Search results for: Bacillus subtilis strain LB5
55 The Efficacy of Andrographis paniculata and Chromolaena odorata Plant Extract against Malaria Parasite
Authors: Funmilola O. Omoya, Abdul O. Momoh
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Malaria constitutes one of the major health problems in Nigeria. One of the reasons attributed for the upsurge was the development of resistance of Plasmodium falciparum and the emergence of multi-resistant strains of the parasite to anti-malaria drugs. A continued search for other effective, safe and cheap plantbased anti-malaria agents thus becomes imperative in the face of these difficulties. The objective of this study is therefore to evaluate the in vivo anti-malarial efficacy of ethanolic extracts of Chromolaena odorata and Androgaphis paniculata leaves. The two plants were evaluated for their anti-malaria efficacy in vivo in a 4-day curative test assay against Plasmodium berghei strain in mice. The group treated with 500mg/ml dose of ethanolic extract of A. paniculata plant showed parasite suppression with increase in Packed Cell Volume (PCV) value except day 3 which showed a slight decrease in PCV value. During the 4-day curative test, an increase in the PCV values, weight measurement and zero count of Plasmodium berghei parasite values was recorded after day 3 of drug administration. These results obtained in group treated with A. paniculata extract showed anti-malarial efficacy with higher mortality rate in parasitaemia count when compared with Chromolaena odorata group. These results justify the use of ethanolic extracts of A. paniculata plant as medicinal herb used in folklore medicine in the treatment of malaria.
Keywords: Anti-malaria, Curative, Plant-based anti-malaria agents.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 205654 Micromechanics Modeling of 3D Network Smart Orthotropic Structures
Authors: E. M. Hassan, A. L. Kalamkarov
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Two micromechanical models for 3D smart composite with embedded periodic or nearly periodic network of generally orthotropic reinforcements and actuators are developed and applied to cubic structures with unidirectional orientation of constituents. Analytical formulas for the effective piezothermoelastic coefficients are derived using the Asymptotic Homogenization Method (AHM). Finite Element Analysis (FEA) is subsequently developed and used to examine the aforementioned periodic 3D network reinforced smart structures. The deformation responses from the FE simulations are used to extract effective coefficients. The results from both techniques are compared. This work considers piezoelectric materials that respond linearly to changes in electric field, electric displacement, mechanical stress and strain and thermal effects. This combination of electric fields and thermo-mechanical response in smart composite structures is characterized by piezoelectric and thermal expansion coefficients. The problem is represented by unitcell and the models are developed using the AHM and the FEA to determine the effective piezoelectric and thermal expansion coefficients. Each unit cell contains a number of orthotropic inclusions in the form of structural reinforcements and actuators. Using matrix representation of the coupled response of the unit cell, the effective piezoelectric and thermal expansion coefficients are calculated and compared with results of the asymptotic homogenization method. A very good agreement is shown between these two approaches.
Keywords: Asymptotic Homogenization Method, Effective Piezothermoelastic Coefficients, Finite Element Analysis, 3D Smart Network Composite Structures.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 209053 Polyvinyl Alcohol Processed Templated Polyaniline Films: Preparation, Characterization and Assessment of Tensile Strength
Authors: J. Subbalakshmi, G. Dhruvasamhith, S. M. Hussain
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Polyaniline (PANI) is one of the most extensively studied material among the conducting polymers due to its simple synthesis by chemical and electrochemical routes. PANIs have advantages of chemical stability and high conductivity making their commercial applications quite attractive. However, to our knowledge, very little work has been reported on the tensile strength properties of templated PANIs processed with polyvinyl alcohol and also, detailed study has not been carried out. We have investigated the effect of small molecule and polymers as templates on PANI. Stable aqueous colloidal suspensions of trisodium citrate (TSC), poly(ethylenedioxythiophene)-polystyrene sulfonate (PEDOT-PSS), and polyethylene glycol (PEG) templated PANIs were prepared through chemical synthesis, processed with polyvinyl alcohol (PVA) and were fabricated into films by solution casting. Absorption and infra-red spectra were studied to gain insight into the possible molecular interactions. Surface morphology was studied through scanning electron microscope and optical microscope. Interestingly, tensile testing studies revealed least strain for pure PVA when compared to the blends of templated PANI. Furthermore, among the blends, TSC templated PANI possessed maximum elasticity. The ultimate tensile strength for PVA processed, PEG-templated PANI was found to be five times more than other blends considered in this study. We establish structure–property correlation with morphology, spectral characterization and tensile testing studies.Keywords: Processed films, polyvinyl alcohol, spectroscopy, surface morphology, templated polyanilines, tensile test.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 116952 Evaluation of Seismic Behavior of Steel Shear Wall with Opening with Hardener and Beam with Reduced Cross Section under Cycle Loading with Finite Element Analysis Method
Authors: Masoud Mahdavi
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During an earthquake, the structure is subjected to seismic loads that cause tension in the members of the building. The use of energy dissipation elements in the structure reduces the percentage of seismic forces on the main members of the building (especially the columns). Steel plate shear wall, as one of the most widely used types of energy dissipation element, has evolved today, and regular drilling of its inner plate is one of the common cases. In the present study, using a finite element method, the shear wall of the steel plate is designed as a floor (with dimensions of 447 × 6/246 cm) with Abacus software and in three different modes on which a cyclic load has been applied. The steel shear wall has a horizontal element (beam) with a reduced beam section (RBS). The hole in the interior plate of the models is created in such a way that it has the process of increasing the area, which makes the effect of increasing the surface area of the hole on the seismic performance of the steel shear wall completely clear. In the end, it was found that with increasing the opening level in the steel shear wall (with reduced cross-section beam), total displacement and plastic strain indicators increased, structural capacity and total energy indicators decreased and the Mises Monson stress index did not change much.
