Search results for: Spandan Sahu

Authors: Sandeep Santosh, O. P. Sahu

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In this paper, performance comparison of wideband covariance matrix sparse representation (W-CMSR) method with other existing wideband Direction of Arrival (DOA) estimation methods has been made.W-CMSR relies less on a priori information of the incident signal number than the ordinary subspace based methods.Consider the perturbation free covariance matrix of the wideband array output. The diagonal covariance elements are contaminated by unknown noise variance. The covariance matrix of array output is conjugate symmetric i.e its upper right triangular elements can be represented by lower left triangular ones.As the main diagonal elements are contaminated by unknown noise variance,slide over them and align the lower left triangular elements column by column to obtain a measurement vector.Simulation results for W-CMSR are compared with simulation results of other wideband DOA estimation methods like Coherent signal subspace method (CSSM), Capon, l1-SVD, and JLZA-DOA. W-CMSR separate two signals very clearly and CSSM, Capon, L1-SVD and JLZA-DOA fail to separate two signals clearly and an amount of pseudo peaks exist in the spectrum of L1-SVD.

Keywords: W-CMSR, wideband direction of arrival (DOA), covariance matrix, electrical and computer engineering

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Authors: Lipsa Dash, Gyanendra Sahu

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Intelligence is the cornerstone of humans, and now they have created a counterpart of themselves artificially. Our understanding of the word intelligence is a very perspective based and mostly superior understanding of what we read, write, perceive and understand the adversities around better. A wide range of industrial sectors have also started involving the technology to perceive, reason and act. Similarly, intellectual property is the product of human intelligence and creativity. The World Intellectual Property Organisation is currently working on technology trends across the globe, and AI tops the list in the digital frontier that will have a profound impact on the world, transforming the way we live and work. Coming to Intellectual Property, patents and creations of the AI’s itself have constantly been in question. This paper explores whether AI’s can fit in the flexibilities of Trade Related Intellectual Property Studies and gaps in the existing IP laws or rthere is a need of amendment to include them in the ambit. The researcher also explores the right of AI’s who create things out of their intelligence and whether they could qualify to be legal persons making the other laws applicable on them. Differentiation between AI creations and human creations are explored in the paper, and the need of amendments to determine authorship, ownership, inventorship, protection, and identification of beneficiary for remuneration or even for determining liability. The humans and humanoids are all indulged in matters related to Privacy, and that attracts another constitutional legal issue to be addressed. The authors will be focusing on the legal conundrums of AI, transhumanism, and the Internet of things.

Keywords: artificial intelligence, humanoids, healthcare, privacy, legal conundrums, transhumanism

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Authors: Kritika Kulhari, Aparna Sahu

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Test anxiety results from concerns about failure in examinations or evaluative situations. Attentional biases are known to pronounce the symptomatic expression of test anxiety. In recent times, the inattentional blindness (IB) paradigm has shown promise as an attention bias modification treatment (ABMT) for anxiety by overcoming practice and expectancy effects which preexisting paradigms fail to counter. The IB paradigm assesses the inability of an individual to attend to a stimulus that appears suddenly while indulging in a perceptual discrimination task. The present study incorporated an IB task with three critical items (book, face, and triangle) appearing randomly in the perceptual discrimination task. Attentional biases were assessed as detection and identification of the critical item. The sample (N = 50) consisted of low test anxiety (LTA) and high test anxiety (HTA) groups based on the reactions to tests scale scores. Test threat manipulation was done with pre- and post-test assessment of test anxiety using the State Test Anxiety Inventory. A mixed factorial design with gender, test anxiety, presence or absence of test threat, and critical items was conducted to assess their effects on attentional biases. Results showed only a significant main effect for test anxiety on detection with higher accuracy of detection of the critical item for the LTA group. The study presents promising results in the realm of ABMT for test anxiety.

Keywords: attentional bias, attentional bias modification treatment, inattentional blindness, test anxiety

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Authors: Suresh Sahu, Abhijeet M. Vaidya, Naresh K. Maheshwari

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The efforts to understand the heat transfer behavior of supercritical water in supercritical water cooled reactor (SCWR) are ongoing worldwide to fulfill the future energy demand. The higher thermal efficiency of these reactors compared to a conventional nuclear reactor is one of the driving forces for attracting the attention of nuclear scientists. In this work, a solution procedure has been described for solving supercritical fluid flow problems in complex geometries. The solution procedure is based on non-staggered grid. All governing equations are discretized by finite volume method (FVM) in curvilinear coordinate system. Convective terms are discretized by first-order upwind scheme and central difference approximation has been used to discretize the diffusive parts. k-ε turbulence model with standard wall function has been employed. SIMPLE solution procedure has been implemented for the curvilinear coordinate system. Based on this solution method, 3-D Computational Fluid Dynamics (CFD) code has been developed. In order to demonstrate the capability of this CFD code in supercritical fluid flows, heat transfer to supercritical water in circular tubes has been considered as a test problem. Results obtained by code have been compared with experimental results reported in literature.

Keywords: curvilinear coordinate, body-fitted mesh, momentum interpolation, non-staggered grid, supercritical fluids

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Authors: Tripta Dixit, Smita Sahu, William Selvamurthy, Sadhana Srivastava

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The translation of healthcare technologies is an expensive, complex affair, current healthcare challenges in Asian countries and their efforts to meet Millennium Development Goals (MDGs), necessitates continuous technology advancement to save countless lives, improve the quality of life and for socio-economic development. India’s consistently improving global innovation index (57) demonstrates its innovation potential, but access to health care is asymmetric and lacks priority in India. Therefore, there is utmost need of a robust translation system for point-of-care (POC) solutions, inexpensive, low-maintenance, reliable, and easy-to-use diagnostic technologies. Few cases of POC technologies viz. Elisa based diagnostic kits for regional viral disease, a device for detection of cancerous lesions were studied to understand the process and challenges involved in their translation. Accordingly, the entire translation ecosystem was summarized proposing a nexus of various actors such as technology developer, technology transferor technology receiver, funding entities, government/regulatory bodies and their effect on translation of different medical technologies. This study highlights the role and concerns pertaining to these actors for POC such as unsystematic and unvalidated research roadmap, low profit preposition, unfocused approach of up-scaling, low market acceptability and multiple window regulatory framework, etc. This provides an opportunity to devise solutions to overcome problem areas in translation path.

