Since PAs contains amino instead of hydroxyl functional groups, they are able to trigger ionic interactions with negatively charged proteins. Bitter vetch (Vicia ervilia; BV) is an ancient grain legume crop, originated in the Mediterranean region, which can be found today in many countries around the world. This annual Vicia genus shows several favorable features, being their seeds a cheap and abundant protein source. The main objectives of this study were to investigate the effect of different concentrations of SPD on the mechanical and permeability properties of films prepared with native or heat denatured BV proteins in the presence of different concentrations of SPD and/or GLY. Therefore, a BV seed protein concentrate (BVPC), containing about 77% proteins, was used to prepare film forming solutions (FFSs), whereas GLY and SPD were added as film plasticizers, either singly or in combination, at various concentrations. Since a primary plasticizer is generally defined as a molecule that when added to a material makes it softer, more flexible and easier to be processed, our findings lead to consider SPD as a possible primary plasticizer of protein-based films. In fact, the addition of millimolar concentrations of SPD to BVPC FFS allowed obtaining handleable biomaterials with improved properties. Moreover, SPD can be also considered as a secondary plasticizer, namely an 'extender', because of its ability even to enhance the plasticizing performance of GLY. In conclusion, our studies indicate that innovative edible protein-based films and coatings can be obtained by using PAs as new plasticizers.

Keywords: edible films, glycerol, plasticizers, polyamines, spermidine

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Authors: Hafiza Javaria Ashraf, Xinghong Wang, Zhanghong Shi, Youming Hou

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Insect’s innate immunity depends on a variety of defense responses for the recognition of invading pathogens. Pathogen recognition involves particular proteins known as pattern recognition receptors (PRRs). These PRRs interact with pathogen-associated molecular patterns (PAMPs) present on the surface of pathogens to distinguish between self and non-self. C-type lectins (CTLs) belong to a superfamily of PPRs which involved in insect immunity and defense mechanism. Rhynchophorus ferrugineus Olivier is a devastating pest of Palm cultivations in China. Although studies on R. ferrugineus immune mechanism and host defense have conducted, however, the role of CTL in immune responses of R. ferrugineus remains elusive. Here, we report RfCTL, which is a secreted protein containing a single-CRD domain. The open reading frame (ORF) of CTL is 226 bp, which encodes a putative protein of 168 amino acids. Transcript expression analysis revealed that RfCTL highly expressed in immune-related tissues, i.e., hemolymph and fat body. The abundance of RfCTL in the gut and fat body dramatically increased upon Staphylococcus aureus and Escherichia coli bacterial challenges, suggesting a role in defense against gram-positive and gram-negative bacterial infection. Taken together, we inferred that RfCTL might be involved in the immune defense of R. ferrugineus and established a solid foundation for future studies on R. ferrugineus CTL domain proteins for better understanding of insect immunity.

Keywords: biological invasion, c-type lectin, insect immunity, Rhynchophorus ferrugineus Oliver

Procedia PDF Downloads 145

Authors: Ewa Rudnik

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Poster shows a comparison of hydrometallurgical routes of copper recovery from low-grade e-waste. Electronic scrap was smelted to produce Cu–Zn–Ag alloy. The alloy was then treated in the following ways: (a) anodic dissolution with simultaneous metal electrodeposition using ammoniacal and sulfuric acid solutions. This resulted in the separation of metals, where lead, silver and tin accumulated mainly in the slimes, while copper was transferred to the electrolyte and then recovered on the cathode. The best conditions of the alloy treatment were obtained in the sulfuric acid, where the final product was metal of high purity (99% Cu) at the current efficiency of 90%. (b) leaching in ammoniacal solutions of various compositions and then copper electrowinning. Alloy was leached in chloride, carbonate, sulfate and thiosulfate baths. This resulted in the separation of the metals, wherein copper and zinc were transferred to the electrolyte, while metallic tin and silver as well as lead salts remained in the slimes. Copper was selectively recovered from the ammoniacal solutions by the electrolysis, leaving zinc ions in the electrolyte. The best conditions of the alloy treatment were obtained in the ammonia-carbonate system, where the final product was copper of high purity (99.9%) at the current efficiency of 60%. Thiosulfate solution was not applicable for the leaching of the copper alloy due to secondary reactions of the formation of copper (I) thiosulfate complexes and precipitation of copper (I) sulfide.

