Search results for: soil enzyme activity
8159 Enzymatic Synthesis of Olive-Based Ferulate Esters: Optimization by Response Surface Methodology
Authors: S. Mat Radzi, N. J. Abd Rahman, H. Mohd Noor, N. Ariffin
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Ferulic acid has widespread industrial potential by virtue of its antioxidant properties. However, it is partially soluble in aqueous media, limiting their usefulness in oil-based processes in food, cosmetic, pharmaceutical, and material industry. Therefore, modification of ferulic acid should be made by producing of more lipophilic derivatives. In this study, a preliminary investigation of lipase-catalyzed trans-esterification reaction of ethyl ferulate and olive oil was investigated. The reaction was catalyzed by immobilized lipase from Candida antarctica (Novozym 435), to produce ferulate ester, a sunscreen agent. A statistical approach of Response surface methodology (RSM) was used to evaluate the interactive effects of reaction temperature (40-80°C), reaction time (4-12 hours), and amount of enzyme (0.1-0.5 g). The optimum conditions derived via RSM were reaction temperature 60°C, reaction time 2.34 hours, and amount of enzyme 0.3 g. The actual experimental yield was 59.6% ferulate ester under optimum condition, which compared well to the maximum predicted value of 58.0%.Keywords: ferulic acid, enzymatic synthesis, esters, RSM
Procedia PDF Downloads 3308158 Biochar-induced Metals Immobilization in the Soil as Affected by Citric Acid
Authors: Md. Shoffikul Islam, Hongqing Hu
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Reducing trace elements' mobility and bioavailability through amendment addition, especially biochar (BC), is a cost-effective and efficient method to address their toxicity in the soil environment. However, the low molecular weight organic acids (LMWOAs) in the rhizosphere could affect BC's efficiency to stabilize trace metals as the LMWOAs could either mobilize or fix metals in the soils. Therefore, understanding the BC's and LMWOAs' interaction mechanisms on metals stabilization in the rhizosphere is crucial. The present study explored the impact of BC derived from rice husk and citric acid (CA) and the combination of BC and CA on the redistribution of cadmium (Cd), lead (Pb), and zinc (Zn) among their geochemical forms through incubation experiment. The changes of zeta potential and X-ray diffraction (XRD) pattern of BC and BC-amended soils to investigate the probable mechanisms of trace elements' immobilization by BC under the CA attack were also examined. The rice husk BC at 5% (w/w) was mixed with the air-dry soil (an Anthrosols) contaminated with Cd, Pb, and Zn in the plastic pot. The 2, 5, 10, and 20 mM kg-1 (w/v) of CA were added separately into the pot. All the ingredients were mixed thoroughly with the soil. A control (CK) treatment was also prepared without BC and CA addition. After 7, 15, and 60 days of incubation with 60% (w/v) moisture level at 25 °C, the incubated soils were determined for pH and EC and were sequentially extracted to assess the metals' transformation in soil. The electronegative charges and XRD peaks of BC and BC-amended soils were also measured. Compared to CK, the application of BC, low level of CA (2 mM kg-1 soil) (CA2), and BC plus the low concentration of CA (BC-CA2) considerably declined the acid-soluble Cd, Pb, and Zn in which BC-CA2 was found to be the most effective treatment. The reversed trends were observed concerning the high levels of CA (>5-20 mM kg-1 soil) and the BC plus high concentrations of CA treatments. BC-CA2 changed the highest amounts of acid-soluble and reducible metals to the oxidizable and residual forms with time. The most increased electronegative charges of BC-CA2 indicate its (BC-CA2) highest Cd, Pb, and Zn immobilizing efficiency, probably through metals adsorption and fixation with the negative charge sites. The XRD study revealed the presence of P, O, CO32-, and Cl1- in BC, which might be responsible for the precipitation of CdCO3, pyromorphite, and hopeite in the case of Cd, Pb, and Zn immobilization, respectively. The findings depicted that the low concentration of CA increased metals' stabilization, whereas the high levels of CA enhanced their mobilization. The BC-CA2 emerged as the best amendment among treatments for metals stabilization in contaminated soils.Keywords: Biochar, citric acid, immobilization, trace elements contaminated soil
Procedia PDF Downloads 818157 Effect of Problem Based Learning (PBL) Activities to Thai Undergraduate Student Teachers Attitude and Their Achievement
Authors: Thanawit Tongmai, Chatchawan Saewor
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Learning management is very important for students’ development. To promote students’ potential, the teacher should design appropriate learning activity that brings their students potential out. Problem based learning has been using worldwide and it has presented numerous of success. This research aims to study third year students’ attitude and their achievement in scientific research course. To find the results, mix method was used to design research conduction. The researcher used PBL and reflection activity in the class. The students had to choose a topic, reviewed information, designed experimental, wrote academic report and presented their research by themselves. The researcher was only a facilitator. Reflection activity was used to progressing and consulting their research. The data was collected along with research conduction by questionnaire and test, including attitude, opinion and their achievement. The result of this study showed that 74.71% from all of students (n = 87) benefited from PBL and reflection activity, while 25.19% were just satisfied. 100% of students had a positive reflection toward PBL activity and they believed that PBL was the best pedagogy method for scientific research course. The achievements of these students were higher than the previous study (P < 0.05). The student’s learning achievement, A, B+ and B, was 48.28, 28.74 and 22.98% respectively. Therefore, it can conclude that PBL activity is appropriate for scientific research course and it can also promote student’s achievement.Keywords: reflection, attitude, learning, achievement, PBL
Procedia PDF Downloads 2808156 Experimental Studies of the Response of Single Piles Under Torsional and Vertical Combined Loads in Contaminated Sand
Authors: Ahmed Mohamed Nasr, Waseim Ragab Azzam, Nada Osama Ramadan
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Contaminated soil can weaken the stability of buildings and infrastructure, posing serious risks to their structural integrity. Therefore, this study aims to understand how oil contamination affects the torsion behavior of model steel piles at different soil densities. This research is crucial for evaluating the structural integrity and stability of piles in oil-contaminated environments. Clean sand samples and heavy motor oil were mixed in amounts ranging from 0 to 6% of the soil's dry weight. The mixture was thoroughly mixed to ensure uniform distribution of the oil throughout the sandy soil for simulating the field conditions. In these investigations, the relative densities (Dr), pile slenderness ratio (Lp/Dp), oil content (O.C%), and contaminated sand layer thickness (LC) were all different. Also, the paper presents an analysis of piles that are loaded both vertically and torsionally. The findings demonstrated that the pre-applied torsion load led to a decrease in the vertical bearing ability of the pile. Also, at Dr = 80%, the ultimate vertical load under combined load at constant torsional load T = (1/3Tu, 2/3Tu, and Tu) in the cases of (Lc/Lp) = 0.5 and (Lp/Dp) =13.3 was found to be reduced by (1.48, 2.78, and 4.15%) less than piles under independent vertical load, respectively so it is crucial to consider the torsion load during pile design.Keywords: torsion-vertical load, oil-contaminated sand, twist angle, steel pile