Keywords: healthcare technologies, point-of-care solutions, public health, translation

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Authors: Neeraj Sahu, Ahmad Saadiq

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Anaerobic sulphate reduction has the potential for being effective and economically viable over conventional treatment methods for the treatment of sulphate-rich wastewater. However, a major challenge in anaerobic sulphate reduction is the diversion of a fraction of organic carbon towards methane production and some minor problem such as odour problems, corrosion, and increase of effluent chemical oxygen demand. A high-rate anaerobic technology has encouraged researchers to extend its application to the treatment of complex wastewaters with relatively low cost and energy consumption compared to physicochemical methods. Therefore, the aim of this study was to investigate the effects of COD/SO₄²⁻ ratio on the performance of lab scale UASB reactor. A lab-scale upflow anaerobic sludge blanket (UASB) reactor was operated for 170 days. In which first 60 days, for successful start-up with acclimation under methanogenesis and sulphidogenesis at COD/SO₄²⁻ of 18 and were operated at COD/SO₄²⁻ ratios of 12, 8, 4 and 1 to evaluate the effects of the presence of sulfate on the reactor performance. The reactor achieved maximum COD removal efficiency and biogas evolution at the end of acclimation (control). This phase lasted 53 days with 89.5% efficiency. The biogas was 0.6 L/d at (OLR) of 1.0 kg COD/m³d when it was treating synthetic wastewater with effective volume of reactor as 2.8 L. When COD/SO₄²⁻ ratio changed from 12 to 1, slight decrease in COD removal efficiencies (76.8–87.4%) was observed, biogas production decreased from 0.58 to 0.32 L/d, while the sulfate removal efficiency increased from 42.5% to 72.7%.

Keywords: anaerobic, chemical oxygen demand, organic loading rate, sulphate, up-flow anaerobic sludge blanket reactor

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Authors: Arpit K. Shrivastava, Nirmal K. Mohakud, Subrat Kumar, Priyadarshi S. Sahu

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Acute diarrhea is one of the major causes of morbidity and mortality among children less than five years of age. Multiple etiologies have been implicated for infectious gastroenteritis causing acute diarrhea. In our study fecal samples (n=165) were collected from children (<5 years) presenting with symptoms of acute diarrhea. Samples were screened for viral, bacterial, and parasitic etiologies such as Rotavirus, Adenovirus, Diarrhoeagenic Escherichia coli (EPEC, EHEC, STEC, O157, O111), Shigella spp., Salmonella spp., Vibrio cholera, Cryptosporidium spp., and Giardia spp. The overall results from our study showed that 57% of children below 5 years of age with acute diarrhea were positive for at least one infectious etiology. Diarrhoeagenic Escherichia coli was detected to be the major etiological agent (29.09%) followed by Rotavirus (24.24%), Shigella (21.21%), Adenovirus (5.45%), Cryptosporidium (2.42%), and Giardia (0.60%). Among the different DEC strains, EPEC was detected significantly higher in <2 years children in comparison to >2 years age group (p =0.001). Concurrent infections with two or more pathogens were observed in 47 of 160 (28.48%) cases with a predominant incidence particularly in <2-year-old children (66.66%) compared to children of 2 to 5 years age group. Co-infection of Rotavirus with Shigella was the most frequent combination, which was detected in 17.94% cases, followed by Rotavirus with EPEC (15.38%) and Shigella with STEC (12.82%). Detection of multiple infectious etiologies and diagnosis of the right causative agent(s) can immensely help in better management of acute childhood diarrhea. In future more studies focusing on the detection of cases with concurrent infections must be carried out, as we believe that the etiological agents might be complementing each other’s strategies of pathogenesis resulting in severe diarrhea.

Keywords: children, co-infection, infectious diarrhea, Odisha

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Authors: Atma Sahu

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After 9/11, there will only be cyberwars. The cyberwars increase in intensity the country's cybersecurity workforce's hiring and retention issues. Currently, many organizations have unfilled cybersecurity positions, and to a lesser degree, their cybersecurity teams are understaffed. Therefore, there is a critical need to develop a new program to help meet the market demand for cybersecurity engineers (CYSE) and personnel. Coppin State University in the United States was responsible for developing a cybersecurity engineering BS degree program. The CYSE curriculum design methodology consisted of three parts. First, the ACM Cross-Cutting Concepts standard's pervasive framework helped curriculum designers and students explore connections among the core courses' knowledge areas and reinforce the security mindset conveyed in them. Second, the core course context was created to assist students in resolving security issues in authentic cyber situations involving cyber security systems in various aspects of industrial work while adhering to the NIST standards framework. The last part of the CYSE curriculum design aspect was the institutional student learning outcomes (SLOs) integrated and aligned in content courses, representing more detailed outcomes and emphasizing what learners can do over merely what they know. The CYSE program's core courses express competencies and learning outcomes using action verbs from Bloom's Revised Taxonomy. This aspect of the CYSE BS degree program's design is based on these three pillars: the ACM, NIST, and SLO standards, which all CYSE curriculum designers should know. This unique CYSE curriculum design methodology will address how students and the CYSE program will be assessed and evaluated. It is also critical that educators, program managers, and students understand the importance of staying current in this fast-paced CYSE field.

Keywords: cyber security, cybersecurity engineering, systems engineering, NIST standards, physical systems

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Authors: Satpal Yadav, Ashish Phulkar, Krishna K. Sahu

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Aim: The research aims at doing a 3 Dimension biomechanical analysis in the shot put techniques of International throwers to evaluate the performance. Research Method: The researcher adopted the descriptive method and the data was subjected to calculate by using Pearson’s product moment correlation for the correlation of the biomechanical parameters with the performance of shot put throw. In all the analyses, the 5% critical level (p ≤ 0.05) was considered to indicate statistical significance. Research Sample: Eight (N=08) international shot putters using rotational/glide technique in male category was selected as subjects for the study. The researcher used the following methods and tools to obtain reliable measurements the instrument which was used for the purpose of present study namely the tesscorn slow-motion camera, specialized motion analyzer software, 7.260 kg Shot Put (for a male shot-putter) and steel tape. All measurement pertaining to the biomechanical variables was taken by the principal investigator so that data collected for the present study was considered reliable. Results: The finding of the study showed that negative significant relationship between the angular velocity right shoulder, acceleration distance at pre flight (-0.70), (-0.72) respectively were obtained, the angular displacement of knee, angular velocity right shoulder and acceleration distance at flight (0.81), (0.75) and (0.71) respectively were obtained, the angular velocity right shoulder and acceleration distance at transition phase (0.77), (0.79) respectively were obtained and angular displacement of knee, angular velocity right shoulder, release velocity shot, angle of release, height of release, projected distance and measured distance as the values (0.76), (0.77), (-0.83), (-0.79), (-0.77), (0.99) and (1.00) were found higher than the tabulated value at 0.05 level of significance. On the other hand, there exists an insignificant relationship between the performance of shot put and acceleration distance [m], angular displacement shot, C.G at release and horizontal release distance on the technique of shot put.