Keywords: alloy, electrolysis, e-waste, leaching

Procedia PDF Downloads 359

Authors: T. N. Girish, B. E. Pradeep, Ravi Parkash

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Insects from tropical wet or dry seasons are likely to cope starvation stress through seasonal phenotypic plasticity in energy metabolites. Accordingly, we analyzed such plastic changes in Zaprionus indianus flies reared under wet or dry season-specific conditions; and also after adult acclimation at 32℃ for 1 to 6 days; and to low (40% RH) or high (70% RH) humidity. Both thermal or humidity acclimation revealed significant accumulation of body lipids for wet season flies but low humidity acclimation did not change the level of body lipids in dry season flies. Developmental and adult acclimation showed sex specific differences i.e., starvation resistance and body lipids were higher in the males of dry season but in females of wet season. We found seasonal and sex specific differences in the relative level for body lipids at death; and in the rates of accumulation or utilization of energy metabolites (body lipids, carbohydrates and proteins). Body lipids constitute the preferred energy source under starvation for flies of both the seasons. However, utilization of carbohydrates (~20% to 30%) and proteins (~20% to 25%) was evident only in dry season flies. Higher starvation resistance after thermal or humidity acclimation is achieved by increased accumulation of lipids. Adult acclimation of wet or dry season flies revealed plastic changes in mean daily fecundity despite reduction in fecundity under starvation. Thus, thermal or humidity induced plastic responses in body lipids support starvation resistance under wet or dry seasons.

Keywords: heat or humidity acclimation, plastic changes in body lipids and starvation resistance, tropical drosophilid, Wet- or Dry seasons, Zaprionus indianus

Procedia PDF Downloads 147

Authors: Reza Babajanivalashedi, Stefania Lo Feudo, Jean-Luc Dion

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Liquid sloshing is a crucial problem for the dynamic of moving containers in the packaging industries. Sloshing issues have been so far mainly modeled within the framework of fluid dynamics or by using equivalent mechanical models with different kinds of movements and shapes of containers. Nevertheless, these approaches do not allow to determinate the shape of the free surface of the liquid in case of the irregular shape of the moving containers, so that experimental measurements may be required. If there is too much slosh in the moving tank, the liquid can be splashed out on the packages. So, the free surface oscillation must be controlled/reduced to eliminate the splashing. The purpose of this research is to propose an optimization algorithm for finding an optimum command law to reduce surface elevation. In the first step, the free surface of the liquid is simulated based on the separation variable and weak formulation models. Then Genetic and Gradient algorithms are developed for finding the optimum command law. The optimum command law is compared with existing command laws, and the results show that there is a significant difference in surface oscillation between optimum and existing command laws. This algorithm is applicable for different varieties of bottles in case of using the camera for detecting the liquid elevation, and it can produce new command laws for different kinds of tanks to reduce the surface oscillation and remove the splashing phenomenon.

Keywords: sloshing phenomenon, separation variables, weak formulation, optimization algorithm, command law