Keywords: Steel plate shear wall with opening, cyclic loading, reduced cross-section beam, finite element method, Abaqus Software.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 61151 Substantial Fatigue Similarity of a New Small-Scale Test Rig to Actual Wheel-Rail System
Authors: Meysam Naeimi, Zili Li, Roumen Petrov, Rolf Dollevoet, Jilt Sietsma, Jun Wu
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The substantial similarity of fatigue mechanism in a new test rig for rolling contact fatigue (RCF) has been investigated. A new reduced-scale test rig is designed to perform controlled RCF tests in wheel-rail materials. The fatigue mechanism of the rig is evaluated in this study using a combined finite element-fatigue prediction approach. The influences of loading conditions on fatigue crack initiation have been studied. Furthermore, the effects of some artificial defects (squat-shape) on fatigue lives are examined. To simulate the vehicle-track interaction by means of the test rig, a threedimensional finite element (FE) model is built up. The nonlinear material behaviour of the rail steel is modelled in the contact interface. The results of FE simulations are combined with the critical plane concept to determine the material points with the greatest possibility of fatigue failure. Based on the stress-strain responses, by employing of previously postulated criteria for fatigue crack initiation (plastic shakedown and ratchetting), fatigue life analysis is carried out. The results are reported for various loading conditions and different defect sizes. Afterward, the cyclic mechanism of the test rig is evaluated from the operational viewpoint. The results of fatigue life predictions are compared with the expected number of cycles of the test rig by its cyclic nature. Finally, the estimative duration of the experiments until fatigue crack initiation is roughly determined.
Keywords: Fatigue, test rig, crack initiation, life, rail, squats.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 216150 Dry Needling Treatment in 38 Cases of Chronic Sleep Disturbance
Authors: P. Gao, Z. Q. Li, Y. G. Jin
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In the past 10 years, computers and cellphones have become one of the most important factors in our lives, and one which has a tremendously negative impact on our muscles. Muscle tension may be one of the causes of sleep disturbance. Tension in the shoulders and neck can affect blood circulation to the muscles. This research uses a dry needling treatment to reduce muscle tension in order to determine if the strain in the head and shoulders can influence sleep duration. All 38 patients taking part in the testing suffered from tinnitus and have been experiencing disturbed sleep for at least one to five years. Even after undergoing drug therapy treatments and traditional acupuncture therapies, their sleep disturbances have not shown any improvement. After five to 10 dry needling treatments, 24 of the patients reported an improvement in their sleep duration. Five patients considered themselves to be completely recovered, while 12 patients experienced no improvement. This study investigated these pathogenic and therapeutic problems. The standard treatment for sleep disturbances is drug-based therapy; the results of most standard treatments are unfortunately negative. The result of this clinical research has demonstrated that: The possible cause of sleep disturbance for a lot of patients is the result of tensions in the neck and shoulder muscles. Blood circulation to those muscles is also influenced by the duration of sleep. Hypertonic neck and shoulder muscles are considered to impact sleeping patterns and lead to disturbed sleep. Poor posture, often adopted while speaking on the phone, is one of the main causes of hypertonic neck and shoulder muscle problems. The dry needling treatment specifically focuses on the release of muscle tension.
Keywords: Dry needling, sleep disturbance, sleep duration, muscle tension, trigger points.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 213549 The Expression of Lipoprotein Lipase Gene with Fat Accumulations and Serum Biochemical Levels in Betong (KU Line) and Broiler Chickens
Authors: W. Loongyai, N. Saengsawang, W. Danvilai, C. Kridtayopas, P. Sopannarath, C. Bunchasak
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Betong chicken is a slow growing and a lean strain of chicken, while the rapid growth of broiler is accompanied by increased fat. We investigated the growth performance, fat accumulations, lipid serum biochemical levels and lipoprotein lipase (LPL) gene expression of female Betong (KU line) at the age of 4 and 6 weeks. A total of 80 female Betong chickens (KU line) and 80 female broiler chickens were reared under open system (each group had 4 replicates of 20 chicks per pen). The results showed that feed intake and average daily gain (ADG) of broiler chicken were significantly higher than Betong (KU line) (P < 0.01), while feed conversion ratio (FCR) of Betong (KU line) at week 6 were significantly lower than broiler chicken (P < 0.01) at 6 weeks. At 4 and 6 weeks, two birds per replicate were randomly selected and slaughtered. Carcass weight did not significantly differ between treatments; the percentage of abdominal fat and subcutaneous fat yield was higher in the broiler (P < 0.01) at 4 and 6 week. Total cholesterol and LDL level of broiler were higher than Betong (KU line) at 4 and 6 weeks (P < 0.05). Abdominal fat samples were collected for total RNA extraction. The cDNA was amplified using primers specific for LPL gene expression and analysed using real-time PCR. The results showed that the expression of LPL gene was not different when compared between Betong (KU line) and broiler chickens at the age of 4 and 6 weeks (P > 0.05). Our results indicated that broiler chickens had high growth rate and fat accumulation when compared with Betong (KU line) chickens, whereas LPL gene expression did not differ between breeds.
Keywords: Lipoprotein lipase gene, Betong (KU line), broiler, abdominal fat, gene expression.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 94748 Optimization of Lead Bioremediation by Marine Halomonas sp. ES015 Using Statistical Experimental Methods
Authors: Aliaa M. El-Borai, Ehab A. Beltagy, Eman E. Gadallah, Samy A. ElAssar
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Bioremediation technology is now used for treatment instead of traditional metal removal methods. A strain was isolated from Marsa Alam, Red sea, Egypt showed high resistance to high lead concentration and was identified by the 16S rRNA gene sequencing technique as Halomonas sp. ES015. Medium optimization was carried out using Plackett-Burman design, and the most significant factors were yeast extract, casamino acid and inoculums size. The optimized media obtained by the statistical design raised the removal efficiency from 84% to 99% from initial concentration 250 ppm of lead. Moreover, Box-Behnken experimental design was applied to study the relationship between yeast extract concentration, casamino acid concentration and inoculums size. The optimized medium increased removal efficiency to 97% from initial concentration 500 ppm of lead. Immobilized Halomonas sp. ES015 cells on sponge cubes, using optimized medium in loop bioremediation column, showed relatively constant lead removal efficiency when reused six successive cycles over the range of time interval. Also metal removal efficiency was not affected by flow rate changes. Finally, the results of this research refer to the possibility of lead bioremediation by free or immobilized cells of Halomonas sp. ES015. Also, bioremediation can be done in batch cultures and semicontinuous cultures using column technology.