Keywords: biomechanics, analysis, shot put, international throwers

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Authors: Pabitra Sahu, M. Manohari, S. Priyadharshini, R. Santhanam, S. Chandrasekaran, B. Venkatraman

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Radiation workers of reprocessing plants have a potential for internal exposure due to actinides and fission products. Radionuclides like Americium, lead, Polonium and Europium are bone seekers and get accumulated in the skeletal part. As the major skeletal content is in the skull (13%) and knee (22%), measurements of old intake have to be carried out in the skull and knee. At the Indira Gandhi Centre for Atomic Research, a twin HPGe-based actinide monitor is used for the measurement of actinides present in bone. Efficiency estimation, which is one of the prerequisites for the quantification of radionuclides, requires anthropomorphic phantoms. Such phantoms are very limited. Hence, in this study, efficiency curves for a Twin HPGe-based actinide monitoring system are established theoretically using the FLUKA Monte Carlo method and ICRP adult male voxel phantom. In the case of skull measurement, the detector is placed over the forehead, and for knee measurement, one detector is placed over each knee. The efficiency values of radionuclides present in the knee and skull vary from 3.72E-04 to 4.19E-04 CPS/photon and 5.22E-04 to 7.07E-04 CPS/photon, respectively, for the energy range 17 to 3000keV. The efficiency curves for the measurement are established, and it is found that initially, the efficiency value increases up to 100 keV and then starts decreasing. It is found that the skull efficiency values are 4% to 63% higher than that of the knee, depending on the energy for all the energies except 17.74 keV. The reason is the closeness of the detector to the skull compared to the knee. But for 17.74 keV the efficiency of the knee is more than the skull due to the higher attenuation caused in the skull bones because of its greater thickness. The Minimum Detectable Activity (MDA) for 241Am present in the skull and knee is 9 Bq. 239Pu has a MDA of 950 Bq and 1270 Bq for knee and skull, respectively, for a counting time of 1800 sec. This paper discusses the simulation method and the results obtained in the study.

Keywords: FLUKA Monte Carlo Method, ICRP adult male voxel phantom, knee, Skull.

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Authors: Malay K. Das, Bhanu P. Sahu

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Furosemide is a weakly acidic diuretic indicated for treatment of edema and hypertension. It has very poor solubility but high permeability through stomach and upper gastrointestinal tract (GIT). Due to its limited solubility it has poor and variable oral bioavailability of 10-90%. The aim of this study was to enhance the oral bioavailability of furosemide by preparation of nanosuspensions. The nanosuspensions were prepared by nanoprecipitation with sonication using DMSO (dimethyl sulfoxide) as a solvent and water as an antisolvent (NA). The prepared nanosuspensions were sterically stabilized with polyvinyl acetate (PVA).These were characterized for particle size, ζ potential, polydispersity index, scanning electron microscopy (SEM), differential scanning calorimetry (DSC), X-ray diffraction (XRD) pattern and release behavior. The effect of nanoprecipitation on oral bioavailability of furosemide nanosuspension was studied by in vitro dissolution and in vivo absorption study in rats and compared to pure drug. The stable nanosuspension was obtained with average size range of the precipitated nanoparticles between 150-300 nm and was found to be homogenous showing a narrow polydispersity index of 0.3±0.1. DSC and XRD studies indicated that the crystalline furosemide drug was converted to amorphous form upon precipitation into nanoparticles. The release profiles of nanosuspension formulation showed up to 81.2% release in 4 h. The in vivo studies on rats revealed a significant increase in the oral absorption of furosemide in the nanosuspension compared to pure drug. The AUC0→24 and Cmax values of nanosuspension were approximately 1.38 and 1.68-fold greater than that of pure drug, respectively. Furosemide nanosuspension showed 20.06±0.02 % decrease in systolic blood pressure compared to 13.37±0.02 % in plain furosemide suspension, respectively. The improved oral bioavailability and pharmacodynamics effect of furosemide may be due to the improved dissolution of furosemide in simulated gastric fluid which results in enhanced oral systemic absorption of furosemide from stomach region where it has better permeability.

Keywords: furosemide, nanosuspension, bioavailability enhancement, nanoprecipitation, oral drug delivery

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Authors: Abhilash Sahu, Amit Priyadarshi

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In this paper, we will discuss the existence of weak solutions of the Kirchhoff type boundary value problem on the Sierpinski gasket. Where S denotes the Sierpinski gasket in R² and S₀ is the intrinsic boundary of the Sierpinski gasket. M: R → R is a positive function and h: S × R → R is a suitable function which is a part of our main equation. ∆p denotes the p-Laplacian, where p > 1. First of all, we will define a weak solution for our problem and then we will show the existence of at least two solutions for the above problem under suitable conditions. There is no well-known concept of a generalized derivative of a function on a fractal domain. Recently, the notion of differential operators such as the Laplacian and the p-Laplacian on fractal domains has been defined. We recall the result first then we will address the above problem. In view of literature, Laplacian and p-Laplacian equations are studied extensively on regular domains (open connected domains) in contrast to fractal domains. In fractal domains, people have studied Laplacian equations more than p-Laplacian probably because in that case, the corresponding function space is reflexive and many minimax theorems which work for regular domains is applicable there which is not the case for the p-Laplacian. This motivates us to study equations involving p-Laplacian on the Sierpinski gasket. Problems on fractal domains lead to nonlinear models such as reaction-diffusion equations on fractals, problems on elastic fractal media and fluid flow through fractal regions etc. We have studied the above p-Laplacian equations on the Sierpinski gasket using fibering map technique on the Nehari manifold. Many authors have studied the Laplacian and p-Laplacian equations on regular domains using this Nehari manifold technique. In general Euler functional associated with such a problem is Frechet or Gateaux differentiable. So, a critical point becomes a solution to the problem. Also, the function space they consider is reflexive and hence we can extract a weakly convergent subsequence from a bounded sequence. But in our case neither the Euler functional is differentiable nor the function space is known to be reflexive. Overcoming these issues we are still able to prove the existence of at least two solutions of the given equation.

