Search results for: Isha Shukla

Authors: Premjit K. Sanjram, Gagan Jakhotiya, Apoorv Goyal, Shanu Shukla

Abstract:

In psychological research, laboratory experiments often face the trade-off issue between experimental control and mundane realism. With the advent of Immersive Virtual Environment Technology (IVET), this issue seems to be at bay. However there is a growing challenge within the IVET itself to design and develop system or software that captures the psychological phenomenon of everyday lives. One such phenomena that is of growing interest is ‘media-multitasking’ To aid laboratory researches in media-multitasking this paper introduces Query Task Modulator (QTM), a computerized experimentation system to study media-multitasking behavior in a controlled laboratory environment. The system provides a computerized platform in conducting an experiment for experimenters to study media-multitasking in which participants will be involved in a query task. The system has Instant Messaging, E-mail, and Voice Call features. The answers to queries are provided on the left hand side information panel where participants have to search for it and feed the information in the respective communication media blocks as fast as possible. On the whole the system will collect multitasking behavioral data. To analyze performance there is a separate output table that records the reaction times and responses of the participants individually. Information panel and all the media blocks will appear on a single window in order to ensure multi-modality feature in media-multitasking and equal emphasis on all the tasks (thus avoiding prioritization to a particular task). The paper discusses the development of QTM in the light of current techniques of studying media-multitasking.

Keywords: experimentation system, human performance, media-multitasking, query-task

Procedia PDF Downloads 557

Authors: Suruchi Shukla, Shantanu Prakash, Amita Jain

Abstract:

Introduction: Arboviral diseases are one of the most common causes of viral hemorrhagic fever (VHF). Of which, Dengue and Chikungunya pose a significant health problem in India. Arbovirus has a tendency to cross the territories and emerge in the new region. Considering the above issues, in the current study active surveillance was conducted among viral hemorrhagic fever (VHF) cases reported from Uttar Pradesh (UP), India. We studied the arboviral etiology of VHF; mainly Dengue, Chikungunya, and ZIKA. Methods: Clinical samples of 465 suspected VHF cases referred to tertiary care referral center of UP, India were enrolled in the study during a period from 15th May 2016 to 9th March 2018. Serum specimens were collected and analyzed for the presence of Dengue, Chikungunya, and ZIKA either by serology and/or by molecular assays. Results: Of all tested, 165 (35.4%) cases were positive for either Dengue or Chikungunya. Dengue (21.2%) was found to be the most prevalent, followed by Chikungunya, (6.6%). None of the cases tested positive for ZIKA virus. Serum samples of 35 (7.5%) cases were positive for both Dengue and Chikungunya. DEN-2 serotype was the most predominant serotype. Phylogenetic and sequence analysis of DEN-2 strains showed 100% clustering with the Cosmopolitan genotype strain. Bleeding from several sites, jaundice, abdominal pain, arthralgia, haemoconcentration, and thrombocytopenia were significantly higher in dengue hemorrhagic cases. However, the rash was significantly more common in Chikungunya patients. Most of the Dengue and Chikungunya positive cases (Age group 6-40 years) were seen in post monsoon season (September to November). Conclusion: Only one-third of total VHF cases are positive for either Dengue/Chikungunya or both. This necessitates the screening of other etiologies capable of causing hemorrhagic manifestations.

Keywords: viral hemorrhagic fever, dengue, chikungunya, zika, India

Procedia PDF Downloads 156

Authors: Amita Jain, Shantanu Prakash, Suruchi Shukla

Abstract:

Introduction: Acute encephalitis syndrome (AES) is a complex multi etiology syndrome posing a great public health problem in the northern part of India. Japanese encephalitis (JE) virus is an established etiology of AES in this region. Recently, Scrub typhus (ST) is being recognized as an emerging aetiology of AES in JE endemic belt. This study was conducted to establish the direct evidence of Central nervous system invasion by Orientia tsutsugamushi leading to AES. Methodology: A total of 849 cases with clinical diagnosis of AES were enrolled from six districts (Deoria and its adjoining area) of the traditional north Indian Japanese encephalitis (JE) belt. Serum and Cerebrospinal fluid samples were collected and tested for major agent causing acute encephalitis. AES cases either positive for anti-ST IgM antibodies or negative for all tested etiologies were investigated for ST-DNA by real-time PCR. Results: Of these 505 cases, 250 patients were laboratory confirmed for O. tsutsugamushi infection either by anti-ST IgM antibodies positivity (n=206) on serum sample or by ST-DNA detection by real-time PCR assay on CSF sample (n=2) or by both (n=42).Total 29 isolate could be sequenced for 56KDa gene. Conclusion: All the strains were found to cluster with Gilliam strains. The majority of the isolates showed a 97–99% sequence similarity with Thailand and Cambodian strains. Gilliam strain of O.tsusugamushi is an emerging as one of the major aetiologies leading to AES in northern part of India.

Keywords: acute encephalitis syndrome, O. tsutsugamushi, Gilliam strain, North India, cerebrospinal fluid

Procedia PDF Downloads 252

Authors: Saurabh Shukla, G. N. Pandey

Abstract:

In recent times, oil exploration and monitoring in on-shore areas have gained much importance considering the fact that in India the oil import is 62 percent of the total imports. Thus, architectural model like wireless sensor network to monitor on-shore deep sea oil well is being developed to get better estimate of the oil prospects. The problem we are facing nowadays that we have very few restricted areas of oil left today. Countries like India don’t have much large areas and resources for oil and this problem with most of the countries that’s why it has become a major problem when we are talking about oil exploration in on-shore areas also the increase of oil prices has further ignited the problem. For this the use of wireless network system having relative simplicity, smallness in size and affordable cost of wireless sensor nodes permit heavy deployment in on-shore places for monitoring oil wells. Deployment of wireless sensor network in large areas will surely reduce the cost it will be very much cost effective. The objective of this system is to send real time information of oil monitoring to the regulatory and welfare authorities so that suitable action could be taken. This system architecture is composed of sensor network, processing/transmission unit and a server. This wireless sensor network system could remotely monitor the real time data of oil exploration and monitoring condition in the identified areas. For wireless sensor networks, the systems are wireless, have scarce power, are real-time, utilize sensors and actuators as interfaces, have dynamically changing sets of resources, aggregate behaviour is important and location is critical. In this system a communication is done between the server and remotely placed sensors. The server gives the real time oil exploration and monitoring conditions to the welfare authorities.

