Search results for: Ashish Ranjan

Authors: Maneesh Kumar, Roshan Kamal Topno, Manas Ranjan Dikhit, Vahab Ali, Ganesh Chandra Sahoo, Bhawana, Major Madhukar, Rishikesh Kumar, Krishna Pandey, Pradeep Das

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Chandipura vesiculovirus is an emerging (-) ssRNA viral entity belonging to the genus Vesiculovirus of the family Rhabdoviridae, associated with fatal encephalitis in tropical regions. The multi-functionally active viral RNA-dependent RNA polymerase (vRdRp) that has been incorporated with conserved amino acid residues in the pathogens, assigned to synthesize distinct viral polypeptides. The lack of proofreading ability of the vRdRp produces many mutated variants. Here, we have performed the evolutionary analysis of 20 viral protein sequences of vRdRp of different strains of Chandipura vesiculovirus along with other viral species from genus Vesiculovirus inferred in MEGA6.06, employing the Neighbour-Joining method. The p-distance algorithmic method has been used to calculate the optimum tree which showed the sum of branch length of about 1.436. The percentage of replicate trees in which the associated taxa are clustered together in the bootstrap test (1000 replicates), is shown next to the branches. No mutation was observed in the Indian strains of Chandipura vesiculovirus. In vRdRp, 1230(His) and 1231(Arg) are actively participated in catalysis and, are found conserved in different strains of Chandipura vesiculovirus. Both amino acid residues were also conserved in the other viral species from genus Vesiculovirus. Many isolates exhibited maximum number of mutations in catalytic regions in strains of Chandipura vesiculovirus at position 26(Ser→Ala), 47 (Ser→Ala), 90(Ser→Tyr), 172(Gly→Ile, Val), 172(Ser→Tyr), 387(Asn→Ser), 1301(Thr→Ala), 1330(Ala→Glu), 2015(Phe→Ser) and 2065(Thr→Val) which make them variants under different tropical conditions from where they evolved. The result clarifies the actual concept of RNA evolution using vRdRp to develop as an evolutionary marker. Although, a limited number of vRdRp protein sequence similarities for Chandipura vesiculovirus and other species. This might endow with possibilities to identify the virulence level during viral multiplication in a host.

Keywords: Chandipura, (-) ssRNA, viral RNA-dependent RNA polymerase, neighbour-joining method, p-distance algorithmic, evolutionary marker

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Authors: Nishaben Desai Dholakiya, Anirban Roy, Ranjan Dey

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Understanding ocean thermodynamics has become increasingly critical as Earth's oceans serve as the primary planetary heat regulator, absorbing approximately 93% of excess heat energy from anthropogenic greenhouse gas emissions. This investigation presents a comprehensive analysis of Arabian Sea surface seawater thermodynamics, focusing specifically on heat capacity (Cp) and thermal expansion coefficient (α) - parameters fundamental to global heat distribution patterns. Through high-precision experimental measurements of ultrasonic velocity and density across varying temperature (293.15-318.15K) and salinity (0.5-35 ppt) conditions, it characterize critical thermophysical parameters including specific heat capacity, thermal expansion, and isobaric and isothermal compressibility coefficients in natural seawater systems. The study employs advanced machine learning frameworks - Random Forest, Gradient Booster, Stacked Ensemble Machine Learning (SEML), and AdaBoost - with SEML achieving exceptional accuracy (R² > 0.99) in heat capacity predictions. the findings reveal significant temperature-dependent molecular restructuring: enhanced thermal energy disrupts hydrogen-bonded networks and ion-water interactions, manifesting as decreased heat capacity with increasing temperature (negative ∂Cp/∂T). This mechanism creates a positive feedback loop where reduced heat absorption capacity potentially accelerates oceanic warming cycles. These quantitative insights into seawater thermodynamics provide crucial parametric inputs for climate models and evidence-based environmental policy formulation, particularly addressing the critical knowledge gap in thermal expansion behavior of seawater under varying temperature-salinity conditions.

Keywords: climate change, arabian sea, thermodynamics, machine learning

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Authors: Abhinav Bajpayee, Shubham Damke, Rupal Ranjan, Neha Bharti

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Polymer flooding is a dramatic improvement in water flooding and quickly becoming one of the EOR technologies. Used for improving oil recovery. With the increasing energy demand and depleting oil reserves EOR techniques are becoming increasingly significant .Since most oil fields have already begun water flooding, chemical EOR technique can be implemented by using fewer resources than any other EOR technique. Polymer helps in increasing the viscosity of injected water thus reducing water mobility and hence achieves a more stable displacement .Polymer flooding helps in increasing the injection viscosity as has been revealed through field experience. While the injection of a polymer solution improves reservoir conformance the beneficial effect ceases as soon as one attempts to push the polymer solution with water. It is most commonly applied technique because of its higher success rate. In polymer flooding, a water-soluble polymer such as Polyacrylamide is added to the water in the water flood. This increases the viscosity of the water to that of a gel making the oil and water greatly improving the efficiency of the water flood. It also improves the vertical and areal sweep efficiency as a consequence of improving the water/oil mobility ratio. Polymer flooding plays an important role in oil exploitation, but around 60 million ton of wastewater is produced per day with oil extraction together. Therefore the treatment and reuse of wastewater becomes significant which can be carried out by electro dialysis technology. This treatment technology can not only decrease environmental pollution, but also achieve closed-circuit of polymer flooding wastewater during crude oil extraction. There are three potential ways in which a polymer flood can make the oil recovery process more efficient: (1) through the effects of polymers on fractional flow, (2) by decreasing the water/oil mobility ratio, and (3) by diverting injected water from zones that have been swept. It has also been suggested that the viscoelastic behavior of polymers can improve displacement efficiency Polymer flooding may also have an economic impact because less water is injected and produced compared with water flooding. In future we need to focus on developing polymers that can be used in reservoirs of high temperature and high salinity, applying polymer flooding in different reservoir conditions and also combine polymer with other processes (e.g., surfactant/ polymer flooding).

Keywords: fractional flow, polymer, viscosity, water/oil mobility ratio

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Authors: Prangya Ranjan Rout, Puspendu Bhunia, Rajesh Roshan Dash

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In a developing country like India, providing reliable and affordable wastewater treatment facilities in rural areas is a huge challenge. With the aim of enhancing the nutrient removal from rural domestic wastewater while reducing the cost of treatment process, a novel, integrated treatment system consisting of a multistage bio-filter with drop aeration and a post positioned attached growth carbonaceous denitrifying-bioreactor was designed and developed in this work. The bio-filter was packed with ‘dolochar’, a sponge iron industry waste, as an adsorbent mainly for phosphate removal through physiochemical approach. The Denitrifying bio-reactor was packed with many waste organic solid substances (WOSS) as carbon sources and substrates for biomass attachment, mainly to remove nitrate in biological denitrification process. The performance of the modular system, treating real domestic wastewater was monitored for a period of about 60 days and the average removal efficiencies during the period were as follows: phosphate, 97.37%; nitrate, 85.91%, ammonia, 87.85%, with mean final effluent concentration of 0.73, 9.86, and 9.46 mg/L, respectively. The multistage bio-filter played an important role in ammonium oxidation and phosphate adsorption. The multilevel drop aeration with increasing oxygenation, and the special media used, consisting of certain oxides were likely beneficial for nitrification and phosphorus removal, respectively, whereas the nitrate was effectively reduced by biological denitrification in the carbonaceous bioreactor. This treatment system would allow multipurpose reuse of the final effluent. Moreover, the saturated dolochar can be used as nutrient suppliers in agricultural practices and the partially degraded carbonaceous substances can be subjected to composting, and subsequently used as an organic fertilizer. Thus, the system displays immense potential for treating domestic wastewater significantly decreasing the concentrations of nutrients and more importantly, facilitating the conversion of the waste materials into usable ones.