Procedia PDF Downloads 708155 Caged Compounds as Light-Dependent Initiators for Enzyme Catalysis Reactions
Authors: Emma Castiglioni, Nigel Scrutton, Derren Heyes, Alistair Fielding
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By using light as trigger, it is possible to study many biological processes, such as the activity of genes, proteins, and other molecules, with precise spatiotemporal control. Caged compounds, where biologically active molecules are generated from an inert precursor upon laser photolysis, offer the potential to initiate such biological reactions with high temporal resolution. As light acts as the trigger for cleaving the protecting group, the ‘caging’ technique provides a number of advantages as it can be intracellular, rapid and controlled in a quantitative manner. We are developing caging strategies to study the catalytic cycle of a number of enzyme systems, such as nitric oxide synthase and ethanolamine ammonia lyase. These include the use of caged substrates, caged electrons and the possibility of caging the enzyme itself. In addition, we are developing a novel freeze-quench instrument to study these reactions, which combines rapid mixing and flashing capabilities. Reaction intermediates will be trapped at low temperatures and will be analysed by using electron paramagnetic resonance (EPR) spectroscopy to identify the involvement of any radical species during catalysis. EPR techniques typically require relatively long measurement times and very often, low temperatures to fully characterise these short-lived species. Therefore, common rapid mixing techniques, such as stopped-flow or quench-flow are not directly suitable. However, the combination of rapid freeze-quench (RFQ) followed by EPR analysis provides the ideal approach to kinetically trap and spectroscopically characterise these transient radical species. In a typical RFQ experiment, two reagent solutions are delivered to the mixer via two syringes driven by a pneumatic actuator or stepper motor. The new mixed solution is then sprayed into a cryogenic liquid or surface, and the frozen sample is then collected and packed into an EPR tube for analysis. The earliest RFQ instrument consisted of a hydraulic ram unit as a drive unit with direct spraying of the sample into a cryogenic liquid (nitrogen, isopentane or petroleum). Improvements to the RFQ technique have arisen from the design of new mixers in order to reduce both the volume and the mixing time. In addition, the cryogenic isopentane bath has been coupled to a filtering system or replaced by spraying the solution onto a surface that is frozen via thermal conductivity with a cryogenic liquid. In our work, we are developing a novel RFQ instrument which combines the freeze-quench technology with flashing capabilities to enable the studies of both thermally-activated and light-activated biological reactions. This instrument also uses a new rotating plate design based on magnetic couplings and removes the need for mechanical motorised rotation, which can otherwise be problematic at cryogenic temperatures.Keywords: caged compounds, freeze-quench apparatus, photolysis, radicals
Procedia PDF Downloads 2078154 Toxic Activity of the Entomopathogenic Fungus Beauveria bassiana on the Wistar Rat Rattus norvegicus
Authors: F. Haddadj, S. Hamdi, M. Khames, A. Kadi, S. Zenia, A. Smai, H. Saadi, B. Doumandji-Mitiche
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The use of a biopesticide based on a microorganism scale requires particular care including safety against the useful auxiliary fauna and mammals among other human beings. Due to its persistence in soil and its apparent human and animal safety, Beauveria bassiana is a cryptogram used for controlling pests organizations, particularly in the locust where its effectiveness has been proven by several highly studies. This fungus is also called for greater respect for biotic communities and the environment. Indeed, biopesticides have several environmental benefits: biodegradability, their activity and selectivity decrease unintended non-target species effects, decreased resistance to some of them. It is in this sense that we contribute by presenting our work on the safety of B. bassiana against mammals. For this we conducted a toxicological study of this fungus strain on Wistar rats Rattus norvegicus, first its effect on weight gain. In a second time were performed histological target organ is the liver. After 20 days of treatment, the results of the toxicological studies have shown that B. bassiana caused no change in the physiological state of rats or weight gain, behavior and diet. On cuts in liver histology revealed no disturbance on the organ.Keywords: entomopathogenic fungus, B. bassiana, Rattus norvegicus, toxicological study, environment
Procedia PDF Downloads 3128153 Heavy Metal Contamination in Soils: Detection and Assessment Using Machine Learning Algorithms Based on Hyperspectral Images
Authors: Reem El Chakik
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The levels of heavy metals in agricultural lands in Lebanon have been witnessing a noticeable increase in the past few years, due to increased anthropogenic pollution sources. Heavy metals pose a serious threat to the environment for being non-biodegradable and persistent, accumulating thus to dangerous levels in the soil. Besides the traditional laboratory and chemical analysis methods, Hyperspectral Imaging (HSI) has proven its efficiency in the rapid detection of HMs contamination. In Lebanon, a continuous environmental monitoring, including the monitoring of levels of HMs in agricultural soils, is lacking. This is due in part to the high cost of analysis. Hence, this proposed research aims at defining the current national status of HMs contamination in agricultural soil, and to evaluate the effectiveness of using HSI in the detection of HM in contaminated agricultural fields. To achieve the two main objectives of this study, soil samples were collected from different areas throughout the country and were analyzed for HMs using Atomic Absorption Spectrophotometry (AAS). The results were compared to those obtained from the HSI technique that was applied using Hyspex SWIR-384 camera. The results showed that the Lebanese agricultural soils contain high contamination levels of Zn, and that the more clayey the soil is, the lower reflectance it has.Keywords: agricultural soils in Lebanon, atomic absorption spectrophotometer, hyperspectral imaging., heavy metals contamination
Procedia PDF Downloads 1108152 Probabilistic Models to Evaluate Seismic Liquefaction In Gravelly Soil Using Dynamic Penetration Test and Shear Wave Velocity
