Search results for: Rishabh Shukla

Authors: Anupam Saxena, Achin Agrawal, Rishabh Shukla, S. Mandal

Abstract:

It is quite important that the properties of structural elements do not change significantly before and after cracking, and if they do, it adversely affects the structure. Corrosion in rebars causes cracking in concrete which can lead to the change in properties of beam. In the present study, two RC beams with same flexural strength but with different reinforcement arrangements are considered and modelling of cracks of RC beams has been done at different degrees of corrosion in the case of delamination using boundary conditions of Three Point Bending Test. Finite Element Analysis (FEA) has been done at different degree of corrosion to observe the variation of different parameters like modal frequency, Elasticity and Flexural strength in case of delamination. Also, the comparison between two different RC arrangements is made to conclude which one of them is more suitable.

Keywords: delamination, elasticity, FEA, flexural strength, modal frequency, RC beam

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Authors: Rishabh Shukla, Achin Agrawal, Anupam Saxena, S. Mandal

Abstract:

The existence of a crack, affects the mechanical behaviour and various properties of a structure to a great degree. This paper focuses on recognizing the parameters that gets changed due to the formation of cracks and have a great impact on the performance of the structure. Spalling is a major concern as it leaves the reinforcement bars more susceptible to environmental attacks. Beams of cross section 300 mm × 500 mm are designed and for a calculated area of steel, two different arrangements of reinforced bars are analysed. Results are prepared for different stages of cracking for each arrangement of rebars. The parameters for both arrangements are then compared. The Finite Element Analysis (FEA) is carried out and changes in the properties like flexural strength, Elasticity and modal frequency are reported. The conclusions have been drawn by comparing the results.

Keywords: cracks, elasticity, spalling, FEA

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Authors: Seyed Mohammad Nasir Mousavi, Amir Mashayekh, Pasha Hejazi, Sanaz Kanani Zadeh Khalkhali

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To determine the stability of the oil percent canola varieties, an experiment was done in a randomized complete block design with four replications in four research stations of the country Shahrood, Esfahan, Kermanshah, Varamin. Analysis of variance showed that there is cultivars considerable variability in the percentage of oil. The results showed that the coefficient of variation of oil Hyola 401 and Hyola308 stability and flexibility are high. Cultivars Cooper and Likord are minimum variance Shukla that stable for the percentage of oil Based on the chart AMMI 1, cultivars Zarfam and Hyola 401 are of oil percentage than other varieties had higher stability. On the chart AMMI2, cultivars Karun and Hyola 308 are identified as stable, also location Isfahan is stable

Keywords: canola, stability, AMMI, variance Shukla

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Authors: Rishabh Beri, Sahil Shah

Abstract:

We have applied a range of filters and processing in order to extract out the various features of the football game, like the field lines of a football field. Another important aspect was the detection of the players in the field and tagging them according to their teams distinguished by their jersey colours. This extracted information combined about the players and field helped us to create a virtual field that consists of the playing field and the players mapped to their locations in it.

Keywords: Detect, Football, Players, Virtual

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Authors: Rishabh Sharma

Abstract:

The main design parameter of WSN (wireless sensor network) is the energy consumption. To compensate this parameter, hierarchical clustering is a technique that assists in extending duration of the networks life by efficiently consuming the energy. This paper focuses on dealing with the WSNs and the FIS (fuzzy interface system) which are deployed to enhance the BEENISH protocol. The node energy, mobility, pause time and density are considered for the selection of CH (cluster head). The simulation outcomes exhibited that the projected system outperforms the traditional system with regard to the energy utilization and number of packets transmitted to sink.

Keywords: wireless sensor network, sink, sensor node, routing protocol, fuzzy rule, fuzzy inference system

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Authors: Rishabh Garg

Abstract:

Traditionally, business parties build trust with a centralized operating mechanism, such as payment by letter of credit. However, the increase in cyber-attacks and malicious hacking has jeopardized business operations and finance practices. Emerging markets, owing to their higher banking risks and bigger presence of digital financing, are looking forward to technology-driven solutions, financial inclusion and innovative working paradigms. Blockchain has the potential to enhance transaction transparency and supply chain traceability. It has captured a vast landscape with 200 million crypto users worldwide. Fintech and blockchain products are popping up across brokerage, digital wallets, exchanges, post-trade clearance, settlement, middleware, infrastructure, and base protocols.

