Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 22

Search results for: Trilok Mathur

22 Caputo-Type Fuzzy Fractional Riccati Differential Equations with Fuzzy Initial Conditions

Authors: Trilok Mathur, Shivi Agarwal

Abstract:

This paper deals with the solutions of fuzzy-fractional-order Riccati equations under Caputo-type fuzzy fractional derivatives. The Caputo-type fuzzy fractional derivatives are defined based on Hukuhura difference and strongly generalized fuzzy differentiability. The Laplace-Adomian-Pade method is used for solving fractional Riccati-type initial value differential equations of fractional order. Moreover, we also displayed some examples to illustrate our methods.

Keywords: Caputo-type fuzzy fractional derivative, Fractional Riccati differential equations, Laplace-Adomian-Pade method, Mittag Leffler function

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21 Fuzzy Total Factor Productivity by Credibility Theory

Authors: Shivi Agarwal, Trilok Mathur

Abstract:

This paper proposes the method to measure the total factor productivity (TFP) change by credibility theory for fuzzy input and output variables. Total factor productivity change has been widely studied with crisp input and output variables, however, in some cases, input and output data of decision-making units (DMUs) can be measured with uncertainty. These data can be represented as linguistic variable characterized by fuzzy numbers. Malmquist productivity index (MPI) is widely used to estimate the TFP change by calculating the total factor productivity of a DMU for different time periods using data envelopment analysis (DEA). The fuzzy DEA (FDEA) model is solved using the credibility theory. The results of FDEA is used to measure the TFP change for fuzzy input and output variables. Finally, numerical examples are presented to illustrate the proposed method to measure the TFP change input and output variables. The suggested methodology can be utilized for performance evaluation of DMUs and help to assess the level of integration. The methodology can also apply to rank the DMUs and can find out the DMUs that are lagging behind and make recommendations as to how they can improve their performance to bring them at par with other DMUs.

Keywords: chance-constrained programming, credibility theory, data envelopment analysis, fuzzy data, Malmquist productivity index

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20 Policy Innovation and its Determinants: A Literature Review

Authors: Devasheesh Mathur

Abstract:

The presentation reviews the literature on the phenomenon of policy innovation. Policy innovation refers to a shift in the way policy is made or executed. The paper covers comprehensively on the definition and also the various types of policy innovations. The emphasis is on the antecedents or the determinants of innovation in policies. The author has then made an effort to discover the knowledge gap in the field of policy innovation so as to identify the future scope of research. The objective is to lend more clarity in the area of policy innovation and help in creating a framework for policy-makers as well as academics.

Keywords: literature review, policy innovation, determinants, antecedents

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19 Modified Active (MA) Algorithm to Generate Semantic Web Related Clustered Hierarchy for Keyword Search

Authors: G. Leena Giri, Archana Mathur, S. H. Manjula, K. R. Venugopal, L. M. Patnaik

Abstract:

Keyword search in XML documents is based on the notion of lowest common ancestors in the labelled trees model of XML documents and has recently gained a lot of research interest in the database community. In this paper, we propose the Modified Active (MA) algorithm which is an improvement over the active clustering algorithm by taking into consideration the entity aspect of the nodes to find the level of the node pertaining to a particular keyword input by the user. A portion of the bibliography database is used to experimentally evaluate the modified active algorithm and results show that it performs better than the active algorithm. Our modification improves the response time of the system and thereby increases the efficiency of the system.

Keywords: keyword matching patterns, MA algorithm, semantic search, knowledge management

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18 Graphical User Interface Testing by Using Deep Learning

Authors: Akshat Mathur, Sunil Kumar Khatri

Abstract:

This paper presents brief about how the use of Artificial intelligence in respect to GUI testing can reduce workload by using DL-fueled method. This paper also discusses about how graphical user interface and event driven software testing can derive benefits from the use of AI techniques. The use of AI techniques not only reduces the task and work load but also helps in getting better output than manual testing. Although results are same, but the use of Artifical intelligence techniques for GUI testing has proven to provide ideal results. DL-fueled framework helped us to find imperfections of the entire webpage and provides test failure result in a score format between 0 and 1which signifies that are test meets it quality criteria or not. This paper proposes DL-fueled method which helps us to find the genuine GUI bugs and defects and also helped us to scale the existing labour-intensive and skill-intensive methodologies.