Keywords: Euler functional, p-Laplacian, p-energy, Sierpinski gasket, weak solution

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Authors: S. K. Dewangan, K. R. Sahu, S. Soni

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Unemployment is today’s basic socio-economic problem eroding national income and living standards, aggravating national development and poverty alleviation. The farmers are encouraged to take up non-agriculture practices which are integrated with Sericulture. Sericulture is one of the primary occupations for livelihood of poor people in tribal area. Most of tribal are involved in Sericulture. Tasar, Eri are the main forest-based cultivation. Among these sericultures is the major crop adopted by the Tribal’s and practiced in respective areas. Out of the 6, 38,588 villages in India, sericultures are practiced in about 69000 villages providing employment to about 7.85 million people. Sericulture is providing livelihood for 9, 47,631 families. India continues to be the second largest producer of silk in the World. Among the four varieties of silk produced, as in 2012-13, Mulberry accounts for 18,715 MT, Eri 3116 MT, Tasar 1729 MT and Muga 119MT of the total raw silk production in the country. Sericulture with its unique features plays an important role in upgrading the socio-economic conditions of the rural folk and with employment opportunities to the educated rural youth and women. In view of the importance of sericulture enterprise for the biodiversity conservation as well as its cultural bondage, the paper tries to enlighten and discuss the significance of sericulture and strategies to be taken for the employment generation in Indian sericulture industry. The present paper explores the possible employment opportunities derived from problem analysis and strategies to be adopted aiming at revolutionary biodiversity conservation in the study area. The paper highlights the sericulture is a way for biodiversity conservation, employment generation in Raigarh district, their utilization and needs as they act as a tool for socio-economic change for tribal. The study concludes with some suggestions to improve the feasibility of sericulture in long term.

Keywords: bio-diversity, employment, sericulture, tribal, income, socio-economic

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Authors: Arun Kharate, Sarata Kumar Sahu, Susen Kumar Panda, Niranjan Sahoo, H. K. Panda

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In the present study, an attempt has been made for isolation and identification of feline panleukopaenia virus (FPLV) from wild felids of Nandankanan zoo, Odisha, India, along with prevalence study of FPLV. Fecal samples collected from wild felids (26 tigers, 22 lions, 5 leopards, 3 hyenas, 1 jaguar, 2 foxes and 1 wild cat) were subjected to hemagglutinnation test and fluorescent antibody test. In hemagglutinnation test 13 (50%) samples from tiger, 14 (63.63%) samples from lions, 1 (20%) sample from leopards, 1 (50%) from fox, 3 (100%) samples from hyenas and 1 (100%) sample from wild cat were positive. On fluorescent antibody test (FAT), 15 (57.69%) samples from tiger, 18 (81.81%) from lions, 2 (40%) from leopards, 1 (50%) from fox, 3 (100%) from hyenas and 1 (100%) from wild cat were positive. FPLV was isolated using MDBK cell line and preliminary characterization was done on the basis of characteristic cytopathic effect. The virus samples were quantified through titration in MDBK cells. Serological confirmation of FPLV isolates was carried out by HI test, micro-SNT and indirect-ELISA. Physico-chemical characters like pH and temperature resistance along molecular identification using specific FPLV primers was carried out. Seroprevalence study of 36 serum samples employing HI test, micro SNT and indirect-ELISA revealed prevalence of 38.8, 44.4 and 72.2% respectively. During study period an adult tigress and a tiger cub died suspected of feline panleukopenia. The necropsy findings in both animals showed hemorrhagic gastroenteritis. The cytological examination revealed presence of intranuclear inclusion bodies in the intestinal epithelial cells. Spleen, mesenteric lymph node and intestine were positive for feline panleukopenia by FAT. The investigation revealed that feline panleukopenia was prevalent in wild felines of Nandankanan zoo.

Keywords: Feline panleukopenia, fluorescent antibody test, hemagglutination test, indirect-ELISA, Nandankanan zoo

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Authors: Ranjit Kumar Sahu, Ramakar Jha

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Urban Water Management is the practice of managing freshwater, waste water, and storm water as components of a basin-wide management plan. It builds on existing water supply and sanitation considerations within an urban settlement by incorporating urban water management within the scope of the entire river basin. The pervasive problems generated by urban development have prompted, in the present work, to study the spatial extent of urbanization in Golden Triangle of Odisha connecting the cities Bhubaneswar (20.2700° N, 85.8400° E), Puri (19.8106° N, 85.8314° E) and Konark (19.9000° N, 86.1200° E)., and patterns of periodic changes in urban development (systematic/random) in order to develop future plans for (i) urbanization promotion areas, and (ii) urbanization control areas. Remote Sensing, using USGS (U.S. Geological Survey) Landsat8 maps, supervised classification of the Urban Sprawl has been done for during 1980 - 2014, specifically after 2000. This Work presents the following: (i) Time series analysis of Hydrological data (ground water and rainfall), (ii) Application of SWMM (Storm Water Management Model) and other soft computing techniques for Urban Water Management, and (iii) Uncertainty analysis of model parameters (Urban Sprawl and correlation analysis). The outcome of the study shows drastic growth results in urbanization and depletion of ground water levels in the area that has been discussed briefly. Other relative outcomes like declining trend of rainfall and rise of sand mining in local vicinity has been also discussed. Research on this kind of work will (i) improve water supply and consumption efficiency (ii) Upgrade drinking water quality and waste water treatment (iii) Increase economic efficiency of services to sustain operations and investments for water, waste water, and storm water management, and (iv) engage communities to reflect their needs and knowledge for water management.

Keywords: Storm Water Management Model (SWMM), uncertainty analysis, urban sprawl, land use change

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Authors: Baikunthanath Sahoo, Santosh Kumar Sahu, Krishna Malakar

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With the essence of global environmental sustainability and green business management, the wind of business research moved towards Corporate Social Responsibility. In addition to international and national treaties, businesses have also started realising environmental protection and energy efficiency through CSR as part of business strategy in response to climate change. Considering the ambitious emission reduction target and rapid economic development of India, this study is an attempt to explore the effect of CSR on the energy efficiency management of manufacturing firms in India. By using firm-level data, the panel fixed effect model shows that the CSR dummy variable is negatively influencing the energy intensity or technically, they are energy efficient. The result demonstrates that in the presence of CSR, all the production economic variables are significant. The result also shows that doing environmental expenditure does not improve energy efficiency might be because very few firms are motivated to do such expenditure and also not common to all sectors. The interactive effect model result conforms that without considering CSR dummy as an intervening variable only Manufacturers of Chemical and Chemical products, Manufacturers of Pharmaceutical, medical chemical, and botanical products firms energy intensity low but after considering CSR in their business practices all six sub-sector firms become energy efficient. The empirical result also validate that firms are continuously engaged in CSR activities they are highly energy efficient. It is an important motivational factor for firms to become economically and environmentally sustainable in the corporate world. This analysis would help business practitioners to know how to manage today’s profitability and tomorrow’s sustainability to achieve a comparative advantage in the emerging market economy. The paper concludes that reducing energy consumption as part of their social responsibility to care for the environment, will need collaborative efforts of business society and policy bodies.