Keywords: Bioremediation, lead, Box–Behnken, Halomonas sp. ES015, loop bioremediation, Plackett-Burman.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 100847 Improvement of Short Channel Effects in Cylindrical Strained Silicon Nanowire Transistor
Authors: Fatemeh Karimi, Morteza Fathipour, Hamdam Ghanatian, Vala Fathipour
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In this paper we investigate the electrical characteristics of a new structure of gate all around strained silicon nanowire field effect transistors (FETs) with dual dielectrics by changing the radius (RSiGe) of silicon-germanium (SiGe) wire and gate dielectric. Indeed the effect of high-κ dielectric on Field Induced Barrier Lowering (FIBL) has been studied. Due to the higher electron mobility in tensile strained silicon, the n-type FETs with strained silicon channel have better drain current compare with the pure Si one. In this structure gate dielectric divided in two parts, we have used high-κ dielectric near the source and low-κ dielectric near the drain to reduce the short channel effects. By this structure short channel effects such as FIBL will be reduced indeed by increasing the RSiGe, ID-VD characteristics will be improved. The leakage current and transfer characteristics, the threshold-voltage (Vt), the drain induced barrier height lowering (DIBL), are estimated with respect to, gate bias (VG), RSiGe and different gate dielectrics. For short channel effects, such as DIBL, gate all around strained silicon nanowire FET have similar characteristics with the pure Si one while dual dielectrics can improve short channel effects in this structure.Keywords: SNWT (silicon nanowire transistor), Tensile Strain, high-κ dielectric, Field Induced Barrier Lowering (FIBL), cylindricalnano wire (CW), drain induced barrier lowering (DIBL).
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 199446 Comparative Study of Tensile Properties of Cast and Hot Forged Alumina Nanoparticle Reinforced Composites
Authors: S. Ghanaraja, Subrata Ray, S. K. Nath
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Particle reinforced Metal Matrix Composite (MMC) succeeds in synergizing the metallic matrix with ceramic particle reinforcements to result in improved strength, particularly at elevated temperatures, but adversely it affects the ductility of the matrix because of agglomeration and porosity. The present study investigates the outcome of tensile properties in a cast and hot forged composite reinforced simultaneously with coarse and fine particles. Nano-sized alumina particles have been generated by milling mixture of aluminum and manganese dioxide powders. Milled particles after drying are added to molten metal and the resulting slurry is cast. The microstructure of the composites shows good distribution of both the size categories of particles without significant clustering. The presence of nanoparticles along with coarser particles in a composite improves both strength and ductility considerably. Delay in debonding of coarser particles to higher stress is due to reduced mismatch in extension caused by increased strain hardening in presence of the nanoparticles. However, higher addition of powder mix beyond a limit results in deterioration of mechanical properties, possibly due to clustering of nanoparticles. The porosity in cast composite generally increases with the increasing addition of powder mix as observed during process and on forging it has got reduced. The base alloy and nanocomposites show improvement in flow stress which could be attributed to lowering of porosity and grain refinement as a consequence of forging.
Keywords: Aluminum, alumina, nanoparticle reinforced composites, porosity.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 146645 Clinical Signs of Neonatal Calves in Experimental Colisepticemia
Authors: Samad Lotfollahzadeh
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Escherichia coli (E. coli) is the most isolated bacteria from blood circulation of septicemic calves. Given the prevalence of septicemia in animals and its economic importance in veterinary practice, better understanding of changes in clinical signs following disease, may contribute to early detection of disorder. The present study has been carried out to detect changes of clinical signs in induced sepsis in calves with E. coli. Colisepticemia has been induced in 10 twenty-day old healthy Holstein- Frisian calves with intravenous injection of 1.5 X 109 colony forming units (cfu) of O111:H8 strain of E. coli. Clinical signs including rectal temperature, heart rate, respiratory rate, shock, appetite, sucking reflex, feces consistency, general behavior, dehydration and standing ability were recorded in experimental calves during 24 hours after induction of colisepticemia. Blood culture was also carried out from calves four times during experiment. ANOVA with repeated measure is used to see changes of calves’ clinical signs to experimental colisepticemia, and values of P≤ 0.05 was considered statistically significant. Mean values of rectal temperature and heart rate as well as median values of respiratory rate, appetite, suckling reflex, standing ability and feces consistency of experimental calves increased significantly during study (P<0.05). In the present study median value of shock score was not significantly increased in experimental calves (P> 0.05). The results of present study showed that total score of clinical signs in calves with experimental colisepticemia increased significantly, although score of some clinical signs such as shock did not change significantly.Keywords: Calves, Clinical signs scoring, E. coli O111:H8, Experimental colisepticemia,
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 209144 Computational Identification of Bacterial Communities
Authors: Eleftheria Tzamali, Panayiota Poirazi, Ioannis G. Tollis, Martin Reczko
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Stable bacterial polymorphism on a single limiting resource may appear if between the evolved strains metabolic interactions take place that allow the exchange of essential nutrients [8]. Towards an attempt to predict the possible outcome of longrunning evolution experiments, a network based on the metabolic capabilities of homogeneous populations of every single gene knockout strain (nodes) of the bacterium E. coli is reconstructed. Potential metabolic interactions (edges) are allowed only between strains of different metabolic capabilities. Bacterial communities are determined by finding cliques in this network. Growth of the emerged hypothetical bacterial communities is simulated by extending the metabolic flux balance analysis model of Varma et al [2] to embody heterogeneous cell population growth in a mutual environment. Results from aerobic growth on 10 different carbon sources are presented. The upper bounds of the diversity that can emerge from single-cloned populations of E. coli such as the number of strains that appears to metabolically differ from most strains (highly connected nodes), the maximum clique size as well as the number of all the possible communities are determined. Certain single gene deletions are identified to consistently participate in our hypothetical bacterial communities under most environmental conditions implying a pattern of growth-condition- invariant strains with similar metabolic effects. Moreover, evaluation of all the hypothetical bacterial communities under growth on pyruvate reveals heterogeneous populations that can exhibit superior growth performance when compared to the performance of the homogeneous wild-type population.