Keywords: graphical user interface, GUI, artificial intelligence, deep learning, ML technology

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17 Competitive Advantage: Sustainable or Transient

Authors: Pallavi Thacker, H. P. Mathur

Abstract:

This paper tries to find out from the available literature the status of Competitive Advantage. It has been stated a number of times that firms must strive to attain sustainable competitive advantage; but is the concept of sustainability of advantage still valid in this new diversified and too-rapidly changing world? The paper reaches a conclusion that the answer is “no”. Gone is the time when once attained position could easily be retained forever or at-least for a substantial amount of time. We live in a time which is very much globalised. We are used to a high level of competition from all directions. Technological advances, developed human capital, flexibility and end number of factors make the sustenance of competitive advantage difficult. This paper analyses competitive advantage from the view points of Michael Porter (who talks about sustainability) and Rita Gunther McGrath (who says competitive advantage can no more be sustained). It uses many examples and evidences from papers, journals and news. A research in this area is very much required (especially in a developing country like India) so that industries, firms and people can find out the suitable strategies that match with the changing times.

Keywords: competitive advantage, sustainable, transient, globalisation

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16 Rolling Contact Fatigue Failure Analysis of Ball Bearing in Gear Box

Authors: Piyas Palit, Urbi Pal, Jitendra Mathur, Santanu Das

Abstract:

Bearing is an important machinery part in the industry. When bearings fail to meet their expected life the consequences are increased downtime, loss of revenue and missed the delivery. This article describes the failure of a gearbox bearing in rolling contact fatigue. The investigation consists of visual observation, chemical analysis, characterization of microstructures using optical microscopes and hardness test. The present study also considers bearing life as well as the operational condition of bearings. Surface-initiated rolling contact fatigue, leading to a surface failure known as pitting, is a life-limiting failure mode in many modern machine elements, particularly rolling element bearings. Metallography analysis of crack propagation, crack morphology was also described. Indication of fatigue spalling in the ferrography test was also discussed. The analysis suggested the probable reasons for such kind of failure in operation. This type of spalling occurred due to (1) heavier external loading condition or (2) exceeds its service life.

Keywords: bearing, rolling contact fatigue, bearing life

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15 Synthesis and Characterization of Nickel and Sulphur Sensitized Zinc Oxide Structures

Authors: Ella C. Linganiso, Bonex W. Mwakikunga, Trilock Singh, Sanjay Mathur, Odireleng M. Ntwaeaborwa

Abstract:

The use of nanostructured semiconducting material to catalyze degradation of environmental pollutants still receives much attention to date. One of the desired characteristics for pollutant degradation under ultra-violet visible light is the materials with extended carrier charge separation that allows for electronic transfer between the catalyst and the pollutants. In this work, zinc oxide n-type semiconductor vertically aligned structures were fabricated on silicon (100) substrates using the chemical bath deposition method. The as-synthesized structures were treated with nickel and sulphur. X-ray diffraction, scanning electron microscopy, energy dispersive X-ray spectroscopy were used to characterize the phase purity, structural dimensions and elemental composition of the obtained structures respectively. Photoluminescence emission measurements showed a decrease in both the near band edge emission as well as the defect band emission upon addition of nickel and sulphur with different concentrations. This was attributed to increased charger-carrier-separation due to the presence of Ni-S material on ZnO surface, which is linked to improved charge transfer during photocatalytic reactions.

Keywords: Carrier-charge-separation, nickel, photoluminescence, sulphur, zinc oxide

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14 Introduction of Knowledge Management in a Public Sector Organization in India

Authors: Siddharth Vashisth, Varun Mathur

Abstract:

This review provides an overview of the impact that implementation of various Knowledge Management (KM) strategies has had on the growth of a department in a Public Sector Company in India. In a regulated utility controlled by the government, the growth of an organization such as Hindustan Petroleum Corporation Limited (HPCL) had depended largely on the efficiencies of the systems and its people. However, subsequent to the de-regularization & to the entry of the private competition, the need for a ‘systematic templating’ of knowledge was recognized. This necessitated the introduction of Knowledge Management Centre (KMC). Projects & Pipelines Department (P&P) of HPCL introduced KMC that contributed significantly towards KM by adopting various strategies such as standardization, leveraging information system, competency enhancement, and improvements & innovations. These strategies gave both tangible as well as intangible benefits towards KM. Knowledge, technology & people are the three pillars that need to be catered for effective knowledge management in any organization. In HPCL, the initiative of KMC has served as an intermediary between these three major pillars as each activity of the strategy was centered on them and contributed significantly to their growth and up-gradation, ensuring overall growth of KM in the department.