Authors: Nima Pirhadi, Shao Yong Bo, Xusheng Wan, Jianguo Lu, Jilei Hu
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Although gravels and gravelly soils are assumed to be non-liquefiable because of high conductivity and small modulus; however, the occurrence of this phenomenon in some historical earthquakes, especially recently earthquakes during 2008 Wenchuan, Mw= 7.9, 2014 Cephalonia, Greece, Mw= 6.1 and 2016, Kaikoura, New Zealand, Mw = 7.8, has been promoted the essential consideration to evaluate risk assessment and hazard analysis of seismic gravelly soil liquefaction. Due to the limitation in sampling and laboratory testing of this type of soil, in situ tests and site exploration of case histories are the most accepted procedures. Of all in situ tests, dynamic penetration test (DPT), Which is well known as the Chinese dynamic penetration test, and shear wave velocity (Vs) test, have been demonstrated high performance to evaluate seismic gravelly soil liquefaction. However, the lack of a sufficient number of case histories provides an essential limitation for developing new models. This study at first investigates recent earthquakes that caused liquefaction in gravelly soils to collect new data. Then, it adds these data to the available literature’s dataset to extend them and finally develops new models to assess seismic gravelly soil liquefaction. To validate the presented models, their results are compared to extra available models. The results show the reasonable performance of the proposed models and the critical effect of gravel content (GC)% on the assessment.Keywords: liquefaction, gravel, dynamic penetration test, shear wave velocity
Procedia PDF Downloads 1998151 Pain Intensity, Functional Disability and Physical Activity among Elderly Individuals with Chronic Mechanical Low Back Pain
Authors: Adesola Odole, Nse Odunaiya, Samuel Adewale
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Chronic Mechanical Low Back Pain (CMLBP) is prevalent in the aging population; some studies have documented the association among pain intensity, functional disability and physical activity in the general population but very few studies in the elderly. This study was designed to investigate the association among pain intensity, functional disability and physical activity of elderly individuals with CMLBP in the University College Hospital (UCH), Ibadan, Nigeria and also to determine the difference in physical activity, pain intensity and functional disability between males and females. A total of 96 participants diagnosed with CMLBP participated in this cross-sectional survey. They were conveniently sampled from selected units in the UCH, Ibadan, Nigeria. Data on sex, marital status, occupation and duration of onset of pain of participants were obtained from the participants. The Physical Activity Scale for the Elderly, Visual Analogue Scale and Oswestry Disability Questionnaire were used to measure the physical activity, pain intensity and functional disability of the participants respectively. Data was analysed using Spearman correlation, independent t-test; and α was set at 0.05. Participants (25 males, 71 females) were aged 69.64±7.43 years. The majority (76.0%) of the participants were married, and over half (55.2%) were retirees. Participants’ mean pain intensity score was 5.21±2.03 and mean duration of onset of low back pain was 63.63 ± 90.01 months. The majority (67.6%) of the participants reported severe to crippled functional disability. Their mean functional disability was 46.91 ± 13.99. Participants’ mean physical activity score was 97.47 ± 82.55. There was significant association between physical activity and pain intensity (r = -0.21, p = 0.04). There was significant association between physical activity and functional disability (r = -0.47, p = 0.00). Male (87.26 ± 79.94) and female (101.07 ± 83.71) participants did not differ significantly in physical activity (t = 0.00, p = 0.48). In addition, male (5.48 ± 2.06) and female (5.11 ± 2.02) participants’ pain intensity were comparable (t = 0.26, p = 0.44). There was also no significant difference in functional disability (t = 0.05, p = 0.07) between male (42.56 ±13.85) and female (48.45 ± 13.81) participants. It can be concluded from this study that majority of the elderly individuals with chronic mechanical low back pain had a severe to crippled functional disability. Those who reported increased physical activity had reduced pain intensity and functional disability. Male and female elderly individuals with chronic mechanical low back pain are comparable in their pain intensity, functional disability, and physical activity. Elderly individuals with CMLBP should be educated on the importance of participating in physical activity which could reduce their pain symptoms and improve functional disability.Keywords: elderly, functional disability, mechanical low back pain, pain intensity, physical activity
Procedia PDF Downloads 3178150 Conservation Agriculture in North America
Authors: Ying Chen
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Conservation Agriculture in a sustainable way of farming, as it brings many benefits, such as preventing soil from erosion and degradation, improving soil health, conserving energy, and sequestrating carbon. However, adoption of conservation agriculture has been progressing slowly in some part of the world due to some challenges. Among them, seeding in heavy crop residue is challenging, especially in corn production systems. Weed control is also challenging in conservation agriculture. This research aimed to investigate some technologies that can address these challenges. For crop residue management, vertical tillage and vertical seeding have been studied in multiple research projects. Results showed that vertical tillage and seeding were able to deal with crop residue through cutting residue into small segments, which would not plug seeder in the sub-sequent seeding. Vertical tillage is a conservation tillage system, as it leaves more than 30% crop residue on soil surface while incorporating some residue into the shallow soil layer for fast residue decomposition. For weed control, mechanical weeding can reduce chemical inputs in crop production. A tine weeder was studied for weed control during the early growing season of several field crops (corn, soybean, flax, and pea). Detail results of these studies will be shared at the conference.Keywords: tillage, seeding, mechanical weeding, crop residue
Procedia PDF Downloads 738149 Effect of Hand Grip Strength on Shoulder Muscles Activity in Patients with Subacromial Impingement
Authors: Mohamed E. Abdelrahamn, Mahmoud Aly Hassan, Mohamed Sarhan
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Subacromial impingement syndrome (SIS) is a common shoulder disorder. Patients often complain from a decrease in electromyography (EMG) activity of the rotator cuff muscles especially the supraspinatus muscle during glenohumeral elevation. Objective: The purpose of the study is to assess the effect of applying 50% of maximum voluntary contraction of hand grip strength on the EMG activity of the shoulder muscles in patients with SIS. Methods: Thirty male and female patients participated in this study. Their ages ranged from 25 to 40 years. EMG activity of supraspinatus muscle and middle deltoid muscle was assessed without and with applying 50% of maximum voluntary contraction (MVC). Results: A significant difference was found for both supraspinatus and middle deltoid muscles, indicating that the gripping resulted in increasing muscle activity. Conclusion: Applying 50% MVC of hand grip strength could increase the supraspinatus and middle deltoid muscles activity in patients of SIS. This might be useful in the development and monitoring of shoulder rehabilitation strategies.Keywords: electromyography, supraspinatus muscle, deltoid muscle, subacromial impingement syndrome