Keywords: CSR, Energy Efficiency, Indian manufacturing Sector, Business strategy

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Authors: Pushkar Lal Sharma, Sanjana Singh, Umesh Kumar Sahu

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This paper helps to understand the need of Customer Relationship Management in Libraries and why Librarians should implement the marketing concept of Customer Relationship Management in their libraries. As like any industry, libraries too face growing challenges to continuously meet customer expectations, and attract and retain users in light of overflowing competition. The ability to understand customers, build relationships and market diverse services is essential when considering ways to expand service offerings and improve Return on Investment. Since Library is service oriented Enterprise, hence the Customer/User/ Reader/Patron are the most important element of Library & Information System to whom and for whom library offers various services. How to provide better and most efficient services to its users is the main concern of every Library & Information centre in the present era. The basic difference between Business Enterprise and Library Information System is that ‘in Business System ‘the efficiency is measured in terms of ’profit’ or ‘monetary gains’; whereas in a Library & Information System, the efficiency is measured in terms of ‘services’ and therefore the goals that are set in Business Enterprise are’ profit oriented’ whereas goals set in the Library & Information Centre are ‘Service-oriented’. With the explosion of information and advancement of technology readers have so many choices to get information rather than visiting a library. Everything is available at the click of a mouse, library customers have become more knowledgeable and demanding in an era marked by abundance of information resources and services. With this explosion of information in every field of knowledge and choice in selection of service, satisfying user has become a challenge now a day for libraries. Accordingly, Libraries have to build good relationship with its users by adopting Customer relationship Management. CRM refers to the methods and tools which help an organization to manage its relationship with its customers in an organized way. The Customer Relationship Management (CRM) combines business strategy and technology to identify, acquire and retain good customer relationship. The goal of CRM is to optimize management of customer information needs & interests and increase customer satisfaction and loyalty. Implementing CRM in Libraries can improve customer data and process management, customer loyalty, retention and satisfaction.

Keywords: customer relationship management, CRM, CRM tools, customer satisfaction

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Authors: Anamika Sahu

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The Himalayas, like many mountainous regions, is susceptible to multiple hazards. In recent times, the frequency of such disasters is continuously increasing due to extreme weather phenomena. These natural hazards are responsible for irreparable human and economic loss. The Indian Himalayas has repeatedly been ruptured by great earthquakes in the past and has the potential for a future large seismic event as it falls under the seismic gap. Damages caused by earthquakes are different in different localities. It is well known that, during earthquakes, damage to the structure is associated with the subsurface conditions and the quality of construction materials. So, for sustainable mountain development, prior estimation of site characterization will be valuable for designing and constructing the space area and for efficient mitigation of the seismic risk. Both geotechnical and geophysical investigation of the subsurface is required to describe the subsurface complexity. In mountainous regions, geophysical methods are gaining popularity as areas can be studied without disturbing the ground surface, and also these methods are time and cost-effective. The MASW method is used to calculate the Vs30. Vs30 is the average shear wave velocity for the top 30m of soil. Shear wave velocity is considered the best stiffness indicator, and the average of shear wave velocity up to 30 m is used in National Earthquake Hazards Reduction Program (NEHRP) provisions (BSSC,1994) and Uniform Building Code (UBC), 1997 classification. Parameters obtained through geotechnical investigation have been integrated with findings obtained through the subsurface geophysical survey. Joint interpretation has been used to establish inter-relationships among mineral constituents, various textural parameters, and unconfined compressive strength (UCS) with shear wave velocity. It is found that results obtained through the MASW method fitted well with the laboratory test. In both conditions, mineral constituents and textural parameters (grain size, grain shape, grain orientation, and degree of interlocking) control the petrophysical and mechanical properties of rocks and the behavior of shear wave velocity.

Keywords: MASW, mechanical, petrophysical, site characterization

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Authors: Arpit Kumar Shrivastava, Subrat Kumar, Rajani Kanta Mohapatra, Priyadarshi Soumyaranjan Sahu

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Computational approaches to predict structure, function and other biological characteristics of proteins are becoming more common in comparison to the traditional methods in drug discovery. Cryptosporidiosis is a major zoonotic diarrheal disease particularly in children, which is caused primarily by Cryptosporidium hominis and Cryptosporidium parvum. Currently, there are no vaccines for cryptosporidiosis and recommended drugs are not effective. With the availability of complete genome sequence of C. hominis, new targets have been recognized for the development of effective and better drugs and/or vaccines. We identified a unique hypothetical epitopic protein in C. hominis genome through BLASTP analysis. A 3D model of the hypothetical protein was generated using I-Tasser server through threading methodology. The quality of the model was validated through Ramachandran plot by PROCHECK server. The functional annotation of the hypothetical protein through DALI server revealed structural similarity with human Transportin 3. Phylogenetic analysis for this hypothetical protein also showed C. hominis hypothetical protein (CUV04613) was the closely related to human transportin 3 protein. The 3D protein model is further subjected to virtual screening study with inhibitors from the Zinc Database by using Dock Blaster software. Docking study reported N-(3-chlorobenzyl) ethane-1,2-diamine as the best inhibitor in terms of docking score. Docking analysis elucidated that Leu 525, Ile 526, Glu 528, Glu 529 are critical residues for ligand–receptor interactions. The molecular dynamic simulation was done to access the reliability of the binding pose of inhibitor and protein complex using GROMACS software at 10ns time point. Trajectories were analyzed at each 2.5 ns time interval, among which, H-bond with LEU-525 and GLY- 530 are significantly present in MD trajectories. Furthermore, antigenic determinants of the protein were determined with the help of DNA Star software. Our study findings showed a great potential in order to provide insights in the development of new drug(s) or vaccine(s) for control as well as prevention of cryptosporidiosis among humans and animals.

Keywords: cryptosporidium hominis, hypothetical protein, molecular docking, molecular dynamics simulation

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Authors: S. Vaibhav Kant Sahu, Surabhi Bipin Seth

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Human well-being has multiple constituents including the basic material for a good life, freedom and choice, health, good social relations, and security. Poverty is also multidimensional and has been defined as the pronounced deprivation of well-being. Dhurwa tribe of Bastar (India) have symbiotic relation with nature, it provisions ecosystem service such as food, fuel and fiber; regulating services such as climate regulation and non-material benefits such as spiritual or aesthetic benefits and they are managing their forest from ages. The demand for ecosystem services is now so great that trade-off among services become rule. Aim of study to explore evidences for linkages between ecosystem services and well-being of indigenous community, how much it helps them in poverty reduction and interaction between them. Objective of study was to find drivers of change and evidence concerning link between ecosystem, human development and sustainability, evidence in decision making does it opt for multi sectoral objectives. Which means human well-being as the central focus for assessment, while recognizing that biodiversity and ecosystems also have intrinsic value. Ecosystem changes that may have little impact on human well-being over days or weeks may have pronounced impacts over years or decades; so assessments needed to be conducted at spatial and temporal scales under social, political, economic scales to have high-resolution data. Researcher used framework developed by Millennium ecosystem assessment; since human action now directly or unknowingly virtually alter ecosystem. Researcher used ethnography study to get primary qualitative data, secondary data collected from panchayat office. The responses were transcribed and translated into English, as interview held in Hindi and local indigenous language. Focus group discussion were held with group of 10 women at Tiriya village. Researcher concluded with well-being is not just gap between ecosystem service supply but also increases vulnerability. Decision can have consequences external to the decision framework these consequences are called externalities because they are not part of the decision-making calculus.