Keywords: nutrient removal, denitrifying bioreactor, multi-stage bio-filter, dolochar, waste organic solid substances

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Authors: Mahendran Ganesan, Sanjeet Kumar Verma, Zafar Iqbal, Ashish Chandran, Zakir Husain, Shama Afroz, Sana Shahid, Laiq Ur Rahman

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Highly efficient in vitro regeneration system has been developed for Swertia chirayita (Roxb. ex Fleming) H. Karst, a high prized traditional medicinal plant to treat numerous ailments such as liver disorders, malaria and diabetes and are reported to have a wide spectrum of pharmacological properties. Its medicinal usage is well-documented in Indian pharmaceutical codex, the British and the American pharmacopeias, and in different traditional medicine such as the Ayurveda, Unani and Siddha medical systems. Nodal explants were cultured on MS medium supplemented with various phytohormones for multiple shoot induction. The nodal segments failed to respond in growth regulator free medium. All the concentrations of BAP, Kin and TDZ facilitated shoot bud break and multiple shoot induction. Among the various cytokinins tested, BAP was found to be more effective with respect to initiation and subsequent development of shoots. Of the various concentrations BAP tested, BAP at 4.0 mg/L showed the higher average number of shoot regeneration (10.80 shoots per explant). Kin at 4 mg/L and TDZ at 4 mg/L induced 5.70 and 04.5+0 shoots per explant, respectively. Further increase in concentration did not favour an increase in the number of shoots. However, these shoots failed to elongate further. Hence, addition of GA₃ (1 mg/L) was added to the above medium. This treatment resulted in the elongation of shoots (2.50 cm) and a further increase in the number of microshoots (34.20 shoots/explant). Roots were also induced in the same medium containing BAP (4 mg/L) + GA₃ (1 mg/L) + NAA (0.5 mg/L). In vitro derived plantlets with well-developed roots were transferred to the potting media containing garden soil: sand: vermicompost (2:1:1). Plantlets were covered with a polyethylene bag and irrigated with water. The pots were maintained at 25 ± 2ºC, and then the polyethylene cover was gradually loosened, thus dropping the humidity (65–70%). This procedure subsequently resulted in in vitro hardening of the plantlet.

Keywords: micropropagation, nodal explant, plant growth regulators, Swertia chirayita

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Authors: Ujala, Diksha Sharma, Mahinder Partap, Ashish R. Warghat, Bhavya Bhargava

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In soil-less agriculture, hydroponic is considered a potential farming system for the production of uniform quality plant material in significantly less time. Therefore, for the first time, the current investigation corroborates the effect of different cultivation conditions (open-field, poly-house, and hydroponic) on morpho-physiological traits, phenolic content, and essential oil components analysis in three flower color variants (yellow, scarlet red, and orange) of Tagetes patula. The results revealed that the maximum plant height, number of secondary branches, number of flowers, photosynthesis, stomatal conductance, and transpiration rate were observed under the hydroponic system as compared to other conditions. However, the maximum content of gallic acid (0.82 mg/g DW), syringic acid (3.98 mg/g DW), epicatechin (0.48 mg/g DW), p-coumaric acid (7.28 mg/g DW), protocatechuic acid (0.59 mg/g DW), ferulic acid (2.58 mg/g DW), and luteolin (8.24 mg/g DW) were quantified maximally under open-field conditions. However, under hydroponic conditions, the higher content of vanillic acid (0.43 mg/g DW), caffeic acid (0.49 mg/g DW), and quercetin (0.92 mg/g DW) were quantified. Moreover, a total of nineteen volatile components were identified in the essential oil of different flower color variants of T. patula cultivated under different conditions. The major reported volatile components in essential oil were (-)-caryophyllene oxide, trans-β-caryophyllene, trans-geraniol, 3 methyl-benzyl alcohol, and 2,2’:5’,2”-terthiophene. It has also been observed that the volatile component percentage range in all variants was observed in open-field (70.85 % to 90.54 %), poly-house (59.03 % to 77.93 %), and hydroponic (68.78 % to 89.41 %). In conclusion, the research highlighted that morpho-physiological performance with flower production was enhanced in the hydroponic system. However, phenolic content and volatile components were maximally observed in open-field conditions. However, significant results have been reported under hydroponic conditions in all studied parameters, so it could be a potential strategy for quality biomass production in T. patula.

Keywords: Tagetes patula, cultivation conditions, hydroponic, morpho-physiology

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Authors: Swati Srivastava, Ashish Kumar Upadhyay

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Background: According to United Nation Children’s Fund, it has been estimated that worldwide about 165 million children were stunted in 2012 and India alone accounts for 38% of global burden of stunting. In terms of incidence, India is home of more than 60 million stunted children worldwide. Our study aims to examine the effect of pregnancy intention and maternal postnatal depressive symptoms on early childhood stunting in India. We hypothesized that effect of pregnancy intention and postnatal maternal depressive symptoms were mediated by social support. Methods: We used data from first wave of Young Lives Study India. Out of 2011 children recruited in original cohort, 1833 children had complete information on pregnancy intention, maternal depression and other variables. A series of multivariate logistic regression model were used to examine the effect of pregnancy intention and postnatal depressive symptoms on early childhood stunting. Results: Bivariate result indicates that a higher percent of children born after unintended pregnancy (40%) were stunted than children of intended pregnancy (26%). Likewise, proportion of stunted children was also higher among women of high postnatal depressive symptoms (35%) than low level of depression (24%). Results of multivariate logistic regression model indicate that children born after unintended pregnancy were significantly more likely to be stunted than children born after intended pregnancy (Coefficient: 1.70, CI: 1.17, 2.48). Likewise, early childhood stunting was also associated with maternal postnatal depressive symptoms among women (Coefficient: 1.48, CI: 1.16, 1.88). The effect of pregnancy intention and postnatal depressive symptoms on early childhood stunting remains unchanged after controlling for social support and other variables. Conclusions: The findings of this study provide conclusive evidence regarding consequences of pregnancy intention and postnatal depressive symptoms on early childhood stunting in India. Therefore, there is need to identify the women with unintended pregnancy and incorporate the promotion of mental health into their national reproductive and child health programme.