Keywords: blockchain, distributed ledger technology, decentralized applications, ethereum, smart contracts, trade finance

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Authors: Rishabh Srivastav, Divyam Sharma

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We propose an algorithm to cluster categorical data named as ‘Genetic algorithm initialized rough set based fuzzy K-Modes for categorical data’. We propose an amalgamation of the simple K-modes algorithm, the Rough and Fuzzy set based K-modes and the Genetic Algorithm to form a new algorithm,which we hypothesise, will provide better Centroid Model clustering results, than existing standard algorithms. In the proposed algorithm, the initialization and updation of modes is done by the use of genetic algorithms while the membership values are calculated using the rough set and fuzzy logic.

Keywords: categorical data, fuzzy logic, genetic algorithm, K modes clustering, rough sets

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Authors: R. C. Sharma, Rishabh Bansal, Prajwal Menon, Manoj K. Sharma

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Silicon-air battery is highly promising for electric vehicles due to its high theoretical energy density (8470 Whkg⁻¹) and its discharge products are non-toxic. For the first time, pure silicon and germanium powders are used as anode material. Nickel wire meshes embedded with charcoal and manganese dioxide powder as cathode and concentrated potassium hydroxide is used as electrolyte. Voltage-time curves have been presented in this study for pure silicon and germanium powder and 5% and 10% germanium with silicon powder. Silicon powder cell assembly gives a stable voltage of 0.88 V for ~20 minutes while Si-Ge provides cell voltage of 0.80-0.76 V for ~10-12 minutes, and pure germanium cell provides cell voltage 0.80-0.76 V for ~30 minutes. The cell voltage is higher for concentrated (10%) sodium hydroxide solution (1.08 V) and it is stable for ~40 minutes. A sharp decrease in cell voltage beyond 40 min may be due to rapid corrosion.

Keywords: Silicon-air battery, Germanium-air battery, voltage-time curve, open circuit voltage, Anodic corrosion

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Authors: Vishal H. Shukla, Sanjay B. Deshmukh

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Traffic engineering, at its center, is the ability of moving traffic approximately so that traffic from a congested link is moved onto the unused capacity on another link. Traffic Engineering ensures the best possible use of the resources. Now to support traffic engineering in the today’s network, Multiprotocol Label Switching (MPLS) is being used which is very helpful for reliable packets delivery in an ongoing internet services. Here a topology is been implemented on GNS3 to focus on the analysis of the communication take place from one site to other through the ISP. The comparison is made between the IP network & MPLS network based on Bandwidth & Jitter which are one of the performance parameters using JPERF simulator.

Keywords: GNS3, JPERF, MPLS, traffic engineering, VMware

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Authors: Sheo K. Mishra, Rajneesh K. Srivastava, R. K. Shukla

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In the present work, we report on the optical properties including UV-vis absorption and photoluminescence (PL) of ZnO nanostructures synthesized by sol-gel method. Structural and morphological investigations have been performed by X-ray diffraction method (XRD) and scanning electron microscopy (SEM). The XRD result confirms the formation of hexagonal wurtzite phase of ZnO nanostructures. The presence of various diffraction peaks suggests polycrystalline nature. The XRD pattern exhibits no additional peak due to by-products such as Zn(OH)2. The average crystallite size of prepared ZnO sample corresponding to the maximum intensity peaks is to be ~38.22 nm. The SEM micrograph shows different nanostructures of pure ZnO. Photoluminescence (PL) spectrum shows several emission peaks around 353 nm, 382 nm, 419 nm, 441 nm, 483 nm and 522 nm. The obtained results suggest that the prepared phosphors are quite suitable for optoelectronic applications.

Keywords: ZnO, sol-gel, XRD, PL

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Authors: Ila Shukla, Lubna Azmi, Shyam Sunder Gupta, C. V. Rao

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Treatments against Hepatitis-C virus (HCV) are already available, but the current high cost of such treatments limit them to wealthy patients only. Hence our current study is aimed at the rectification of HCV infection by using Lichen rangiferinus (LRE) extract in in vitro cultures. Anti-HCV activity of the given extract was evaluated using the virus grown in cell culture (HCVcc). Two control inhibitors, erlotinib and telaprevir, were systematically included in each experiment. At the end of the incubation period, we evaluated cell viability and viral replication. The LRE inhibited the growth of HCV in a dose dependent manner.