Keywords: sensor, wireless sensor network, oil, sensor, on-shore level

Procedia PDF Downloads 447

Authors: Prakash Chittoor

Abstract:

In the recent past, it is realised especially in third sector that employing social innovation is crucial for achieving viable and long lasting social transformation. In this context, education is one among many sectors that have opened up itself for such move where employing social innovation emerges as key for reaching out to the excluded sections who are often failed to get support from either policy or market interventions. In fact, education is being as a crucial factor for social development is well understood at both academic and policy level. In order to move forward to achieve better results, interventions from multiple sectors may be required as its reach cultivates capabilities and skill of the deprived in order to ensure both market and social participation in the long run. Despite state’s intervention, it is found that still millions of children are out of school due to lack of political will, lapses in policy implementation and neoliberal intervention of marketization. As a result, universalisation of elementary education became as an elusive goal to poor and marginalised sections where state obtain constant pressure by corporate sector to withdraw from education sector that led convince in providing quality education. At this juncture, the role of third sector organizations plays is quite remarkable. Especially, it has evolved as a key player in education sector to reach out to the poor and marginalised in the far-flung areas. These organisations work in resources constrain environment, yet, in order to achieve larger social impact they adopt various social innovations from time to time to reach out to the unreached. Their attempts not only limited to just approaching the unreached children but to retain them for long-time in the schooling system in order to ripe the results for their families and communities. There is a need to highlight various innovative ways adopted and practiced by the third sector organisations in India to achieve the elusive goal of universal access of primary education with quality. With this background, the paper primarily attempts to present an in-depth understanding about innovative practices employed by third sectors organisations like Isha Vidya through government schools adoption programme in India where it engages itself with government and build capabilities among the government teachers to promote state run schooling with quality and better infrastructure. Further, this paper assess whether such innovative attempts succeeded in to achieving universal quality education in the areas where it operates and draws implications for State policy.

Keywords: school education, third sector organisations, social innovation, market domination

Procedia PDF Downloads 263

Authors: Sateesh Kumar Pisini, Swetha Priya Darshini, Sanjay Kumar Shukla

Abstract:

This paper seeks to present a research study on sustainability in construction of retaining wall using geosynthetics. Sustainable construction is a way for the building and infrastructure industry to move towards achieving sustainable development, taking into account environmental, socioeconomic and cultural issues. Geotechnical engineering, being very resource intensive, warrants an environmental sustainability study, but a quantitative framework for assessing the sustainability of geotechnical practices, particularly at the planning and design stages, does not exist. In geotechnical projects, major economic issues to be addressed are in the design and construction of stable slopes and retaining structures within space constraints. In this paper, quantitative indicators for assessing the environmental sustainability of retaining wall with geosynthetics are compared with conventional concrete retaining wall through life cycle assessment (LCA). Geosynthetics can make a real difference in sustainable construction techniques and contribute to development in developing countries in particular. Their imaginative application can result in considerable cost savings over the use of conventional designs and materials. The acceptance of geosynthetics in reinforced retaining wall construction has been triggered by a number of factors, including aesthetics, reliability, simple construction techniques, good seismic performance, and the ability to tolerate large deformations without structural distress. Reinforced retaining wall with geosynthetics is the best cost-effective and eco-friendly solution as compared with traditional concrete retaining wall construction. This paper presents an analysis of the theme of sustainability applied to the design and construction of traditional concrete retaining wall and presenting a cost-effective and environmental solution using geosynthetics.

Keywords: sustainability, retaining wall, geosynthetics, life cycle assessment

Procedia PDF Downloads 2061

Authors: Deepshikha Shukla, C. H. Solanki, Mayank Desai

Abstract:

In the last few years, it has been observed that earthquake is proving a threat to the scientist across the world. With a large number of earthquakes occurring in day to day life, the threat to life and property has increased manifolds which call for an urgent attention of all the researchers globally to carry out the research in the field of Earthquake Engineering. Any hazard related to the earthquake and seismicity is considered to be seismic hazards. The common forms of seismic hazards are Ground Shaking, Structure Damage, Structural Hazards, Liquefaction, Landslides, Tsunami to name a few. Among all the natural hazards, the most devastating and damaging is the earthquake as all other hazards are triggered only after the occurrence of an earthquake. In order to quantify and estimate the seismicity and seismic hazards, many methods and approaches have been proposed in the past few years. Such approaches are Mathematical, Conventional and Computational. Convex Set Theory, Empirical Green’s Function are some of the Mathematical Approaches whereas the Deterministic and Probabilistic Approaches are the Conventional Approach for the estimation of the seismic Hazards. Ground response and Ground Shaking of a particular area or region plays an important role in the damage caused due to the earthquake. In this paper, seismic study using Deterministic Approach and 1 D Ground Response Analysis has been carried out for Madhya Pradesh Tourism Office in Indore Region in Madhya Pradesh in Central India. Indore lies in the seismic zone III (IS: 1893, 2002) in the Seismic Zoning map of India. There are various faults and lineament in this area and Narmada Some Fault and Gavilgadh fault are the active sources of earthquake in the study area. Deepsoil v6.1.7 has been used to perform the 1 D Linear Ground Response Analysis for the study area. The Peak Ground Acceleration (PGA) of the city ranges from 0.1g to 0.56g.