Keywords: pregnancy intention, postnatal depressive symptoms, social support, childhood stunting, young lives study, India

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Authors: Shankha Sanyal, Archi Banerjee, Sayan Biswas, Sourya Sengupta, Sayan Nag, Ranjan Sengupta, Dipak Ghosh

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The term ‘gestalt’ has been widely used in the field of psychology which defined the perception of human mind to group any object not in part but as a 'unified' whole. Music, in general, is polyphonic - i.e. a combination of a number of pure tones (frequencies) mixed together in a manner that sounds harmonious. The study of human brain response due to different frequency groups of the acoustic signal can give us an excellent insight regarding the neural and functional architecture of brain functions. Hence, the study of music cognition using neuro-biosensors is becoming a rapidly emerging field of research. In this work, we have tried to analyze the effect of different frequency bands of music on the various frequency rhythms of human brain obtained from EEG data. Four widely popular Rabindrasangeet clips were subjected to Wavelet Transform method for extracting five resonant frequency bands from the original music signal. These frequency bands were initially analyzed with Detrended/Adaptive Fractal analysis (DFA/AFA) methods. A listening test was conducted on a pool of 100 respondents to assess the frequency band in which the music becomes non-recognizable. Next, these resonant frequency bands were presented to 20 subjects as auditory stimulus and EEG signals recorded simultaneously in 19 different locations of the brain. The recorded EEG signals were noise cleaned and subjected again to DFA/AFA technique on the alpha, theta and gamma frequency range. Thus, we obtained the scaling exponents from the two methods in alpha, theta and gamma EEG rhythms corresponding to different frequency bands of music. From the analysis of music signal, it is seen that loss of recognition is proportional to the loss of long range correlation in the signal. From the EEG signal analysis, we obtain frequency specific arousal based response in different lobes of brain as well as in specific EEG bands corresponding to musical stimuli. In this way, we look to identify a specific frequency band beyond which the music becomes non-recognizable and below which in spite of the absence of other bands the music is perceivable to the audience. This revelation can be of immense importance when it comes to the field of cognitive music therapy and researchers of creativity.

Keywords: AFA, DFA, EEG, gestalt in music, Hurst exponent

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Authors: Nishaben Dholakiya, Anirban Roy, Ranjan Dey

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The unprecedented rise in global temperatures has triggered complex changes in marine ecosystems, necessitating a deeper understanding of seawater's thermophysical properties by experimentally measuring ultrasonic velocity and density at varying temperatures and salinity. This study investigates the critical relationship between temperature variations and molecular-level interactions in Arabian Sea surface waters, specifically focusing on internal pressure (π) and cohesion energy density (CED) as key indicators of ecosystem disruption. Our experimental findings reveal that elevated temperatures significantly reduce internal pressure, weakening the intermolecular forces that maintain seawater's structural integrity. This reduction in π correlates directly with decreased habitat stability for marine organisms, particularly affecting pressure-sensitive species and their physiological processes. Similarly, the observed decline in cohesion energy density at higher temperatures indicates a fundamental shift in water molecule organization, impacting the dissolution and distribution of vital nutrients and gases. These molecular-level changes cascade through the ecosystem, affecting everything from planktonic organisms to complex food webs. By employing advanced machine learning techniques, including Stacked Ensemble Machine Learning (SEML) and AdaBoost (AB), we developed highly accurate predictive models (>99% accuracy) for these thermophysical parameters. The results provide crucial insights into the mechanistic relationship between climate warming and marine ecosystem degradation, offering valuable data for environmental policymaking and conservation strategies. The novelty of this research serves as no such thermodynamic investigation has been conducted before in literature, whereas this research establishes a quantitative framework for understanding how molecular-level changes in seawater properties directly influence marine ecosystem stability, emphasizing the urgent need for climate change mitigation efforts.

Keywords: thermophysical properties, Arabian Sea, internal pressure, cohesion energy density, machine learning

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Authors: Ranjan Banerjee, W. D. Gunawardana

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The free vibration and flutter analysis plays a major part in aircraft design which is indeed, a mandatory requirement. In particular, high aspect ratio transport airliner wings are prone to free vibration and flutter problems that must be addressed during the design process as demanded by the airworthiness authorities. The purpose of this paper is to carry out a detailed free vibration and flutter analysis for a wide range of high aspect ratio aircraft wings and generate design curves to provide useful visions and understandings of aircraft design from an aeroelastic perspective. In the initial stage of the investigation, the bending and torsional stiffnesses of a number of transport aircraft wings are looked at and critically examined to see whether it is possible to express the stiffness distributions in polynomial form, but in a sufficiently accurate manner. A similar attempt is made for mass and mass moment of inertia distributions of the wing. Once the choice of stiffness and mass distributions in polynomial form is made, the high aspect ratio wing is idealised by a series of bending-torsion coupled beams from a structural standpoint. Then the dynamic stiffness method is applied to compute the natural frequencies and mode shape of the wing. Next the wing is idealised aerodynamically and to this end, unsteady aerodynamic of Theodorsen type is employed to represent the harmonically oscillating wing. Following this step, a normal mode method through the use of generalised coordinates is applied to formulate the flutter problem. In essence, the generalised mass, stiffness and aerodynamic matrices are combined to obtain the flutter matrix which is subsequently solved in the complex domain to determine the flutter speed and flutter frequency. In the final stage of the investigation, an exhaustive parametric study is carried out by varying significant wing parameters to generate design curves which help to predict the free vibration and flutter behaviour of high aspect ratio transport aircraft wings in a generic manner. It is in the aeroelastic context of aircraft design where the results are expected to be most useful.

Keywords: high-aspect ratio wing, flutter, dynamic stiffness method, free vibration, aeroelasticity

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Authors: Krupali Mehta, Sandip Bhatt, Umesh Trivedi, Bhavesh Bharatiya, Mukesh Ranjan, Atindra D. Shukla

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Owing to the dominating surface forces and large-scale surface interactions, the nano-scale particles face difficulties in getting suspended in various media. Magnetic nanoparticles of iron oxide offer a great deal of promise due to their ease of preparation, reasonable magnetic properties, low cost and environmental compatibility. We intend to modify the surface of magnetic Fe₂O₃ nanoparticles with selected surface modifying agents using simple and effective single-step chemical reactions in order to enhance dispersibility of magnetic nanoparticles in non-polar media. Magnetic particles were prepared by hydrolysis of Fe²⁺/Fe³⁺ chlorides and their subsequent oxidation in aqueous medium. The dried particles were then treated with Octadecyl quaternary ammonium silane (Terrasil™), stearic acid and gallic acid ester of stearyl alcohol in ethanol separately to yield S-2 to S-4 respectively. The untreated Fe₂O₃ was designated as S-1. The surface modified nanoparticles were then analysed with Dynamic Light Scattering (DLS), Fourier Transform Infrared spectroscopy (FTIR), X-Ray Diffraction (XRD), Thermogravimetric Gravimetric Analysis (TGA) and Scanning Electron Microscopy and Energy dispersive X-Ray analysis (SEM-EDAX). Characterization reveals the particle size averaging 20-50 nm with and without modification. However, the crystallite size in all cases remained ~7.0 nm with the diffractogram matching to Fe₂O₃ crystal structure. FT-IR suggested the presence of surfactants on nanoparticles’ surface, also confirmed by SEM-EDAX where mapping of elements proved their presence. TGA indicated the weight losses in S-2 to S-4 at 300°C onwards suggesting the presence of organic moiety. Hydrophobic character of modified surfaces was confirmed with contact angle analysis, all modified nanoparticles showed super hydrophobic behaviour with average contact angles ~129° for S-2, ~139.5° for S-3 and ~151° for S-4. This indicated that surface modified particles are super hydrophobic and they are easily dispersible in non-polar media. These modified particles could be ideal candidates to be suspended in oil-based fluids, polymer matrices, etc. We are pursuing elaborate suspension/sedimentation studies of these particles in various oils to establish this conjecture.

