Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 20

Search results for: Sapan Bhatnagar

20 Determination of Air Quality Index Using Respirable Dust Sampler

Authors: Asif Ekbal, Salman Ahmed, Sapan Bhatnagar, Danish Akhtar, Gufran Beig

Abstract:

Particulates are the solid and liquid droplets present in the atmosphere, they have serious negative effects on human health and environment. PM10 and PM2.5 are so small that they can penetrate deep into our lungs through the respiratory system. Determination of the amount of particulates present in the atmosphere per cubic meter is necessary to monitor, regulate and model atmospheric particulate levels. Air Quality Index is an index tells us how clean or polluted our air is, and what associated health effects might be a concern for us. The AQI focuses on health affects you may experience within a few hours or days after breathing polluted air. The quality rating for each pollutant was calculated. The geometric mean of these quality ratings gives the Air Quality Index. The existing concentrations of pollutants were compared with ambient air quality standards.

Keywords: air quality index, particulate, respirable dust sampler, dust sampler

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19 An Investigation of Sustainability: Scope of Eco Denim Fashion

Authors: Sneha Bhatnagar, Sachin Bhatnagar

Abstract:

Denim presently is the most widely accepted textile product and shows its hold even in future with its growing popularity. Denim today is no longer restricted to only a pair of jeans but has diversified in all different product categories. Although denim is considered as an expression of youth and demonstrates durability and comfort, denim raises issues of sustainability. Through an exploratory research, the researcher aims at addressing the possibilities of denim fashion promoting environmental sustainability by means of creativity, awareness, recycle and artisan appreciation. It also touches on how eco conscious fashion brands involve in development in terms of ideation and modification of denim as a fabric or product into diversified sustainable fashion. In conclusion, it is shown that blue denim fashion continues to evolve and shows eventual transformation in becoming green denim in future, nurturing values of both quality and sustainability.

Keywords: Arts, Sustainability, Creativity, Fashion, recycle, denim, craft

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18 Characterization of Atmospheric Aerosols by Developing a Cascade Impactor

Authors: Sapan Bhatnagar

Abstract:

Micron size particles emitted from different sources and produced by combustion have serious negative effects on human health and environment. They can penetrate deep into our lungs through the respiratory system. Determination of the amount of particulates present in the atmosphere per cubic meter is necessary to monitor, regulate and model atmospheric particulate levels. Cascade impactor is used to collect the atmospheric particulates and by gravimetric analysis, their concentration in the atmosphere of different size ranges can be determined. Cascade impactors have been used for the classification of particles by aerodynamic size. They operate on the principle of inertial impaction. It consists of a number of stages each having an impaction plate and a nozzle. Collection plates are connected in series with smaller and smaller cutoff diameter. Air stream passes through the nozzle and the plates. Particles in the stream having large enough inertia impact upon the plate and smaller particles pass onto the next stage. By designing each successive stage with higher air stream velocity in the nozzle, smaller diameter particles will be collected at each stage. Particles too small to be impacted on the last collection plate will be collected on a backup filter. Impactor consists of 4 stages each made of steel, having its cut-off diameters less than 10 microns. Each stage is having collection plates, soaked with oil to prevent bounce and allows the impactor to function at high mass concentrations. Even after the plate is coated with particles, the incoming particle will still have a wet surface which significantly reduces particle bounce. The particles that are too small to be impacted on the last collection plate are then collected on a backup filter (microglass fiber filter), fibers provide larger surface area to which particles may adhere and voids in filter media aid in reducing particle re-entrainment.

Keywords: Environment, particulates, aerodynamic diameter, cascade, re-entrainment

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17 Electronic Structure and Optical Properties of YNi₄Si-Type GdNi₅: A Coulomb Corrected Local-Spin Density Approximation Study

Authors: Sapan Mohan Saini

Abstract:

In this work, we report the calculations on the electronic and optical properties of YNi₄Si-type GdNi₅ compound. Calculations are performed using the full-potential augmented plane wave (FPLAPW) method in the framework of density functional theory (DFT). The Coulomb corrected local-spin density approximation (LSDA+U) in the self-interaction correction (SIC) has been used for exchange-correlation potential. Spin polarised calculations of band structure show that several bands cross the Fermi level (EF) reflect the metallic character. Analysis of density of states (DOS) demonstrates that spin up Gd-f states lie around 7.5 eV below EF and spin down Gd-f lie around 4.5 eV above EF. We found Ni-3d states mainly contribute to DOS from -5.0 eV to the EF. Our calculated results of optical conductivity agree well with the experimental data.