Procedia PDF Downloads 3008148 Validation of Contemporary Physical Activity Tracking Technologies through Exercise in a Controlled Environment
Authors: Reem I. Altamimi, Geoff D. Skinner
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Extended periods engaged in sedentary behavior increases the risk of becoming overweight and/or obese which is linked to other health problems. Adding technology to the term ‘active living’ permits its inclusion in promoting and facilitating habitual physical activity. Technology can either act as a barrier to, or facilitate this lifestyle, depending on the chosen technology. Physical Activity Monitoring Technologies (PAMTs) are a popular example of such technologies. Different contemporary PAMTs have been evaluated based on customer reviews; however, there is a lack of published experimental research into the efficacy of PAMTs. This research aims to investigate the reliability of four PAMTs: two wristbands (Fitbit Flex and Jawbone UP), a waist-clip (Fitbit One), and a mobile application (iPhone Health Application) for recording a specific distance walked on a treadmill (1.5km) at constant speed. Physical activity tracking technologies are varied in their recordings, even while performing the same activity. This research demonstrates that Jawbone UP band recorded the most accurate distance compared to Fitbit One, Fitbit Flex, and iPhone Health Application.Keywords: Fitbit, jawbone up, mobile tracking applications, physical activity tracking technologies
Procedia PDF Downloads 3218147 Systems of Liquid Organic Fertilizer Application with Respect to Environmental Impact
Authors: Hidayatul Fitri, Petr Šařec
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The use of organic fertilizer is increasing nowadays, and the application must be conducted accurately to provide the right benefits for plants and maintain soil health. Improper application of fertilizers can cause problems for both plants and the environment. This study investigated the liquid organic fertilizer application, particularly digestate, varied into different application doses concerning mitigation of adverse environmental impacts, improving water infiltration ability, and crop yields. The experiment was established into eight variants with different digestate doses, conducted on emission monitoring and soil physical properties. As a result, the digestate application with shallow injection (5 cm in depth) was confirmed as an appropriate technique for applying liquid fertilizer into the soil. Gas emissions resulted in low concentration and declined gradually over time, obviously proved from the experiment conducted under two measurements immediately after application and the next day. Applied various doses of liquid digestate fertilizer affected the emission concentrations of NH3 volatilization, differing significantly and decreasing about 40% from the first to second measurement. In this study, winter wheat crop production significantly increases under digestate application with additional N fertilizer. This study suggested the long-term application of digestate to obtain more alteration of soil properties such as bulk density, penetration resistance, and hydraulic conductivity.Keywords: liquid organic fertilizer, digestate, application, ammonia, emission
Procedia PDF Downloads 2858146 Sludge and Compost Amendments in Tropical Soils: Impact on Coriander (Coriandrum sativum) Nutrient Content
Authors: M. López-Moreno, L. Lugo Avilés, F. Román, J. Lugo Rosas, J. Hernández-Viezcas Jr., Peralta-Videa, J. Gardea-Torresdey
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Degradation of agricultural soils has increased rapidly during the last 20 years due to the indiscriminate use of pesticides and other anthropogenic activities. Currently, there is an urgent need of soil restoration to increase agricultural production. Utilization of sewage sludge or municipal solid waste is an important way to recycle nutrient elements and improve soil quality. With these amendments, nutrient availability in the aqueous phase might be increased and production of healthier crops can be accomplished. This research project aimed to achieve sustainable management of tropical agricultural soils, specifically in Puerto Rico, through the amendment of water treatment plant sludge’s. This practice avoids landfill disposal of sewage sludge and at the same time results cost-effective practice for recycling solid waste residues. Coriander sativum was cultivated in a compost-soil-sludge mixture at different proportions. Results showed that Coriander grown in a mixture of 25% compost+50% Voladora soi+25% sludge had the best growth and development. High chlorophyll content (33.01 ± 0.8) was observed in Coriander plants cultivated in 25% compost+62.5% Coloso soil+ 12.5% sludge compared to plants grown with no sludge (32.59 ± 0.7). ICP-OES analysis showed variations in mineral element contents (macro and micronutrients) in coriander plant grown I soil amended with sludge and compost.Keywords: compost, Coriandrum sativum, nutrients, waste sludge
Procedia PDF Downloads 4058145 Synthesis, Biological Evaluation and Molecular Modeling Studies on Chiral Chloroquine Analogues as Antimalarial Agents
Authors: Srinivasarao Kondaparla, Utsab Debnath, Awakash Soni, Vasantha Rao Dola, Manish Sinha, Kumkum Kumkum Srivastava, Sunil K. Puri, Seturam B. Katti
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In a focused exploration, we have designed synthesized and biologically evaluated chiral conjugated new chloroquine (CQ) analogs with substituted piperazines as antimalarial agents. In vitro as well as in vivo studies revealed that compound 7c showed potent activity [for in vitro IC₅₀= 56.98nM (3D7), 97.76nM (K1); for in vivo (up to at the dose of 12.5 mg/kg); SI = 3510] as a new lead of antimalarial agent. Other compounds 6b, 6d, 7d, 7h, 8c, 8d, 9a, and 9c are also showing moderate activity against CQ-sensitive (3D7) strain and superior activity against resistant (K1) strain of P. falciparum. Furthermore, we have carried out docking and 3D-QSAR studies of all in-house data sets (168 molecules) of chiral CQ analogs to explain the structure activity relationships (SAR). Our new findings specified the significance of H-bond interaction with the side chain of heme for biological activity. In addition, the 3D-QSAR study against 3D7 strain indicated the favorable and unfavorable sites of CQ analogs for incorporating steric, hydrophobic and electropositive groups to improve the antimalarial activity.Keywords: piperazines, CQ-sensitive strain-3D7, in-vitro and in-vivo assay, docking, 3D-QSAR
Procedia PDF Downloads 1698144 Development of Functional Dandelion (Tarazacum officinale) Beverage Using Lactobacillus acidophilus F46 with Cinnamoyl Esterase Activity