Keywords: iron nanoparticles, modification, hydrophobic, dispersion

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Authors: Ashish Trivedi, Amol Singh

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In the recent years, the magnitude and frequency of disasters have increased at an alarming rate. Every year, more than 400 natural disasters affect global population. A large-scale disaster leads to destruction or damage to houses, thereby rendering a notable number of residents homeless. Since humanitarian response and recovery process takes considerable time, temporary establishments are arranged in order to provide shelter to affected population. These shelter areas are vital for an effective humanitarian relief; therefore, they must be strategically planned. Choosing the locations of temporary shelter areas for accommodating homeless people is critical to the quality of humanitarian assistance provided after a large-scale emergency. There has been extensive research on the facility location problem both in theory and in application. In order to deliver sufficient relief aid within a relatively short timeframe, humanitarian relief organisations pre-position warehouses at strategic locations. However, such approaches have received limited attention from the perspective of providing shelters to disaster-affected people. In present research work, this aspect of humanitarian logistics is considered. The present work proposes a hybrid decision support model to determine relative preference of potential shelter locations by assessing them based on key subjective criteria. Initially, the factors that are kept in mind while locating potential areas for establishing temporary shelters are identified by reviewing extant literature and through consultation from a panel of disaster management experts. In order to determine relative importance of individual criteria by taking into account subjectivity of judgements, a hybrid approach of fuzzy sets and Analytic Hierarchy Process (AHP) was adopted. Further, Technique for order preference by similarity to ideal solution (TOPSIS) was applied on an illustrative data set to evaluate potential locations for establishing temporary shelter areas for homeless people in a disaster scenario. The contribution of this work is to propose a range of possible shelter locations for a humanitarian relief organization, using a robust multi criteria decision support framework.

Keywords: AHP, disaster preparedness, fuzzy set theory, humanitarian logistics, TOPSIS, temporary shelters

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Authors: Selvaprakash Ramalingam, Rabi N. Sahoo, Dharmendra Saraswat, A. Kumar, Rajeev Ranjan, Joydeep Mukerjee, Viswanathan Chinnasamy, K. K. Chaturvedi, Sanjeev Kumar

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Rice is one of the most important staple food crops in the world. Among the various diseases that affect rice crops, rice blast is particularly significant, causing crop yield and economic losses. While the plant has defense mechanisms in place, such as chemical indicators (proteins, salicylic acid, jasmonic acid, ethylene, and azelaic acid) and resistance genes in certain varieties that can protect against diseases, susceptible varieties remain vulnerable to these fungal diseases. Early prediction of rice blast (RB) disease is crucial, but conventional techniques for early prediction are time-consuming and labor-intensive. Hyperspectral remote sensing techniques hold the potential to predict RB disease at its asymptomatic stage. In this study, we aimed to demonstrate the prediction of RB disease at the asymptomatic stage using non-imaging hyperspectral ASD spectroradiometer under controlled laboratory conditions. We applied statistical spectral discrimination theory to identify unknown spectra of M. Oryzae, the fungus responsible for rice blast disease. The infrared (IR) region was found to be significantly affected by RB disease. These changes may result in alterations in the absorption, reflection, or emission of infrared radiation by the affected plant tissues. Our research revealed that the protein spectrum in the IR region is impacted by RB disease. In our study, we identified strong correlations in the region (Amide group - I) around X 1064 nm and Y 1300 nm with the Lambda / Lambda derived spectra methods for protein detection. During the stages when the disease is developing, typically from day 3 to day 5, the plant's defense mechanisms are not as effective. This is especially true for the PB-1 variety of rice, which is highly susceptible to rice blast disease. Consequently, the proteins in the plant are adversely affected during this critical time. The spectral contour plot reveals the highly correlated spectral regions 1064 nm and Y 1300 nm associated with RB disease infection. Based on these spectral sensitivities, we developed new spectral disease indices for predicting different stages of disease emergence. The goal of this research is to lay the foundation for future UAV and satellite-based studies aimed at long-term monitoring of RB disease.

Keywords: rice blast, asymptomatic stage, spectral sensitivity, IR

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Authors: Mrinalini Ranjan, Sudheesh Chethil

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Epilepsy is a common neurological disorder characterised by the recurrence of seizures. Electroencephalogram (EEG) signals are complex biomedical signals which exhibit nonlinear and nonstationary behavior. We use two methods 1) Detrended Fluctuation Analysis (DFA) and 2) Recurrence Plots (RP) to capture this complex behavior of EEG signals. DFA considers fluctuation from local linear trends. Scale invariance of these signals is well captured in the multifractal characterisation using detrended fluctuation analysis (DFA). Analysis of long-range correlations is vital for understanding the dynamics of EEG signals. Correlation properties in the EEG signal are quantified by the calculation of a scaling exponent. We report the existence of two scaling behaviours in the epileptic EEG signals which quantify short and long-range correlations. To illustrate this, we perform DFA on extant ictal (seizure) and interictal (seizure free) datasets of different patients in different channels. We compute the short term and long scaling exponents and report a decrease in short range scaling exponent during seizure as compared to pre-seizure and a subsequent increase during post-seizure period, while the long-term scaling exponent shows an increase during seizure activity. Our calculation of long-term scaling exponent yields a value between 0.5 and 1, thus pointing to power law behaviour of long-range temporal correlations (LRTC). We perform this analysis for multiple channels and report similar behaviour. We find an increase in the long-term scaling exponent during seizure in all channels, which we attribute to an increase in persistent LRTC during seizure. The magnitude of the scaling exponent and its distribution in different channels can help in better identification of areas in brain most affected during seizure activity. The nature of epileptic seizures varies from patient-to-patient. To illustrate this, we report an increase in long-term scaling exponent for some patients which is also complemented by the recurrence plots (RP). RP is a graph that shows the time index of recurrence of a dynamical state. We perform Recurrence Quantitative analysis (RQA) and calculate RQA parameters like diagonal length, entropy, recurrence, determinism, etc. for ictal and interictal datasets. We find that the RQA parameters increase during seizure activity, indicating a transition. We observe that RQA parameters are higher during seizure period as compared to post seizure values, whereas for some patients post seizure values exceeded those during seizure. We attribute this to varying nature of seizure in different patients indicating a different route or mechanism during the transition. Our results can help in better understanding of the characterisation of epileptic EEG signals from a nonlinear analysis.

