Search results for: Rupinder Bakshi

Authors: Rupinder Bakshi, Geeta Walia, Anita Gupta

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Introduction: Urinary tract infections (UTI) are considered to be one of the most common bacterial infections with an estimated annual global incidence of 150 million. Antimicrobial drug resistance is one of the major threats due to widespread usage of uncontrolled antibiotics. Materials and Methods: A total number of 9149 urine samples were collected from R.H Patiala and processed in the Department of Microbiology G.M.C Patiala. Urine samples were inoculated on MacConkey’s and blood agar plates by using calibrated loop delivering 0.001 ml of sample and incubated at 37 °C for 24 hrs. The organisms were identified by colony characters, gram’s staining and biochemical reactions. Antimicrobial susceptibility of the isolates was determined against various antimicrobial agents (Hi – Media Mumbai India) by Kirby-Bauer disk diffusion method on Muller Hinton agar plates. Results: Maximum patients were in the age group of 21-30 yrs followed by 31-40 yrs. Males (34%) are less prone to urinary tract infections than females (66%). Out of 9149 urine sample, the culture was positive in 25% (2290) samples. Esch. coli was the most common isolate 60.3% (n = 1378) followed by Klebsiella pneumoniae 13.5% (n = 310), Proteus spp. 9% (n = 209), Staphylococcus aureus 7.6 % (n = 173), Pseudomonas aeruginosa 3.7% (n = 84), Citrobacter spp. 3.1 % (70), Staphylococcus saprophyticus 1.8 % (n = 142), Enterococcus faecalis 0.8%(n=19) and Acinetobacter spp. 0.2%(n=5). Gram negative isolates showed higher sensitivity towards, Piperacillin +Tazobactum (67%), Amikacin (80%), Nitrofurantoin (82%), Aztreonam (100%), Imipenem (100%) and Meropenam (100%) while gram positive showed good response towards Netilmicin (69%), Nitrofurantoin (79%), Linezolid (98%), Vancomycin (100%) and Teicoplanin (100%). 465 (23%) isolates were resistant to Penicillins, 1st generation and 2nd generation Cehalosporins which were further tested by double disk approximation test and combined disk method for ESBL production. Out of 465 isolates, 375 were ESBLs consisting of n 264 (70.6%) Esch.coli and 111 (29.4%) Klebsiella pneumoniae. Susceptibility of ESBL producers to Imipenem, Nitrofurantoin and Amikacin were found to be 100%, 76%, and 75% respectively. Conclusion: Uropathogens are increasingly showing resistance to many antibiotics making empiric management of outpatients UTIs challenging. Ampicillin, Cotrimoxazole, and Ciprofloxacin should not be used in empiric treatment. Nitrofurantoin could be used in lower urinary tract infection. Knowledge of uropathogens and their antimicrobial susceptibility pattern in a geographical region will help inappropriate and judicious antibiotic usage in a health care setup.

Keywords: Urinary Tract Infection, UTI, antibiotic susceptibility pattern, ESBL

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Authors: Rupinder Pal Kaur, Vishal Goyal

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In this research, a systematic and quantitative engineering-based approach is followed by applying well-known international standards and guidelines to develop a web quality model (PH.WQT- Punjabi and Hindi Website Quality Tester) to measure external quality for websites of government domain that are developed in Punjabi and Hindi. Correspondingly, the model can be used for websites developed in other languages also. The research is valuable to researchers and practitioners interested in designing, implementing and managing websites of government domain Also, by implementing PH.WQT analysis and comparisons among web sites of government domain can be performed in a consistent way.

Keywords: external quality, PH.WQT, indian languages, punjabi and hindi, quality model, websites of government

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Authors: Rupinder Kaur, Harjot Kaur

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The objective of this paper is to present the study of the effect of varying thickness on rotating composite disks made from Al-SiC_P having different particle sizes. Mathematical modeling is used to calculate the effect of varying thickness with different particle sizes on rotating composite disks in radial as well as tangential directions with thermal gradients. In comparison to various particle sizes with varied thicknesses, long-term deformation occurs. The results are displayed visually, demonstrating how creep deformation decreases with changing particle size and thickness.