Authors: Yong Geun Yun, Jong Hui kim, Sang Ho Baik
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This study was carried out to develop a fermented dandelion (Tarazacum officinale) beverage using lactic acid bacteria with cinnamoyl esterase (CE) activity isolated from human feces. Lactic acid bacteria were screened based on bacterial survival ability in dandelion extract and CE activity. Dandelion extract fermented by Lactobacillus acidophilus F-46 (LA-F46) maintained approximately 105-106 log CFU/mL over an 8 days period. After fermented dandelion beverage (FDB) with LA-46 for 8 days at 37oC the pH was decreased from pH 7.0 to 3.5. Antioxidant activity by using DPPH radical scavenging activity of the prepared FDB was significantly increased compared to that of non-fermented dandelion beverage (NFDB). Moreover, CE activity was significantly enhanced during fermentation and showed the approximately 4.3 times increased concentration of caffeic acid up to 9.91 mg/100 mL after 8 days of incubation compared to NFDB. Therefore, it concluded that dandelion can be a good source for preparing a functional beverage and fermentation by LA-F46 enhanced the food functionality with enhanced caffeic acids.Keywords: cinnamoyl esterase, dandelion, fermented beverage, lactic acid bacteria
Procedia PDF Downloads 4018143 Role of Matric Suction in Mechanics behind Swelling Characteristics of Expansive Soils
Authors: Saloni Pandya, Nikhil Sharma, Ajanta Sachan
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Expansive soils in the unsaturated state are part of vadose zone and encountered in several arid and semi-arid parts of the world. Influence of high temperature, low precipitation and alternate cycles of wetting and drying are responsible for the chemical weathering of rocks, which results in the formation of expansive soils. Shrinkage-swelling (expansive) soils cover a substantial portion of area in India. Damages caused by expansive soils to various geotechnical structures are alarming. Matric suction develops in unsaturated soil due to capillarity and surface tension phenomena. Matric suction influences the geometric arrangement of soil skeleton, which induces the volume change behaviour of expansive soil. In the present study, an attempt has been made to evaluate the role of matric suction in the mechanism behind swelling characteristics of expansive soil. Four different soils have been collected from different parts of India for the current research. Soil sample S1, S2, S3 and S4 were collected from Nagpur, Bharuch, Bharuch-Dahej highway and Ahmedabad respectively. DFSI (Differential Free Swell Index) of these soils samples; S1, S2, S3, and S4; were determined to be 134%, 104%, 70% and 30% respectively. X-ray diffraction analysis of samples exhibited that percentage of Montmorillonite mineral present in the soils reduced with the decrease in DFSI. A series of constant volume swell pressure tests and in-contact filter paper tests were performed to evaluate swelling pressure and matric suction of all four soils at 30% saturation and 1.46 g/cc dry density. Results indicated that soils possessing higher DFSI exhibited higher matric suction as compared to lower DFSI expansive soils. Significant influence of matric suction on swelling pressure of expansive soils was observed with varying DFSI values. Higher matric suction of soil might govern the water uptake in the interlayer spaces of Montmorillonite mineral present in expansive soil leading to crystalline swelling.Keywords: differential free swell index, expansive soils, matric suction, swelling pressure
Procedia PDF Downloads 1628142 Molecular Insights into the 5α-Reductase Inhibitors: Quantitative Structure Activity Relationship, Pre-Absorption, Distribution, Metabolism, and Excretion and Docking Studies
Authors: Richa Dhingra, Monika, Manav Malhotra, Tilak Raj Bhardwaj, Neelima Dhingra
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5-Alpha-reductases (5AR), a membrane bound, NADPH dependent enzyme and convert male hormone testosterone (T) into more potent androgen dihydrotestosterone (DHT). DHT is the required for the development and function of male sex organs, but its overproduction has been found to be associated with physiological conditions like Benign Prostatic Hyperplasia (BPH). Thus the inhibition of 5ARs could be a key target for the treatment of BPH. In present study, 2D and 3D Quantitative Structure Activity Relationship (QSAR) pharmacophore models have been generated for 5AR based on known inhibitory concentration (IC₅₀) values with extensive validations. The four featured 2D pharmacophore based PLS model correlated the topological interactions (–OH group connected with one single bond) (SsOHE-index); semi-empirical (Quadrupole2) and physicochemical descriptors (Mol. wt, Bromines Count, Chlorines Count) with 5AR inhibitory activity, and has the highest correlation coefficient (r² = 0.98, q² =0.84; F = 57.87, pred r² = 0.88). Internal and external validation was carried out using test and proposed set of compounds. The contribution plot of electrostatic field effects and steric interactions generated by 3D-QSAR showed interesting results in terms of internal and external predictability. The well validated 2D Partial Least Squares (PLS) and 3D k-nearest neighbour (kNN) models were used to search novel 5AR inhibitors with different chemical scaffold. To gain more insights into the molecular mechanism of action of these steroidal derivatives, molecular docking and in silico absorption, distribution, metabolism, and excretion (ADME) studies were also performed. Studies have revealed the hydrophobic and hydrogen bonding of the ligand with residues Alanine (ALA) 63A, Threonine (THR) 60A, and Arginine (ARG) 456A of 4AT0 protein at the hinge region. The results of QSAR, molecular docking, in silico ADME studies provide guideline and mechanistic scope for the identification of more potent 5-Alpha-reductase inhibitors (5ARI).Keywords: 5α-reductase inhibitor, benign prostatic hyperplasia, ligands, molecular docking, QSAR
Procedia PDF Downloads 1618141 Activity of Malate Dehydrogenase in Cell Free Extracts from S. proteamaculans, A. hydrophila, and K. pneumoniae
Authors: Mohamed M. Bumadian, D. James Gilmour
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Three bacterial species were isolated from the River Wye (Derbyshire, England) and identified using 16S rRNA gene sequencing as Serratia proteamaculans, Aeromonas hydrophila and Klebsiella pneumoniae. Respiration rates of the strains were measured in order to determine the metabolic activity under salt stress. The highest respiration rates of all three strains were found at 0.17 M and 0.5 M NaCl and then the respiration rate decreased with increasing concentrations of NaCl. In addition, the effect of increasing concentrations of NaCl on malate dehydrogenase activity was determined using cell-free extracts of the three strains. Malate dehydrogenase activity was stimulated at NaCl concentrations up to 0.5 M, and a small level of activity remained even at 3.5 M NaCl. The pH optimum of the malate dehydrogenase in cell-free extracts of all strains was higher than pH 7.5.Keywords: fresh water, halotolerant pathogenic bacteria, 16S rRNA gene, cell-free extracts, respiration rates, malate dehydrogenase