Keywords: seismicity, seismic hazards, deterministic, probabilistic methods, ground response analysis

Procedia PDF Downloads 167

Authors: Nirali Shukla, Neel Shah

Abstract:

The World Bank Institute (WBI) is undertaking a review of government interventions aimed at facilitating sustainable investment in public private partnerships (PPPs) in various under developed countries. The study presents best practice for applying financial model to make PPPs financially viable. The lessons presented here, if properly implemented, can help countries use limited funds to attract more private investment, get more infrastructure built and, as a result, achieve greater economic growth. The four countries Brazil, Colombia, Mexico, and India in total develop an average of nearly US$50 billion in PPPs per year. There are a range of policies and institutional arrangements governments use to provide subsidies to PPPs. For example, some countries have created dedicated agencies, or ‘funds’, capitalized with money from the national budget to manage and allocate subsidies. Other countries have established well-defined policies for appropriating subsidies on an ad hoc basis through an annual budget process. In this context, subsidies are direct fiscal contributions or grants paid by the government to a project when revenues from user fees are insufficient to cover all capital and operating costs while still providing private investors with a reasonable rate of return. Without subsidies, some infrastructure projects that would provide economic or social gains, but are not financially viable, would go undeveloped. But the Financial model of BOLT (PPP) model described in this study suggests that it is most feasible option rather than going for subsidy schemes for making infrastructure projects financially viable. The major advantage for implementing this model is the government money is saved and can be used for other projects as well as the private investors are getting better rate of return than subsidized schemes.

Keywords: PPP, BOLT, subsidy schemes, financial model

Procedia PDF Downloads 770

Authors: Abhaydeep Pandey, Shweta Shukla, Saptamita Goswami, Bhaswati Bandyopadhyay, Vishnampettai Ramachandran, Sudhanshu Vrati, Arup Banerjee

Abstract:

Background: Long non-coding RNAs (lncRNAs) are the important regulators of gene expression and play important role in viral replication and disease progression. The role of lncRNA genes in the pathogenesis of Dengue virus-mediated pathogenesis is currently unknown. Methods: To gain additional insights, we utilized an unbiased RNA sequencing followed by in silico analysis approach to identify the differentially expressed lncRNA and genes that are associated with dengue disease progression. Further, we focused our study on lncRNAs NEAT1 (Nuclear Paraspeckle Assembly Transcript 1) as it was found to be differentially expressed in PBMC of dengue infected patients. Results: The expression of lncRNAs NEAT1, as compared to dengue infection (DI), was significantly down-regulated as the patients developed the complication. Moreover, pairwise analysis on follow up patients confirmed that suppression of NEAT1 expression was associated with rapid fall in platelet count in dengue infected patients. Severe dengue patients (DS) (n=18; platelet count < 20K) when recovered from infection showing high NEAT1 expression as it observed in healthy donors. By co-expression network analysis and subsequent validation, we revealed that coding gene; IFI27 expression was significantly up-regulated in severe dengue cases and negatively correlated with NEAT1 expression. To discriminate DI from dengue severe, receiver operating characteristic (ROC) curve was calculated. It revealed sensitivity and specificity of 100% (95%CI: 85.69 – 97.22) and area under the curve (AUC) = 0.97 for NEAT1. Conclusions: Altogether, our first observations demonstrate that monitoring NEAT1and IFI27 expression in dengue patients could be useful in understanding dengue virus-induced disease progression and may be involved in pathophysiological processes.

Keywords: dengue, lncRNA, NEAT1, transcriptome

Procedia PDF Downloads 310

Authors: Palak J. Shukla, Atul K. Desai, Chentankumar D. Modhera

Abstract:

For any country in the world, it has become a priority to protect the critical infrastructure from looming risks of terrorism. In any infrastructure system, the structural elements like lower floors, exterior columns, walls etc. are key elements which are the most susceptible to damage due to blast load. The present study revisits the state of art review of the design and analysis of reinforced concrete panels subjected to blast loading. Various aspects in association with blast loading on structure, i.e. estimation of blast load, experimental works carried out previously, the numerical simulation tools, various material models, etc. are considered for exploring the current practices adopted worldwide. Discussion on various parametric studies to investigate the effect of reinforcement ratios, thickness of slab, different charge weight and standoff distance is also made. It was observed that for the simulation of blast load, CONWEP blast function or equivalent numerical equations were successfully employed by many researchers. The study of literature indicates that the researches were carried out using experimental works and numerical simulation using well known generalized finite element methods, i.e. LS-DYNA, ABAQUS, AUTODYN. Many researchers recommended to use concrete damage model to represent concrete and plastic kinematic material model to represent steel under action of blast loads for most of the numerical simulations. Most of the studies reveal that the increase reinforcement ratio, thickness of slab, standoff distance was resulted in better blast resistance performance of reinforced concrete panel. The study summarizes the various research results and appends the present state of knowledge for the structures exposed to blast loading.

Keywords: blast phenomenon, experimental methods, material models, numerical methods

Procedia PDF Downloads 158

Authors: Shahnaz Haque, Anand Shukla, Sunita Singh, Anil Kem

Abstract:

Introduction: Diabetes Mellitus is a chronic metabolic disease affecting millions worldwide. Metformin is the most commonly prescribed first line oral hypoglycemic drug for type 2 diabetes mellitus, but due to progressive worsening of blood glucose control during the natural history of type 2 diabetes, combination therapy usually becomes necessary. Objective: This study was designed to assess the treatment satisfaction between Sitagliptin versus Pioglitazone added to Metformin in patients with type 2 diabetes mellitus (T2DM). Methods: We conducted a prospective, open label, randomized, parallel group study in SIMS, Hapur, U.P. Eligible patients fulfilling inclusion criteria were randomized into two groups having 25 patients in each group using tab Sitagliptin 100mg, tab Pioglitazone 30mg added to ongoing tab Metformin (500mg) therapy for 16 weeks. The follow-up visits were on weeks 4,12 and 16. Result: 16 weeks later, addition of Sitagliptin 100mg compared to that of Pioglitazone 30 mg to ongoing Metformin therapy provided similar glycosylated hemoglobin (HbA1c) lowering efficacy in patients with T2DM with inadequate glycemic control on metformin monotherapy. Change in HbA1c in group1 was -0.656±0.21%(p<0.0001) whereas in group2 was -0.748±0.35%(p<0.0001). Hence decrease in HbA1c from baseline was more in group2. Both treatments were well tolerated with negligible risk of hypoglycaemia. Weight loss was observed with Sitagliptin in contrast to weight gain seen in Pioglitazone. Conclusion: In this study, Sitagliptin 100 mg along with metformin therapy in comparison to pioglitazone 30 mg plus metformin therapy was both effective, well-tolerated and improved glycemic control in both the groups. Addition of pioglitazone had cause oedema and weight gain to the patients whereas sitagliptin caused weight loss in its patients.