Keywords: knowledge, knowledge management, public sector organization, standardization, technology, people, skill, information system, innovation, competency, impact

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13 Design and Development of the Force Plate for the Study of Driving-Point Biodynamic Responses

Authors: Vikas Kumar, V. H. Saran, Arpit Mathur, Avik Kathuria

Abstract:

The evaluation of biodynamic responses of the human body to whole body vibration exposure is necessary to quantify the exposure effects. A force plate model has been designed with the help of CAD software, which was investigated by performing the modal, stress and strain analysis using finite element approach in the software. The results of the modal, stress and strain analysis were under the limits for measurements of biodynamic responses to whole body vibration. The physical model of the force plate was manufactured and fixed to the vibration simulator and further used in the experimentation for the evaluation of apparent mass responses of the ten recruited subjects standing in an erect posture exposed to vertical whole body vibration. The platform was excited with sinusoidal vibration at vibration magnitude: 1.0 and 1.5 m/s2 rms at different frequency of 2, 3, 4, 5, 6, 8, 10, 12.5, 16 and 20 Hz. The results of magnitude of normalised apparent mass have shown the trend observed in the many past studies. The peak in the normalised apparent mass has been observed at 4 & 5 Hz frequency of vertical whole body vibration. The nonlinearity with respect to vibration magnitude has been also observed in the normalised apparent mass responses.

Keywords: whole body vibration, apparent mass, modeling, force plate

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12 Acoustic Analysis for Comparison and Identification of Normal and Disguised Speech of Individuals

Authors: Surbhi Mathur, J. M. Vyas

Abstract:

Although the rapid development of forensic speaker recognition technology has been conducted, there are still many problems to be solved. The biggest problem arises when the cases involving disguised voice samples come across for the purpose of examination and identification. Such type of voice samples of anonymous callers is frequently encountered in crimes involving kidnapping, blackmailing, hoax extortion and many more, where the speaker makes a deliberate effort to manipulate their natural voice in order to conceal their identity due to the fear of being caught. Voice disguise causes serious damage to the natural vocal parameters of the speakers and thus complicates the process of identification. The sole objective of this doctoral project is to find out the possibility of rendering definite opinions in cases involving disguised speech by experimentally determining the effects of different disguise forms on personal identification and percentage rate of speaker recognition for various voice disguise techniques such as raised pitch, lower pitch, increased nasality, covering the mouth, constricting tract, obstacle in mouth etc by analyzing and comparing the amount of phonetic and acoustic variation in of artificial (disguised) and natural sample of an individual, by auditory as well as spectrographic analysis.

Keywords: forensic, speaker recognition, voice, speech, disguise, identification

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11 Molecular Portraits: The Role of Posttranslational Modification in Cancer Metastasis

Authors: Navkiran Kaur, Apoorva Mathur, Abhishree Agarwal, Sakshi Gupta, Tuhin Rashmi

Abstract:

Aim: Breast cancer is the most common cancer in women worldwide, and resistance to the current therapeutics, often concurrently, is an increasing clinical challenge. Glycosylation of proteins is one of the most important post-translational modifications. It is widely known that aberrant glycosylation has been implicated in many different diseases due to changes associated with biological function and protein folding. Alterations in cell surface glycosylation, can promote invasive behavior of tumor cells that ultimately lead to the progression of cancer. In breast cancer, there is an increasing evidence pertaining to the role of glycosylation in tumor formation and metastasis. In the present study, an attempt has been made to study the disease associated sialoglycoproteins in breast cancer by using bioinformatics tools. The sequence will be retrieved from UniProt database. A database in the form of a word document was made by a collection of FASTA sequences of breast cancer gene sequence. Glycosylation was studied using yinOyang tool on ExPASy and Differential genes expression and protein analysis was done in context of breast cancer metastasis. The number of residues predicted O-glc NAc threshold containing 50 aberrant glycosylation sites or more was detected and recorded for individual sequence. We found that the there is a significant change in the expression profiling of glycosylation patterns of various proteins associated with breast cancer. Differential aberrant glycosylated proteins in breast cancer cells with respect to non-neoplastic cells are an important factor for the overall progression and development of cancer.