Keywords: Bacterial polymorphism, clique identification, dynamic FBA, evolution, metabolic interactions.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 136943 Structural Characteristics of HPDSP Concrete on Beam Column Joints
Authors: Sushil Kumar Swar, Sanjay Kumar Sharma, Hari Krishan Sharma, Sushil Kumar
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The seriously damaged structures during earthquakes show the need and importance of design of reinforced concrete structures with high ductility. Reinforced concrete beam-column joints have an important function in all structures. Under seismic excitation, the beam column joint region is subjected to horizontal and vertical shear forces whose magnitude is many times higher than the adjacent beam and column. Strength and ductility of structures depends mainly on proper detailing of the reinforcement in beamcolumn joints and the old structures were found ductility deficient. DSP materials are obtained by using high quantities of super plasticizers and high volumes of micro silica. In the case of High Performance Densified Small Particle Concrete (HPDSPC), since concrete is dense even at the micro-structure level, tensile strain would be much higher than that of the conventional SFRC, SIFCON & SIMCON. This in turn will improve cracking behaviour, ductility and energy absorption capacity of composites in addition to durability. The fine fibers used in our mix are 0.3mm diameter and 10 mm which can be easily placed with high percentage. These fibers easily transfer stresses and act as a composite concrete unit to take up extremely high loads with high compressive strength. HPDSPC placed in the beam column joints helps in safety of human life due to prolonged failure.Keywords: High Performance Densified Small Particle Concrete (HPDSPC), Steel Fıber Reinforced Concrete (SFRC), Slurry Infiltrated Concrete (SIFCON), Slurry Infiltrated Mat Concrete (SIMCON).
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 215342 Stress Relaxation of Date at Different Temperature and Moisture Content of Product: A New Approach
Authors: D. Zare, M. Alirezaei, S.M. Nassiri
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Iran is one of the greatest producers of date in the world. However due to lack of information about its viscoelastic properties, much of the production downgraded during harvesting and postharvesting processes. In this study the effect of temperature and moisture content of product were investigated on stress relaxation characteristics. Therefore, the freshly harvested date (kabkab) at tamar stage were put in controlled environment chamber to obtain different temperature levels (25, 35, 45, and 55 0C) and moisture contents (8.5, 8.7, 9.2, 15.3, 20, 32.2 %d.b.). A texture analyzer TAXT2 (Stable Microsystems, UK) was used to apply uniaxial compression tests. A chamber capable to control temperature was designed and fabricated around the plunger of texture analyzer to control the temperature during the experiment. As a new approach a CCD camera (A4tech, 30 fps) was mounted on a cylindrical glass probe to scan and record contact area between date and disk. Afterwards, pictures were analyzed using image processing toolbox of Matlab software. Individual date fruit was uniaxially compressed at speed of 1 mm/s. The constant strain of 30% of thickness of date was applied to the horizontally oriented fruit. To select a suitable model for describing stress relaxation of date, experimental data were fitted with three famous stress relaxation models including the generalized Maxwell, Nussinovitch, and Pelege. The constant in mentioned model were determined and correlated with temperature and moisture content of product using non-linear regression analysis. It was found that Generalized Maxwell and Nussinovitch models appropriately describe viscoelastic characteristics of date fruits as compared to Peleg mode.Keywords: Stress relaxation, Viscoelastic properties, Date, Texture analyzer.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 189941 Nonlinear and Chaotic Motions for a Shock Absorbing Structure Supported by Nonlinear Springs with Hysteresis Using Fast FEA
Authors: T. Yamaguchi, Y. Kurosawa, S. Maruyama, K. Tobita, Y. Hirano, K. Yokouchi, K. Kihara, T. Sunaga
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This paper describes dynamic analysis using proposed fast finite element method for a shock absorbing structure including a sponge. The structure is supported by nonlinear concentrated springs. The restoring force of the spring has cubic nonlinearity and linear hysteresis damping. To calculate damping properties for the structures including elastic body and porous body, displacement vectors as common unknown variable are solved under coupled condition. Under small amplitude, we apply asymptotic method to complex eigenvalue problem of this system to obtain modal parameters. And then expressions of modal loss factor are derived approximately. This approach was proposed by one of the authors previously. We call this method as Modal Strain and Kinetic Energy Method (MSKE method). Further, using the modal loss factors, the discretized equations in physical coordinate are transformed into the nonlinear ordinary coupled equations using normal coordinate corresponding to linear natural modes. This transformation yields computation efficiency. As a numerical example of a shock absorbing structures, we adopt double skins with a sponge. The double skins are supported by nonlinear concentrated springs. We clarify influences of amplitude of the input force on nonlinear and chaotic responses.