Keywords: detrended fluctuation, epilepsy, long range correlations, recurrence plots

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Authors: Bikash Ranjan Mishra, Lopamudra D. Satpathy

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In the wake of the phenomenal surge in international business in the last decades or more, both the developed and developing economies around the world are in massive competition to attract more and more FDI flows. While the developed countries have marched ahead in the race, the developing countries, especially those of Asian economies, have followed them at a rapid pace. While most of the previous studies have analysed the role of institutional quality in the promotion of FDI flows in developing countries, very few studies have taken an integrated approach of examining the comprehensive impact of institutional quality, globalization pattern and domestic financial development on FDI flows. In this context, the paper contributes to the literature in two important ways. Firstly, two composite indices of institutional quality and domestic financial development for the Asian countries are constructed in comparison to earlier studies that resort to a single variable for indicating the institutional quality and domestic financial development. Secondly, the impact of these variables on FDI flows through their interaction with geographical region is investigated. The study uses panel data covering the time period of 1996 to 2012 by selecting twenty Asian developing countries by emphasizing the quality of institutions from the geographical regions of eastern, south-eastern, southern and western Asia. Control of corruption, better rule of law, regulatory quality, effectiveness of the government, political stability and voice and accountability are used as indicators of institutional quality. Besides these, the study takes into account the domestic credits in the hands of public, private sectors and in stock markets as domestic financial indicators. First in the specification of model, a factor analysis is performed to reduce the vast determinants, which are highly correlated with each other, to a manageable size. Afterwards, a reduced version of the model is estimated with the extracted factors in the form of index as independent variables along with a set of control variables. It is found that the institutional quality index and index of globalization exert a significant effect on FDI inflows of the host countries; in contrast, the domestic financial index does not seem to play much worthy role. Finally, some robustness tests are performed to make sure that the results are not sensitive to temporal and spatial unobserved heterogeneity. On the basis of the above study, one general inference can be drawn from the policy prescription point of view that the government of these developing countries should strengthen their domestic institution, both financial and non-financial. In addition to these, welfare policies should also target for rapid globalization. If the financial and non-financial institutions of these developing countries become sound and grow more globalized in the economic, social and political domain, then they can appeal to more amounts of FDI inflows that will subsequently result in advancement of these economies.

Keywords: Asian developing economies, FDI, institutional quality, panel data

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Authors: Rajesh Ranjan, Marimuthu Palaniswami, A. A. Hashmi

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With the changing lifestyle and environment around us, the prevalence of the critical and incurable disease has proliferated. One such condition is the neurological disorder which is rampant among the old age population and is increasing at an unstoppable rate. Most of the neurological disorder patients suffer from some movement disorder affecting the movement of their body parts. Tremor is the most common movement disorder which is prevalent in such patients that infect the upper or lower limbs or both extremities. The tremor symptoms are commonly visible in Parkinson’s disease patient, and it can also be a pure tremor (essential tremor). The patients suffering from tremor face enormous trouble in performing the daily activity, and they always need a caretaker for assistance. In the clinics, the assessment of tremor is done through a manual clinical rating task such as Unified Parkinson’s disease rating scale which is time taking and cumbersome. Neurologists have also affirmed a challenge in differentiating a Parkinsonian tremor with the pure tremor which is essential in providing an accurate diagnosis. Therefore, there is a need to develop a monitoring and assistive tool for the tremor patient that keep on checking their health condition by coordinating them with the clinicians and caretakers for early diagnosis and assistance in performing the daily activity. In our research, we focus on developing a system for automatic classification of tremor which can accurately differentiate the pure tremor from the Parkinsonian tremor using a wearable accelerometer-based device, so that adequate diagnosis can be provided to the correct patient. In this research, a study was conducted in the neuro-clinic to assess the upper wrist movement of the patient suffering from Pure (Essential) tremor and Parkinsonian tremor using a wearable accelerometer-based device. Four tasks were designed in accordance with Unified Parkinson’s disease motor rating scale which is used to assess the rest, postural, intentional and action tremor in such patient. Various features such as time-frequency domain, wavelet-based and fast-Fourier transform based cross-correlation were extracted from the tri-axial signal which was used as input feature vector space for the different supervised and unsupervised learning tools for quantification of severity of tremor. A minimum covariance maximum correlation energy comparison index was also developed which was used as the input feature for various classification tools for distinguishing the PT and ET tremor types. An automatic system for efficient classification of tremor was developed using feature extraction methods, and superior performance was achieved using K-nearest neighbors and Support Vector Machine classifiers respectively.

Keywords: machine learning approach for neurological disorder assessment, automatic classification of tremor types, feature extraction method for tremor classification, neurological movement disorder, parkinsonian tremor, essential tremor

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Authors: Rahul Ranjan

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Using gender as a category to cull out historical analysis, feminist scholars have produced plethora of literature on the sexual symbolics and carnal practices of modern European empires. At a symbolic level, the penetration and conquest of faraway lands was charged with sexual significance and intrigue. The white male’s domination and possession of dark and fertile lands in Africa, Asia and the Americas offered, in Anne McClintock’s words, ‘a fantastic magic lantern of the mind onto which Europe projected its forbidden sexual desires and fears’. The politics of rape were also symbolically a question significant to the politics of empire. To the colonized subject, rape was a fearsome factor, a language that spoke of violent and voracious nature of imperial exploitation. The colonized often looked at rape as an act which colonizers used as tool of oppression. The rape as act of violence got encoded into the legal structure under the helm of Lord Macaulay in the so called ‘Age of Reform’ in 1860 under IPC (Indian penal code). Initially Lord Macaulay formed Indian Law Commission in 1837 in which he drafted a bill and defined the ‘crime of rape as sexual intercourse by a man to a woman against her will and without her consent , except in cases involving girls under nine years of age where consent was immaterial’. The modern English law of rape formulated under the colonial era introduced twofold issues to the forefront. On the one hand it deployed ‘technical experts’ who wrote textbooks of medical jurisprudence that were used as credential citation to make case more ‘objective’, while on the other hand the presumptions about barbaric subjects, the colonized women’s body that was docile which is prone to adultery reflected in cases. The untrustworthiness of native witness also remained an imperative for British jurists to put extra emphasis making ‘objective’ and ‘presumptuous’. This sort of formulation put women down on the pedestrian of justice because it disadvantaged her doubly through British legality and their thinking about the rape. The Imperial morality that acted as vanguards of women’s chastity coincided language of science propagated in the post-enlightenment which not only annulled non-conformist ideas but also made itself a hegemonic language, was often used as a tool and language in encoding of law. The medico-legal understanding of rape in the colonial India has its clear imprints in the post-colonial legality. The onus on the part of rape’s victim was dictated for the longest time and still continues does by widely referred idea that ‘there should signs, marks of resistance on the body of the victim’ otherwise it is likely to be considered consensual. Having said so, this paper looks at the textual continuity that had prolonged the colonial construct of women’s body and the self.