Keywords: breast cancer, bioinformatics, cancer, metastasis, glycosylation

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10 Pathological Observations of Intestinal Coccidiosis in Camel (Camelus dromedarius)

Authors: Abhilasha Dadhich, Manisha Mathur, Sanjay Kumar, Hemant Dadhich

Abstract:

The camel (Camelus dromedarius) is an important animal component of the fragile desert eco-system of India. Apart from others, impaired milk and meat production decrease in performance and even death are some of the major consequences of parasitic disease like coccidiosis in camel. Coccidiosis which is an acute invasion and destruction of intestinal mucosa by protozoa of the genera Eimeria or isospora spp. Post-Mortem examinations of 5 carcasses of dromedary of different age groups aged from 2 to 5 years were conducted. The history indicated that the camels were suffering from diarrhoea, dysentery, pyrexia, inappetence, weight loss, and emaciation. Post mortem examinations showed macroscopic and microscopic alterations in the small intestine, particularly in jejunum and ileum regions. The mucosae were congested, and haemorrhagic on which there were numerous whitish-grey nodular foci were observed. The affected intestinal tissue specimens were preserved in 10% formal saline and processed mechanically for paraffin embedding by acetone and benzene technique. The sections were stained with haematoxylin and eosin method of staining for histopathological examinations. Histologically, typical lesions such as congestion and haemorrhages were present. The intestinal villi were oedematous; mucosa degenerated and desquamated, along with infiltration of eosinophils and macrophages. Crypts of lieberkuhn were obliterated due to presence of schizonts in lamina propria. Older camels served as the source of spread of coccidial infection and were also predisposed to secondary infections.

Keywords: camel, coccidiosis, Eimeria, histopathology

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9 Failure of Cable Reel Flat Spring of Crane: Beyond Fatigue Life Use

Authors: Urbi Pal, Piyas Palit, Jitendra Mathur, Abhay Chaturvedi, Sandip Bhattacharya

Abstract:

The hot rolled slab lifting crane cable reel drum (CRD) failed due to failure of cable reel flat spring which are inside the cassette of CRD. CRD is used for the movement of tong cable. Stereoscopic observation revealed beach marks and Scanning Electron Microscopy showed striations confirming fatigue mode of failure. Chemical composition should be spring steel (Cr-Mo-V) as per IS 3431:1982 instead of C-Mn steel. To find out the reason of fatigue failure, the theoretical fatigue life of flat spiral spring has been calculated. The calculation of number of fatigue cycles included bending moment, maximum stress on the spring, ultimate tensile strength and alternative stress. The bending moment determination has been taken account with various parameters like Young’s Modulus, width, thickness, outer diameter, arbor diameter, pay out the length and angular deflection in rotations. With all the required data, the calculated fatigue life turned to be 10000 cycles, but the spring served 15000 cycles which clearly indicated beyond fatigue life usage. Different UTS values have been plotted with respect to the number of fatigue cycles and clearly showed that the increase in UTS by 40% increases fatigue life by 50%. The significance of higher UTS lied here, and higher UTS depends on modified chemistry with proper tempered martensite microstructure. This kind of failure can be easily avoided by changing the crane spring maintenance schedule from 2 years to 1.5 years considering 600 cycles per month. The plant has changed changing the schedule of cable reel spring and procured new flat reel spring made of 50CrV2 steel.