Keywords: Bastar, Dhurwa tribe, ecosystem services, millennium ecosystem assessment, sustainability

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Authors: Turin Datta, Kisor K. Sahu

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Structural materials used in today’s oil and gas exploration and drilling of both onshore and offshore oil and gas wells must possess superior tensile properties, excellent resistance to corrosive degradation that includes general, localized (pitting and crevice) and environment assisted cracking such as stress corrosion cracking and hydrogen embrittlement. The High Pressure and High Temperature (HPHT) wells are typically operated at temperature and pressure that can exceed 300-3500F and 10,000psi (69MPa) respectively which necessitates the use of exotic materials in these exotic sources of natural resources. This research investigation is focussed on the evaluation of tensile properties and corrosion behavior of AISI 4140 High-Strength Low Alloy Steel (HSLA) possessing tempered martensitic microstructure and Duplex 2205 Stainless Steel (DSS) having austenitic and ferritic phase. The selection of this two alloys are primarily based on economic considerations as 4140 HSLA is cheaper when compared to DSS 2205. Due to the harsh aggressive chemical species encountered in deep oil and gas wells like chloride ions (Cl-), carbon dioxide (CO2), hydrogen sulphide (H2S) along with other mineral organic acids, DSS 2205, having a dual-phase microstructure can mitigate the degradation resulting from the presence of both chloride ions (Cl-) and hydrogen simultaneously. Tensile properties evaluation indicates a ductile failure of DSS 2205 whereas 4140 HSLA exhibit quasi-cleavage fracture due to the phenomenon of ‘tempered martensitic embrittlement’. From the potentiodynamic polarization testing, it is observed that DSS 2205 has higher corrosion resistance than 4140 HSLA; the former exhibits passivity signifying resistance to localized corrosion while the latter exhibits active dissolution in all the environmental parameters space that was tested. From the Scanning Electron Microscopy (SEM) evaluation, it is understood that stable pits appear in DSS 2205 only when the temperature exceeds the critical pitting temperature (CPT). SEM observation of the corroded 4140 HSLA specimen tested in aqueous 3.5 wt.% NaCl solution reveals intergranular cracking which appears due to the adsorption and diffusion of hydrogen during polarization, thus, causing hydrogen-induced cracking/hydrogen embrittlement. General corrosion testing of DSS 2205 in acidic brine (pH~3.0) solution at ambient temperature using coupons indicate no weight loss even after three months whereas the corrosion rate of AISI 4140 HSLA is significantly higher after one month of testing.

Keywords: DSS 2205, polarization, pitting, SEM

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Authors: Neha Sahu, Awantika Singh, Brijesh Kumar, K. R. Arya

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Taraxacum officinale Weber or dandelion (Asteraceae) is an important Indian traditional herb used to treat liver detoxification, digestive problems, spleen, hepatic and kidney disorders, etc. The plant is well known to possess important phenolic and flavonoids to serve as a potential source of antioxidative and chemoprotective agents. Biosynthesis of bioactive compounds through in vitro cultures is a requisite for natural resource conservation and to provide an alternative source for pharmaceutical applications. Thus an efficient and reproducible protocol was developed for in vitro biosynthesis of bioactive antioxidative compounds from leaf derived callus and in vitro regenerated cultures of Taraxacum officinale using MS media fortified with various combinations of auxins and cytokinins. MS media containing 0.25 mg/l 2, 4-D (2, 4-Dichloro phenoxyacetic acid) with 0.05 mg/l 2-iP [N6-(2-Isopentenyl adenine)] was found as an effective combination for the establishment of callus with 92 % callus induction frequency. Moreover, 2.5 mg/l NAA (α-Naphthalene acetic acid) with 0.5 mg/l BAP (6-Benzyl aminopurine) and 1.5 mg/l NAA showed the optimal response for in vitro plant regeneration with 80 % regeneration frequency and rooting respectively. In vitro regenerated plantlets were further transferred to soil and acclimatized. Quantitative variability of accumulated bioactive compounds in cultures (in vitro callus, plantlets and acclimatized) were determined through UPLC-MS/MS (ultra-performance liquid chromatography-triple quadrupole-linear ion trap mass spectrometry) and compared with wild plants. The phytochemical determination of in vitro and wild grown samples showed the accumulation of 6 compounds. In in vitro callus cultures and regenerated plantlets, two major antioxidative compounds i.e. chlorogenic acid (14950.0 µg/g and 4086.67 µg/g) and umbelliferone (10400.00 µg/g and 2541.67 µg/g) were found respectively. Scopoletin was found to be highest in vitro regenerated plants (83.11 µg/g) as compared to wild plants (52.75 µg/g). Notably, scopoletin is not detected in callus and acclimatized plants, but quinic acid (6433.33 µg/g) and protocatechuic acid (92.33 µg/g) were accumulated at the highest level in acclimatized plants as compared to other samples. Wild grown plants contained highest content (948.33 µg/g) of flavonoid glycoside i.e. luteolin-7-O-glucoside. Our data suggests that in vitro callus and regenerated plants biosynthesized higher content of antioxidative compounds in controlled conditions when compared to wild grown plants. These standardized cultural conditions may be explored as a sustainable source of plant materials for enhanced production and adequate supply of oxidative polyphenols.