Keywords: Dynamic response, Nonlinear and chaotic motions, Finite Element analysis, Numerical analysis.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 196040 Bio-Surfactant Production and Its Application in Microbial EOR
Authors: A. Rajesh Kanna, G. Suresh Kumar, Sathyanaryana N. Gummadi
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There are various sources of energies available worldwide and among them, crude oil plays a vital role. Oil recovery is achieved using conventional primary and secondary recovery methods. In-order to recover the remaining residual oil, technologies like Enhanced Oil Recovery (EOR) are utilized which is also known as tertiary recovery. Among EOR, Microbial enhanced oil recovery (MEOR) is a technique which enables the improvement of oil recovery by injection of bio-surfactant produced by microorganisms. Bio-surfactant can retrieve unrecoverable oil from the cap rock which is held by high capillary force. Bio-surfactant is a surface active agent which can reduce the interfacial tension and reduce viscosity of oil and thereby oil can be recovered to the surface as the mobility of the oil is increased. Research in this area has shown promising results besides the method is echo-friendly and cost effective compared with other EOR techniques. In our research, on laboratory scale we produced bio-surfactant using the strain Pseudomonas putida (MTCC 2467) and injected into designed simple sand packed column which resembles actual petroleum reservoir. The experiment was conducted in order to determine the efficiency of produced bio-surfactant in oil recovery. The column was made of plastic material with 10 cm in length. The diameter was 2.5 cm. The column was packed with fine sand material. Sand was saturated with brine initially followed by oil saturation. Water flooding followed by bio-surfactant injection was done to determine the amount of oil recovered. Further, the injection of bio-surfactant volume was varied and checked how effectively oil recovery can be achieved. A comparative study was also done by injecting Triton X 100 which is one of the chemical surfactant. Since, bio-surfactant reduced surface and interfacial tension oil can be easily recovered from the porous sand packed column.
Keywords: Bio-surfactant, Bacteria, Interfacial tension, Sand column.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 277139 Prophylactic Effects of Dairy Kluyveromyces marxianus YAS through Overexpression of BAX, CASP 3, CASP 8 and CASP 9 on Human Colon Cancer Cell Lines
Authors: Amir Saber Gharamaleki, Beitollah Alipour, Zeinab Faghfoori, Ahmad YariKhosroushahi
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Colorectal cancer (CRC) is one of the most prevalent cancers and intestinal microbial community plays an important role in colorectal tumorigenesis. Probiotics have recently been assessed as effective anti-proliferative agents and thus this study was performed to examine whether CRC undergo apoptosis by treating with isolated Iranian native dairy yeast, Kluyveromyces marxianus YAS, secretion metabolites. The cytotoxicity assessments on cells (HT-29, Caco-2) were accomplished through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as well as qualitative DAPI (4',6-diamidino-2-phenylindole staining) and quantitative (flow cytometry assessments) evaluations of apoptosis. To evaluate the main mechanism of apoptosis, Real time PCR method was applied. Kluyveromyces marxianus YAS secretions (IC50) showed significant cytotoxicity against HT-29 and Caco-2 cancer cell lines (66.57 % and 66.34 % apoptosis) similar to 5-Fluorouracil (5-FU) while apoptosis only was developed in 27.57 % of KDR normal cells. The prophylactic effects of Kluyveromyces marxianus (PTCC 5195), as a reference yeast, was not similar to Kluyveromyces marxianus YAS indicating strain dependency of bioactivities on CRC disease prevention. Based on real time PCR results, the main cytotoxicity is related to apoptosis phenomenon and the core related mechanism is depended on the overexpression of BAX, CASP 9, CASP 8 and CASP 3 inducing apoptosis genes. However, several investigations should be conducted to precisely determine the effective compounds to be used as anticancer therapeutics in the future.Keywords: Anticancer, anti-proliferative, apoptosis, cytotoxicity, yeast.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 165138 Assessing Storage of Stability and Mercury Reduction of Freeze-Dried Pseudomonas putida within Different Types of Lyoprotectant
Authors: A. A. M. Azoddein, Y. Nuratri, A. B. Bustary, F. A. M. Azli, S. C. Sayuti
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Pseudomonas putida is a potential strain in biological treatment to remove mercury contained in the effluent of petrochemical industry due to its mercury reductase enzyme that able to reduce ionic mercury to elementary mercury. Freeze-dried P. putida allows easy, inexpensive shipping, handling and high stability of the product. This study was aimed to freeze dry P. putida cells with addition of lyoprotectant. Lyoprotectant was added into the cells suspension prior to freezing. Dried P. putida obtained was then mixed with synthetic mercury. Viability of recovery P. putida after freeze dry was significantly influenced by the type of lyoprotectant. Among the lyoprotectants, tween 80/ sucrose was found to be the best lyoprotectant. Sucrose able to recover more than 78% (6.2E+09 CFU/ml) of the original cells (7.90E+09CFU/ml) after freeze dry and able to retain 5.40E+05 viable cells after 4 weeks storage in 4oC without vacuum. Polyethylene glycol (PEG) pre-treated freeze dry cells and broth pre-treated freeze dry cells after freeze-dry recovered more than 64% (5.0 E+09CFU/ml) and >0.1% (5.60E+07CFU/ml). Freeze-dried P. putida cells in PEG and broth cannot survive after 4 weeks storage. Freeze dry also does not really change the pattern of growth P. putida but extension of lag time was found 1 hour after 3 weeks of storage. Additional time was required for freeze-dried P. putida cells to recover before introduce freeze-dried cells to more complicated condition such as mercury solution. The maximum mercury reduction of PEG pre-treated freeze-dried cells after freeze dry and after storage 3 weeks was 56.78% and 17.91%. The maximum of mercury reduction of tween 80/sucrose pre-treated freeze-dried cells after freeze dry and after storage 3 weeks were 26.35% and 25.03%. Freeze dried P. putida was found to have lower mercury reduction compare to the fresh P. putida that has been growth in agar. Result from this study may be beneficial and useful as initial reference before commercialize freeze-dried P. putida.
Keywords: Pseudomonas putida, freeze-dry, PEG, Tween80/Sucrose, mercury, cell viability.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 110737 Consequential Influences of Work-Induced Emotions on the Work-Induced Happiness of Frontline Workers in Finance-Oriented Firms
Authors: Mohammed-Aminu Sanda, Emmanuel K. Mawuena
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Frontline workers performing client service duties in finance-oriented firms in most sub-Saharan African countries, such as Ghana, are known to be challenged in the conduct of their activities. The challenge is attributed to clients’ continued demand for real-time services from such workers, despite the introduction of technological interventions to offset the situation. This has caused such frontline workers to experience increases in their work-induced emotions with consequential effects on their work-induced happiness. This study, therefore, explored the effect of frontline workers’ work-induced emotions on their worked-induced happiness when providing tellering services to clients. A cross-sectional design and quantitative technique were used. Data were collected from a sample of 280 frontline workers using questionnaire. Based on the analysis, it was found that an increase in the frontline workers’ work-induced emotions, caused by their feelings of strain, burnout, frustration, and hard work, had consequential effect on their work-induced happiness. This consequential effect was also found to be aggravated by the workers’ senses of being stretched beyond limit, being emotionally drained, and being used up by their work activities. It is concluded that frontline workers in finance-oriented firms can provide quality real-time services to clients without increases in their work-induced emotions, but with enhanced work-induced happiness, when the psychological and physiological emotional factors associated with the challenged work activities are understood and remedied. Management of the firms can use such understanding to redesign the activities of their frontline workers and improve the quality of their service delivery interactivity with clients.