Keywords: cable reel spring, fatigue life, stress, spring steel

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8 Multi-Criteria Optimal Management Strategy for in-situ Bioremediation of LNAPL Contaminated Aquifer Using Particle Swarm Optimization

Authors: Deepak Kumar, Jahangeer, Brijesh Kumar Yadav, Shashi Mathur

Abstract:

In-situ remediation is a technique which can remediate either surface or groundwater at the site of contamination. In the present study, simulation optimization approach has been used to develop management strategy for remediating LNAPL (Light Non-Aqueous Phase Liquid) contaminated aquifers. Benzene, toluene, ethyl benzene and xylene are the main component of LNAPL contaminant. Collectively, these contaminants are known as BTEX. In in-situ bioremediation process, a set of injection and extraction wells are installed. Injection wells supply oxygen and other nutrient which convert BTEX into carbon dioxide and water with the help of indigenous soil bacteria. On the other hand, extraction wells check the movement of plume along downstream. In this study, optimal design of the system has been done using PSO (Particle Swarm Optimization) algorithm. A comprehensive management strategy for pumping of injection and extraction wells has been done to attain a maximum allowable concentration of 5 ppm and 4.5 ppm. The management strategy comprises determination of pumping rates, the total pumping volume and the total running cost incurred for each potential injection and extraction well. The results indicate a high pumping rate for injection wells during the initial management period since it facilitates the availability of oxygen and other nutrients necessary for biodegradation, however it is low during the third year on account of sufficient oxygen availability. This is because the contaminant is assumed to have biodegraded by the end of the third year when the concentration drops to a permissible level.

Keywords: groundwater, in-situ bioremediation, light non-aqueous phase liquid, BTEX, particle swarm optimization

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7 Dealing with Buckling Effect in Snorkel by Finite Element Analysis: A Life Enhancement Approach in CAS-OB Operation

Authors: Subodh Nath Patel, Raja Raman, Mananshi Adhikary, Jitendra Mathur, Sandip Bhattacharyya

Abstract:

The composition adjustment by sealed argon bubbling–oxygen blowing (CAS-OB) process is a process designed for adjusting steel composition and temperature during secondary metallurgy. One of the equipment in the said process is a snorkel or bell, fixed to a movable bracket. Snorkel serves the purpose of feeding ferroalloys into the liquid metal simultaneously removing gases to the gas cleaning system through its port at its top. The bell-shaped snorkel consists of two parts. The upper part has an inside liner, and the lower part is lined on both side with high-alumina castable reinforced with 2% stainless steel needles. Both the parts are coupled with a flange bolt system. These flanges were found to get buckled during operation, and the gap was generating between them. This problem was chronic since its. It was expected to give a life of 80 heats, but it was failing within 45-50 heats. After every 25-30 heats, it had to be repaired by changing and/or tightening its nuts and bolts. Visual observation, microstructural analysis through optical microscopes and SEM, hardness measurement and thermal strain calculation were carried out to find out the root cause of this problem. The calculated thermal strain was compared with actual thermal strain; comparison of the two revealed that thermal strain was responsible for buckling. Finite element analysis (FEA) was carried out to reaffirm the effect temperature on the flanges. FEA was also used in the modification in the design of snorkel flange to accommodate thermal strain. Thermal insulation was also recommended which increased its life from 45 heats to 65 heats, impacting business process positively.

Keywords: CAS OB process, finite element analysis, snorkel, thermal strain

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6 A Feasibility Study of Waste (d) Potential: Synergistic Effect Evaluation by Co-digesting Organic Wastes and Kinetics of Biogas Production

Authors: Kunwar Paritosh, Sanjay Mathur, Monika Yadav, Paras Gandhi, Subodh Kumar, Nidhi Pareek, Vivekanand Vivekanand

Abstract:

A significant fraction of energy is wasted every year managing the biodegradable organic waste inadequately as development and sustainability are the inherent enemies. The management of these waste is indispensable to boost its optimum utilization by converting it to renewable energy resource (here biogas) through anaerobic digestion and to mitigate greenhouse gas emission. Food and yard wastes may prove to be appropriate and potential feedstocks for anaerobic co-digestion for biogas production. The present study has been performed to explore the synergistic effect of co-digesting food waste and yard trimmings from MNIT campus for enhanced biogas production in different ratios in batch tests (37±10C, 90 rpm, 45 days). The results were overwhelming and showed that blending two different organic waste in proper ratio improved the biogas generation considerably, with the highest biogas yield (2044±24 mLg-1VS) that was achieved at 75:25 of food waste to yard waste ratio on volatile solids (VS) basis. The yield was 1.7 and 2.2 folds higher than the mono-digestion of food or yard waste (1172±34, 1016±36mLg-1VS) respectively. The increase in biogas production may be credited to optimum C/N ratio resulting in higher yield. Also Adding TiO2 nanoparticles showed virtually no effect on biogas production as sometimes nanoparticles enhance biogas production. ICP-MS, FTIR analysis was carried out to gain an insight of feedstocks. Modified Gompertz and logistics models were applied for the kinetic study of biogas production where modified Gompertz model showed goodness-of-fit (R2=0.9978) with the experimental results.