Keywords: anti-oxidative compounds, in vitro cultures, Taraxacum officinale, UPLC-MS/MS

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Authors: Ahmad Saadiq, Neeraj Sahu

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Sediment, which comprises of solid particles of mineral and organic material are transported by water. In river systems, the amount of sediment transported is controlled by both the transport capacity of the flow and the supply of sediment. The transport of sediment in rivers is important with respect to pollution, channel navigability, reservoir ageing, hydroelectric equipment longevity, fish habitat, river aesthetics and scientific interests. The sediment load transported in a river is a very complex hydrological phenomenon. Hence, sediment transport has attracted the attention of engineers from various aspects, and different methods have been used for its estimation. So, several experimental equations have been submitted by experts. Though the results of these methods have considerable differences with each other and with experimental observations, because the sediment measures have some limits, these equations can be used in estimating sediment load. In this present study, two black box models namely, an SRC (Sediment Rating Curve) and ANN (Artificial Neural Network) are used in the simulation of the suspended sediment load. The study is carried out for Seonath subbasin. Seonath is the biggest tributary of Mahanadi river, and it carries a vast amount of sediment. The data is collected for Jondhra hydrological observation station from India-WRIS (Water Resources Information System) and IMD (Indian Meteorological Department). These data include the discharge, sediment concentration and rainfall for 10 years. In this study, sediment load is estimated from the input parameters (discharge, rainfall, and past sediment) in various combination of simulations. A sediment rating curve used the water discharge to estimate the sediment concentration. This estimated sediment concentration is converted to sediment load. Likewise, for the application of these data in ANN, they are normalised first and then fed in various combinations to yield the sediment load. RMSE (root mean square error) and R² (coefficient of determination) between the observed load and the estimated load are used as evaluating criteria. For an ideal model, RMSE is zero and R² is 1. However, as the models used in this study are black box models, they don’t carry the exact representation of the factors which causes sedimentation. Hence, a model which gives the lowest RMSE and highest R² is the best model in this study. The lowest values of RMSE (based on normalised data) for sediment rating curve, feed forward back propagation, cascade forward back propagation and neural network fitting are 0.043425, 0.00679781, 0.0050089 and 0.0043727 respectively. The corresponding values of R² are 0.8258, 0.9941, 0.9968 and 0.9976. This implies that a neural network fitting model is superior to the other models used in this study. However, a drawback of neural network fitting is that it produces few negative estimates, which is not at all tolerable in the field of estimation of sediment load, and hence this model can’t be crowned as the best model among others, based on this study. A cascade forward back propagation produces results much closer to a neural network model and hence this model is the best model based on the present study.

Keywords: artificial neural network, Root mean squared error, sediment, sediment rating curve

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Authors: Jonathan Sahu, Jill Aylott

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Individuals with Autism, Intellectual and Developmental disabilities (AIDD) are one of the most vulnerable groups in society, hampered not only by their own limitations to understand and interact with the wider society, but also societal limitations in perception and understanding. Communication to express their needs and wishes is fundamental to enable such individuals to live and prosper in society. This research project was designed as an organisational case study, in a large secondary health care hospital within the National Health Service (NHS), to assess the quality of care provided to people with AIDD and to review the role of advocacy to reduce health inequalities in these individuals. Methods: The research methodology adopted was as an “insider researcher”. Data collection included both quantitative and qualitative data i.e. a mixed method approach. A semi-structured interview schedule was designed and used to obtain qualitative and quantitative primary data from a wide range of interdisciplinary frontline health care workers to assess their understanding and awareness of systems, processes and evidence based practice to offer a quality service to people with AIDD. Secondary data were obtained from sources within the organisation, in keeping with “Case Study” as a primary method, and organisational performance data were then compared against national benchmarking standards. Further data sources were accessed to help evaluate the effectiveness of different types of advocacy that were present in the organisation. This was gauged by measures of user and carer experience in the form of retrospective survey analysis, incidents and complaints. Results: Secondary data demonstrate near compliance of the Organisation with the current national benchmarking standard (Monitor Compliance Framework). However, primary data demonstrate poor knowledge of the Mental Capacity Act 2005, poor knowledge of organisational systems, processes and evidence based practice applied for people with AIDD. In addition there was poor knowledge and awareness of frontline health care workers of advocacy and advocacy schemes for this group. Conclusions: A significant amount of work needs to be undertaken to improve the quality of care delivered to individuals with AIDD. An operational strategy promoting the widespread dissemination of information may not be the best approach to deliver quality care and optimal patient experience and patient advocacy. In addition, a more robust set of standards, with appropriate metrics, needs to be developed to assess organisational performance which will stand the test of professional and public scrutiny.

Keywords: advocacy, autism, health inequalities, intellectual developmental disabilities, quality of care

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Authors: Jyoti Sahu, Vinay A. Juvekar

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Electrolytes are often used to bring about salting-in and salting-out of organic molecules and polymers (e.g. polyethylene glycols/proteins) from the aqueous solutions. For quantification of these phenomena, a thermodynamic model which can accurately predict activity coefficient of electrolyte as a function of temperature is needed. The thermodynamics models available in the literature contain a large number of empirical parameters. These parameters are estimated using lower/upper critical solution temperature of the solution in the electrolyte/organic molecule at different temperatures. Since the number of parameters is large, inaccuracy can bethe creep in during their estimation, which can affect the reliability of prediction beyond the range in which these parameters are estimated. Cloud point of solution is related to its free energy through temperature and composition derivative. Hence, the Cloud point measurement can be used for accurate estimation of the temperature and composition dependence of parameters in the model for free energy. Hence, if we use a two pronged procedure in which we first use cloud point of solution to estimate some of the parameters of the thermodynamic model and determine the rest using osmotic coefficient data, we gain on two counts. First, since the parameters, estimated in each of the two steps, are fewer, we achieve higher accuracy of estimation. The second and more important gain is that the resulting model parameters are more sensitive to temperature. This is crucial when we wish to use the model outside temperatures window within which the parameter estimation is sought. The focus of the present work is to prove this proposition. We have used electrolyte (NaCl/Na2CO3)-water-organic molecule (Iso-propanol/ethanol) as the model system. The model of Robinson-Stokes-Glukauf is modified by incorporating the temperature dependent Flory-Huggins interaction parameters. The Helmholtz free energy expression contains, in addition to electrostatic and translational entropic contributions, three Flory-Huggins pairwise interaction contributions viz., and (w-water, p-polymer, s-salt). These parameters depend both on temperature and concentrations. The concentration dependence is expressed in the form of a quadratic expression involving the volume fractions of the interacting species. The temperature dependence is expressed in the form .To obtain the temperature-dependent interaction parameters for organic molecule-water and electrolyte-water systems, Critical solution temperature of electrolyte -water-organic molecules is measured using cloud point measuring apparatus The temperature and composition dependent interaction parameters for electrolyte-water-organic molecule are estimated through measurement of cloud point of solution. The model is used to estimate critical solution temperature (CST) of electrolyte water-organic molecules solution. We have experimentally determined the critical solution temperature of different compositions of electrolyte-water-organic molecule solution and compared the results with the estimates based on our model. The two sets of values show good agreement. On the other hand when only osmotic coefficients are used for estimation of the free energy model, CST predicted using the resulting model show poor agreement with the experiments. Thus, the importance of the CST data in the estimation of parameters of the thermodynamic model is confirmed through this work.