Keywords: Erlotinib, Hepatitis C, Lichen rangiferinus, Telaprevir

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Authors: M. S. Thakur, Mohit Shukla

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A channel/tunnel, which carries the water to the penstock/pressure shaft is called headrace tunnel (HRT). It is necessary to know the general topography, geology of the area, state of stress and other mechanical properties of the strata. For this certain topographical and geological investigations, in-situ and laboratory tests, and observations are required to be done. These investigations play an important role in a tunnel design as these help in deciding the optimum layout, shape and size and support requirements of the tunnel. The paper includes inputs from Nathpa Jhakri Hydeoelectric project which is India’s highest capacity (1500 MW) operating hydroelectric project. The paper would help the design engineers with various new concepts and preparedness against geological surprises.

Keywords: tunnelling, geology, HRT, rockmass

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Authors: Nikhil Verma, Akshay Kant Mishra, Rishabh Rastogi, Bikarama Prasad Yadav

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Emerging fire incidents are the protuberant contributor out turning into life loss, property damage and importantly firefighters. It insinuates that a firefighting and rescue operation of the existing equipment or apparatus and their proficiency is limited, particularly in annihilating firefighting environments. The proposed methodology will help in developing a technology which can be useful in minimizing the risks and losses due to fire. In this paper, design and development of combat mini vehicle comprising of multi-purpose nozzle system is proposed which can target diverse fires simultaneously at distinct time and location. Basically, the system is semi-automated type protection system which can be manoeuvred by controller. Designing of robust vehicle based on semi-automated protection type system is consummated using SolidWorks platform. Concept of developing a robust vehicle will help to fight fires in multiple directions reducing the time required to douse multiple fires.

Keywords: fire douse vehicle, multiple fires, multi-purpose nozzle, semi-automated system

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Authors: Shyam Shukla

Abstract:

A brand is the identity of a specific product, service or business. A brand can take many forms, including a name, sign, symbol, color, combination or slogan. The word brand began simply as a way to tell one person's identity from another by means of a hot iron stamp. A legally protected brand name is called a trademark. The word brand has continued to evolve to encompass identity - it affects the personality of a product, company or service. A concept brand is a brand that is associated with an abstract concept, like AIDS awareness or environmentalism, rather than a specific product, service, or business. A commodity brand is a brand associated with a commodity1. In this paper, it is tried to explore the significance of an intellectual capital for the branding of an Institution.

Keywords: brand, commodity, consumer, cultural values, intellectual capital, zonal cluster

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Authors: Hiral D. Trivedi, Dinesh S. Patel, Sachin P. Shukla

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We have immobilized alcohol dehydrogenase on k-carrageenan, which is a natural polysaccharide obtained from seaweeds by entrapment and on copolymer of acrylamide and 2-hydroxy ethylmethaacrylate by covalent coupling. We have optimized all the immobilization parameters and also carried the comparison studies of both. In case of copolymer of acrylamide and 2-hydroxy ethylmethaacrylate, we have activated both the amino and hydroxyl group individually and simultaneously using different activating agents and obtained some interesting results. We have found that covalently bound enzyme was found to be better under all tested conditions. The reaction on ethanol was carried out using these immobilized systems.

Keywords: alcohol dehydrogenase, acrylamide-co-2-hydroxy ethylmethaacrylate, ethanol, k-carrageenan

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Authors: Sandeep K. Shukla, Roli Jain, Soumitra S. Pande, Archna Pandey

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Sulfur nanoparticles were prepared by different methods with different sizes and shapes. When the sulfur is present as nanoparticles they have many practical applications in our life. This research discusses sulfur nanoparticles synthesis, characterizations and applications. With dandruff being a common everyday problem and the market is loaded with antidandruff shampoos and such skin care products, it is obvious to assume resourceful research into this area would be both objective to present scenario and potentially lucrative. Nanoparticles are frequently in use in some very powerful antimicrobial, antifungal cosmetics nowadays, especially silver. To check its antidandruff activity, experiments have been conducted on Malassezia furfur the causal organism for seborrheaic dermatitis or dandruff, which have been cultured for such study in our lab.

Keywords: CTAB surfactant SEM, sulfur nanoparticles (S-NPs), XRD, polymeric surfactant

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Authors: Rishabh Agrawal, S. C. Kaushik, T. S. Bhatti

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In order to reduce energy consumption for sustainable development, continuous energy consumption tracking of building energy systems are essential. In this paper an assessment study has been done to identify the energy consumption & energy conservation potential for commercial buildings sector in Karnataka state, India. There are a total of 326 commercial buildings in the state of Karnataka who has qualified as designated consumers (i.e., having a Contract Demand ≥ 600 KVA), was consider for the study. It has estimated that the annual electricity sale to commercial sector is 3.62 Billion Units (BU) in alone Karnataka State, India, which is an account for 9.57 % of the total electricity sold. The commercial sector constitutes Government & private establishments, hospitals, hotels, restaurants, educational institutions, malls etc. Total 326 commercial buildings in the state accounting for annual energy consumption of 1295.72 Million Units (MU) which works out to about 35% of the sectoral consumption. The annual energy savings potential for 326 commercial buildings is assessed to be 0.25 BU.