Keywords: creep, varying thickness, particle size, stresses and strain rates

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Authors: Pooja Bakshi

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In the following paper, an attempt would be made to engage with the relationship between the state and the imperatives of security from a gendered lens. This will be juxtaposed with the feminist engagement with International Law. Theorizations from the literature on South Asian politics and Global politics would be applied to the manner in which the Indian state has defined and proposed to deal with concerns of internal security pertaining to the ‘Left Wing Extremism’ in 2010-2011. It would be argued that the state needs to be disaggregated into the legislature, executive and the judiciary; since there are times when some institutional parts of the state provide space for progressive democratic engagement whilst other institutions don’t. The specific contours of violence faced by women and children at the hands of the state, in the above-mentioned discourse would also be examined. In the end, implications of the security state discourse on debates in International Law would be elaborated.

Keywords: feminist engagement, human rights, state response to left extremism, security studies in South Asia

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Authors: K. C. Iyer, Yogita Manan Bindal, Sumit Kumar Bakshi

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Disputes and litigation are becoming inherent to the construction industry in India, and despite construction being one of the major drivers of growth, there have not been many reforms in the government construction contracts. Many of the disputes arising from the government contracts, can be avoided by the proper drafting of contracts and their administration. This study aims to 1) identify the best practices in the construction contract as reviewed from the research papers and additional literature on contract management, 2) obtain perspectives from the industry experts on the implementation of these best practices with regards to likely challenges and relative benefits for implementing the best practices in construction contracts. The best practices for disputes arising due to delay events have been identified through extensive literature survey. The industry perspective is gathered by way of a questionnaire survey to understand the applicability of the identified best practices, the benefits that are likely to be obtained and the challenges that are likely to be faced in the implementation of these practices. The study concludes with the recommended best practices that can be implemented based on the perspectives obtained from the survey. The findings of the study can be used by the industry professionals while drafting construction contracts with a view to avoid disputes related to delay events.

Keywords: best practices, construction contract, delay, dispute avoidance

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Authors: K. C. Iyer, Yogita Manan Bindal, Sumit Kumar Bakshi

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The construction industry is plagued with disputes and litigation in India with many stalled projects seeking dispute resolution. This has an adverse effect on the performance and overall project delivery and impacts future investments within the industry. While construction industry is the major driver of growth, there has not been major reforms in the government construction contracts. The study is aimed at identifying the proactive means of dispute avoidance, focusing on reforms required within the construction contracts, by studying 49 arbitration awards of construction disputes. The claims presented in the awards are aggregated to study the causes linked to the contract document and are referred against the prospective recommendation and practices as surveyed from literature review of research papers. Within contract administration, record keeping has been a major concern as they are required by the parties to substantiate the claims or the counterclaims and therefore are essential in any dispute redressal process. The study also observes that the right judgment is inhibited when the record keeping is improper and due to lack of coherence between documents, the dispute resolution period is also prolonged. The finding of the research will be relevant to industry practitioners in contract drafting with a view to avoid disputes.

Keywords: construction contract, contract administration, contract management, dispute avoidance

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Authors: Surjeet Bakshi, Surabhi Sharma

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Objective: To assess the impact of educational sessions by reputed regional faculties on knowledge of primary care physicians on evidence based diabetes management methods and practice. Study Design: Retrospective pre-post intervention study. Methodology: Nine cities in Kerala from August to October, 2012 were selected for the study. 125 MBBS doctors participated in the study. 11 regional faculties provided six educational sessions throughout the period. Validated questionnaires were used to evaluate the knowledge of the participants on evidence based diabetes management methods before and after the intervention. Results: The mean score on pre-test was 8 and the mean score on post-test was 9. A paired t-test was conducted on participant’s pre- and post test score and the results were statistically significant (p<0.001). Conclusion: Even though the general attitude to and level of knowledge of diabetes management is good among the primary care physicians in India, there do exist some knowledge gaps which might influence their future practices when it comes to counselling and information on diabetes management methods. In the present study, the performance and awareness level of the participants have expressively improved among primary care physicians. There is a significant improvement in the test score and the training conducted. It seems that if such study programmes are included in the students study programme, it will give higher score in the knowledge and attitude towards diabetes management.

