Search results for: Mukhopadayay Asish K.
8 Characterization of Enterotoxigenic Escherichia coli CS6 Promoter
Authors: Mondal Indranil, Bhakat Debjyoti, Mukhopadayay Asish K., Chatterjee Nabendu S.
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CS6 is the prevalent CF in our region and deciphering its molecular regulators would play a pivotal role in reducing the burden of ETEC pathogenesis. In prokaryotes, most of the genes are under the control of one operon and the promoter present upstream of the gene regulates the transcription of that gene. Here the promoter of CS6 was characterized by computational method and further analyzed by β-galactosidase assay and sequencing. Promoter constructs and deletions were prepared as required to analyze promoter activity. The effect of different additives on the CS6 promoter was analysed by the β-galactosidase assay. Bioinformatics analysis done by Softberry/BPROM predicted fur, lrp, and crp boxes, -10 and -35 region upstream of the CS6 gene. The promoter construction in no promoter plasmid pTL61T showed that region -573 to +1 is actually the promoter region as predicted. Sequential deletion of the region upstream of CS6 revealed that promoter activity remains the same when -573bp to -350bp is deleted. But after the deletion of the upstream region -350 bp to -255bp, promoter expression decreases drastically to 26%. Further deletion also decreases promoter activity up to a little range. So the region -355bp to -255bp holds the promoter sequence for the CS6 gene. Additives like iron, NaCl, etc., modulate promoter activity in a dose-dependent manner. From the promoter analysis, it can be said that the minimum region lies between -254 and +1. Important region(s) lies between -350 bp to -255 bp upstream in the promoter, which might have important elements needed to control CS6 gene expression.Keywords: microbiology, promoter, colonization factor, ETEC
Procedia PDF Downloads 1627 Understanding the Prevalence and Expression of Virulence Factors Harbored by Enterotoxigenic Escherichia Coli
Authors: Debjyoti Bhakat, Indranil Mondal, Asish K. Mukhopadayay, Nabendu S. Chatterjee
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Enterotoxigenic Escherichia coli is one of the leading causes of diarrhea in infants and travelers in developing countries. Colonization factors play an important role in pathogenesis and are one of the main targets for Enterotoxigenic Escherichia coli (ETEC) vaccine development. However, ETEC vaccines had poorly performed in the past, as the prevalence of colonization factors is region-dependent. There are more than 25 classical colonization factors presently known to be expressed by ETEC, although all are not expressed together. Further, there are other multiple non-classical virulence factors that are also identified. Here the presence and expression of common classical and non-classical virulence factors were studied. Further studies were done on the expression of prevalent colonization factors in different strains. For the prevalence determination, multiplex polymerase chain reaction (PCR) was employed, which was confirmed by simplex PCR. Quantitative RT-PCR was done to study the RNA expression of these virulence factors. Strains negative for colonization factors expression were confirmed by SDS-PAGE. Among the clinical isolates, the most prevalent toxin was est+elt, followed by est and elt, while the pattern was reversed in the control strains. There were 29% and 40% strains negative for any classical colonization factors (CF) or non-classical virulence factors (NCVF) among the clinical and control strains, respectively. Among CF positive ETEC strains, CS6 and CS21 were the prevalent ones in the clinical strains, whereas in control strains, CS6 was the predominant one. For NCVF genes, eatA was the most prevalent among the clinical isolates and etpA for control. CS6 was the most expressed CF, and eatA was the predominantly expressed NCVF for both clinical and controlled ETEC isolates. CS6 expression was more in strains having CS6 alone. Different strains express CS6 at different levels. Not all strains expressed their respective virulence factors. Understanding the prevalent colonization factor, CS6, and its nature of expression will contribute to designing an effective vaccine against ETEC in this region of the globe. The expression pattern of CS6 also will help in examining the relatedness between the ETEC subtypes.Keywords: classical virulence factors, CS6, diarrhea, enterotoxigenic escherichia coli, expression, non-classical virulence factors
Procedia PDF Downloads 1546 Effect of Incentives on Knowledge Sharing and Learning: Evidence from the Indian IT Sector
Authors: Asish O. Mathew, Lewlyn L. R. Rodrigues
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The organizations in the knowledge economy era have recognized the importance of building knowledge assets for sustainable growth and development. In comparison to other industries, Information Technology (IT) enterprises, holds an edge in developing an effective Knowledge Management (KM) program, thanks to their in-house technological abilities. This paper tries to study the various knowledge-based incentive programs and its effect on Knowledge Sharing and Learning in the context of the Indian IT sector. A conceptual model is developed linking KM incentives, knowledge sharing, and learning. A questionnaire study is conducted to collect primary data from the knowledge workers of the IT organizations located in India. The data was analysed using Structural Equation Modeling using Partial Least Square method. The results show a strong influence of knowledge management incentives on knowledge sharing and an indirect influence on learning.Keywords: knowledge management, knowledge management incentives, knowledge sharing, learning