Procedia PDF Downloads 138

Authors: Shuang Chen, Zongqian Shi, Jiajia Sun, Mingjia Li

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Microfluidics is a technology that small amounts of fluids are manipulated using channels with dimensions of tens to hundreds of micrometers. At present, this significant technology is required for several applications in some fields, including disease diagnostics, genetic engineering, and environmental monitoring, etc. Among these fields, manipulation of microparticles and cells in microfluidic device, especially separation, have aroused general concern. In magnetic field, the separation methods include positive and negative magnetophoresis. By comparison, negative magnetophoresis is a label-free technology. It has many advantages, e.g., easy operation, low cost, and simple design. Before the separation of particles or cells, focusing them into a single tight stream is usually a necessary upstream operation. In this work, the focusing of diamagnetic particles in ferrofluid microflows with a single set of overhead permanent magnets is investigated numerically. The geometric model of the simulation is based on the configuration of previous experiments. The straight microchannel is 24mm long and has a rectangular cross-section of 100μm in width and 50μm in depth. The spherical diamagnetic particles of 10μm in diameter are suspended into ferrofluid. The initial concentration of the ferrofluid c₀ is 0.096%, and the flow rate of the ferrofluid is 1.8mL/h. The magnetic field is induced by five identical rectangular neodymium−iron− boron permanent magnets (1/8 × 1/8 × 1/8 in.), and it is calculated by equivalent charge source (ECS) method. The flow of the ferrofluid is governed by the Navier–Stokes equations. The trajectories of particles are solved by the discrete phase model (DPM) in the ANSYS FLUENT program. The positions of diamagnetic particles are recorded by transient simulation. Compared with the results of the mentioned experiments, our simulation shows consistent results that diamagnetic particles are gradually focused in ferrofluid under magnetic field. Besides, the diamagnetic particle focusing is studied by varying the flow rate of the ferrofluid. It is in agreement with the experiment that the diamagnetic particle focusing is better with the increase of the flow rate. Furthermore, it is investigated that the diamagnetic particle focusing is affected by other factors, e.g., the width and depth of the microchannel, the concentration of the ferrofluid and the diameter of diamagnetic particles.

Keywords: diamagnetic particle, focusing, microfluidics, permanent magnet

Procedia PDF Downloads 124

Authors: Ebrahim Rahmani, Siyamak Moazeni

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According to the Iraq's first constitutional law (approved 2005), Iraq is an independent with sovereignty and its governmental structure is parliamentary democratic republic and federal. Even in reforms in 2009, this article of law did not changed at all. But considering the existence of this emphasis and clarity which is mentioned in the law, different and sometimes contradictory interpretations and positions are expressed about federalism in the way that we can say, considering the importance of the matter, federalism is a focus point to create and expansion of the cold war among leaders of different groups of the country. Iraq's today political and security position has granted the suitable opportunity to Iraq's Kurdistan in appearing of the recent security crisis to increase its share from the central political power or to achieve to its independent dream. The federalism the weakest point of Iraq's territorial integrity in a way that if different groups do not come to a consensus about it and do not think about a mechanism which is accepted by all of them, this can effect on and Iraq's political stability and security. Iraq's Kurdistan follows the option of disintegration and separation under the shadow of political and security changes, even with existence of some Iraqi groups' hopes regarding the improvement of situation after parliament election and also considering Masoud Barezani's power will for separation from Iraq as well as regarding special international changes and disintegration of Karime from Ukraine and ISIS crises; concerns have been created among regional and international powers and interior players. In this article, a paradox due constitutional law about federalism, Iraq's central government view and its politicians to the matter and the regional effect of this action on region's geopolitics are reviewed as well.

Keywords: constitutional law, federalism, decomposition, Iraq's Kurdistan

Procedia PDF Downloads 318

Authors: Dhruv J. Limaye, Hanga Galfalvy, Cheick A. Sissoko, Yung-yu Huang, Chunanning Tang, Ying Liu, Shu-Chi Hsiung, Andrew J. Dwork, Gorazd B. Rosoklija, Victoria Arango, Lewis Brown, J. John Mann, Maura Boldrini

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Memory and emotion require hippocampal cell viability and connectivity and are disrupted in major depressive disorder (MDD). Applying shotgun proteomics and stereological quantification of neural progenitor cells (NPCs), intermediate neural progenitors (INPs), and mature granule neurons (GNs), to postmortem human hippocampus, identified differentially expressed proteins (DEPs), and fewer NPCs, INPs and GNs, in untreated MDD (uMDD) compared with non-psychiatric controls (CTRL) and antidepressant-treated MDD (MDDT). DEPs lower in uMDD vs. CTRL promote mitosis, differentiation, and prevent apoptosis. DEPs higher in uMDD vs. CTRL inhibit the cell cycle, and regulate cell adhesion, neurite outgrowth, and DNA repair. DEPs lower in MDDT vs. uMDD block cell proliferation. We observe group-specific correlations between numbers of NPCs, INPs, and GNs and an abundance of proteins regulating mitosis, differentiation, and apoptosis. Altered protein expression underlies hippocampus cellular and volume loss in uMDD, supports a trophic effect of antidepressants, and offers new treatment targets.