Keywords: diabetes, management, primary care physicians, evidence base, improvement score, knowledge

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Authors: Parmjit S. Panesar, Rupinder Kaur, Ram S. Singh

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Enzymes are the biocatalysts which catalyze the biochemical processes and thus have a wide variety of applications in the industrial sector. β-Galactosidase (E.C. 3.2.1.23) also known as lactase, is one of the prime enzymes, which has significant potential in the dairy and food processing industries. It has the capability to catalyze both the hydrolytic reaction for the production of lactose hydrolyzed milk and transgalactosylation reaction for the synthesis of prebiotics such as lactulose and galactooligosaccharides. These prebiotics have various nutritional and technological benefits. Although, the enzyme is naturally present in almonds, peaches, apricots and other variety of fruits and animals, the extraction of enzyme from these sources increases the cost of enzyme. Therefore, focus has been shifted towards the production of low cost enzyme from the microorganisms such as bacteria, yeast and fungi. As compared to yeast and bacteria, fungal β-galactosidase is generally preferred as being extracellular and thermostable in nature. Keeping the above in view, the present study was carried out for the isolation of the β-galactosidase producing fungal strain from the food as well as the agricultural wastes. A total of more than 100 fungal cultures were examined for their potential in enzyme production. All the fungal strains were screened using X-gal and IPTG as inducers in the modified Czapek Dox Agar medium. Among the various isolated fungal strains, the strain exhibiting the highest enzyme activity was chosen for further phenotypic and genotypic characterization. The strain was identified as Rhizomucor pusillus on the basis of 5.8s RNA gene sequencing data.

Keywords: beta-galactosidase, enzyme, fungal, isolation

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Authors: Lopamudra Bakshi Basu

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Transport is one of the most important characteristic features of Indian cities. The physical and societal requirements determine the selection of a particular transport system along with the uniqueness of road networks. Kolkata has a mixed traffic of which Paratransit system plays a crucial role. It is an indispensable transport system in Kolkata mainly because of its size and service flexibility which has led to a unique network character. The paratransit system, mainly the autorickshaws, is the most favoured mode of transport in the city. Its fast movement and comfortability make it a vital transport system of the city. Since the inception of the autorickshaws in Kolkata in 1981, this mode has gained popularity and presently serves nearly 80 to 90 percent of the total passenger trips. This employment generating mode of transport has increased its number rapidly affecting the city’s traffic. Minimal check on their growth by the authority has led to traffic snarls along many streets of Kolkata. Indiscipline behavior, violation of traffic rules and rash driving make situations even worse. The rise in the number and increasing popularity of the autorickshaws make it an interesting study area. Autorickshaws as a paratransit mode play its role as a leader or a follower. However, it is informal in its planning and operations, which makes it a problem area for the city. The entire research work deals with the growth and expansion of the number of vehicles and the routes within the city. The development of transport system has been interesting in the city, which has been studied. The growth of the paratransit modes in the city has been rapid. The network pattern of the paratransit mode within Kolkata has been analysed.

Keywords: growth, informal, network characteristics, paratransit, service flexibility

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Authors: Rupinder Kaur, Parmjit S. Panesar, Ram S. Singh

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Whey is the lactose rich by-product of the dairy industry, having good amount of nutrient reservoir. Most abundant nutrients are lactose, soluble proteins, lipids and mineral salts. Disposing of whey by most of milk plants which do not have proper pre-treatment system is the major issue. As a result of which, there can be significant loss of potential food and energy source. Thus, whey has been explored as the substrate for the synthesis of different value added products such as enzymes. β-galactosidase is one of the important enzymes and has become the major focus of research due to its ability to catalyze both hydrolytic as well as transgalactosylation reaction simultaneously. The enzyme is widely used in dairy industry as it catalyzes the transformation of lactose to glucose and galactose, making it suitable for the lactose intolerant people. The enzyme is intracellular in both bacteria and yeast, whereas for molds, it has an extracellular location. The present work was carried to utilize the whey for the production of β-galactosidase enzyme using both yeast and fungal cultures. The yeast isolate Kluyveromyces marxianus WIG2 and various fungal strains have been used in the present study. Different disruption techniques have also been investigated for the extraction of the enzyme produced intracellularly from yeast cells. Among the different methods tested for the disruption of yeast cells, SDS-chloroform showed the maximum β-galactosidase activity. In case of the tested fungal cultures, Aureobasidium pullulans NCIM 1050, was observed to be the maximum extracellular enzyme producer.