Procedia PDF Downloads 4618140 Antimicrobial Activity of the Cyanobacteria spp. against Fish Pathogens in Aquaculture
Authors: I. Tulay Cagatay
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Blue-green microalgae cyanobacteria, which are important photosynthetic organisms of aquatic ecosystems, are the primary sources of many bioactive compounds such as proteins, carbohydrates, lipids, vitamins and enzymes that can be used as antimicrobial and antiviral agents. Some of these organisms are nowadays used directly in the food, cosmetic and pharmaceutical industry, or in aquaculture and biotechnological approaches like biofuel or drug therapy. Finding the effective, environmental friendly chemotropic and antimicrobial agents to control fish pathogens are crucial in a country like Turkey which has a production capacity of about 240 thousand tons of cultured fish and has 2377 production farms and which is the second biggest producer in Europe. In our study, we tested the antimicrobial activity of cyanobacterium spp. against some fish pathogens Aeromonas hydrophila and Yersinia ruckeri that are important pathogens for rainbow trout farms. Agar disk diffusion test method was used for studying antimicrobial activity on pathogens. Both tested microorganisms have shown antimicrobial activity positively as the inhibition zones were 0.45 mm and 0.40 mm respectively.Keywords: fish pathogen, cyanobacteria, antimicrobial activity, trout
Procedia PDF Downloads 1628139 A Numerical Study for Mixing Depth and Applicability of Partial Cement Mixing Method Utilizing Geogrid and Fixing Unit
Authors: Woo-seok Choi, Eun-sup Kim, Nam-Seo Park
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The demand for new technique in soft ground improvement continuously increases as general soft ground methods like PBD and DCM have a application problem in soft grounds with deep depth and wide distribution in Southern coast of Korea and Southeast. In this study, partial cement mixing method utilizing geogrid and fixing unit(CMG) is suggested and Finite element analysis is performed for analyzing the depth of surface soil and deep soil stabilization and comparing with DCM method. In the result of the experiment, the displacement in DCM method were lower than the displacement in CMG, it's because the upper load is transferred to deep part soil not treated by cement in CMG method case. The differential settlement in DCM method was higher than the differential settlement in CMG, because of the effect load transfer effect by surface part soil treated by cement and geogrid. In conclusion, CMG method has the advantage of economics and constructability in embankment road, railway, etc in which differential settlement is the important consideration.Keywords: soft ground, geogrid, fixing unit, partial cement mixing, finite element analysis
Procedia PDF Downloads 3778138 Geological Engineering Mapping Approach to Know Factor of Safety Distribution and Its Implication to Landslide Potential at Muria Mountain, Kudus, Central Java Province, Indonesia
Authors: Sony Hartono, Azka Decana, Vilia Yohana, Annisa Luthfianihuda, Yuni Faizah, Tati Andriani, Dewi Kania, Fachri Zulfiqar, Sugiar Yusup, Arman Nugraha
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Landslide is a geological hazard that is quite common in some areas in Indonesia and have disadvantages impact for public around. Due to the high frequency of landslides in Indonesia, and extensive damage, landslides should be specifically noted. Landslides caused by a soil or rock unit that has been in a state of unstable slopes and not in ideal state again, so the value of ground resistance or the rock been passed by the value of the forces acting on the slope. Based on this fact, authors held a geological engineering mapping at Muria Mountain, Kudus, Central Java province which is known as an agriculture and religion tourism area. This geological engineering mapping is performed to determine landslides potential at Muria Mountain. Slopes stability will be illustrated by a number called the “factor of safety” where the number can describe how much potential a slope to fall. Slopes stability can be different depending on the physical and mechanical characteristics of the soil and slope conditions. Testing of physical and mechanical characteristics of the soil conducted in the geotechnical laboratory. The characteristics of the soil must be same when sampled as well as in the test laboratory. To meet that requirement, authors used "undisturb sample" method that will be guarantee sample will not be distracted by environtment influences. From laboratory tests on soil physical and mechanical properties obtained characteristics of the soil on a slope, and then inserted into a Geological Information Software that would generate a value of factor of safety and give a visualization slope form area of research. Then, as a result of the study, obtained a map of the ground movement distribution map and i is implications for landslides potential areas.Keywords: factor of safety, geological engineering mapping, landslides, slope stability, soil
Procedia PDF Downloads 4188137 Evaluation of Moroccan Microalgae Spirulina platensis as a Potential Source of Natural Antioxidants
Authors: T. Ould Bellahcen, A. Amiri, I. Touam, F. Hmimid, A. El Amrani, M. Cherki
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The antioxidant activity of three extracts (water, lipidic and ethanolic) prepared from the microalgae Spirulina platensis isolated from Moroccan lake, using 2, 2 diphenyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis ethylbenzthiazoline-6-sulfonic acid (ABTS) radical assay, was studied and compared. The obtained results revealed that the IC₅₀ found using DPPH were lower than that of ABTS for all extracts from these planktonic blue-green algae. The high levels of phenolic and flavonoid content were found in the ethanolic extract 0,33 ± 0,01 mg GAE/g dw and 0,21 ± 0,01 mg quercetin/g dw respectively. In addition, using DPPH, the highest activity with IC₅₀ = 0,449 ± 0,083 mg/ml, was found for the ethanolic extract, followed by that of lipidic extract (IC₅₀ = 0,491 ± 0,059 mg/ml). The lowest activity was for the aqueous extract (IC₅₀ = 4,148 ± 0,132 mg/ml). For ABTS, the highest activity was observed for the lipidic extract with IC₅₀ = 0,740 ± 0,012 mg/ml, while, the aqueous extract recorded the lowest activity (IC₅₀ = 6,914 ± 0, 0067 mg/ml). A moderate activity was showed for the ethanolic extract (IC₅₀ = 5,852 ± 0, 0171 mg/ml). It can be concluded from this first study that Spirulina platensis extracts show an interesting antioxidant and antiradicals properties suggesting that this alga could be used as a potential source of antioxidants. A qualitative and quantitative analysis of polyphenol and flavonoids in the extracts using HPLC is in progress so as to study the correlation between the antioxidant activity and chemical composition.Keywords: Spirulina platensis, antioxidant, DPPH, ABTS
Procedia PDF Downloads 1648136 Ability of Gastric Enzyme Extract of Adult Camel to Clot Bovine Milk
Authors: Boudjenah-Haroun Saliha, Isselnane Souad, Nouani Abdelwahab, Baaissa Babelhadj, Mati Abderrahmane