Procedia PDF Downloads 4755 A Comparative Study on Achievement Motivation and Sports Competition Anxiety among the Students of Different Tier of Academic Hierarchy
Authors: Nitai Biswas, Prasenjit Kapas, Arumay Jana, Asish Paul
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Introduction: Motivation is basic drive for all kinds of action. It has direct influence on academic achievement and sports performance that builds urge to incentive values of success. In other words, it can be defined as the need for success to attain excellence. Anxiety in pre competition especially in sports formulates positive inward settings in mind to overcome the challenge. There is a tendency to perceive competitive situations as some threatening issues and to respond them with feelings of apprehension and tension. Aim: Aim of the study was to compare the achievement motivation and competition anxiety among three different classes of students. Methods and Materials: To conduct the study the researcher has taken 131 male subjects from three different classes as Extra Department, Bachelor of Physical Education-I and Master of Physical EducationII, aged 19-28 years. Achievement motivation and sports competition anxiety were measured by the questionnaire. To analyze the data mean, standard deviation for each parameter as descriptive statistics and one way analysis of variance as inferential statistics were employed. Results: From the result of the study in achievement motivation (p ≥ 0.05) and competition anxiety (p ≥ 0.05) no significant differences were found among the said three groups. Conclusion: The study concluded that all three groups had almost the same state of achievement motivation and sports competition anxiety.Keywords: anxiety, sports psychology, sports competition anxiety, achievement motivation, academic hierarchy, E.D., B.P.Ed., M.P.Ed
Procedia PDF Downloads 1434 Association of Preoperative Pain Catastrophizing with Postoperative Pain after Lower Limb Trauma Surgery
Authors: Asish Subedi, Krishna Pokharel, Birendra Prasad Sah, Pashupati Chaudhary
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Objectives: To evaluate an association between preoperative Nepali pain catastrophizing scale (N-PCS) scores and postoperative pain intensity and total opioid consumption. Methods: In this prospective cohort study we enrolled 135 patients with an American Society of Anaesthesiologists physical status I or II, aged between 18 and 65 years, and scheduled for surgery for lower-extremity fracture under spinal anaesthesia. Maximum postoperative pain reported during the 24 h was classified into two groups, no-mild pain group (Numeric rating scale [NRS] scores 1 to 3) and a moderate-severe pain group (NRS 4-10). The Spearman correlation coefficient was used to compare the association between the baseline N-PCS scores and outcome variables, i.e., the maximum NRS pain score and the total tramadol consumption within the first 24 h after surgery. Logistic regression models were used to identify the predictors for the intensity of postoperative pain. Results: As four patients violated the protocol, the data of 131 patients were analysed. Mean N-PCS scores reported by the moderate-severe pain group was 27.39 ±9.50 compared to 18.64 ±10 mean N-PCS scores by the no-mild pain group (p<0.001). Preoperative PCS scores correlated positively with postoperative pain intensity (r =0.39, [95% CI 0.23-0.52], p<0.001) and total tramadol consumption (r =0.32, [95% CI 0.16-0.47], p<0.001). An increase in catastrophizing scores was associated with postoperative moderate-severe pain (odds ratio, 1.08 [95% confidence interval, 1.02-1.15], p=0.006) after adjusting for gender, ethnicity and preoperative anxiety. Conclusion: Patients who reported higher pain catastrophizing preoperatively were at increased risk of experiencing moderate-severe postoperative pain.Keywords: nepali, pain catastrophizing, postoperative pain, trauma
Procedia PDF Downloads 1203 High Prevalence of Multi-drug Resistant Diarrheagenic Escherichia coli among Hospitalised Diarrheal Patients in Kolkata, India
Authors: Debjani Ghosh, Goutam Chowdhury, Prosenjit Samanta, Asish Kumar Mukhopadhyay