Keywords: Electronic Structure, Optical Properties, FPLAPW method, YNi₄Si-type GdNi₅

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16 Unified Gas-Kinetic Scheme for Gas-Particle Flow in Shock-Induced Fluidization of Particles Bed

Authors: Zhao Wang, Hong Yan

Abstract:

In this paper, a unified-gas kinetic scheme (UGKS) for the gas-particle flow is constructed. UGKS is a direct modeling method for both continuum and rarefied flow computations. The dynamics of particle and gas are described as rarefied and continuum flow, respectively. Therefore, we use the Bhatnagar-Gross-Krook (BGK) equation for the particle distribution function. For the gas phase, the gas kinetic scheme for Navier-Stokes equation is solved. The momentum transfer between gas and particle is achieved by the acceleration term added to the BGK equation. The new scheme is tested by a 2cm-in-thickness dense bed comprised of glass particles with 1.5mm in diameter, and reasonable agreement is achieved.

Keywords: gas-particle flow, unified gas-kinetic scheme, momentum transfer, shock-induced fluidization

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15 Hydrogen Storage in Carbonized Coconut Meat (Kernel)

Authors: Viney Dixit, Rohit R. Shahi, Ashish Bhatnagar, P. Jain, T. P. Yadav, O. N. Srivastava

Abstract:

Carbons are being widely investigated as hydrogen storage material owing to their light weight, fast hydrogen absorption kinetics and low cost. However, these materials suffer from low hydrogen storage capacity at room temperature. The aim of the present study is to synthesize carbon based material which shows moderate hydrogen storage at room temperature. For this purpose, hydrogenation characteristics of natural precursor coconut kernel is studied in this work. The hydrogen storage measurement reveals that the as-synthesized materials have good hydrogen adsorption and desorption capacity with fast kinetics. The synthesized material absorbs 8 wt.% of hydrogen at liquid nitrogen temperature and 2.3 wt.% at room temperature. This could be due to the presence of certain elements (KCl, Mg, Ca) which are confirmed by TEM.

Keywords: hydrogenation, carbonization, coconut kernel, KCl

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14 Comparative Assessment of Bus Rapid Transit System in India

Authors: Sapan Tiwari, Namrata Ghosh

Abstract:

Public transport service plays an important role in people's transportation needs in urban areas. Bus Rapid Transit System (BRTS) is a transport service that provides passengers with a quick and efficient mode of transport. It is developed by changing the existing infrastructure, vehicles, route, or by developing a new dedicated corridor for the bus route. This dedicated lanes transport passengers to their destination quickly and efficiently and flexible in meeting demand. However, with rapid urbanization and increasing population density in Indian cities, traffic congestion has become a significant issue. In a few Indian cities, the BRTS concept is implemented to address the issue of traffic congestion that eventually resulted in less road congestion. The research aims to provide a literature review on the overall outlook of the BRTS system and its practical implementation in mass urban transit. First, it reflects a literature review on the concept of the BRTS system in both developed and developing countries. Afterward, comparative analysis of BRTS, hindrances associated with the permanent integrated system, and the need for establishing the Bus Rapid Transit System in Indian cities is demonstrated. The research concludes with some recommendations that could help in improving the loopholes in the existing system.

Keywords: public transport, Traffic Congestion, bus rapid transit system(BRTS), dedicated corridor

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13 Trend Detection Using Community Rank and Hawkes Process

Authors: W. Wilfred Godfrey, Shashank Bhatnagar

Abstract:

We develop in this paper, an approach to find the trendy topic, which not only considers the user-topic interaction but also considers the community, in which user belongs. This method modifies the previous approach of user-topic interaction to user-community-topic interaction with better speed-up in the range of [1.1-3]. We assume that trend detection in a social network is dependent on two things. The one is, broadcast of messages in social network governed by self-exciting point process, namely called Hawkes process and the second is, Community Rank. The influencer node links to others in the community and decides the community rank based on its PageRank and the number of users links to that community. The community rank decides the influence of one community over the other. Hence, the Hawkes process with the kernel of user-community-topic decides the trendy topic disseminated into the social network.