Keywords: commercial buildings, connected load, energy conservation studies, energy savings, energy efficiency, energy conservation strategy, energy efficiency, thermal energy, HVAC system

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Authors: S. K. Tiwari, Dinesh Kumar Shukla, R. Chandra

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Friction stir welding is a solid state joining process. High strength aluminum alloys are widely used in aircraft and marine industries. Generally, the mechanical properties of fusion-welded aluminum joints are poor. As friction stir welding occurs in the solid state, no solidification structures are created thereby eliminating the brittle and eutectic phases common in fusion welding of high strength aluminum alloys. In this review, the process parameters, microstructural evolution and effect of friction stir welding on the properties of weld specific to aluminum alloys have been discussed.

Keywords: aluminum alloys, friction stir welding (FSW), microstructure, Properties.

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Authors: Ashish Shukla, K. P. Mishra, Pushplata Tripathi

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This paper investigates the production and optimization of β-galactosidase enzyme using synthetic medium by isolated wild strains (S1, S2) mutated strains (M1, M2) through SSF and SmF. Among the different cell disintegration methods used, the highest specific activity was obtained when the cells were permeabilized using isoamyl alcohol. Wet lab experiments were performed to investigate the effects of carbon and nitrogen substrates present in Vogel’s medium on β-galactosidase enzyme activity using S1, S2, and M1, M2 strains through SSF. SmF experiments were performed for effects of carbon and nitrogen sources in YLK2Mg medium on β-galactosidase enzyme activity using S1, S2 and M1, M2 strains. Effect of pH on β-galactosidase enzyme production was also done using S1, S2, and M1, M2 strains. Results were found to be very appreciable in all the cases.

Keywords: β-galactosidase, cell disintegration, permeabilized, SSF, SmF

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Authors: Shivendra Pratap Singh, Rishabh Sharma

Abstract:

The widespread adoption of mobile technology in recent years has revolutionized the way we communicate and access information. The evolution of mobile systems has been rapid and impactful, shaping our lives and changing the way we live and work. However, despite its significant influence, the history and development of mobile technology are not well understood by the general public. This research paper aims to examine the history, technology and future of mobile systems, exploring their evolution from early mobile phones to the latest smartphones and beyond. The study will analyze the technological advancements and innovations that have shaped the mobile industry, from the introduction of mobile internet and multimedia capabilities to the integration of artificial intelligence and 5G networks. Additionally, the paper will also address the challenges and opportunities facing the future of mobile technology, such as privacy concerns, battery life, and the increasing demand for high-speed internet. Finally, the paper will also provide insights into potential future developments and innovations in the mobile sector, such as foldable phones, wearable technology, and the Internet of Things (IoT). The purpose of this research paper is to provide a comprehensive overview of the history, technology, and future of mobile systems, shedding light on their impact on society and the challenges and opportunities that lie ahead.

Keywords: mobile technology, artificial intelligence, networking, iot, technological advancements, smartphones

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Authors: P. Shrivastava, A. Shukla, K. Verma, S. Rungta

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Parkinson's disease is a chronic neurological disorder that directly affects human gait. It leads to slowness of movement, causes muscle rigidity and tremors. Gait serve as a primary outcome measure for studies aiming at early recognition of disease. Using gait techniques, this paper implements efficient binary bat algorithm for an early detection of Parkinson's disease by selecting optimal features required for classification of affected patients from others. The data of 166 people, both fit and affected is collected and optimal feature selection is done using PSO and Bat algorithm. The reduced dataset is then classified using neural network. The experiments indicate that binary bat algorithm outperforms traditional PSO and genetic algorithm and gives a fairly good recognition rate even with the reduced dataset.