Keywords: body, politics, textual construct, phallocentric

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Authors: Shreeyam, Ranjan Kumar Sah, Shivangi

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Recognizing and solving handwritten mathematical expressions can be a challenging task, particularly when certain characters are segmented and classified. This project proposes a solution that uses Convolutional Neural Network (CNN) and image processing techniques to accurately solve various types of equations, including arithmetic, quadratic, and trigonometric equations, as well as logical operations like logical AND, OR, NOT, NAND, XOR, and NOR. The proposed solution also provides a graphical solution, allowing users to visualize equations and their solutions. In addition to equation solving, the platform, called CNNCalc, offers a comprehensive learning experience for students. It provides educational content, a quiz platform, and a coding platform for practicing programming skills in different languages like C, Python, and Java. This all-in-one solution makes the learning process engaging and enjoyable for students. The proposed methodology includes horizontal compact projection analysis and survey for segmentation and binarization, as well as connected component analysis and integrated connected component analysis for character classification. The compact projection algorithm compresses the horizontal projections to remove noise and obtain a clearer image, contributing to the accuracy of character segmentation. Experimental results demonstrate the effectiveness of the proposed solution in solving a wide range of mathematical equations. CNNCalc provides a powerful and user-friendly platform for solving equations, learning, and practicing programming skills. With its comprehensive features and accurate results, CNNCalc is poised to revolutionize the way students learn and solve mathematical equations. The platform utilizes a custom-designed Convolutional Neural Network (CNN) with image processing techniques to accurately recognize and classify symbols within handwritten equations. The compact projection algorithm effectively removes noise from horizontal projections, leading to clearer images and improved character segmentation. Experimental results demonstrate the accuracy and effectiveness of the proposed solution in solving a wide range of equations, including arithmetic, quadratic, trigonometric, and logical operations. CNNCalc features a user-friendly interface with a graphical representation of equations being solved, making it an interactive and engaging learning experience for users. The platform also includes tutorials, testing capabilities, and programming features in languages such as C, Python, and Java. Users can track their progress and work towards improving their skills. CNNCalc is poised to revolutionize the way students learn and solve mathematical equations with its comprehensive features and accurate results.

Keywords: AL, ML, hand written equation solver, maths, computer, CNNCalc, convolutional neural networks

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Authors: Debraj Gangopadhyay, Moumita Das, Ranjan K. Singh, Poonam Tandon

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Creatinine is a toxic chemical waste generated from muscle metabolism. Abnormally high levels of creatinine in the body fluid indicate possible malfunction or failure of the kidneys. This leads to a condition termed as creatinine induced nephrotoxicity. N-acetylcysteine is an antioxidant drug which is capable of preventing creatinine induced nephrotoxicity and is helpful to treat renal failure in its early stages. Taurine is another antioxidant drug which serves similar purpose. The kidneys have a natural power that whenever reactive oxygen species radicals increase in the human body, the kidneys make an antioxidant shell so that these radicals cannot harm the kidney function. Taurine plays a vital role in increasing the power of that shell such that the glomerular filtration rate can remain in its normal level. Thus taurine protects the kidneys against several diseases. However, taurine also has some negative effects on the body as its chloramine derivative is a weak oxidant by nature. N-acetylcysteine is capable of inhibiting the residual oxidative property of taurine chloramine. Therefore, N-acetylcysteine is given to a patient along with taurine and this combination is capable of suppressing the negative effect of taurine. Both N-acetylcysteine and taurine being affordable, safe, and widely available medicines, knowledge of the mechanism of their combined effect on creatinine, the favored route of administration, and the proper dose may be highly useful in their use for treating renal patients. Raman spectroscopy is a precise technique to observe minor structural changes taking place when two or more molecules interact. The possibility of formation of a complex between a drug molecule and an analyte molecule in solution can be explored by analyzing the changes in the Raman spectra. The formation of a stable complex of creatinine with N-acetylcysteinein vitroin aqueous solution has been observed with the help of Raman spectroscopic technique. From the Raman spectra of the mixtures of aqueous solutions of creatinine and N-acetylcysteinein different molar ratios, it is observed that the most stable complex is formed at 1:1 ratio of creatinine andN-acetylcysteine. Upon drying, the complex obtained is gel-like in appearance and reddish yellow in color. The complex is hygroscopic and has much better water solubility compared to creatinine. This highlights that N-acetylcysteineplays an effective role in reducing the toxic effect of creatinine by forming this water soluble complex which can be removed through urine. Since the drug taurine is also known to be useful in reducing nephrotoxicity caused by creatinine, the aqueous solution of taurine with those of creatinine and N-acetylcysteinewere mixed in different molar ratios and were investigated by Raman spectroscopic technique. It is understood that taurine itself does not undergo complexation with creatinine as no additional changes are observed in the Raman spectra of creatinine when it is mixed with taurine. However, when creatinine, N-acetylcysteine and taurine are mixed in aqueous solution in molar ratio 1:1:3, several changes occurring in the Raman spectra of creatinine suggest the diminishing toxic effect of creatinine in the presence ofantioxidant drugs N-acetylcysteine and taurine.

Keywords: creatinine, creatinine induced nephrotoxicity, N-acetylcysteine, taurine

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Authors: Ashish Sinha, Pushpend Agrawal

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Background: Patients with undiagnosed pulmonary TB predominantly act as reservoirs for its transmission through 10-15 secondary infections in the next 1-5 Yrs. Delays in the diagnosis and treatment may worsen the disease with increase the risk of death. Factors responsible for such delays by tracking patient pathways to treatment may help in planning better interventions. The provision of ‘free diagnosis and treatment’ forms the cornerstone of the National Tuberculosis Elimination Programme (NTEP). OOPE is defined as the money spent by the patient during TB care other than public health facilities. Free TB care at all health facilities could reduce out-of-pocket expenses to the minimum possible levels. Material and Methods: This cross-sectional study was conducted among randomly selected 252 TB patients from Nov – Oct 2022 by taking in-depth interviews following informed verbal consent. We documented their journey from initial symptoms until they reached the public health facility, along with their ‘out-of-pocket expenditure’ (OOPE) pertaining to TB care. Results: Total treatment delay was 91±72 days on average (median: 77days, IQR: 45-104 days), while the isolated patient delay was 31±45 days (median: 15 days, IQR: 0 days to 43 days); diagnostic delay; 57±60 days (median: 42days, IQR 14-78 days), treatment delay 19 ± 18 days (median: 15days, IQR: 11-19 days). A patient delay (> 30 days) was significantly associated with ignorance about classic symptoms of pulmonary TB, adoption of self-medication, illiteracy, and middle and lower social class. Diagnostic delay was significantly higher among those who contacted private health facilities, were unaware of signs and symptoms, had >2 consultations, and not getting an appropriate referral for TB care. Most (97%) of the study participants interviewed claimed to have incurred some expenditure.Median total expenses were 6155(IQR: 2625-15175) rupees. More than half 141 (56%) of the study participants had expenses >5000 rupees. Median transport expenses were 525(IQR: 200-1012) rupees; Median consultation expenses were 700(IQR: 200-1600) rupees; Median investigation expenses were 1000(IQR: 0-3025) rupees and the Median medicine expenses were 3350(IQR: 1300-7525).OOPE for consultation, investigation, and medicine was observed to be significantly higher among patients who ignored classical signs& symptoms of TB, repeated visits to private health facilities, and due to self-medication practices. Transport expenses and delays in seeking care at facilities were observed to have an upward trend with OOP Expenses (r =1). Conclusion: Delay in TB care due to low awareness about signs and symptoms of TB and poor seeking care, lack of proper consultation, and appropriate referrals reported by the study subjects indicate the areas which need proper attention by the program managers. Despite a centrally sponsored programme, the financial burden on TB patients is still in the unacceptable range. OOPE could be reduced as low as possible by addressing the responsible factors linked to it.