Keywords: community detection, Trend Detection, community rank, Hawkes process, influencer node, pagerank

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12 Electronic and Optical Properties of YNi4Si-Type DyNi4Si Compound: A Full Potential Study

Authors: Sapan Mohan Saini, Dinesh Kumar Maurya

Abstract:

A theoretical formalism to calculate the structural, electronic and optical properties of orthorhombic crystals from first principle calculations is described. This is applied first time to new YNi4Si-type DyNi4Si compound. Calculations are performed using full-potential augmented plane wave (FPLAPW) method in the framework of density functional theory (DFT). The Coulomb corrected local-spin density approximation (LSDA+U) in the self-interaction correction (SIC) has been used for exchange-correlation potential. Our optimized results of lattice parameters show good agreement to the previously reported experimental study. Analysis of the calculated band structure of DyNi4Si compound demonstrates their metallic character. We found Ni-3d states mainly contribute to density of states from -5.0 eV to the Fermi level while the Dy-f states peak stands tall in comparison to the small contributions made by the Ni-d and R-d states above Fermi level, which is consistent with experiment, in DNi4Si compound. Our calculated optical conductivity compares well with the experimental data and the results are analyzed in the light of band-to-band transitions. We also report the frequency-dependent refractive index n(ω) and the extinction coefficient k(ω) of the compound.

Keywords: Optical Properties, density of states, band structure, LSDA+U approximation, YNi4Si- type DyNi4Si compound

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11 Surface-Quenching Induced Cell Opening Technique in Extrusion of Thermoplastic Foamed Sheets

Authors: Abhishek Gandhi, Naresh Bhatnagar

Abstract:

In this article, a new technique has been developed to manufacture open cell extruded thermoplastic foamed sheets with the aid of extrudate surface-quenching phenomenon. As the extrudate foam exits the die, its surface is rapidly quenched which results in freezing of cells on the surface, while the cells at the core continue to grow and leads to development of open-cellular microstructure at the core. Influence of chill roll temperature was found to be extremely significant in developing porous morphological attributes. Subsequently, synergistic effect of blowing agent content and chill roll temperature was examined for their expansion ratio and open-cell microstructure. Further, chill roll rotating speed was found extremely significant in obtaining open-cellular foam structures. This study intends to enhance the understanding of researchers working in the area of open-cell foam processing.

Keywords: Morphology, Microscopy, Composite, foams, Porous Materials, open-cell foams

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10 Vehicle Maneuverability on Horizontal Curves on Hilly Terrain: A Study on Shillong Highway

Authors: Sapan Tiwari, Surendra Choudhary

Abstract:

The driver has two fundamental duties i) controlling the position of the vehicle along the longitudinal and lateral direction of movement ii) roadway width. Both of these duties are interdependent and are concurrently referred to as two-dimensional driver behavior. One of the main problems facing driver behavior modeling is to identify the parameters for describing the exemplary driving conduct and car maneuver under distinct traffic circumstances. Still, to date, there is no well-accepted theory that can comprehensively model the 2-D driver conduct (longitudinal and lateral). The primary objective of this research is to explore the vehicle's lateral longitudinal behavior in the heterogeneous condition of traffic on horizontal curves as well as the effect of road geometry on dynamic traffic parameters, i.e., car velocity and lateral placement. In this research, with their interrelationship, a thorough assessment of dynamic car parameters, i.e., speed, lateral acceleration, and turn radius. Also, horizontal curve road parameters, i.e., curvature radius, pavement friction, are performed. The dynamic parameters of the various types of car drivers are gathered using a VBOX GPS-based tool with high precision. The connection between dynamic car parameters and curve geometry is created after the removal of noise from the GPS trajectories. The major findings of the research are that car maneuvers with higher than the design limits of speed, acceleration, and lateral deviation on the studied curves of the highway. It can become lethal if the weather changes from dry to wet.