Keywords: parkinson, gait, feature selection, bat algorithm

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Authors: Reena Behal, D. P. Shukla

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In this paper, we investigate Bulk Viscous Bianchi Type V Cosmological Model with Time dependent gravitational constant and cosmological constant in general Relativity by assuming ξ(t)=ξ_(0 ) p^m where ξ_(0 ) and m are constants. We also assume a variation law for Hubble parameter as H(R) = a (R^(-n)+1), where a>0, n>1 being constant. Two universe models were obtained, and their physical behavior has been discussed. When n=1 the Universe starts from singular state whereas when n=0 the cosmology follows a no singular state. The presence of bulk viscosity increase matter density’s value.

Keywords: Bulk Viscous Bianchi Type V Cosmological Model, hubble constants, gravitational constant, cosmological constants

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Authors: Umesh Kumar, Arun Kumar Shukla, B. V. V. Ravindra Babu

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The paper presents a compact tri band Curved U-Slot patch antenna with improved bandwidth and isolation characteristics. The proposed antenna excited by coaxial feed resonates at tri band of 2.8 GHz, 4.1 GHz and 5.7 GHz for VSWR ≤ 1.5 with an improved bandwidth of 99.7% and also for getting high gain antenna of 11.31 dB. A 2×2 MIMO is developed using the proposed antenna giving an excellent isolation of 28 dB between the two antennas. The simulation results of return loss, Mutual Coupling, Gain, VSWR, Surface Current Distribution and Electrical Distribution are presented. By keeping the substrate thickness constant over various dielectric constants, simulations were carried out using MATLAB® and HFSS (High Frequency Structure Simulator) software.

Keywords: performance analysis, curved U-slot patch, antenna with enhanced bandwidth, isolation for mimo systems

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Authors: Swetha Priya Darshini Thammadi, Sateesh Kumar Pisini, Sanjay Kumar Shukla

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Source apportionment using Dispersion model depends primarily on the quality of Emission Inventory. In the present study, a CMB receptor model has been used to identify the sources of PM_2.5, while the AERMOD dispersion model has been used to account for missing sources of PM_2.5 in the Emission Inventory. A statistical approach has been developed to quantify the missing sources not considered in the Emission Inventory. The inventory of each grid was improved by adjusting emissions based on road lengths and deficit in measured and modelled concentrations. The results showed that in CMB analyses, fugitive sources - soil and road dust - contribute significantly to ambient PM_2.5 pollution. As a result, AERMOD significantly underestimated the ambient air concentration at most locations. The revised Emission Inventory showed a significant improvement in AERMOD performance which is evident through statistical tests.

Keywords: CMB, GIS, AERMOD, PM₂.₅, fugitive, emission inventory

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Authors: Rishabh Pandey, Umang Kumar Yadav

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In today’s era of technological advancement, we come across incalculable innovations, almost every day. No doubt that the society has developed a lot in learning and technology, but we should also take into account the problems and inflictions that are occurring. Focusing on the petroleum sector a trending global concern is toward lowering fuel consumption and emissions. It is well known that gasoline is non-renewable source of energy and its burning produces harmful emissions which are adversely affecting the environment, such issues are motivating us to seek alternative solutions that would not require much modification in engine design and help us come out with an outcome. Keeping in mind the importance of environment and human race, we present a factious idea of use of oxyhydrogen gas or HHO gas in place of gasoline in the vehicles and petroleum industry. This technology is prospering, highly efficient, could be used economically and safe, and it will be responsible for changing the future of oil and gas sector in accordance with protection to the environment. In the coming future, we will check the compatibility of HHO generator with fuel engine for production of oxyhydrogen gas with use of water and effect of introducing HHO gas to the combustion on both thermal efficiency and specific fuel consumption. We will also work on the comparison of HHO gas and commercially available gasoline fuel in support of their chemical structures; ignition rate; octane rating; knocking properties; storage; transportation and cost effectiveness and it is trusted that use of HHO gas will be ecofriendly as no harmful emissions are produced, rather the only emission is water. Additionally, this paper will include the use of HHO cell in fuel engines and challenges faced in installing it in the current period and provide effective solutions for the same.

Keywords: fuel, gas, generator, water

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Authors: Mohammed Affanuddin H. Siddique, Jayesh S. Shukla, Chetan B. Meshram

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The neural networks are one of the power tools of machine learning. After the invention of perceptron in early 1980's, the neural networks and its application have grown rapidly. Neural networks are a technique originally developed for pattern investigation. The structure of a neural network consists of neurons connected through synapse. Here, we have investigated the different algorithms and cost function reduction techniques for optimization of vertical axis wind turbine (VAWT) rotor blades. The aerodynamic force coefficients corresponding to the airfoils are stored in a database along with the airfoil coordinates. A forward propagation neural network is created with the input as aerodynamic coefficients and output as the airfoil co-ordinates. In the proposed algorithm, the hidden layer is incorporated into cost function having linear and non-linear error terms. In this article, it is observed that the ANNs (Artificial Neural Network) can be used for the VAWT’s optimization.