Keywords: proteomics, hippocampus, depression, mitosis, migration, differentiation, mitochondria, apoptosis, antidepressants, human brain

Procedia PDF Downloads 95

Authors: Mahsa Taghavi

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In the treatment of breast cancer, tamoxifen is a regularly prescribed medication. The effect of tamoxifen on breast cancer patients' EMT pathways was studied. In this study to see if it had any effect on the cancer cells' resistance to tamoxifen and to look for specific miRNAs associated with EMT. In this work, we used continuous and integrated bioinformatics analysis to choose the optimal GEO datasets. Once we had sorted the gene expression profile, we looked at the mechanism of signaling, the ontology of genes, and the protein interaction of each gene. In the end, we used the GEPIA database to confirm the candidate genes. after that, I investigated critical miRNAs related to candidate genes. There were two gene expression profiles that were categorized into two distinct groups. Using the expression profile of genes that were lowered in the EMT pathway, the first group was examined. The second group represented the polar opposite of the first. A total of 253 genes from the first group and 302 genes from the second group were found to be common. Several genes in the first category were linked to cell death, focal adhesion, and cellular aging. Two genes in the second group were linked to cell death, focal adhesion, and cellular aging. distinct cell cycle stages were observed. Finally, proteins such as MYLK, SOCS3, and STAT5B from the first group and BIRC5, PLK1, and RAPGAP1 from the second group were selected as potential candidates linked to tamoxifen's influence on the EMT pathway. hsa-miR-1204 and hsa-miR-639 have a very close relationship with the candidates genes according to the node degrees and betweenness index. With this, the action of tamoxifen on the EMT pathway was better understood. It's important to learn more about how tamoxifen's target genes and proteins work so that we can better understand the drug.

Keywords: tamoxifen, breast cancer, bioinformatics analysis, EMT, miRNAs

Procedia PDF Downloads 124

Authors: Dorothea Voß, Tobias Esser, Michael Huber, Jakob Albert

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The growing world population and the associated increase in demand for energy and consumer goods, as well as increasing waste production, requires the development of sustainable processes. In addition, the increasing environmental awareness of our society is a driving force for the requirement that processes must be as resource and energy efficient as possible. In this context, the use of polyoxometalate catalysts (POMs) has emerged as a promising approach for the development of green processes. POMs are bifunctional polynuclear metal-oxo-anion cluster characterized by a strong Brønsted acidity, a high proton mobility combined with fast multi-electron transfer and tunable redox potential. In addition, POMs are soluble in many commonly known solvents and exhibit resistance to hydrolytic and oxidative degradation. Due to their structure and excellent physicochemical properties, POMs are efficient acid and oxidation catalysts that have attracted much attention in recent years. Oxidation processes with molecular oxygen are worth mentioning here. However, the fact that the POM catalysts are homogeneous poses a challenge for downstream processing of product solutions and recycling of the catalysts. In this regard, nanofiltration membranes have gained increasing interest in recent years, particularly due to their relative sustainability advantage over other technologies and their unique properties such as increased selectivity towards multivalent ions. In order to establish an efficient downstream process for the highly selective separation of homogeneous POM catalysts from aqueous solutions using nanofiltration membranes, a laboratory-scale membrane system was designed and constructed. By varying various process parameters, a sensitivity analysis was performed on a model system to develop an optimized method for the recovery of POM catalysts. From this, process-relevant key figures such as the rejection of various system components were derived. These results form the basis for further experiments on other systems to test the transferability to serval separation tasks with different POMs and products, as well as for recycling experiments of the catalysts in processes on laboratory scale.

Keywords: downstream processing, nanofiltration, polyoxometalates, homogeneous catalysis, green chemistry