Keywords: sitagliptin, pioglitazone, metformin, type 2 diabetes mellitus

Procedia PDF Downloads 304

Authors: Lubna Azmi, Ila Shukla, Shyam Sundar Gupta, Padam Kant, Ch. V. Rao

Abstract:

Gastric ulcer is a major global health threat, and it is the leading cause of stomach cancer death worldwide. Helicobacter pylori bacteriumis the most important etiologic factor for gastric ulcer. This infection is highly pervasive in South Asian developing countries, especially in India, Nepal, Srilanka etc. due to diversification in geographic area. Pathophysiology of gastric mucosal damage associated with non-invasive bacterium has not justified in detail, but it leads to change in histopathology, immunochemistry of the gastric and duodenal reason of host. The mechanism responsible for bacteria tissue tropism and mucosal damage in stomach variance during the disease is not clearly described and understood scientifically in treatment and control of pathogenic organisms. Polyphenols are secondary metabolites of plants and are generally involved in defense against aggression by pathogens. 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxychromen-4-one and 1-hydroxy-5,7-dimethoxy-2-naphthalene-carboxaldehyde are polyphenolic compound obtained from popular Indian medicinal plants ghavpatta (ArgeriaspeciosaLinn.f) andBael (Aeglemarmelos) have long been used in traditional Ayurvedic Indian medicine for various diseases. They have promising effects on ulcer, as detailed investigation has made in our laboratory. Therefore, the aim of present study is to explore membrane –dependent morphogenesis of H. pylori and associated apoptosis-mediated cell death. Based on this we analyzed immune gene expression in stomach of experimental animals with H. pylori, using quantitative reverse transcription polymerase chain reaction(q RT-PCR). This revealed rapid induction of prostaglandin, interferon I (INF-I), interferon II (INF-II) and INF-I associated genes in the infected animal. Ultrastructural changes associated with H. pylori will be taken for advanced studies. This investigation shows that the biomarkers eradicate H. pylori bacterium caused gastric ulcer which is a major risk factor for gastric cancer.

Keywords: gastric ulcer, Helicobacter pylori, immunochemistry, polyphenols

Procedia PDF Downloads 372

Authors: Abhaydeep Pandey, Shweta Shukla, Saptamita Goswami, Bhaswati Bandyopadhyay, Vishnampettai Ramachandran, Sudhanshu Vrati, Arup Banerjee

Abstract:

Dengue virus infection is now considered as one of the most important mosquito-borne infection in human. The virus is known to promote vascular permeability, cerebral edema leading to Dengue hemorrhagic fever (DHF) or Dengue shock syndrome (DSS). Dengue infection has known to be endemic in India for over two centuries as a benign and self-limited disease. In the last couple of years, the disease symptoms have changed, manifesting severe secondary complication. So far, Delhi has experienced 12 outbreaks of dengue virus infection since 1997 with the last reported in 2014-15. Without specific antivirals, the case management of high-risk dengue patients entirely relies on supportive care, involving constant monitoring and timely fluid support to prevent hypovolemic shock. Nonetheless, the diverse clinical spectrum of dengue disease, as well as its initial similarity to other viral febrile illnesses, presents a challenge in the early identification of this high-risk group. WHO recommends the use of warning signs to identify high-risk patients, but warning signs generally appear during, or just one day before the development of severe illness, thus, providing only a narrow window for clinical intervention. The ability to predict which patient may develop DHF and DSS may improve the triage and treatment. With the recent discovery of high throughput RNA sequencing allows us to understand the disease progression at the genomic level. Here, we will collate the results of RNA-Sequencing data obtained recently from PBMC of different categories of dengue patients from India and will discuss the possible role of deregulated genes and long non-coding RNAs NEAT1 for development of disease progression.

Keywords: long non-coding RNA (lncRNA), dengue, peripheral blood mononuclear cell (PBMC), nuclear enriched abundant transcript 1 (NEAT1), dengue hemorrhagic fever (DHF), dengue shock syndrome (DSS)

Procedia PDF Downloads 309

Authors: Sumanta Meher, Gaurav Shukla

Abstract:

The torments of the victims of terrorism have not only confined to loss of life and limp but also includes the physiological trauma to the innocent victims. The physical wounds may heal, but the trauma remains in the mind and heart of the victims and their loved ones; however, one should not deny that these terrorist activities affect to a major extent to their livelihood. To protect their human rights and restore the shattered lives of the victims of terrorism all the Nations beyond their differences have to show solidarity and frame a comprehensive restorative policy with an effective implementing mechanism. The General Assembly of United Nations, through its several resolutions, has appealed Nations to show solidarity and also committed to helping the Members State to frame the law and policy to support the victims of terrorism. To achieve the objectives of the resolutions adopted by the United Nations, the Indian legislators in 2008 amended the Code of Criminal Procedure, 1973 and incorporated Section 357A to provide financial assistance to the victims of terrorism. In India, the contemporary developments in the victims’ oriented studies have increased the dimension of the traditional criminal justice systems to protect the rights of the victims. In this regard, the paper has ascertained the Indian legal framework in respect to the restorative justice to the victims of terrorism and also addressed the question as to whether the statutory provisions and enforcement mechanisms are efficient enough to protect the human rights of the victims of terrorism. For that purpose, the paper has analyzed the International instruments and the reports with regard to the compensation to the victims of terrorist attacks, with that, the article also evaluates the initiatives of United Nations to help Members State to frame the law and policies to support the victims of terrorism. The study also made an attempt to critically analyze the legal provisions of compensation and rehabilitation of the victims of terrorist attacks in India and whether they are in alignment with the International standards. While concluding, the paper has made an endeavor for a robust legal framework towards the restorative justice for the victims of terrorism in India.