Keywords: Client-service activity, finance industrial sector, frontline workers, work-induced emotion, work-induced happiness.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 74736 Plasma Spraying of 316 Stainless Steel on Aluminum and Investigation of Coat/Substrate Interface
Authors: P. Abachi, T. W. Coyle, P. S. Musavi Gharavi
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By applying coating onto a structural component, the corrosion and/or wear resistance requirements of the surface can be fulfilled. Since the layer adhesion of the coating influences the mechanical integrity of the coat/substrate interface during the service time, it should be examined accurately. At the present work, the tensile bonding strength of the 316 stainless steel plasma sprayed coating on aluminum substrate was determined by using tensile adhesion test, TAT, specimen. The interfacial fracture toughness was specified using four-point bend specimen containing a saw notch and modified chevron-notched short-bar (SB) specimen. The coating microstructure and fractured specimen surface were examined by using scanning electron- and optical-microscopy. The investigation of coated surface after tensile adhesion test indicates that the failure mechanism is mostly cohesive and rarely adhesive type. The calculated value of critical strain energy release rate proposes relatively good interface status. It seems that four-point bending test offers a potentially more sensitive means for evaluation of mechanical integrity of coating/substrate interfaces than is possible with the tensile test. The fracture toughness value reported for the modified chevron-notched short-bar specimen testing cannot be taken as absolute value because its calculation is based on the minimum stress intensity coefficient value which has been suggested for the fracture toughness determination of homogeneous parts in the ASTM E1304-97 standard.
Keywords: Bonding strength, four-point bend test, interfacial fracture toughness, modified chevron-notched short-bar specimen, plasma sprayed coating.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 157835 Survey of Potato Viral Infection Using Das-Elisa Method in Georgia
Authors: Maia Kukhaleishvili, Ekaterine Bulauri, Iveta Megrelishvili, Tamar Shamatava, Tamar Chipashvili
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Plant viruses can cause loss of yield and quality in a lot of important crops. Symptoms of pathogens are variable depending on the cultivars and virus strain. Selection of resistant potato varieties would reduce the risk of virus transmission and significant economic impact. Other way to avoid reduced harvest yields is regular potato seed production sampling and testing for viral infection. The aim of this study was to determine the occurrence and distribution of viral diseases according potato cultivars for further selection of virus-free material in Georgia. During the summer 2015- 2016, 5 potato cultivars (Sante, Laura, Jelly, Red Sonia, Anushka) at 5 different farms located in Akhalkalaki were tested for 6 different potato viruses: Potato virus A (PVA), Potato virus M (PVM), Potato virus S (PVS), Potato virus X (PVX), Potato virus Y (PVY) and potato leaf roll virus (PLRV). A serological method, Double Antibody Sandwich-Enzyme linked Immunosorbent Assay (DASELISA) was used at the laboratory to analyze the results. The result showed that PVY (21.4%) and PLRV (19.7%) virus presence in collected samples was relatively high compared to others. Researched potato cultivars except Jelly and Laura were infected by PVY with different concentrations. PLRV was found only in three potato cultivars (Sante, Jelly, Red Sonia) and PVM virus (3.12%) was characterized with low prevalence. PVX, PVA and PVS virus infection was not reported. It would be noted that 7.9% of samples were containing PVY/PLRV mix infection. Based on the results it can be concluded that PVY and PLRV infections are dominant in all research cultivars. Therefore significant yield losses are expected. Systematic, long-term control of potato viral infection, especially seed-potatoes, must be regarded as the most important factor to increase seed productivity.Keywords: Diseases, infection, potato, virus.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 92734 Fuzzy Optimization in Metabolic Systems
Authors: Feng-Sheng Wang, Wu-Hsiung Wu, Kai-Cheng Hsu
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The optimization of biological systems, which is a branch of metabolic engineering, has generated a lot of industrial and academic interest for a long time. In the last decade, metabolic engineering approaches based on mathematical optimizations have been used extensively for the analysis and manipulation of metabolic networks. In practical optimization of metabolic reaction networks, designers have to manage the nature of uncertainty resulting from qualitative characters of metabolic reactions, e.g., the possibility of enzyme effects. A deterministic approach does not give an adequate representation for metabolic reaction networks with uncertain characters. Fuzzy optimization formulations can be applied to cope with this problem. A fuzzy multi-objective optimization problem can be introduced for finding the optimal engineering interventions on metabolic network systems considering the resilience phenomenon and cell viability constraints. The accuracy of optimization results depends heavily on the development of essential kinetic models of metabolic networks. Kinetic models can quantitatively capture the experimentally observed regulation data of metabolic systems and are often used to find the optimal manipulation of external inputs. To address the issues of optimizing the regulatory structure of metabolic networks, it is necessary to consider qualitative effects, e.g., the resilience phenomena and cell viability constraints. Combining the qualitative and quantitative descriptions for metabolic networks makes it possible to design a viable strain and accurately predict the maximum possible flux rates of desired products. Considering the resilience phenomena in metabolic networks can improve the predictions of gene intervention and maximum synthesis rates in metabolic engineering. Two case studies will present in the conference to illustrate the phenomena.