Keywords: Geometry, Terrain, maneuverability, trajectory, VBOX

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9 Sinusoidal Roughness Elements in a Square Cavity

Authors: Muhammad Yousaf, Shoaib Usman

Abstract:

Numerical studies were conducted using Lattice Boltzmann Method (LBM) to study the natural convection in a square cavity in the presence of roughness. An algorithm basedon a single relaxation time Bhatnagar-Gross-Krook (BGK) model of Lattice Boltzmann Method (LBM) was developed. Roughness was introduced on both the hot and cold walls in the form of sinusoidal roughness elements. The study was conducted for a Newtonian fluid of Prandtl number (Pr) 1.0. The range of Ra number was explored from 103 to 106 in a laminar region. Thermal and hydrodynamic behavior of fluid was analyzed using a differentially heated square cavity with roughness elements present on both the hot and cold wall. Neumann boundary conditions were introduced on horizontal walls with vertical walls as isothermal. The roughness elements were at the same boundary condition as corresponding walls. Computational algorithm was validated against previous benchmark studies performed with different numerical methods, and a good agreement was found to exist. Results indicate that the maximum reduction in the average heat transfer was16.66 percent at Ra number 105.

Keywords: natural convection, roughness, Rayleigh number, nusselt number, lattice Boltzmann method

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8 Review on Crew Scheduling of Bus Transit: A Case Study in Kolkata

Authors: Sapan Tiwari, Namrata Ghosh

Abstract:

In urban mass transit, crew scheduling always plays a significant role. It deals with the formulation of work timetables for its staff so that an organization can meet the demand for its products or services. The efficient schedules of a specified timetable have an enormous impact on staff demand. It implies that an urban mass transit company's financial outcomes are strongly associated with planning operations in the region. The research aims to demonstrate the state of the crew scheduling studies and its practical implementation in mass transit businesses in metropolitan areas. First, there is a short overview of past studies in the field. Subsequently, the restrictions and problems with crew scheduling and some models, which have been developed to solve the related issues with their mathematical formulation, are defined. The comments are completed by a description of the solution opportunities provided by computer-aided scheduling program systems for operational use and exposures from urban mass transit organizations. Furthermore, Bus scheduling is performed using the Hungarian technique of problem-solving tasks and mathematical modeling. Afterward, the crew scheduling problem, which consists of developing duties using predefined tasks with set start and end times and places, is resolved. Each duty has to comply with a set line of work. The objective is to minimize a mixture of fixed expenses (number of duties) and varying costs. After the optimization of cost, the outcome of the research is that the same frequency can be provided with fewer buses and less workforce.

Keywords: duty, crew scheduling, optimization of cost, urban mass transit

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7 Design Thinking and Creative Problem Solving for Undergraduate Engineering Education in India: Relevance and Student's Reactions

Authors: Tigmanshu Bhatnagar, Petra Badke-Schaub

Abstract:

Facilitating Design Thinking (DT) and Creative Problem Solving (CPS) in engineering education could benefit students by aiding them to think creatively and meaningfully in their education and future profession. A study in the pseudonym of a ‘popup class’ was conducted for a week at the Indian Institute of Technology, Delhi (IITD) to have an indication for the perceived relevance, benefits and challenges of DT and CPS from the perspective of engineering students in India. 30 third year Bachelor of Technology students from various technical fields participated in the study. They were introduced to the notion of DT and CPS via a mix of theoretical lectures, case discussions and practical workshops. Their reactions were identified on the basis of silent observations made during the course and responses were recorded through a questionnaire, which was filled after the course. All the respondents felt that DT and CPS are relevant to their education. It was perceived by them that there is a subtle improvement in the quality, quantity and approach of solutions to open ended problems. 90% responded positively to the induction of such an exercise in their education and reasoned it by stating that it’s important for engineers to know, how to solve open-ended real world problems in a meaningful and innovative way.