Keywords: anaerobic co-digestion, biogas, kinetics, nanoparticle, organic waste

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5 Methicillin Resistant Staphylococcus aureus Specific Bacteriophage Isolation from Sewage Treatment Plant and in vivo Analysis of Phage Efficiency in Swiss Albino Mice

Authors: Pratibha Goyal, Nupur Mathur, Anuradha Singh

Abstract:

Antibiotic resistance is the worldwide threat to human health in this century. Excessive use of antibiotic after their discovery in 1940 makes certain bacteria to become resistant against antibiotics. Most common antibiotic-resistant bacteria include Staphylococcus aureus, Salmonella typhi, E.coli, Klebsiella pneumonia, and Streptococcus pneumonia. Among all Staphylococcus resistant strain called Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for several lives threatening infection in human commonly found in the hospital environment. Our study aimed to isolate bacteriophage against MRSA from the hospital sewage treatment plant and to analyze its efficiency In Vivo in Swiss albino mice model. Sewage sample for the isolation of bacteriophages was collected from SDMH hospital sewage treatment plant in Jaipur. Bacteriophages isolated by the use of enrichment technique and after characterization, isolated phages used to determine phage treatment efficiency in mice. Mice model used to check the safety and suitability of phage application in human need which in turn directly support the use of natural bacteriophage rather than synthetic chemical to kill pathogens. Results show the plaque formation in-vitro and recovery of MRSA infected mice during the experiment. Favorable lytic efficiency determination of MRSA and Salmonella presents a natural way to treat lethal infections caused by Multidrug-resistant bacteria by using their natural host-pathogen relationship.

Keywords: antibiotic resistance, bacteriophages, methicillin resistance Staphylococcus aureus, pathogens, phage therapy, Salmonella typhi

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4 Exploring the Prebiotic Potential of Glucosamine

Authors: Shilpi Malik, Ramneek Kaur, Archita Gupta, Deepshikha Yadav, Ashwani Mathur, Manisha Singh

Abstract:

Glucosamine (GS) is the most abundant naturally occurring amino monosaccharide and is normally produced in human body via cellular glucose metabolism. It is regarded as the building block of cartilage matrix and is also an essential component of cartilage matrix repair mechanism. Besides that, it can also be explored for its prebiotic potential as many bacterial species are known to utilize the amino sugar by acquiring them to form peptidoglycans and lipopolysaccharides in the bacterial cell wall. Glucosamine can therefore be considered for its fermentation by bacterial species present in the gut. Current study is focused on exploring the potential of glucosamine as prebiotic. The studies were done to optimize considerable concentration of GS to reach GI tract and being fermented by the complex gut microbiota and food grade GS was added to various Simulated Fluids of Gastro-Intestinal Tract (GIT) such as Simulated Saliva, Gastric Fluid (Fast and Fed State), Colonic fluid, etc. to detect its degradation. Since it was showing increase in microbial growth (CFU) with time, GS was Further, encapsulated to increase its residential time in the gut, which exhibited improved resistance to the simulated Gut conditions. Moreover, prepared microspehres were optimized and characterized for their encapsulation efficiency and toxicity. To further substantiate the prebiotic activity of Glucosamine, studies were also performed to determine the effect of Glucosamine on the known probiotic bacterial species, i.e. Lactobacillus delbrueckii (MTCC 911) and Bifidobacteriumbifidum (MTCC 5398). Culture conditions for glucosamine will be added in MRS media in anaerobic tube at 0.20%, 0.40%, 0.60%, 0.80%, and 1.0%, respectively. MRS media without GS was included in this experiment as the control. All samples were autoclaved at 118° C for 15 min. Active culture was added at 5% (v/v) to each anaerobic tube after cooling to room temperature and incubated at 37° C then determined biomass and pH and viable count at incubation 18h. The experiment was completed in triplicate and the results were presented as Mean ± SE (Standard error).The experimental results are conclusive and suggest Glucosamine to hold prebiotic properties.