Keywords: Fuzzy multi-objective optimization problem, kinetic model, metabolic engineering.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 200433 The Effects of Organic or Inorganic Zinc and Microbial Phytase, Alone or in Combination, on the Performance, Biochemical Parameters and Nutrient Utilization of Broilers Fed a Diet Low in Available Phosphorus
Authors: Mustafa Midilli, Mustafa Salman, Omer Hakan Muglali, Tülay Ögretmen, Sena Cenesiz, Neslihan Ormanci
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This study examined the effects of zinc (Zn) from different sources and microbial phytase on the broiler performance, biochemical parameters and digestibility of nutrients when they were added to broiler diets containing low available phosphorus. A total of 875, 1-day-old male broilers of the Ross 308 strain were randomly separated into two control groups (positive and negative) and five treatment groups each containing 125 birds; each group was divided into 5 replicates of 25 birds. The positive control (PC) group was fed a diet containing adequate concentration (0.45%) of available phosphorus due to mineral premix (except zinc) and feeds. The negative control (NC) group was fed a basal diet including low concentration (0.30%) of available phosphorus due to mineral premix (except zinc) and feeds. The basal diet was supplemented with 0.30% phosphorus and 500 FTU phytase (PH); 0.30% phosphorus and organic zinc (OZ; 75mg/kg of Zn from Zn-proteinate); 0.30% phosphorus and inorganic zinc (IZ; 75 mg/kg of Zn from ZnSO4); 0.30% phosphorus, organic zinc and 500 FTU phytase (OZ + PH); and 0.30% phosphorus, inorganic zinc and 500 FTU phytase (IZ + PH) in the treatment groups 1, 2, 3, 4 and 5, respectively. The lowest value for mean body weight was in the negative control group on a diet containing low available phosphorus. The use of supplementation with organic and inorganic zinc alone or in combination with microbial phytase significantly (P<0.05) increased the digestibility of Zn in the male broilers. Supplementation of those diets with OZ + PH or IZ + PH was very effective for increasing the body weight, body weight gain and the feed conversion ratio. In conclusion, the effects on broilers of diets with low phosphorus levels may be overcome by the addition of inorganic or organic zinc compounds in combination with microbial phytase.
Keywords: Broiler, Performance, Phytase, Phosphorus, Zinc.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 279432 Shear Modulus Degradation of a Liquefiable Sand Deposit by Shaking Table Tests
Authors: Henry Munoz, Muhammad Mohsan, Takashi Kiyota
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Strength and deformability characteristics of a liquefiable sand deposit including the development of earthquake-induced shear stress and shear strain as well as soil softening via the progressive degradation of shear modulus were studied via shaking table experiments. To do so, a model of a liquefiable sand deposit was constructed and densely instrumented where accelerations, pressures, and displacements at different locations were continuously monitored. Furthermore, the confinement effects on the strength and deformation characteristics of the liquefiable sand deposit due to an external surcharge by placing a heavy concrete slab (i.e. the model of an actual structural rigid pavement) on the ground surface were examined. The results indicate that as the number of seismic-loading cycles increases, the sand deposit softens progressively as large shear strains take place in different sand elements. Liquefaction state is reached after the combined effects of the progressive degradation of the initial shear modulus associated with the continuous decrease in the mean principal stress, and the buildup of the excess of pore pressure takes place in the sand deposit. Finally, the confinement effects given by a concrete slab placed on the surface of the sand deposit resulted in a favorable increasing in the initial shear modulus, an increase in the mean principal stress and a decrease in the softening rate (i.e. the decreasing rate in shear modulus) of the sand, thus making the onset of liquefaction to take place at a later stage. This is, only after the sand deposit having a concrete slab experienced a higher number of seismic loading cycles liquefaction took place, in contrast to an ordinary sand deposit having no concrete slab.
Keywords: Liquefaction, shaking table, shear modulus degradation, earthquake.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 173931 Non-Linear Load-Deflection Response of Shape Memory Alloys-Reinforced Composite Cylindrical Shells under Uniform Radial Load
Authors: Behrang Tavousi Tehrani, Mohammad-Zaman Kabir
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Shape memory alloys (SMA) are often implemented in smart structures as the active components. Their ability to recover large displacements has been used in many applications, including structural stability/response enhancement and active structural acoustic control. SMA wires or fibers can be embedded with composite cylinders to increase their critical buckling load, improve their load-deflection behavior, and reduce the radial deflections under various thermo-mechanical loadings. This paper presents a semi-analytical investigation on the non-linear load-deflection response of SMA-reinforced composite circular cylindrical shells. The cylinder shells are under uniform external pressure load. Based on first-order shear deformation shell theory (FSDT), the equilibrium equations of the structure are derived. One-dimensional simplified Brinson’s model is used for determining the SMA recovery force due to its simplicity and accuracy. Airy stress function and Galerkin technique are used to obtain non-linear load-deflection curves. The results are verified by comparing them with those in the literature. Several parametric studies are conducted in order to investigate the effect of SMA volume fraction, SMA pre-strain value, and SMA activation temperature on the response of the structure. It is shown that suitable usage of SMA wires results in a considerable enhancement in the load-deflection response of the shell due to the generation of the SMA tensile recovery force.
Keywords: Airy stress function, cylindrical shell, Galerkin technique, load-deflection curve, recovery stress, shape memory alloy.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 70430 A Study of Shear Stress Intensity Factor of PP and HDPE by a Modified Experimental Method together with FEM
Authors: Md. Shafiqul Islam, Abdullah Khan, Sharon Kao-Walter, Li Jian
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Shear testing is one of the most complex testing areas where available methods and specimen geometries are different from each other. Therefore, a modified shear test specimen (MSTS) combining the simple uniaxial test with a zone of interest (ZOI) is tested which gives almost the pure shear. In this study, material parameters of polypropylene (PP) and high density polyethylene (HDPE) are first measured by tensile tests with a dogbone shaped specimen. These parameters are then used as an input for the finite element analysis. Secondly, a specially designed specimen (MSTS) is used to perform the shear stress tests in a tensile testing machine to get the results in terms of forces and extension, crack initiation etc. Scanning Electron Microscopy (SEM) is also performed on the shear fracture surface to find material behavior. These experiments are then simulated by finite element method and compared with the experimental results in order to confirm the simulation model. Shear stress state is inspected to find the usability of the proposed shear specimen. Finally, a geometry correction factor can be established for these two materials in this specific loading and geometry with notch using Linear Elastic Fracture Mechanics (LEFM). By these results, strain energy of shear failure and stress intensity factor (SIF) of shear of these two polymers are discussed in the special application of the screw cap opening of the medical or food packages with a temper evidence safety solution.