Keywords: Design Thinking, India, creative problem solving, undergraduate engineering education

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6 Novel Molecular Mechanisms Involved in Macrophage Phenotypic Polarization

Authors: Deepak Bhatnagar, Mansi Srivastava, Uzma Saqib, Adnan Naim, Anjali Roy, Dongfang Liu, Ravinder Ravinder, Mirza S. Baig

Abstract:

Macrophages polarize to proinflammatory M1 or anti-inflammatory M2 states with distinct physiological functions. This transition within the M1 to M2 phenotypes decides the nature, duration, and severity of an inflammatory response. However, inspite of a substantial understanding of the fate of these phenotypes, the underlying molecular mechanisms are not well understood. We have investigated the role of Neuronal nitric oxide synthase (NOS1) mediated regulation of Activator protein 1 (AP-1) transcription factor in macrophages as a critical effector of macrophage phenotypic change. Activator protein 1 (AP-1) is a group of dimeric transcription factors composed of jun, Fos, and ATF family proteins. We determined that NOS1-derived nitric oxide (NO) facilitate Fos and jun interaction which induces IL12 & IL23 expression. Pharmacological inhibition of NOS1 inhibits Fos and jun interaction but increases ATF2 and Fos dimerization. Switching of Fos and jun dimer to ATF2 and jun dimerization switches phenotype from IL–12high IL-23high IL-10low to IL–12low IL-23lowIL-10high phenotype, respectively. Together, these findings highlight a key role of the TLR4-NOS1-AP1 signaling axis in regulating macrophage polarization.

Keywords: Inflammation, macrophage, lipopolysaccharide (LPS), proinflammatory cytokines, activator protein 1 (AP-1), neuronal nitric oxide synthase (NOS1)

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5 Effect of Transit-Oriented Development on Air Quality in Neighborhoods of Delhi

Authors: Smriti Bhatnagar

Abstract:

This study aims to find if the Transit-oriented planning and development approach benefit the quality of air in neighborhoods of New Delhi. Two methodologies, namely the land use regression analysis and the Transit-oriented development index analysis, are being used to explore this relationship. Land Use Regression Analysis makes use of urban form characteristics as obtained for 33 neighborhoods in Delhi. These comprise road lengths, land use areas, population and household densities, number of amenities and distance between amenities. Regressions are run to establish the relationship between urban form variables and air quality parameters (dependent variables). For the Transit-oriented development index analysis, the Transit-oriented Development index is developed as a composite index comprising 29 urban form indicators. This index is developed by assigning weights to each of the 29 urban form data points. Regressions are run to establish the relationship between the Transit-oriented development index and air quality parameters. The thesis finds that elements of Transit-oriented development if incorporated in planning approach, have a positive effect on air quality. Roads suited for non-motorized transport, well connected civic amenities in neighbourhoods, for instance, have a directly proportional relationship with air quality. Transit-oriented development index, however, is not found to have a consistent relationship with air quality parameters. The reason could this, however, be in the way that the index has been constructed.

Keywords: Air quality, New Delhi, land use regression, mixed-use planning, transit-oriented development index

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4 Development of a Computer Based, Nutrition and Fitness Programme and Its Effect on Nutritional Status and Fitness of Obese Adults

Authors: Richa Soni, Vibha Bhatnagar, N. K. Jain

Abstract:

This study was conducted to develop a computer mediated programme for weight management and physical fitness and examining its efficacy in reducing weight and improving physical fitness in obese adults. A user friendly, computer based programme was developed to provide a simple, quick, easy and user-friendly method of assessing energy balance at individual level. The programme had four main sections viz. personal Profile, know about your weight, fitness and food exchange list. The computer programme was developed to provide facilities of creating individual profile, tracking meal and physical activities, suggesting nutritional and exercise requirements, planning calorie specific menus, keeping food diaries and revising the diet and exercise plans if needed. The programme was also providing information on obesity, underweight, physical fitness. An exhaustive food exchange list was also given in the programme to assist user to make right food choice decisions. The developed programme was evaluated by a panel of 15 experts comprising endocrinologists, nutritionists and diet counselors. Suggestions given by the experts were paned down and the entire programme was modified in light of suggestions given by the panel members and was reevaluated by the same panel of experts. For assessing the impact of the programme 22 obese subjects were selected purposively and randomly assigned to intervention group (n=12) and no information control group. (n=10). The programme group was asked to strictly follow the programme for one month. Significant reduction in the intake of energy, fat and carbohydrates was observed while intake of fruits, green leafy vegetables was increased. The programme was also found to be effective in reducing body weight, body fat percent and body fat mass whereas total body water and physical fitness scores improved significantly. There was no significant alteration observed in any parameters in the control group.