Keywords: victims of terrorism, restorative justice, human rights, criminal justice system of India

Procedia PDF Downloads 161

Authors: Lubna Azmi, Ila Shukla, Shyam Sundar Gupta, Padam Kant, C. V. Rao

Abstract:

To investigate the chemoprotective potential of NBRIQU16 chemotype isolated from Argyreia speciosa (Family: Convolvulaceae) on N-methyl-N-nitro-N-nitrosoguanidine (MNNG) and NaCl-induced gastric carcinomas in Wistar rats. Forty-six male 6-week-old Wistar rats were divided into two groups. Thirty rats in group A were fed with a diet supplemented with 8 % NaCl for 20 weeks and simultaneously given N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) in drinking water at a concentration of 100 ug/ml for the first 17 weeks. After administration of the carcinogen, 200 and 400 mg/kg of NBRIQU16 were administered orally once a day throughout the study. From week 18, these rats were given normal water. From week 21, these rats were fed with a normal diet for 15 weeks. Group B containing 16 rats was fed standard diet for thirty-five days. It served as control. Ten rats from group A were sacrificed after 20 weeks. Scarification of remaining animals was conducted after 35 weeks. Entire stomach and some part of the duodenum were incised parallel to the greater curvature, and the samples were collected. After opening the stomach location and size of tumors were recorded. The number of tumors with their locations and sizes were recorded. Expression of survivin was examined by recording the Immunohistochemistry of the specimens. The treatment with NBRIQU16 significantly reduced the nodule incidence and nodule multiplicity in the rats after MNNG administration. Surviving expression in glandular stomachs of normal rats, of rats in middle induction period, in adenocarcinomas and NBRIQU16 treated tissues adjacent to tumor were 0, 42.0 %, 79.3%, and 36.4 %, respectively. Expression of survivin was significantly different as compared to the normal rats. Histological observations of stomach tissues too correlated with the biochemical observations.These finding powerfully supports that NBRIQU16 chemopreventive effect by suppressing the tumor burden and restoring the activities of gastric cancer marker enzymes on MNNG and NaCl-induced gastric carcinomas in Wistar rats.

Keywords: Argyreia speciosa, gastric carcinoma, immunochemistry, NBRIQU16

Procedia PDF Downloads 298

Authors: Tulika Malviya, Ritesh Chandra Shukla, Praveen Kumar Tandon

Abstract:

Traditional methods of synthesis are hazardous for the environment and need nature friendly processes for the treatment of industrial effluents and contaminated water. Use of plant parts for the synthesis provides an efficient alternative method. In this paper, we report an ecofriendly and nonhazardous biobased method to prepare zerovalent iron nanoparticles (ZVINPs) using the liquor of commercially available tea. Tea liquor as the reducing agent has many advantages over other polymers. Unlike other polymers, the polyphenols present in tea extract are nontoxic and water soluble at room temperature. In addition, polyphenols can form complexes with metal ions and thereafter reduce the metals. Third, tea extract contains molecules bearing alcoholic functional groups that can be exploited for reduction as well as stabilization of the nanoparticles. Briefly, iron nanoparticles were prepared by adding 2.0 g of montmorillonite K10 (MMT K10) to 5.0 mL of 0.10 M solution of Fe(NO3)3 to which an equal volume of tea liquor was then added drop wise over 20 min with constant stirring. The color of the mixture changed from whitish yellow to black, indicating the formation of iron nanoparticles. The nanoparticles were adsorbed on montmorillonite K10, which is safe and aids in the separation of hazardous arsenic species simply by filtration. Particle sizes ranging from 59.08±7.81 nm were obtained which is confirmed by using different instrumental analyses like IR, XRD, SEM, and surface area studies. Removal of arsenic was done via batch adsorption method. Solutions of As(III) of different concentrations were prepared by diluting the stock solution of NaAsO2 with doubly distilled water. The required amount of in situ prepared ZVINPs supported on MMT K10 was added to a solution of desired strength of As (III). After the solution had been stirred for the preselected time, the solid mass was filtered. The amount of arsenic [in the form of As (V)] remaining in the filtrate was measured using ion chromatograph. Stirring of contaminated water with zerovalent iron nanoparticles supported on montmorillonite K10 for 30 min resulted in up to 99% removal of arsenic as As (III) from its solution at both high and low pH (2.75 and 11.1). It was also observed that, under similar conditions, montmorillonite K10 alone provided only <10% removal of As(III) from water. Adsorption at low pH with precipitation at higher pH has been proposed for As(III) removal.