Keywords: β-galactosidase, fungus, yeast, whey

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Authors: Harminder Singh, Rupinder Kaur

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Peach is primarily a temperate fruit, but its low chilling cultivars are grown quite successfully in the sub-tropical climate as well. The area under peach cultivation is picking up rapidly in the sub tropics of northern India due to higher return on a unit area basis, availability of suitable peach cultivar and their production technology. Information on the use of different training systems on peach in the sub tropics is inadequate. In this investigation, conducted at Punjab Agricultural University, Ludhiana (Punjab), India, the trees of the Shan-i-Punjab peach were planted at four different spacings i.e. 6.0x3.0m, 6.0x2.5m, 4.5x3.0m and 4.5x2.5m and were trained to central leader system. The total radiation interception and penetration in the upper and lower canopy parts were higher in 6x3.0m and 6x2.5m planted trees as compared to other spacings. Average radiation interception was maximum in the upper part of the tree canopy, and it decreased significantly with the depth of the canopy in all the spacings. Tree planted at wider spacings produced more vegetative (tree height, tree girth, tree spread and canopy volume) and reproductive growth (flower bud density, number of fruits and fruit yield) per tree but productivity was maximum in the closely planted trees. Fruits harvested from the wider spaced trees were superior in fruit quality (size, weight, colour, TSS and acidity) and matured earlier than those harvested from closed spaced trees.

Keywords: quality, radiation, spacings, yield

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Authors: Meenakshi Verma, Mandeep Singh Bakshi, Kultar Singh

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Magnetic nanoparticles have received incredible importance in view of their diverse applications, which arise primarily due to their response to the external magnetic field. The magnetic behaviour of magnetic nanoparticles (NPs) helps them in numerous different ways. The most important amongst them is the ease with which they can be purified and also can be separated from the media in which they are present merely by applying an external magnetic field. This exceptional ease of separation of the magnetic NPs from an aqueous media enables them to use for extracting/removing metal pollutants from complex aqueous medium. Functionalized magnetic NPs can be subjected for the metallic impurities extraction if are favourably adsorbed on the NPs surfaces. We have successfully used the magnetic NPs as vehicles for gold and silver NPs removal from the complex fluids. The NPs loaded with gold and silver NPs pollutant fractions has been easily removed from the aqueous media by using external magnetic field. Similarly, we have used the magnetic NPs for extraction of protein from complex media and then constantly washed with pure water to eliminate the unwanted surface adsorbed components for quantitative estimation. The purified and protein loaded magnetic NPs are best analyzed with SDS Page to not only for characterization but also for separating the protein fractions. A collective review of the results indicates that we have synthesized surfactant coated iron oxide NPs and then functionalized these with selected materials. These surface active magnetic NPs work very well for the extraction of metallic NPs from the aqueous bulk and make the whole process environmentally sustainable. Also, magnetic NPs-Au/Ag/Pd hybrids have excellent protein extracting properties. They are much easier to use in order to extract the magnetic impurities as well as protein fractions under the effect of external magnetic field without any complex conventional purification methods.