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Acute diarrhoea caused by diarrheagenic Escherichia coli (DEC) is one of the major public health problem in developing countries, mainly in Asia and Africa. DEC consists of six pathogroups, but the majority of the cases were associated with the three pathogropus, enterotoxigenic E. coli (ETEC), enteroaggregative E. coli (EAEC), and enteropathogenic E. coli (EPEC). Hence, we studied the prevalence and antimicrobial resistance of these three major DEC pathogroups in hospitalized diarrheal patients in Kolkata, India, during 2012-2019 with a large sample size. 8,891 stool samples were processed, and 7.8% of them was identified as DEC infection screened by multiplex PCR, in which ETEC was most common (47.7%) followed by EAEC (38.4%) and EPEC (13.9%). Clinical patient history suggested that children <5 years of age were mostly affected with ETEC and EAEC, whereas people within >5-14 years of age were significantly associated with EPEC and ETEC infections. Antibiogram profile showed a high prevalence of multidrug resistant (MDR) isolates among DEC (56.9%), in which 9% were resistant to antibiotics of six different antimicrobial classes. Screening of the antibiotic resistance conferring genes in DEC showed the presence of blaCTX-M (30.2%) in highest number followed by blaTEM (27.5%), tetB (18%), sul2 (12.6%), strA (11.8%), aadA1 (9.8%), blaOXA-1 (9%), dfrA1 (1.6%) and blaSHV (1.2%) which indicates the existence of mobile genetic elements in those isolates. Therefore, the presence of MDR DEC strains in higher number alarms the public health authorities to take preventive measures before the upsurge of the DEC caused diarrhea cases in near future.Keywords: diarrheagenic escherichia coli, ETEC, EAEC, EPEC
Procedia PDF Downloads 1602 Genetic and Phenotypic Variability Among the Vibrio Cholerae O1 Isolates of India
Authors: Sreeja Shaw, Prosenjit Samanta, Asish Kumar Mukhopadhyay
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Cholera is still a global public health burden and is caused by Vibrio cholerae O1 and O139 serogroups. Evidence from recent outbreaks in Haiti and Yemen suggested that circulating V. cholerae O1 El Tor variant strains are continuously changing to cause more ruinous outbreaks worldwide, and most of them have emerged from the Indian subcontinents. Therefore, we studied the changing virulence characteristics along with the antibiotic resistance profile of V. cholerae O1strains isolated from seasonal outbreaks in three cholera endemic regions during 2018, Gujarat and Maharashtra in Western India (87 strains), and to compare those features with the isolates of West Bengal in Eastern India (48 strains) collected during the same period. All the strains from Western India were of Ogawa serotype, polymyxin B-sensitive, hemolytic, and contained a large fragment deletion in VSP-II genomic region similar with Yemen outbreak strains and carried more virulent Haitian genetic alleles of major virulence associated genes ctxB, tcpA, and rtxA. Conversely, 14.6% (7/48) of the strains from Eastern India were belong to the Inaba serotype, polymyxin B-resistant, non-hemolytic, harbored intact VSP-II region, classical ctxB, Haitian tcpA, and El Tor rtxA alleles. Interestingly, resistance to tetracycline and chloramphenicol was seen in isolates from both regions, which are not very common among V. cholerae O1 isolates in India. Therefore, this study indicated West Bengal as a diverse region where two different types of El Tor variant hypervirulent strains are co-existed, probably competing for their better environmental survival, which may result in severe irrepressible disease outcome in the future.Keywords: cholera, vibrio cholerae, polymyxin B, Non-hemolytic, ctxB, tcpA, rtxA, VSP-II
Procedia PDF Downloads 1651 Empirical Modeling and Optimization of Laser Welding of AISI 304 Stainless Steel
Authors: Nikhil Kumar, Asish Bandyopadhyay
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Laser welding process is a capable technology for forming the automobile, microelectronics, marine and aerospace parts etc. In the present work, a mathematical and statistical approach is adopted to study the laser welding of AISI 304 stainless steel. A robotic control 500 W pulsed Nd:YAG laser source with 1064 nm wavelength has been used for welding purpose. Butt joints are made. The effects of welding parameters, namely; laser power, scanning speed and pulse width on the seam width and depth of penetration has been investigated using the empirical models developed by response surface methodology (RSM). Weld quality is directly correlated with the weld geometry. Twenty sets of experiments have been conducted as per central composite design (CCD) design matrix. The second order mathematical model has been developed for predicting the desired responses. The results of ANOVA indicate that the laser power has the most significant effect on responses. Microstructural analysis as well as hardness of the selected weld specimens has been carried out to understand the metallurgical and mechanical behaviour of the weld. Average micro-hardness of the weld is observed to be higher than the base metal. Higher hardness of the weld is the resultant of grain refinement and δ-ferrite formation in the weld structure. The result suggests that the lower line energy generally produce fine grain structure and improved mechanical properties than the high line energy. The combined effects of input parameters on responses have been analyzed with the help of developed 3-D response surface and contour plots. Finally, multi-objective optimization has been conducted for producing weld joint with complete penetration, minimum seam width and acceptable welding profile. Confirmatory tests have been conducted at optimum parametric conditions to validate the applied optimization technique.Keywords: ANOVA, laser welding, modeling and optimization, response surface methodology
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