Procedia PDF Downloads 83

Authors: Sandeep Mohan Ahuja, Gulshan Kumar Jawa

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Cyclone separators are crucial components in various industries tasked with efficiently separating particulate matter from gas streams. Achieving optimal performance hinges on a deep understanding of flow dynamics and particle behaviour within these separators. In this investigation, Computational Fluid Dynamics (CFD) simulations are conducted utilizing the Discrete Phase Model (DPM) to dissect the intricate flow patterns, particle trajectories, and separation efficiency within cyclone separators. The study delves into the influence of pivotal parameters like inlet velocity, particle size distribution, and cyclone geometry on separation efficiency. Through numerical simulations, a comprehensive comprehension of fluid-particle interaction phenomena within cyclone separators is attained, allowing for the assessment of solid collection efficiency across diverse operational conditions and geometrical setups. The insights gleaned from this study promise to advance our understanding of the complex interplay between fluid and particle within cyclone separators, thereby enabling optimization across a wide array of industrial applications. By harnessing the power of CFD simulations and the DPM, this research endeavours to furnish valuable insights for designing, operating, and evaluating the performance of cyclone separators, ultimately fostering greater efficiency and environmental sustainability within industrial processes.

Keywords: cyclone separator, computational fluid dynamics, enhancing efficiency, discrete phase model

Procedia PDF Downloads 30

Authors: S. Jain, R. Savalia, V. Saini

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A simple, accurate, precise, sensitive and specific RP-HPLC method was developed and validated for simultaneous estimation of Metoprolol Succinate and Olmesartan Medoxomil in bulk and tablet dosage form. The RP-HPLC method has shown adequate separation for Metoprolol Succinate and Olmesartan Medoxomil from its degradation products. The separation was achieved on a Phenomenex luna ODS C18 (250mm X 4.6mm i.d., 5μm particle size) with an isocratic mixture of acetonitrile: 50mM phosphate buffer pH 4.0 adjusted with glacial acetic acid in the ratio of 55:45 v/v. The mobile phase at a flow rate of 1.0ml/min, Injection volume 20μl and wavelength of detection was kept at 225nm. The retention time for Metoprolol Succinate and Olmesartan Medoxomil was 2.451±0.1min and 6.167±0.1min, respectively. The linearity of the proposed method was investigated in the range of 5-50μg/ml and 2-20μg/ml for Metoprolol Succinate and Olmesartan Medoxomil, respectively. Correlation coefficient was 0.999 and 0.9996 for Metoprolol Succinate and Olmesartan Medoxomil, respectively. The limit of detection was 0.2847μg/ml and 0.1251μg/ml for Metoprolol Succinate and Olmesartan Medoxomil, respectively and the limit of quantification was 0.8630μg/ml and 0.3793μg/ml for Metoprolol and Olmesartan, respectively. Proposed methods were validated as per ICH guidelines for linearity, accuracy, precision, specificity and robustness for estimation of Metoprolol Succinate and Olmesartan Medoxomil in commercially available tablet dosage form and results were found to be satisfactory. Thus the developed and validated stability indicating method can be used successfully for marketed formulations.

Keywords: metoprolol succinate, olmesartan medoxomil, RP-HPLC method, validation, ICH

Procedia PDF Downloads 303

Authors: Kun Liang Ang, Eng Toon Saw, Wei He, Xuecheng Dong, Seeram Ramakrishna

Abstract:

Ester solvents are widely used in pharmaceutical, printing and flavor industry due to their good miscibility, low toxicity, and high volatility. Through pervaporation, these ester solvents can be recovered from industrial wastewater. While metal-doped silicalite 1 zeolite membranes are commonly used in organic solvent recovery in the pervaporation process, these ceramic membranes suffer from low membrane permeation flux, mainly due to the high thickness of the metal-doped zeolite membrane. Herein, a simple method of fabricating an ultrathin tin-silicalite 1 membrane supported on alumina tube is reported. This ultrathin membrane is able to achieve high permeation flux and separation factor for an ester in a diluted aqueous solution. Nanosized tin-Silicalite 1 seeds which are smaller than 500nm has been formed through hydrothermal synthesis. The sn-Silicalite 1 seeds were then seeded onto alumina tube through dip coating, and the tin-Silicalite 1 membrane was then formed by hydrothermal synthesis in an autoclave through secondary growth method. Multiple membrane synthesis factors such as seed size, ceramic substrate surface pore size selection, and secondary growth conditions were studied for their effects on zeolite membrane growth. The microstructure, morphology and the membrane thickness of tin-Silicalite 1 zeolite membrane were examined. The membrane separation performance and stability will also be reported.

Keywords: ceramic membrane, pervaporation, solvent recovery, Sn-MFI zeolite

Procedia PDF Downloads 180