Keywords: arsenic removal, montmorillonite K10, tea liquor, zerovalent iron nanoparticles

Procedia PDF Downloads 130

Authors: Neetika Mahajan, Awanish Shukla

Abstract:

Entrepreneurship is key a driver to economic development. It opens opportunities for business startups and has potential to expand employment opportunities for many. Entrepreneurship gives ‘Purpose thriving’ approach towards society with new technologies and zeal to develop and compete in the market. There are many more advantages of entrepreneurship like freedom to scope of work and independence in setting own goals. Women contribute to nearly 50 percent of India’s population, constitute nearly 10 percent of a total number of entrepreneurs in India. Women are found to be better risk calculators, more ambitious and less prone to self-confidence. However, It is a hard fact that life has not been easy for women aspiring professional success. Gender parity is the biggest threat faced by female aspirant seeking new businesses. More challenges like socio-cultural barriers, insufficient financial assistance, etc. have been faced by the women of our country. To uplift the status of women in the society, a number of initiatives have been taken up by the Government of India. Initiatives like National Mission for Empowerment of Women by (Ministry of Women And Child Development) and SKILL INDIA aim to increase the technical skills and knowledge of women for tapping employment opportunities and self-confidence. Trade Related Entrepreneurship Assistance and Development (TREAD) Scheme and Mahila Coir Yojana are proposed by the Ministry of MSMEs aiming to facilitate employment opportunities for women and entrepreneurship development. This paper will aim to bring out the gaps and barriers, which are still resisting the potential women come upfront and start a new business irrespective of a number of initiatives being put by government of India. The aim is also to identify focus areas where further intervention is required and proposing suitable interventions. The methodology to take forward this research will include primary and secondary data collection from on ground survey to track various kind of challenges faced by aspirant women entrepreneurs. Insight will be put towards initiations by the government of India towards women empowerment and assistance to entrepreneurship. Scientific quantitative tools will be used to analyze collected information. The final output of the research shall focus on achieving the respective aims and objectives.

Keywords: women entrepreneurship, government programs and schemes, key challenges, economic development

Procedia PDF Downloads 251

Authors: Sameer A. Pathan, Biswadev Mitra, Salman Mirza, Umais Momin, Zahoor Ahmed, Lubna G. Andraous, Dharmesh Shukla, Mohammed Y. Shariff, Magid M. Makki, Tinsy T. George, Saad S. Khan, Stephen H. Thomas, Peter A. Cameron

Abstract:

Study objective: Emergency physician’s (EP) ability to identify hydronephrosis on point-of-care ultrasound (POCUS) has been assessed in the past using CT scan as the reference standard. We aimed to assess EP interpretation of POCUS to identify and grade the hydronephrosis in a direct comparison with the consensus-interpretation of POCUS by radiologists, and also to compare the EP and radiologist performance using CT scan as the criterion standard. Methods: Using data from a POCUS databank, a prospective interpretation study was conducted at an urban academic emergency department. All POCUS exams were performed on patients presenting with renal colic to the ED. Institutional approval was obtained for conducting this study. All the analyses were performed using Stata MP 14.0 (Stata Corp, College Station, Texas). Results: A total of 651 patients were included, with paired sets of renal POCUS video clips and the CT scan performed at the same ED visit. Hydronephrosis was reported in 69.6% of POCUS exams by radiologists and 72.7% of CT scans (p=0.22). The κ for consensus interpretation of POCUS between the radiologists to detect hydronephrosis was 0.77 (0.72 to 0.82) and weighted κ for grading the hydronephrosis was 0.82 (0.72 to 0.90), interpreted as good to very good. Using CT scan findings as the criterion standard, Eps had an overall sensitivity of 81.1% (95% CI: 79.6% to 82.5%), specificity of 59.4% (95% CI: 56.4% to 62.5%), PPV of 84.3% (95% CI: 82.9% to 85.7%), and NPV of 53.8% (95% CI: 50.8% to 56.7%); compared to radiologist sensitivity of 85.0% (95% CI: 82.5% to 87.2%), specificity of 79.7% (95% CI: 75.1% to 83.7%), PPV of 91.8% (95% CI: 89.8% to 93.5%), and NPV of 66.5% (95% CI: 61.8% to 71.0%). Testing for a report of moderate or high degree of hydronephrosis, specificity of EP was 94.6% (95% CI: 93.7% to 95.4%) and to 99.2% (95% CI: 98.9% to 99.5%) for identifying severe hydronephrosis alone. Conclusion: EP POCUS interpretations were comparable to the radiologists for identifying moderate to severe hydronephrosis using CT scan results as the criterion standard. Among patients with moderate or high pre-test probability of ureteric calculi, as calculated by the STONE-score, the presence of moderate to severe (+LR 6.3 and –LR 0.69) or severe hydronephrosis (+LR 54.4 and –LR 0.57) was highly diagnostic of the stone disease. Low dose CT is indicated in such patients for evaluation of stone size and location.

Keywords: renal colic, point-of-care, ultrasound, bedside, emergency physician

Procedia PDF Downloads 284

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

Abstract:

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

Procedia PDF Downloads 141

Authors: Sanjay Shukla

Abstract:

Excessive use of antibiotics is a main problem in the treatment of wounds and other chronic infections and antibiotic treatment is frequently non-curative, thus alternative treatment is necessary. Phage therapy is considered one of the most effective approaches to treat multi-drug resistant bacterial pathogens. Infections caused by Staphylococcus aureus are very efficiently controlled with phage cocktails, containing a different individual phages lysate infecting a majority of known pathogenic S. aureus strains. The aim of current study was to investigate the efficiency of a purified phage cocktail for prophylactic as well as therapeutic application in mouse model and in large animals with chronic septic infection of wounds. A total of 150 sewage samples were collected from various livestock farms. These samples were subjected for the isolation of bacteriophage by double agar layer method. A total of 27 sewage samples showed plaque formation by producing lytic activity against S. aureus in double agar overlay method out of 150 sewage samples. In TEM recovered isolates of bacteriophages showed hexagonal structure with tail fiber. In the bacteriophage (ØVS) had an icosahedral symmetry with the head size 52.20 nm in diameter and long tail of 109 nm. Head and tail were held together by connector and can be classified as a member of the Myoviridae family under the order of Caudovirale. Recovered bacteriophage had shown the antibacterial activity against the S. aureus in vitro. Cocktail (ØVS1, ØVS5, ØVS9 and ØVS 27) of phage lysate were tested to know in vivo antibacterial activity as well as the safety profile. Result of mice experiment indicated that the bacteriophage lysate was very safe, did not show any appearance of abscess formation which indicates its safety in living system. The mice were also prophylactically protected against S. aureus when administered with cocktail of bacteriophage lysate just before the administration of S. aureus which indicates that they are good prophylactic agent. The S. aureus inoculated mice were completely recovered by bacteriophage administration with 100% recovery which was very good as compere to conventional therapy. In present study ten chronic cases of wound were treated with phage lysate and follow up of these cases was done regularly up to ten days (at 0, 5 and 10 d). Result indicated that the six cases out of ten showed complete recovery of wounds within 10 d. The efficacy of bacteriophage therapy was found to be 60% which was very good as compared to the conventional antibiotic therapy in chronic septic wounds infections. Thus, the application of lytic phage in single dose proved to be innovative and effective therapy for treatment of septic chronic wounds.