Keywords: magnetic nanoparticles, protein, functionalized, extraction

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Authors: Vibhore Bakshi

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This research paper is a step towards understanding the role of governance in the plan preparation process. It addresses the complexities of the peri-urban, historical constructions, politics and policies of sustainability, and legislative frameworks. The paper reflects on the Delhi NCT as one of the classical cases that have happened to witness different structural changes in the master plan around 1981, 2001, 2021, and Proposed Draft 2041. The Delhi Landsat imageries for 1989 and 2018 show an increase in the built-up areas around the periphery of NCT. The peri-urbanization has been a result of increasing in-migration to peri–urban areas of Delhi. The built-up extraction for years 1981, 1991, 2001, 2011, and 2018 highlights the growing peri-urbanization on scarce land therefore, it becomes equally important to research the history of the land and its legislative measures. It is interesting to understand the streaks of changes that have occurred in the land of Delhi in accordance with the different master plans and land legislative policies. The process of masterplan process in Delhi has experienced a lot of complexities in juxtaposition to other metropolitan regions of the world. The paper identifies the shortcomings in the current master planning process approach in regard to the stage of the planning process, traditional planning approach, and lagging ICT-based interventions. The metropolitan governance systems across the globe and India depict diversity in the organizational setup and varied dissemination of functions. It addresses the complexity of the peri-urban, historical constructions, politics and policies of sustainability, and legislative frameworks.

Keywords: governance, land provisions, built-up areas, in migration, built up extraction, master planning process, legislative policies, metropolitan governance systems

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Authors: Gurlal Singh, Rupinder Singh

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The possible role of the engine brake is to convert a power-producing engine into a power-absorbing retarding mechanism. In this braking system, exhaust gas (EG) from the internal combustion (IC) engines is used to operate air brake in the automobiles. Airbrake is most used braking system in vehicles. In the proposed model, instead of air brake, EG is used to operate the brake lever and stored in a specially designed tank. This pressure of EG is used to operate the pneumatic cylinder and brake lever. Filters used to remove the impurities from the EG, then it is allowed to store in the tank. Pressure relief valve is used to achieve a specific pressure in the tank and helps to avoid further damage to the tank as well as in an engine. The petrol engine is used in the proposed EG braking system. The petrol engine is chosen initially because it produces less impurity in the exhaust than diesel engines. Moreover, exhaust brake system (EBS) for the Diesel engines is composed of gate valve, pneumatic cylinder and exhaust brake valve with the on-off solenoid. Exhaust brake valve which is core component of EBS should have characteristics such as high reliability and long life. In a diesel engine, there is butterfly valve in exhaust manifold connected with solenoid switch which is used to on and off the butterfly valve. When butterfly valve closed partially, then the pressure starts built up inside the exhaust manifold and cylinder that actually resist the movement of piston leads to crankshaft getting stops resulting stopping of the flywheel. It creates breaking effect in a diesel engine. The exhaust brake is a supplementary breaking system to the service brake. It is noted that exhaust brake increased 2-3 fold the life of service brake may be due to the creation of negative torque which retards the speed of the engine. More study may also be warranted for the best suitable design of exhaust brake in a diesel engine.

Keywords: exhaust gas, automobiles, solenoid, airbrake

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Authors: Rupinder Kaler, Faith Ndebele, Nadia Saleem, Hafsa Sheikh

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Background: Workplace related violence and aggression seems to be considered an acceptable occupational hazard for staff in mental health services. There is literature evidence that healthcare workers in mental health settings are at higher risk from aggression from patients. Aggressive behaviours pose a physical and psychological threat to the psychiatric staff and can result in stress, burnout, sickness, and exhaustion. Further evidence informs that health professionals are the most exposed to psychological disorders such as anxiety, depression and post-traumatic stress disorder. Fear that results from working in a dangerous environment and exhaustion can have a damaging impact on patient care and healthcare relationship. Aim: The aim of this study is to investigate the prevalence and impact of aggressive behaviour on staff working at Coventry and Warwickshire Partnership Trust. Methodology: The study methodology included carrying out a manual, anonymised, multi-disciplinary cross-sectional survey questionnaire across all clinical and non-clinical staff at CWPT from both inpatient and community settings. Findings: The unsurprising finding was that of higher prevalence of aggressive behaviours in in-patients in comparison to community staff. Conclusion: There is a high rate of verbal and physical aggression at work and this has a negative impact on the staff emotional and physical well- being. There is also a higher reliance on colleagues for support on an informal basis than formal organisational support systems. Recommendations: A workforce that is well and functioning is the biggest resource for an organisation. Staff safety during working hours is everyone's responsibility and sits with both individual staff members and the organisation. Post-incident organisational support needs to be consolidated, and hands-on, timely support offered to help maintain emotionally well staff on CWPT. The authors recommend development of preventative and practical protocols for aggression with patient and carer involvement. Post-incident organisational support needs to be consolidated, and hands-on, timely support offered to help maintain emotionally well staff on CWPT.