Keywords: Body Composition, Physical Fitness, body weight, computer programme

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3 Application of Lattice Boltzmann Method to Different Boundary Conditions in a Two Dimensional Enclosure

Authors: Jean Yves Trepanier, Sami Ammar, Sagnik Banik

Abstract:

Lattice Boltzmann Method has been advantageous in simulating complex boundary conditions and solving for fluid flow parameters by streaming and collision processes. This paper includes the study of three different test cases in a confined domain using the method of the Lattice Boltzmann model. 1. An SRT (Single Relaxation Time) approach in the Lattice Boltzmann model is used to simulate Lid Driven Cavity flow for different Reynolds Number (100, 400 and 1000) with a domain aspect ratio of 1, i.e., square cavity. A moment-based boundary condition is used for more accurate results. 2. A Thermal Lattice BGK (Bhatnagar-Gross-Krook) Model is developed for the Rayleigh Benard convection for both test cases - Horizontal and Vertical Temperature difference, considered separately for a Boussinesq incompressible fluid. The Rayleigh number is varied for both the test cases (10^3 ≤ Ra ≤ 10^6) keeping the Prandtl number at 0.71. A stability criteria with a precise forcing scheme is used for a greater level of accuracy. 3. The phase change problem governed by the heat-conduction equation is studied using the enthalpy based Lattice Boltzmann Model with a single iteration for each time step, thus reducing the computational time. A double distribution function approach with D2Q9 (density) model and D2Q5 (temperature) model are used for two different test cases-the conduction dominated melting and the convection dominated melting. The solidification process is also simulated using the enthalpy based method with a single distribution function using the D2Q5 model to provide a better understanding of the heat transport phenomenon. The domain for the test cases has an aspect ratio of 2 with some exceptions for a square cavity. An approximate velocity scale is chosen to ensure that the simulations are within the incompressible regime. Different parameters like velocities, temperature, Nusselt number, etc. are calculated for a comparative study with the existing works of literature. The simulated results demonstrate excellent agreement with the existing benchmark solution within an error limit of ± 0.05 implicates the viability of this method for complex fluid flow problems.

Keywords: nusselt, SRT, Prandtl, BGK, Rayleigh

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2 Severe Infestation of Laspeyresia Koenigana Fab. and Alternaria Leaf Spot on Azadirachta Indica (Neem)

Authors: Sangeeta Singh, Shiwani Bhatnagar, K. K. Srivastava, Ameen Ullah Khan, Bundesh Kumar, Lokendra Singh Rathore

Abstract:

From the instigation of the world medicinal plants are treated as part and parcel of human society to fight against diseases. Azadirachta indica (Neem) a herbal plant has been used as an Indian traditional medicine since ages and its products are acknowledged to solve agricultural, forestry and public health related problems, owing to its beneficial medicinal properties. Each part of the neem tree is known for its medicinal property. Bark & leaf extracts of neem have been used to control leprosy, respiratory disorders, constipation and also as blood purifier and a general health tonic. Neem is still regarded as ' rural community dispensary' in India or a tree for solving medical problems. Use of Neem as pesticides for the management of insect pest of agriculture crops and forestry has been seen as a shift in the use of synthetic pesticides to ecofriendly botanicals. Neem oil and seed extracts possess germicidal and anti-bacterial properties which when sprayed on the plant helps in protecting them from foliage pests. Azadirachtin, the main active ingredient found in neem tree, acts as an insect repellent and antifeedant. However the young plants are susceptible to many insect pest and foliar diseases. Recently, in the avenue plantation, planted by Arid Forest Research Institute, Jodhpur, around the premises of IIT Jodhpur, two years old neem plants were found to be severely infested with tip borer Laspeyresia koenigana (Family: Eucosmidae). The adult moth of L. koenigana lays eggs on the tender shoots and the young larvae tunnel into the shoot and feed inside. A small pinhole can be seen at the entrance point, from where the larva enters in to the stem. The severely attached apical shoots exhibit profuse gum exudation resulting in development of a callus structure. The internal feeding causes the stem to wilt and the leaves to dry up from the tips resulting in growth retardation. Alternaria Leaf spot and blight symptoms were also recorded on these neem plants. For the management of tip borer and Alternaria Leaf spot, foliar spray of monocrotophos @0.05% and Dithane M-45 @ 0.15% and powermin @ 2ml/lit were found efficient in managing the insect pest and foliar disease problem. No Further incidence of pest/diseases was noticed.