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Algeria is experiencing significant development of the dairy sector, where consumption of milk and milk products increased by 2.7 million tons in 2008 to 4,400,000 tons in 2013, and cheese production has reached 1640 tons in the year 2014 with average consumption of 0.7 kg/person/year. Although rennet is still the most used coagulating enzyme in cheese, its production has been growing worldwide shortage. This shortage is primarily due to a growing increase in the production and consumption of cheese, and the inability to increase in parallel the production of rennet. This shortage has caused very large fluctuations in its price). To overcome these obstacles, much research has been undertaken to find effective and competitive substitutes used industrially. For this, the selection of a local production of rennet substitute is desirable. It would allow a permanent supply with limited dependence on imports and price fluctuations. Investigations conducted by our research team showed that extracts coagulants from the stomachs of older camels are characterized by a coagulating power than those from younger camels. The objective of this work is to study the possibility of substituting commercial rennet coagulant by gastric enzymes from adult camels for coagulation bovine milk. Excerpts from the raw camel coagulants obtained are characterized through their teneures proteins and clotting and proteolytic activities. Milk clotting conditions by the action of these extracts were optimized. Milk clotting time all treated with enzyme preparations and under different conditions was calculated. Bovine rennet has been used for comparison. The results show that crude extracts from gastric adult camel can be good substituting bovine rennet.Keywords: Algeria, camel, cheese, coagulation, gastric extracts, milk
Procedia PDF Downloads 4388135 Effects of Enzymatic Liquefaction on the Physicochemical Properties and Antioxidant Activity of Zn-Amaranth (Amaranthus viridis) Puree
Authors: M. A. Siti Faridah, K. Muhammad, H. M. Ghazali, Y. A. Yusof
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This study was conducted to investigate the effects of three variables namely types of cell wall degrading enzymes (Viscozyme L, Pectinex Ultra SP-L, Rapidase PAC, Rohament CL and Rohapect PTE) at varying concentrations (0.25-3% v/w) and times (30 min-24 h) on the zinc (Zn-) amaranth purees. Liquefaction treatment of the Zn-amaranth purees with Viscozyme (1% v/w at pH 5 and 45ºC for 3 h) was found to be the best procedure, which produced Zn-amaranth puree with low viscosity (8.60 mPas). Zn-amaranth purees were also found to have the highest metallo-chlorophyll derivative contents (0.16 mg/g), free radical 2, 2-diphenyl-1-picrylhydrazyl (DPPH) values (12.49 mM (TE)/g fresh weight) and ferric reducing antioxidant power (FRAP) values (4.57 mM (TE)/g fresh weight) within 3 h of liquefaction. Other physicochemical properties of the enzyme-liquefied Zn-amaranth purees indicated that lightness (L*) (12.54), greenness a*/b* (-0.30), reducing sugar (103.88 mg/mL) and soluble dietary fibre (5.94%) of the purees were higher compared to that of nonenzyme-liquefied amaranth purees.Keywords: amaranth, antioxidant, chlorophyll derivative, enzymatic liquefaction
Procedia PDF Downloads 1448134 Characterization of Soil Microbial Communities from Vineyard under a Spectrum of Drought Pressures in Sensitive Area of Mediterranean Region
Authors: Gianmaria Califano, Júlio Augusto Lucena Maciel, Olfa Zarrouk, Miguel Damasio, Jose Silvestre, Ana Margarida Fortes
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Global warming, with rapid and sudden changes in meteorological conditions, is one of the major constraints to ensuring agricultural and crop resilience in the Mediterranean regions. Several strategies are being adopted to reduce the pressure of drought stress on grapevines at regional and local scales: improvements in the irrigation systems, adoption of interline cover crops, and adaptation of pruning techniques. However, still, more can be achieved if also microbial compartments associated with plants are considered in crop management. It is known that the microbial community change according to several factors such as latitude, plant variety, age, rootstock, soil composition and agricultural management system. Considering the increasing pressure of the biotic and abiotic stresses, it is of utmost necessity to also evaluate the effects of drought on the microbiome associated with the grapevine, which is a commercially important crop worldwide. In this study, we characterize the diversity and the structure of the microbial community under three long-term irrigation levels (100% ETc, 50% ETc and rain-fed) in a drought-tolerant grapevine cultivar present worldwide, Syrah. To avoid the limitations of culture-dependent methods, amplicon sequencing with target primers for bacteria and fungi was applied to the same soil samples. The use of the DNeasy PowerSoil (Qiagen) extraction kit required further optimization with the use of lytic enzymes and heating steps to improve DNA yield and quality systematically across biological treatments. Target regions (16S rRNA and ITS genes) of our samples are being sequenced with Illumina technology. With bioinformatic pipelines, it will be possible to obtain a characterization of the bacterial and fungal diversity, structure and composition. Further, the microbial communities will be assessed for their functional activity, which remains an important metric considering the strong inter-kingdom interactions existing between plants and their associated microbiome. The results of this study will lay the basis for biotechnological applications: in combination with the establishment of a bacterial library, it will be possible to explore the possibility of testing synthetic microbial communities to support plant resistance to water scarcity.Keywords: microbiome, metabarcoding, soil, vinegrape, syrah, global warming, crop sustainability
Procedia PDF Downloads 1218133 Virtual Screening and in Silico Toxicity Property Prediction of Compounds against Mycobacterium tuberculosis Lipoate Protein Ligase B (LipB)
Authors: Junie B. Billones, Maria Constancia O. Carrillo, Voltaire G. Organo, Stephani Joy Y. Macalino, Inno A. Emnacen, Jamie Bernadette A. Sy
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The drug discovery and development process is generally known to be a very lengthy and labor-intensive process. Therefore, in order to be able to deliver prompt and effective responses to cure certain diseases, there is an urgent need to reduce the time and resources needed to design, develop, and optimize potential drugs. Computer-aided drug design (CADD) is able to alleviate this issue by applying computational power in order to streamline the whole drug discovery process, starting from target identification to lead optimization. This drug design approach can be predominantly applied to diseases that cause major public health concerns, such as tuberculosis. Hitherto, there has been no concrete cure for this disease, especially with the continuing emergence of drug resistant strains. In this study, CADD is employed for tuberculosis by first identifying a key enzyme in the mycobacterium’s metabolic pathway that would make a good drug target. One such potential target is the lipoate protein ligase B enzyme (LipB), which is a key enzyme in the