Keywords: VAWT, ANN, optimization, inverse design

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Authors: Saurabh Shukla, G. N. Pandey

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The fires have become the most serious disasters to forest resources and the human environment. In recent years, due to climate change, human activities and other factors the frequency of forest fires has increased considerably. The monitoring and prevention of forest fires have now become a global concern for forest fire prevention organizations. Currently, the methods for forest fire prevention largely consist of patrols, observation from watch towers. Thus, software like deployment of the wireless sensor network to prevent forest fire is being developed to get a better estimate of the temperature and humidity prospects. Now days, wireless sensor networks are beginning to be deployed at an accelerated pace. It is not unrealistic to expect that in coming years the world will be covered with wireless sensor networks. This new technology has lots of unlimited potentials and can be used for numerous application areas including environmental, medical, military, transportation, entertainment, crisis management, homeland defense, and smart spaces.

Keywords: deployment, sensors, wireless sensor networks, forest fires

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Authors: Ankita Shukla, Tiratha Raj Singh

Abstract:

Colorectal cancer (CRC) imposes serious mortality burden worldwide and it has been increasing for past consecutive years. Continuous efforts have been made so far to diagnose the disease condition and to identify the root cause for it. In this study, we performed the pathway level as well as the differential gene expression studies for CRC. We analyzed the gene expression profile GSE24514 from Gene Expression Omnibus (GEO) along with the gene pathways involved in the CRC. This analysis helps us to understand the behavior of the genes that have shown differential expression through their targeted pathways. Pathway analysis for the targeted genes covers the wider area which therefore decreases the possibility to miss the significant ones. This will prove to be beneficial to expose the ones that have not been given attention so far. Through this analysis, we attempt to understand the various neighboring genes that have close relationship to the targeted one and thus proved to be significantly controlling the CRC. It is anticipated that the identified hub and neighboring genes will provide new directions to look at the pathway level differently and will be crucial for the regulatory processes of the disease.

Keywords: mismatch repair, microsatellite instability, carcinogenesis, morbidity

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Authors: Lubna Azmi, Ila Shukla, Shyam Sundar Gupta, Padam Kant, C. V. Rao

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Elvitegravir is the mainstay of anti-HIV combination therapy in most endemic countries presently. However, it cannot be used alone owing to its long onset time of action. 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxychromen-4-one (Quercetin: QU) is a polyphenolic compound obtained from Argeria speciosa Linn (Family: Convolvulaceae), an anti-HIV candidate. In the present study, a sensitive, simple and rapid high-performance liquid chromatography coupled with positive ion electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method was developed for the simultaneous determination elvitegravir and Quercetin, in rat plasma. The method was linear over a range of 0.2–500 ng/ml. All validation parameters met the acceptance criteria according to regulatory guidelines. LC–MS/MS method for determination of Elvitegravir and Quercetin was developed and validated. Results show the potential of drug–drug interaction upon co-administration this marketed drugs and plant derived secondary metabolite.

Keywords: anti-HIV resistance, extraction, HPLC-ESI-MS-MS, validation

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Authors: Taru Singh, Shukla Das, V. G. Ramachandran

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Objectives: To develop SYBR green based real-time PCR assay to detect carbapenemases (NDM, IMP) genes in E. coli. Methods: A total of 40 E. coli from stool samples were tested. Six were previously characterized as resistant to carbapenems and documented by PCR. The remaining 34 isolates previously tested susceptible to carbapenems and were negative for these genes. Bacterial RNA was extracted using manual method. The real-time PCR was performed using the Light Cycler III 480 instrument (Roche) and specific primers for each carbapenemase target were used. Results: Each one of the two carbapenemase gene tested presented a different melting curve after PCR amplification. The melting temperature (Tm) analysis of the amplicons identified was as follows: blaIMP type (Tm 82.18°C), blaNDM-1 (Tm 78.8°C). No amplification was detected among the negative samples. The results showed 100% concordance with the genotypes previously identified. Conclusions: The new assay was able to detect the presence of two different carbapenemase gene type by real-time PCR.

Keywords: resistance, b-lactamases, E. coli, real-time PCR

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