Keywords: Management, azadirachta indica, alternaria leaf spot, laspeyresia koenigana

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1 Capability of a Single Antigen to Induce Both Protective and Disease Enhancing Antibody: An Obstacle in the Creation of Vaccines and Passive Immunotherapies

Authors: Subrata Sinha, Parul Kulshreshtha, Rakesh Bhatnagar

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This study was conducted by taking B. anthracis as a model pathogen. On infecting a host, B. anthracis secretes three proteins, namely, protective antigen (PA, 83kDa), edema factor (EF, 89 kDa) and lethal factor (LF, 90 kDa). These three proteins are the components of two anthrax toxins. PA binds to the cell surface receptors, namely, tumor endothelial marker (TEM) 8 and capillary morphogenesis protein (CMG) 2. TEM8 and CMG2 interact with LDL-receptor related protein (LRP) 6 for endocytosis of EF and LF. On entering the cell, EF acts as a calmodulin-dependent adenylate cyclase that causes a prolonged increase of cytosolic cyclic adenosine monophosphate (cAMP). LF is a metalloprotease that cleaves most isoforms of mitogen-activated protein kinase kinases (MAPKK/MEK) close to their N-terminus. By secreting these two toxins, B.anthracis ascertains death of the host. Once the systemic levels of the toxins rise, antibiotics alone cannot save the host. Therefore, toxin-specific inhibitors have to be developed. In this wake, monoclonal antibodies have been developed for the neutralization of toxic effects of anthrax toxins. We created hybridomas by using spleen of mice that were actively immunized with rLFn (recombinant N-terminal domain of lethal factor of B. anthracis) to obtain anti-toxin antibodies. Later on, separate group of mice were immunized with rLFn to obtain a polyclonal control for passive immunization studies of monoclonal antibodies. This led to the identification of one cohort of rLFn-immunized mice that harboured disease-enhancing polyclonal antibodies. At the same time, the monoclonal antibodies from all the hybridomas were being tested. Two hybridomas secreted monoclonal antibodies (H8 and H10) that were cross-reactive with EF (edema factor) and LF (lethal factor), while the other two hybridomas secreted LF-specific antibodies (H7 and H11). The protective efficacy of H7, H8, H10 and H11 was investigated. H7, H8 and H10 were found to be protective. H11 was found to have disease enhancing characteristics in-vitro and in mouse model of challenge with B. anthracis. In this study the disease enhancing character of H11 monoclonal antibody and anti-rLFn polyclonal sera was investigated. Combination of H11 with protective monoclonal antibodies (H8 and H10) reduced its disease enhancing nature both in-vitro and in-vivo. But combination of H11 with LETscFv (an scFv with VH and VL identical to H10 but lacking Fc region) could not abrogate the disease-enhancing character of H11 mAb. Therefore it was concluded that for suppression of disease enhancement, Fc portion was absolutely essential for interaction of H10 with H11. Our study indicates that the protective potential of an antibody depends equally on its idiotype/ antigen specificity and its isotype. A number of monoclonal and engineered antibodies are being explored as immunotherapeutics but it is absolutely essential to characterize each one for their individual and combined protective potential. Although new in the sphere of toxin-based diseases, it is extremely important to characterize the disease-enhancing nature of polyclonal as well as monoclonal antibodies. This is because several anti-viral therapeutics and vaccines have failed in the face of this phenomenon. The passive –immunotherapy thus needs to be well formulated to avoid any contraindications.

Keywords: Immunotherapy, polyclonal, monoclonal, antibody-dependent disease enhancement

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