Keywords: patient pathway, delay, pulmonary tuberculosis, out of pocket expenses

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Authors: Ashish Thakur

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This technical paper was written in view of focusing the biomaterials and its various applications in modern industries. Author tires to elaborate not only the medical, infect plenty of application in other industries. The scope of the research area covers the wide range of physical, biological and chemical sciences that underpin the design of biomaterials and the clinical disciplines in which they are used. A biomaterial is now defined as a substance that has been engineered to take a form which, alone or as part of a complex system, is used to direct, by control of interactions with components of living systems, the course of any therapeutic or diagnostic procedure. Biomaterials are invariably in contact with living tissues. Thus, interactions between the surface of a synthetic material and biological environment must be well understood. This paper reviews the benefits and challenges associated with surface modification of the metals in biomedical applications. The paper also elaborates how the surface characteristics of metallic biomaterials, such as surface chemistry, topography, surface charge, and wettability, influence the protein adsorption and subsequent cell behavior in terms of adhesion, proliferation, and differentiation at the biomaterial–tissue interface. The chapter also highlights various techniques required for surface modification and coating of metallic biomaterials, including physicochemical and biochemical surface treatments and calcium phosphate and oxide coatings. In this review, the attention is focused on the biomaterial-associated infections, from which the need for anti-infective biomaterials originates. Biomaterial-associated infections differ markedly for epidemiology, aetiology and severity, depending mainly on the anatomic site, on the time of biomaterial application, and on the depth of the tissues harbouring the prosthesis. Here, the diversity and complexity of the different scenarios where medical devices are currently utilised are explored, providing an overview of the emblematic applicative fields and of the requirements for anti-infective biomaterials. In addition to this, chapter introduces nanomedicine and the use of both natural and synthetic polymeric biomaterials, focuses on specific current polymeric nanomedicine applications and research, and concludes with the challenges of nanomedicine research. Infection is currently regarded as the most severe and devastating complication associated to the use of biomaterials. Osteoporosis is a worldwide disease with a very high prevalence in humans older than 50. The main clinical consequences are bone fractures, which often lead to patient disability or even death. A number of commercial biomaterials are currently used to treat osteoporotic bone fractures, but most of these have not been specifically designed for that purpose. Many drug- or cell-loaded biomaterials have been proposed in research laboratories, but very few have received approval for commercial use. Polymeric nanomaterial-based therapeutics plays a key role in the field of medicine in treatment areas such as drug delivery, tissue engineering, cancer, diabetes, and neurodegenerative diseases. Advantages in the use of polymers over other materials for nanomedicine include increased functionality, design flexibility, improved processability, and, in some cases, biocompatibility.

Keywords: nanomedicine, tissue, infections, biomaterials

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Authors: Md. Samsuddin Ansari, Ashish Arora

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Every year 10 million people fall ill with one of the oldest diseases known as tuberculosis, caused by Mycobacterium tuberculosis. The success of M. tuberculosis as a pathogen is because of its ability to persist in host tissues. Multidrug resistance (MDR) mycobacteria cases increase every day, which is associated with efflux pumps controlled at the level of transcription. The transcription regulators of MDR transporters in bacteria belong to one of the following four regulatory protein families: AraC, MarR, MerR, and TetR. Phenolic acid decarboxylase repressor (PadR), like a family of transcription regulators, is closely related to the MarR family. Phenolic acid decarboxylase repressor (PadR) was first identified as a transcription factor involved in the regulation of phenolic acid stress response in various microorganisms (including Mycobacterium tuberculosis H37Rv). Recently research has shown that the PadR family transcription factors are global, multifunction transcription regulators. Rv0047c is a PadR subfamily-1 protein. We are exploring the biophysical and structural characterization of Rv0047c. The Rv0047 gene was amplified by PCR using the primers containing EcoRI and HindIII restriction enzyme sites cloned in pET-NH6 vector and overexpressed in DH5α and BL21 (λDE3) cells of E. coli following purification with Ni2+-NTA column and size exclusion chromatography. We did DSC to know the thermal stability; the Tm (transition temperature) of protein is 55.29ºC, and ΔH (enthalpy change) of 6.92 kcal/mol. Circular dichroism to know the secondary structure and conformation and fluorescence spectroscopy for tertiary structure study of protein. To understand the effect of pH on the structure, function, and stability of Rv0047c we employed spectroscopy techniques such as circular dichroism, fluorescence, and absorbance measurements in a wide range of pH (from pH-2.0 to pH-12). At low and high pH, it shows drastic changes in the secondary and tertiary structure of the protein. EMSA studies showed the specific binding of Rv0047c with its own 30-bp promoter region. To determine the effect of complex formation on the secondary structure of Rv0047c, we examined the CD spectra of the complex of Rv0047c with promoter DNA of rv0047. The functional role of Rv0047c was characterized by over-expressing the Rv0047c gene under the control of hsp60 promoter in Mycobacterium tuberculosis H37Rv. We have predicted the three-dimensional structure of Rv0047c using the Swiss Model and Modeller, with validity checked by the Ramachandra plot. We did molecular docking of Rv0047c with dnaA, through PatchDock following refinement through FireDock. Through this, it is possible to easily identify the binding hot-stop of the receptor molecule with that of the ligand, the nature of the interface itself, and the conformational change undergone by the protein pattern. We are using X-crystallography to unravel the structure of Rv0047c. Overall the studies show that Rv0047c may have transcription regulation along with providing an insight into the activity of Rv0047c in the pH range of subcellular environment and helps to understand the protein-protein interaction, a novel target to kill dormant bacteria and potential strategy for tuberculosis control.