Keywords: concentrated electrolytes, Debye-Hückel theory, interaction parameters, Robinson-Stokes-Glueckauf model, Flory-Huggins model, critical solution temperature

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Authors: Yi H. Wong, Li L. Chan, Chee O. Leong, Stephen Ambu, Joon W. Mak, Priyadashi S. Sahu

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Background: Acanthamoeba is a free-living opportunistic protist which is ubiquitously distributed in the environment. Virulent Acanthamoeba can cause fatal encephalitis in immunocompromised patients and potential blinding keratitis in immunocompetent contact lens wearers. Approximately 24 species have been identified but only the A. castellanii, A. polyphaga and A. culbertsoni are commonly associated with human infections. Until to date, the precise molecular basis for Acanthamoeba pathogenesis remains unclear. Previous studies reported that Acanthamoeba virulence can be diminished through prolonged axenic culture but revived through serial mouse passages. As no clear explanation on this reversible pathogenesis is established, hereby, we postulate that the epigenetic regulators, DNA-methyltransferases (DNMT) and histone-deacetylases (HDAC), could possibly be involved in granting the virulence plasticity of Acanthamoeba spp. Methods: Four rounds of mouse passages were conducted to revive the virulence potential of the virulence-attenuated Acanthamoeba castellanii strain (ATCC 50492). Briefly, each mouse (n=6/group) was inoculated intraperitoneally with Acanthamoebae cells (2x 105 trophozoites/mouse) and incubated for 2 months. Acanthamoebae cells were isolated from infected mouse organs by culture method and subjected to subsequent mouse passage. In vitro cytopathic, encystment and gelatinolytic assays were conducted to evaluate the virulence characteristics of Acanthamoebae isolates for each passage. PCR primers which targeted on the 2 members (DNMT1 and DNMT2) and 5 members (HDAC1 to 5) of the DNMT and HDAC gene families respectively were custom designed. Quantitative real-time PCR (qPCR) was performed to detect and quantify the relative expression of the two gene families in each Acanthamoeba isolates. Beta-tubulin of A. castellanii (Genbank accession no: XP_004353728) was included as housekeeping gene for data normalisation. PCR mixtures were also analyzed by electrophoresis for amplicons detection. All statistical analyses were performed using the paired one-tailed Student’s t test. Results: Our pathogenicity tests showed that the virulence-reactivated Acanthamoeba had a higher degree of cytopathic effect on vero cells, a better resistance to encystment challenge and a higher gelatinolytic activity which was catalysed by serine protease. qPCR assay showed that DNMT1 expression was significantly higher in the virulence-reactivated compared to the virulence-attenuated Acanthamoeba strain (p ≤ 0.01). The specificity of primers which targeted on DNMT1 was confirmed by sequence analysis of PCR amplicons, which showed a 97% similarity to the published DNA-methyltransferase gene of A. castellanii (GenBank accession no: XM_004332804.1). Out of the five primer pairs which targeted on the HDAC family genes, only HDAC4 expression was significantly difference between the two variant strains. In contrast to DNMT1, HDAC4 expression was much higher in the virulence-attenuated Acanthamoeba strain. Conclusion: Our mouse passages had successfully restored the virulence of the attenuated strain. Our findings suggested that DNA-methyltransferase (DNMT1) and histone deacetylase (HDAC4) expressions are associated with virulence potential of Acanthamoeba spp.

Keywords: acanthamoeba, DNA-methyltransferase, histone deacetylase, virulence-associated proteins

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Authors: Yi-Hao Wong, Li-Li Chan, Chee-Onn Leong, Stephen Ambu, Joon-Wah Mak, Priyasashi Sahu

Abstract:

Background: Acanthamoeba is a genus of amoebae which lives as a free-living in nature or as a human pathogen that causes severe brain and eye infections. Virulence potential of Acanthamoeba is not constant and can change with growth conditions. DNA methylation, an epigenetic process which adds methyl groups to DNA, is used by eukaryotic cells, including several human parasites to control their gene expression. We used qPCR, siRNA gene silencing, and RNA sequencing (RNA-Seq) to study DNA-methyltransferase gene family (DNMT) in order to indicate the possibility of its involvement in programming Acanthamoeba virulence potential. Methods: A virulence-attenuated Acanthamoeba isolate (designation: ATCC; original isolate: ATCC 50492) was subjected to mouse passages to restore its pathogenicity; a virulence-reactivated isolate (designation: AC/5) was generated. Several established factors associated with Acanthamoeba virulence phenotype were examined to confirm the succession of reactivation process. Differential gene expression of DNMT between ATCC and AC/5 isolates was performed by qPCR. Silencing on DNMT gene expression in AC/5 isolate was achieved by siRNA duplex. Total RNAs extracted from ATCC, AC/5, and siRNA-treated (designation: si-146) were subjected to RNA-Seq for comparative transcriptomic analysis in order to identify the genome-wide effect of DNMT in regulating Acanthamoeba gene expression. qPCR was performed to validate the RNA-Seq results. Results: Physiological and cytophatic assays demonstrated an increased in virulence potential of AC/5 isolate after mouse passages. DNMT gene expression was significantly higher in AC/5 compared to ATCC isolate (p ≤ 0.01) by qPCR. si-146 duplex reduced DNMT gene expression in AC/5 isolate by 30%. Comparative transcriptome analysis identified the differentially expressed genes, with 3768 genes in AC/5 vs ATCC isolate; 2102 genes in si-146 vs AC/5 isolate and 3422 genes in si-146 vs ATCC isolate, respectively (fold-change of ≥ 2 or ≤ 0.5, p-value adjusted (padj) < 0.05). Of these, 840 and 1262 genes were upregulated and downregulated, respectively, in si-146 vs AC/5 isolate. Eukaryotic orthologous group (KOG) assignments revealed a higher percentage of downregulated gene expression in si-146 compared to AC/5 isolate, were related to posttranslational modification, signal transduction and energy production. Gene Ontology (GO) terms for those downregulated genes shown were associated with transport activity, oxidation-reduction process, and metabolic process. Among these downregulated genes were putative genes encoded for heat shock proteins, transporters, ubiquitin-related proteins, proteins for vesicular trafficking (small GTPases), and oxidoreductases. Functional analysis of similar predicted proteins had been described in other parasitic protozoa for their survival and pathogenicity. Decreased expression of these genes in si146-treated isolate may account in part for Acanthamoeba reduced pathogenicity. qPCR on 6 selected genes upregulated in AC/5 compared to ATCC isolate corroborated the RNA sequencing findings, indicating a good concordance between these two analyses. Conclusion: To the best of our knowledge, this study represents the first genome-wide analysis of DNA methylation and its effects on gene expression in Acanthamoeba spp. The present data indicate that DNA methylation has substantial effect on global gene expression, allowing further dissection of the genome-wide effects of DNA-methyltransferase gene in regulating Acanthamoeba pathogenicity.

Keywords: Acanthamoeba, DNA methylation, RNA sequencing, virulence

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