Keywords: phage therapy, phage lysate, antimicrobial resistance, S. aureus

Procedia PDF Downloads 119

Authors: Rashmi Shukla, Yamini Bhusan Tripathi

Abstract:

Diabetic nephropathy (DN) is characterized as diabetic kidney disease which involves many pathways e.g. hyperactivated protein kinase c (PKC), polyol pathway, excess production of advanced glycation end product (AGEs) & free radical accumulation etc. All of them results to hypoxia followed by apoptosis of podocytes, glomerulosclerosis, extracellular matrix (ECM) accumulation and fibrosis resulting to irreversible changes in kidney. This is continuously rising worldwide and there are not enough specific drugs, to retard its progress. Due to increasing side effects of allopathic drugs, interest in herbal remedies is growing. Earlier, we have reported that PTY-2 (a phytomedicine, derived from Pueraria tuberosa Linn.) inhibits the accumulation of extracellular matrix (ECM) through activation of MMP-9. Present study exhibited the therapeutic potential of Pueraria tuberosa in the prevention of podocytes apoptosis and modulation of nephrin expression in streptozotocin (STZ) induced DN rats. DN rats were produced by maintaining persistent hyperglycemia for 8 weeks by intra-peritoneal injection of 55 mg/kg streptozotocin (STZ). These rats were randomly divided in 2 groups, i.e. DN control, and DN+ water extract of Pueraria tuberosa (PTW). One group of age-matched normal rats served as non-diabetic control (group-1), The STZ induced DN rats (group-2) and DN+PTW treated rats (group-3). The PTW was orally administered (0.3g/kg) daily to group-2 rats and drug vector (1 ml of 10% tween 20) in control rats. The treatments were continued for 20 days and blood and urine samples were collected. Rats were then sacrificed to investigate the expression Bcl2, Bax and nephroprotective protein i.e. nephrin in kidney glomerulus. The effect of PTW was evaluated, we have found that the PTW significantly(p < .001) reversed the raised serum urea, serum creatinine, urine protein and improved the creatinine clearance in STZ induce diabetic nephropathy in rats and also significantly(p < .001) prevented the rise in urine albumin excretion. The Western blot analysis of kidney tissue homogenate showed increased expression of Bcl2 in PTW treated rats. The RT-PCR showed the increased expression and accumulation of nephrin mRNA. The confocal photomicrographs also supported the reduction of Bax and a simultaneous increase in Bcl2 and nephrin in glomerular podocytes. Hence, our finding suggests that the nephroprotective role of PTW is mediated via restoration of nephrin thus prevents the podocytes apoptosis and ameliorates diabetic nephropathy. The clinical trial of PTW would prove to be a potential food supplement/ drug of alternative medicine for patients with diabetic nephropathy in early stage.

Keywords: Pueraria tuberosa, diabetic nephropathy, anti-apoptosis, nephrin

Procedia PDF Downloads 219

Authors: Sanjay Shukla, A. Nayak, R. K. Sharma, A. P. Singh, S. P. Tiwari

Abstract:

Excessive use of antibiotics is a major problem in the treatment of wounds and other chronic infections, and antibiotic treatment is frequently non-curative, thus alternative treatment is necessary. Phage therapy is considered one of the most promising approaches to treat multi-drug resistant bacterial pathogens. Infections caused by Staphylococcus aureus are very efficiently controlled with phage cocktails, containing a different individual phages lysate infecting a majority of known pathogenic S. aureus strains. The aim of the present study was to evaluate the efficacy of a purified phage cocktail for prophylactic as well as therapeutic application in mouse model and in large animals with chronic septic infection of wounds. A total of 150 sewage samples were collected from various livestock farms. These samples were subjected for the isolation of bacteriophage by the double agar layer method. A total of 27 sewage samples showed plaque formation by producing lytic activity against S. aureus in the double agar overlay method out of 150 sewage samples. In TEM, recovered isolates of bacteriophages showed hexagonal structure with tail fiber. In the bacteriophage (ØVS) had an icosahedral symmetry with the head size 52.20 nm in diameter and long tail of 109 nm. Head and tail were held together by connector and can be classified as a member of the Myoviridae family under the order of Caudovirale. Recovered bacteriophage had shown the antibacterial activity against the S. aureus in vitro. Cocktail (ØVS1, ØVS5, ØVS9, and ØVS 27) of phage lysate were tested to know in vivo antibacterial activity as well as the safety profile. Result of mice experiment indicated that the bacteriophage lysate were very safe, did not show any appearance of abscess formation, which indicates its safety in living system. The mice were also prophylactically protected against S. aureus when administered with cocktail of bacteriophage lysate just before the administration of S. aureuswhich indicates that they are good prophylactic agent. The S. aureusinoculated mice were completely recovered by bacteriophage administration with 100% recovery, which was very good as compere to conventional therapy. In the present study, ten chronic cases of the wound were treated with phage lysate, and follow up of these cases was done regularly up to ten days (at 0, 5, and 10 d). The result indicated that the six cases out of ten showed complete recovery of wounds within 10 d. The efficacy of bacteriophage therapy was found to be 60% which was very good as compared to the conventional antibiotic therapy in chronic septic wounds infections. Thus, the application of lytic phage in single dose proved to be innovative and effective therapy for the treatment of septic chronic wounds.