M. tuberculosis metabolic pathway involved in the biosynthesis of the lipoic acid cofactor. Its expression is considerably up-regulated in patients with multi-drug resistant tuberculosis (MDR-TB) and it has no known back-up mechanism that can take over its function when inhibited, making it an extremely attractive target. Using cutting-edge computational methods, compounds from AnalytiCon Discovery Natural Derivatives database were screened and docked against the LipB enzyme in order to rank them based on their binding affinities. Compounds which have better binding affinities than LipB’s known inhibitor, decanoic acid, were subjected to in silico toxicity evaluation using the ADMET and TOPKAT protocols. Out of the 31,692 compounds in the database, 112 of these showed better binding energies than decanoic acid. Furthermore, 12 out of the 112 compounds showed highly promising ADMET and TOPKAT properties. Future studies involving in vitro or in vivo bioassays may be done to further confirm the therapeutic efficacy of these 12 compounds, which eventually may then lead to a novel class of anti-tuberculosis drugs.Keywords: pharmacophore, molecular docking, lipoate protein ligase B (LipB), ADMET, TOPKAT
Procedia PDF Downloads 4228132 Case Study: Geomat Installation against Slope Erosion
Authors: Serap Kaymakci, Dogan Gundogdu, M. Bugra Yagcioglu
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Erosion (soil erosion) is a phenomenon in which the soil on the slope surface is exposed to natural influences such as wind, rainfall, etc. in open areas. The most natural solution to prevent erosion is to plant surfaces exposed to erosion. However, proper ground and natural conditions must be provided in order for planting to occur. Erosion is prevented in a fast and natural way and the loss of soil is reduced mostly. Lead to allowing plants to hold onto the soil with its three-dimensional and hollow structure are as follows: The types of geomat called MacMat that is used in a case study in Turkey in order to prevent water carry over due to rainfall. The geosynthetic combined with double twisted steel wire mesh. That consists of 95% Zn–5% Al alloy coated double twisted steel wire based that is a reinforced MacMat (geosynthetic three-dimensional erosion control mat) obtained by a polypropylene consisted (mesh type 8x10-Wire diam. 2.70 mm–95% Zn–5% Al alloy coated). That is developed by the progress of the technology. When using reinforced MacMat on top clay liners, fixing pins should not be used as they will rupture the mats. Mats are simply anchored (J Type) in the top trench and, if necessary, in intermediate berm trenches. If the slope angle greater than 20°, it is necessary to use additional rebar depending soil properties also. These applications may have specific technical and installation requirements. In that project, the main purpose is erosion control after that is greening. There is a slope area around the factory which is located in Gebze, İstanbul.Keywords: erosion, GeoMat, geosynthetic, slope
Procedia PDF Downloads 1748131 Effect of Bored Pile Diameter in Sand on Friction Resistance
Authors: Ashraf Mohammed M. Eid, Hossam El Badry
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The bored pile friction resistance may be affected by many factors such as the method of construction, pile length and diameter, the soil properties, as well as the depth below ground level. These factors can be represented analytically to study the influence of diameter on the unit skin friction. In this research, the Egyptian Code of soil mechanics is used to assess the skin friction capacity for either the ordinary pile diameter as well as for the large pile diameter. The later is presented in the code and through the work of some researchers based on the results of investigations adopted for a sufficient number of field tests. The comparative results of these researchers with respect to the Egyptian Code are used to check the adequacy of both methods. Based on the results of this study, the traditional static formula adopted for piles of diameter less than 60 cm may be continually used for larger piles by correlating the analyzed formulae. Accordingly, the corresponding modified angle of internal friction is concluded demonstrating a reduction of shear strength due to soil disturbance along the pile shaft. Based on this research the difference between driven piles and bored piles constructed in same soil can be assessed and a better understanding can be evaluated for the effect of different factors on pile skin friction capacity.Keywords: large piles, static formula, friction piles, sandy soils
Procedia PDF Downloads 4998130 Multi-Walled Carbon Nanotubes Doped Poly (3,4 Ethylenedioxythiophene) Composites Based Electrochemical Nano-Biosensor for Organophosphate Detection
Authors: Navpreet Kaur, Himkusha Thakur, Nirmal Prabhakar
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One of the most publicized and controversial issue in crop production is the use of agrichemicals- also known as pesticides. This is evident in many reports that Organophosphate (OP) insecticides, among the broad range of pesticides are mainly involved in acute and chronic poisoning cases. Therefore, detection of OPs is very necessary for health protection, food and environmental safety. In our study, a nanocomposite of poly (3,4 ethylenedioxythiophene) (PEDOT) and multi-walled carbon nanotubes (MWCNTs) has been deposited electrochemically onto the surface of fluorine doped tin oxide sheets (FTO) for the analysis of malathion OP. The -COOH functionalization of MWCNTs has been done for the covalent binding with amino groups of AChE enzyme. The use of PEDOT-MWCNT films exhibited an excellent conductivity, enables fast transfer kinetics and provided a favourable biocompatible microenvironment for AChE, for the significant malathion OP detection. The prepared PEDOT-MWCNT/FTO and AChE/PEDOT-MWCNT/FTO nano-biosensors were characterized by Fourier transform infrared spectrometry (FTIR), Field emission-scanning electron microscopy (FE-SEM) and electrochemical studies. Electrochemical studies were done using Cyclic Voltammetry (CV) or Differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS). Various optimization studies were done for different parameters including pH (7.5), AChE concentration (50 mU), substrate concentration (0.3 mM) and inhibition time (10 min). The detection limit for malathion OP was calculated to be 1 fM within the linear range 1 fM to 1 µM. The activity of inhibited AChE enzyme was restored to 98% of its original value by 2-pyridine aldoxime methiodide (2-PAM) (5 mM) treatment for 11 min. The oxime 2-PAM is able to remove malathion from the active site of AChE by means of trans-esterification reaction. The storage stability and reusability of the prepared nano-biosensor is observed to be 30 days and seven times, respectively. The application of the developed nano-biosensor has also been evaluated for spiked lettuce sample. Recoveries of malathion from the spiked lettuce sample ranged between 96-98%. The low detection limit obtained by the developed nano-biosensor made them reliable, sensitive and a low cost process.Keywords: PEDOT-MWCNT, malathion, organophosphates, acetylcholinesterase, nano-biosensor, oxime (2-PAM)
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