Keywords: Shear test specimen, Stress intensity factor, Finite Element simulation, Scanning electron microscopy, Screw cap opening.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 291729 An Efficient Approach for Shear Behavior Definition of Plant Stalk
Authors: M. R. Kamandar, J. Massah
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The information of the impact cutting behavior of plants stalk plays an important role in the design and fabrication of plants cutting equipment. It is difficult to investigate a theoretical method for defining cutting properties of plants stalks because the cutting process is complex. Thus, it is necessary to set up an experimental approach to determine cutting parameters for a single stalk. To measure the shear force, shear energy and shear strength of plant stalk, a special impact cutting tester was fabricated. It was similar to an Izod impact cutting tester for metals but a cutting blade and data acquisition system were attached to the end of pendulum's arm. The apparatus was included four strain gages and a digital indicator to show the real-time cutting force of plant stalk. To measure the shear force and also testing the apparatus, two plants’ stalks, like buxus and privet, were selected. The samples (buxus and privet stalks) were cut under impact cutting process at four loading rates 1, 2, 3 and 4 m.s-1 and three internodes fifth, tenth and fifteenth by the apparatus. At buxus cutting analysis: the minimum value of cutting energy was obtained at fifth internode and loading rate 4 m.s-1 and the maximum value of shear energy was obtained at fifteenth internode and loading rate 1 m.s-1. At privet cutting analysis: the minimum value of shear consumption energy was obtained at fifth internode and loading rate: 4 m.s-1 and the maximum value of shear energy was obtained at fifteenth internode and loading rate: 1 m.s-1. The statistical analysis at both plants showed that the increase of impact cutting speed would decrease the shear consumption energy and shear strength. In two scenarios, the results showed that with increase the cutting speed, shear force would decrease.
Keywords: Buxus, privet, impact cutting, shear energy.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 81828 Application of Particle Image Velocimetry in the Analysis of Scale Effects in Granular Soil
Authors: Zuhair Kadhim Jahanger, S. Joseph Antony
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The available studies in the literature which dealt with the scale effects of strip footings on different sand packing systematically still remain scarce. In this research, the variation of ultimate bearing capacity and deformation pattern of soil beneath strip footings of different widths under plane-strain condition on the surface of loose, medium-dense and dense sand have been systematically studied using experimental and noninvasive methods for measuring microscopic deformations. The presented analyses are based on model scale compression test analysed using Particle Image Velocimetry (PIV) technique. Upper bound analysis of the current study shows that the maximum vertical displacement of the sand under the ultimate load increases for an increase in the width of footing, but at a decreasing rate with relative density of sand, whereas the relative vertical displacement in the sand decreases for an increase in the width of the footing. A well agreement is observed between experimental results for different footing widths and relative densities. The experimental analyses have shown that there exists pronounced scale effect for strip surface footing. The bearing capacity factors Nγ rapidly decrease up to footing widths B=0.25 m, 0.35 m, and 0.65 m for loose, medium-dense and dense sand respectively, after that there is no significant decrease in Nγ. The deformation modes of the soil as well as the ultimate bearing capacity values have been affected by the footing widths. The obtained results could be used to improve settlement calculation of the foundation interacting with granular soil.
Keywords: PIV, granular mechanics, scale effect, upper bound analysis.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 99627 Rice cDNA Encoding PROLM is Capable of Rescuing Salt Sensitive Yeast Phenotypes G19 and Axt3K from Salt Stress
Authors: Prasad Senadheera, Younousse Saidi, Frans JM Maathuis
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Rice seed expression (cDNA) library in the Lambda Zap 11® phage constructed from the developing grain 10-20 days after flowering was transformed into yeast for functional complementation assays in three salt sensitive yeast mutants S. cerevisiae strain CY162, G19 and Axt3K. Transformed cells of G19 and Axt3K with pYES vector with cDNA inserts showed enhance tolerance than those with empty pYes vector. Sequencing of the cDNA inserts revealed that they encode for the putative proteins with the sequence homologous to rice putative protein PROLM24 (Os06g31070), a prolamin precursor. Expression of this cDNA did not affect yeast growth in absence of salt. Axt3k and G19 strains expressing the PROLM24 were able to grow upto 400 mM and 600 mM of NaCl respectively. Similarly, Axt3k mutant with PROLM24 expression showed comparatively higher growth rate in the medium with excess LiCl (50 mM). The observation that expression of PROLM24 rescued the salt sensitive phenotypes of G19 and Axt3k indicates the existence of a regulatory system that ameliorates the effect of salt stress in the transformed yeast mutants. However, the exact function of the cDNA sequence, which shows partial sequence homology to yeast UTR1 is not clear. Although UTR1 involved in ferrous uptake and iron homeostasis in yeast cells, there is no evidence to prove its role in Na+ homeostasis in yeast cells. Absence of transmembrane regions in Os06g31070 protein indicates that salt tolerance is achieved not through the direct functional complementation of the mutant genes but through an alternative mechanism.Keywords: Rice seed expression, salt stress, prolamin, salinitytolerance, Oryza sativa
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 189726 Expression of Tissue Plasminogen Activator in Transgenic Tobacco Plants by Signal Peptides Targeting for Delivery to Apoplast, Endoplasmic Reticulum and Cytosol Spaces
Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi
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Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA4404. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.
Keywords: Recombinant tissue plasminogen activator, plant cell comportment, leader signals, transgenic tobacco.
Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 700