Keywords: mycobacterium tuberculosis, phenolic acid decarboxylase repressor, Rv0047c, Circular dichroism, fluorescence spectroscopy, docking, protein-protein interaction

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Authors: Ashish Kumar Dwivedi

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Uttar Pradesh is largest part of Indian Federal system encapsulating twenty two million populations and has huge cultural, economic and religious diversity. The newly elected 18 months old state leadership of Uttar Pradesh has envisaged and initiated various proactive strides for the public grievance redressal and inclusive development schemes for all the sections of population from its very day of assumption of the office by Hon’ble Chief Minster Shri Yogi Adtiyanath. These initiatives also include Departmental responses via social media handles as Twitter, Facebook Page, and Web interaction. In the same course, every department of state government has been guided for the correct usage of verified social media handle separately and in co-ordination with other departments. These guidelines included making new WhatsApp groups to connect technocrats and politicians to communicate on common platform. Minister for Department of Infrastructure and Industrial Development, Shri Satish Mahana is a very popular leader and very intuitive statesman, has thousands of followers on social media and his accounts receive almost three hundred individually mentioned notifications from the various parts of Uttar Pradesh. These notifications primarily include problems related to livelihood and grievances concerned to department. To address these communications, a body of five experts has been set who are actively responding on various levels and increase bureaucratic engagements with marginalized sections of society. With reference to above background, this piece of research was conducted to analyze, categorize and derive effective implementation of public policies via social media platforms. This act of responsiveness has brought positive change in the mindset of population for the government, which was missed earlier. Department of Industrial Development in the Government is also inclined to attract investors aiming to become first trillion-dollar economy of India henceforth department also organized two major successful events in last one year. These events were also frame worked on social media platform to update 2.5 million population of state who is actively using social media in many ways. To analyze change scientifically, this study has been conducted and big data has been collected from October 2017 to September 2018 from the departmental social media handles as Twitter, Facebook, and emails. For this data, a statistical study has been conducted to analyze sentiments and expectations, specific and common requirement of communities, nature of grievances and their effective elucidation fitted into government policies. The control sample has also been taken from previous government activities to analyze the change. The statistical study used tools such as correlation study and principal component analysis. Also in this research communication, the modus operandi of grievance redressal, proliferation of government policies, connections to their beneficiaries and quick response procedure will be discussed.

Keywords: correlation study, principal component analysis, bureaucratic engagements, social media

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Authors: Priyanka Jain, Ashish K. Sharma, Esha Shukla, K. J. Mukherjee

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We propose a paradigm shift in our approach to design improved platforms for recombinant protein production, by addressing system level issues rather than the individual steps associated with recombinant protein synthesis like transcription, translation, etc. We demonstrate that by controlling and modulating the cellular stress response (CSR), which is responsible for feedback control of protein synthesis, we can generate hyper-producing strains. We did transcriptomic profiling of post-induction cultures, expressing different types of protein, to analyze the nature of this cellular stress response. We found significant down-regulation of substrate utilization, translation, and energy metabolism genes due to generation CSR inside the host cell. However, transcription profiling has also shown that many genes are up-regulated post induction and their role in modulating the CSR is unclear. We hypothesized that these up-regulated genes trigger signaling pathways, generating the CSR and concomitantly reduce the recombinant protein yield. To test this hypothesis, we knocked out the up-regulated genes, which did not have any downstream regulatees, and analyzed their impact on cellular health and recombinant protein expression. Two model proteins i.e., GFP and L-Asparaginase were chosen for this analysis. We observed a significant improvement in expression levels, with some knock-outs showing more than 7-fold higher expression compared to control. The 10 best single knock-outs were chosen to make 45 combinations of all possible double knock-outs. A further increase in expression was observed in some of these double knock- outs with GFP levels being highest in a double knock-out ΔyhbC + ΔelaA. However, for L-Asparaginase which is a secretory protein, the best results were obtained using a combination of ΔelaA+ΔcysW knock-outs. We then tested all the knock outs for their ability to enhance the expression of a 'difficult-to-express' protein. The Rubella virus E1 protein was chosen and tagged with sfGFP at the C-terminal using a linker peptide for easy online monitoring of expression of this fusion protein. Interestingly, the highest increase in Rubella-sGFP levels was obtained in the same double knock-out ΔelaA + ΔcysW (5.6 fold increase in expression yield compared to the control) which gave the highest expression for L-Asparaginase. However, for sfGFP alone, the ΔyhbC+ΔmarR knock-out gave the highest level of expression. These results indicate that there is a fair degree of commonality in the nature of the CSR generated by the induction of different proteins. Transcriptomic profiling of the double knock out showed that many genes associated with the translational machinery and energy biosynthesis did not get down-regulated post induction, unlike the control where these genes were significantly down-regulated. This confirmed our hypothesis of these genes playing an important role in the generation of the CSR and allowed us to design a strategy for making better expression hosts by simply knocking out key genes. This strategy is radically superior to the previous approach of individually up-regulating critical genes since it blocks the mounting of the CSR thus preventing the down-regulation of a very large number of genes responsible for sustaining the flux through the recombinant protein production pathway.

Keywords: cellular stress response, GFP, knock-outs, up-regulated genes

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Authors: Ankita Baidya, Vanishri Ganakumar, Ranveer S. Jadon, Piyush Ranjan, Rita Sood

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Septic embolism can have varied presentations and clinical considerations. Infected central venous catheters are commonly associated with septic emboli but peripheral vascular catheters are rarely implicated. We describe a rare case of septic pulmonary emboli related to infected peripheral venous cannulation caused by an unusual etiological agent. A young male presented with complaints of fever, productive cough, sudden onset shortness of breath and cellulitis in both the upper limbs. He was recently hospitalised for dengue fever and administered intravenous fluids through peripheral venous line. The patient was febrile, tachypneic and in respiratory distress, there were multiple pus filled bullae in left hand alongwith swelling and erythema involving right forearm that started at the site of cannulation. Chest examination showed active accessory muscles of respiration, stony dull percussion at the base of right lung and decreased breath sounds at right infrascapular, infraaxillary and mammary area. Other system examination was within normal limits. Chest X-ray revealed bilateral multiple patchy heterogenous peripheral opacities and infiltrates with right-sided pleural effusion. Contrast-enhanced computed tomography (CECT) chest showed feeding vessel sign confirming the diagnosis as septic emboli. Venous Doppler and 2D-echocardiogarm were normal. Laboratory findings showed marked leucocytosis (22000/mm3). Pus aspirate, blood sample, and sputum sample were sent for microbiological testing. The patient was started empirically on ceftriaxone, vancomycin, and clindamycin. The Pus culture and sputum culture showed Klebsiella pneumoniae sensitive to cefoperazone-sulbactum, piperacillin-tazobactum, meropenem and amikacin. The antibiotics were modified accordingly to antimicrobial sensitivity profile to Cefoperazone-sulbactum. Bronchoalveolar lavage (BAL) was done and sent for microbiological investigations. BAL culture showed Klebsiella pneumoniae with same antimicrobial resistance profile. On day 6 of starting cefoperazone-sulbactum, he became afebrile. The skin lesions improved significantly. He was administered 2 weeks of cefoperazone–sulbactum and discharged on oral faropenem for 4 weeks. At the time of discharge, TLC was 11200/mm3 with marked radiological resolution of infection and healed skin lesions. He was kept in regular follow up. Chest X-ray and skin lesions showed complete resolution after 8 weeks. Till date, only couple of case reports of septic emboli through peripheral intravenous line have been reported in English literature. This case highlights that a simple procedure of peripheral intravenous cannulation can lead to catastrophic complication of septic pulmonary emboli and widespread cellulitis if not done with proper care and precautions. Also, the usual pathogens in such clinical settings are gram positive bacteria, but with the history of recent hospitalization, empirical therapy should also cover drug resistant gram negative microorganisms. It also emphasise the importance of appropriate healthcare practices to be taken care during all procedures.

Keywords: antibiotics, cannula, Klebsiella pneumoniae, septic emboli

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