Keywords: gastro intestinal tract, microspheres, peptidoglycans, simulated fluid

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3 An Attenuated Quadruple Gene Mutant of Mycobacterium tuberculosis Imparts Protection against Tuberculosis in Guinea Pigs

Authors: Shubhita Mathur, Ritika Kar Bahal, Priyanka Chauhan, Anil K. Tyagi

Abstract:

Mycobacterium tuberculosis, the causative agent of human tuberculosis, is a major cause of mortality. Bacillus Calmette-Guérin (BCG), the only licensed vaccine available for protection against tuberculosis confers highly variable protection ranging from 0%-80%. Thus, novel vaccine strains need to be evaluated for their potential as a vaccine against tuberculosis. We had previously constructed a triple gene mutant of M. tuberculosis (MtbΔmms), having deletions in genes encoding for phosphatases mptpA, mptpB, and sapM that are involved in host-pathogen interaction. Though vaccination with Mtb∆mms strain induced protection in the lungs of guinea pigs, the mutant strain was not able to control the hematogenous spread of the challenge strain to the spleens. Additionally, inoculation with Mtb∆mms resulted in some pathological damage to the spleens in the early phase of infection. In order to overcome the pathology caused by MtbΔmms in the spleens of guinea pigs and also to control the dissemination of the challenge strain, MtbΔmms was genetically modified by disrupting bioA gene to generate MtbΔmmsb strain. Further, in vivo attenuation of MtbΔmmsb was evaluated, and its protective efficacy was assessed against virulent M. tuberculosis challenge in guinea pigs. Our study demonstrates that Mtb∆mmsb mutant was highly attenuated for growth and virulence in guinea pigs. Vaccination with Mtb∆mmsb mutant generated significant protection in comparison to sham-immunized animals at 4 and 12 weeks post-infection in lungs and spleens of the infected animals. Our findings provide evidence that deletion of genes involved in signal transduction and biotin biosynthesis severely attenuates the pathogen and the single immunization with the auxotroph was able to provide significant protection as compared to sham-immunized animals. The protection imparted by Mtb∆mmsb fell short in comparison to the protection observed in BCG-immunized animals. This study nevertheless indicates the importance of attenuated multiple gene deletion mutants of M. tuberculosis in generating protection against tuberculosis.

Keywords: Mycobacterium tuberculosis, BCG, MtbΔmmsb, bioA, guinea pigs

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2 Clostridium thermocellum DBT-IOC-C19, A Potential CBP Isolate for Ethanol Production

Authors: Nisha Singh, Munish Puri, Collin Barrow, Deepak Tuli, Anshu S. Mathur

Abstract:

The biological conversion of lignocellulosic biomass to ethanol is a promising strategy to solve the present global crisis of exhausting fossil fuels. The existing bioethanol production technologies have cost constraints due to the involvement of mandate pretreatment and extensive enzyme production steps. A unique process configuration known as consolidated bioprocessing (CBP) is believed to be a potential cost-effective process due to its efficient integration of enzyme production, saccharification, and fermentation into one step. Due to several favorable reasons like single step conversion, no need of adding exogenous enzymes and facilitated product recovery, CBP has gained the attention of researchers worldwide. However, there are several technical and economic barriers which need to be overcome for making consolidated bioprocessing a commercially viable process. Finding a natural candidate CBP organism is critically important and thermophilic anaerobes are preferred microorganisms. The thermophilic anaerobes that can represent CBP mainly belong to genus Clostridium, Caldicellulosiruptor, Thermoanaerobacter, Thermoanaero bacterium, and Geobacillus etc. Amongst them, Clostridium thermocellum has received increased attention as a high utility CBP candidate due to its highest growth rate on crystalline cellulose, the presence of highly efficient cellulosome system and ability to produce ethanol directly from cellulose. Recently with the availability of genetic and molecular tools aiding the metabolic engineering of Clostridium thermocellum have further facilitated the viability of commercial CBP process. With this view, we have specifically screened cellulolytic and xylanolytic thermophilic anaerobic ethanol producing bacteria, from unexplored hot spring/s in India. One of the isolates is a potential CBP organism identified as a new strain of Clostridium thermocellum. This strain has shown superior avicel and xylan degradation under unoptimized conditions compared to reported wild type strains of Clostridium thermocellum and produced more than 50 mM ethanol in 72 hours from 1 % avicel at 60°C. Besides, this strain shows good ethanol tolerance and growth on both hexose and pentose sugars. Hence, with further optimization this new strain could be developed as a potential CBP microbe.