Keywords: phage therapy, S aureus, antimicrobial resistance, lytic phage, and bacteriophage

Procedia PDF Downloads 117

Authors: Soumyadeep Bhaduri, Rahul Ghosh, Rahul Shukla, Manaswini Behera

Abstract:

The textile industry consumes a substantial amount of water throughout the processing and production of textile fabrics. The water eventually turns into wastewater, where it acts as an immense damaging nuisance due to its dye content. Wastewater streams contain a percentage ranging from 2.0% to 50.0% of the total weight of dye used, depending on the dye class. The management of dye effluent in textile industries presents a formidable challenge to global sustainability. The current focus is on implementing wastewater treatment technology that enable the recycling of wastewater, reduce energy usage and offset carbon emissions. Microbial fuel cell (MFC) is a device that utilizes microorganisms as a bio-catalyst to effectively treat wastewater while also producing electricity. The MFC harnesses the chemical energy present in wastewater by oxidizing organic compounds in the anodic chamber and reducing an electron acceptor in the cathodic chamber, thereby generating electricity. This research investigates the potential of MFCs to tackle this challenge of azo dye removal with simultaneously generating electricity. Although MFCs are well-established for wastewater treatment, their application in dye decolorization with concurrent electricity generation remains relatively unexplored. This study aims to address this gap by assessing the effectiveness of MFCs as a sustainable solution for treating wastewater containing azo dyes. By harnessing microorganisms as biocatalysts, MFCs offer a promising avenue for environmentally friendly dye effluent management. The performance of MFCs in treating azo dyes and generating electricity was evaluated by optimizing the Chemical Oxygen Demand (COD) and Hydraulic Retention Time (HRT) of influent. COD and HRT values ranged from 1600 mg/L to 2400 mg/L and 5 to 9 days, respectively. Results showed that the maximum open circuit voltage (OCV) reached 648 mV at a COD of 2400 mg/L and HRT of 5 days. Additionally, maximum COD removal of 98% and maximum color removal of 98.91% were achieved at a COD of 1600 mg/L and HRT of 9 days. Furthermore, the study observed a maximum power density of 19.95 W/m3 at a COD of 2400 mg/L and HRT of 5 days. Electrochemical analysis, including linear sweep voltammetry (LSV), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were done to find out the response current and internal resistance of the system. To optimize pH and dye concentration, pH values were varied from 4 to 10, and dye concentrations ranged from 25 mg/L to 175 mg/L. The highest voltage output of 704 mV was recorded at pH 7, while a dye concentration of 100 mg/L yielded the maximum output of 672 mV. This study demonstrates that MFCs offer an efficient and sustainable solution for treating azo dyes in textile industry wastewater, while concurrently generating electricity. These findings suggest the potential of MFCs to contribute to environmental remediation and sustainable development efforts on a global scale.

Keywords: textile wastewater treatment, microbial fuel cell, renewable energy, sustainable wastewater treatment

Procedia PDF Downloads 23

Authors: Mayoorika Shukla, Pramila Jakhar, Tejendra Dixit, I. A. Palani, Vipul Singh

Abstract:

Biosensors are analytical devices with wide range of applications in biological, chemical, environmental and clinical analysis. It comprises of bio-recognition layer which has biomolecules (enzymes, antibodies, DNA, etc.) immobilized over it for detection of analyte and transducer which converts the biological signal into the electrical signal. The performance of biosensor primarily the depends on the bio-recognition layer and therefore it has to be chosen wisely. In this regard, nanostructures of metal oxides such as ZnO, SnO2, V2O5, and TiO2, etc. have been explored extensively as bio-recognition layer. Recently, ZnO has the attracted attention of researchers due to its unique properties like high iso-electric point, biocompatibility, stability, high electron mobility and high electron binding energy, etc. Although there have been many reports on usage of ZnO as bio-recognition layer but to the authors’ knowledge, none has ever observed correlation between optical properties like defect suppression and biosensing capability of the sensor. Here, ZnO nanorods (ZNR) have been synthesized by a low cost, simple and low-temperature hydrothermal growth process, over Platinum (Pt) coated glass substrate. The ZNR have been synthesized in two steps viz. initially a seed layer was coated over substrate (Pt coated glass) followed by immersion of it into nutrient solution of Zinc nitrate and Hexamethylenetetramine (HMTA) with in situ addition of KMnO4. The addition of KMnO4 was observed to have a profound effect over the growth rate anisotropy of ZnO nanostructures. Clustered and powdery growth of ZnO was observed without addition of KMnO4, although by addition of it during the growth, uniform and crystalline ZNR were found to be grown over the substrate. Moreover, the same has resulted in suppression of defects as observed by Normalized Photoluminescence (PL) spectra since KMnO4 is a strong oxidizing agent which provides an oxygen rich growth environment. Further, to explore the correlation between defect suppression and biosensing capability of the ZNR Glucose oxidase (Gox) was immobilized over it, using physical adsorption technique followed by drop casting of nafion. Here the main objective of the work was to analyze effect of defect suppression over biosensing capability, and therefore Gox has been chosen as model enzyme, and electrochemical amperometric glucose detection was performed. The incorporation of KMnO4 during growth has resulted in variation of optical and charge transfer properties of ZNR which in turn were observed to have deep impact on biosensor figure of merits. The sensitivity of biosensor was found to increase by 12-18 times, due to variations introduced by addition of KMnO4 during growth. The amperometric detection of glucose in continuously stirred buffer solution was performed. Interestingly, defect suppression has been observed to contribute towards the improvement of biosensor performance. The detailed mechanism of growth of ZNR along with the overall influence of defect suppression on the sensing capabilities of the resulting enzymatic electrochemical biosensor and different figure of merits of the biosensor (Glass/Pt/ZNR/Gox/Nafion) will be discussed during the conference.

Keywords: biosensors, defects, KMnO4, ZnO nanorods

Procedia PDF Downloads 282

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

Abstract:

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

Procedia PDF Downloads 229