Keywords: Clostridium thermocellum, consolidated bioprocessing, ethanol, thermophilic anaerobes

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1 Amifostine Analogue, Drde-30, Attenuates Radiation-Induced Lung Injury in Mice

Authors: Aastha Arora, Vikas Bhuria, Saurabh Singh, Uma Pathak, Shweta Mathur, Puja P. Hazari, Rajat Sandhir, Ravi Soni, Anant N. Bhatt, Bilikere S. Dwarakanath

Abstract:

Radiotherapy is an effective curative and palliative option for patients with thoracic malignancies. However, lung injury, comprising of pneumonitis and fibrosis, remains a significant clin¬ical complication of thoracic radiation, thus making it a dose-limiting factor. Also, injury to the lung is often reported as part of multi-organ failure in victims of accidental radiation exposures. Radiation induced inflammatory response in the lung, characterized by leukocyte infiltration and vascular changes, is an important contributing factor for the injury. Therefore, countermeasure agents to attenuate radiation induced inflammatory response are considered as an important approach to prevent chronic lung damage. Although Amifostine, the widely used, FDA approved radio-protector, has been found to reduce the radiation induced pneumonitis during radiation therapy of non-small cell lung carcinoma, its application during mass and field exposure is limited due to associated toxicity and ineffectiveness with the oral administration. The amifostine analogue (DRDE-30) overcomes this limitation as it is orally effective in reducing the mortality of whole body irradiated mice. The current study was undertaken to investigate the potential of DRDE-30 to ameliorate radiation induced lung damage. DRDE-30 was administered intra-peritoneally, 30 minutes prior to 13.5 Gy thoracic (60Co-gamma) radiation in C57BL/6 mice. Broncheo- alveolar lavage fluid (BALF) and lung tissues were harvested at 12 and 24 weeks post irradiation for studying inflammatory and fibrotic markers. Lactate dehydrogenase (LDH) leakage, leukocyte count and protein content in BALF were used as parameters to evaluate lung vascular permeability. Inflammatory cell signaling (p38 phosphorylation) and anti-oxidant status (MnSOD and Catalase level) was assessed by Western blot, while X-ray CT scan, H & E staining and trichrome staining were done to study the lung architecture and collagen deposition. Irradiation of the lung increased the total protein content, LDH leakage and total leukocyte count in the BALF, reflecting endothelial barrier dysfunction. These disruptive effects were significantly abolished by DRDE-30, which appear to be linked to the DRDE-30 mediated abrogation of activation of the redox-sensitive pro- inflammatory signaling cascade, the MAPK pathway. Concurrent administration of DRDE-30 with radiation inhibited radiation-induced oxidative stress by strengthening the anti-oxidant defense system and abrogated p38 mitogen-activated protein kinase activation, which was associated with reduced vascular leak and macrophage recruitment to the lungs. Histopathological examination (by H & E staining) of the lung showed radiation-induced inflammation of the lungs, characterized by cellular infiltration, interstitial oedema, alveolar wall thickening, perivascular fibrosis and obstruction of alveolar spaces, which were all reduced by pre-administration of DRDE-30. Structural analysis with X-ray CT indicated lung architecture (linked to the degree of opacity) comparable to un-irradiated mice that correlated well with the lung morphology and reduced collagen deposition. Reduction in the radiation-induced inflammation and fibrosis brought about by DRDE-30 resulted in a profound increase in animal survival (72 % in the combination vs 24% with radiation) observed at the end of 24 weeks following irradiation. These findings establish the potential of the Amifostine analogue, DRDE-30, in reducing radiation induced pulmonary injury by attenuating the inflammatory and fibrotic responses.

Keywords: amifostine, fibrosis, inflammation, lung injury radiation

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