Keywords: safer staff, survey of staff experiences, violence and aggression, mental health

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Authors: Suman Chaudhary, Rupinder K. Kanwar, Jagat R. Kanwar

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Azadirachta indica, also known as Neem belonging to the mahogany family is actively gaining interest in the era of modern day medicine due to its extensive applications in homeopathic medicine such as Ayurveda and Unani. More than 140 phytochemicals have been extracted from neem leaves, seed, bark and flowers for agro-medicinal applications. Among the various components, neem leaf protein (NLP) is currently the most investigated active ingredient, due to its immunomodulatory activities against tumor growth. However, these therapeutic ingredients of neem are susceptible to degradation and cannot withstand the drastic pH changes under physiological environment, and therefore, there is an urgent need of an alternative strategy such as a nano-delivery system to exploit its medicinal benefits. This study hypothesizes that guar gum (GG) derived biodegradable nano-carrier based encapsulation of NLP will improve its stability, specificity and sensitivity, thus facilitating targeted anti-cancer therapeutics. GG is a galactomannan derived from the endosperm of the guar beans seeds. Synthesis of guar nanocapsules (NCs) was performed using nanoprecipitation technique where the GG was encapsulated with NLP. Preliminary experiments conducted to characterize the NCs confirmed spherical morphology with a narrow size distribution of 30-40 nm. Differential scanning colorimetric analysis (DSC) validated the stability of these NCs even at a temperature range of 50-60°C which was well within the physiological and storage conditions. Thermogravimetric (TGA) analysis indicated high decomposition temperature of these NCs ranging upto 350°C. Additionally, Fourier Transform Infrared spectroscopy (FTIR) and the SDS-PAGE data acquired confirmed the successful encapsulation of NLP in the NCs. The anti-cancerous therapeutic property of this NC was tested on colon cancer cells (caco-2) as they are one of the most prevalent form of cancer. These NCs (both NLP loaded and void) were also tested on human intestinal epithelial cells (FHs 74) cells to evaluate their effect on normal cells. Cytotoxicity evaluation of the NCs in the cell lines confirmed that the IC50 for NLP in FHs 74 cells was ~2 fold higher than in caco-2 cells, indicating that this nanoformulation system possessed biocompatible anti-cancerous properties Immunoconfocal microscopy analysis confirmed the time dependent internalization of the NCs within 6h. Recent findings performed using Annexin V and PI staining indicated a significant increase (p ≤ 0.001) in the early and late apoptotic cell population when treated with the NCs signifying the role of NLP in inducing apoptosis in caco-2 cells. This was further validated using Western blot, Polymerase chain reaction (PCR) and Fluorescence activated cell sorter (FACS) aided protein expressional analysis which presented a downregulation of survivin, an anti-apoptotic cell marker and upregulation of Bax/Bcl-2 ratio (pro-apoptotic indicator). Further, both the NLP NC and unencapsulated NLP treatment destabilized the mitochondrial membrane potential subsequently facilitating the release of the pro-apoptotic caspase cascade initiator, cytochrome-c. Future studies will be focused towards granting specificity to these NCs towards cancer cells, along with a comprehensive analysis of the anti-cancer potential of this naturally occurring compound in different cancer and in vivo animal models, will validate the clinical application of this unprecedented protein therapeutic.

Keywords: anti-tumor, guar gum, nanocapsules, neem leaf protein

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Authors: Rajeswari Raguraman, Sowmya Parameswaran, Krishnakumar Subramanian, Jagat Kanwar, Rupinder Kanwar

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Background: Tumor microenvironment has been implicated in several cancers to regulate cell growth, invasion and metastasis culminating in outcome of therapy. Tumor stroma consists of multiple cell types that are in constant cross-talk with the tumor cells to favour a pro-tumorigenic environment. Not much is known about the existence of tumor microenvironment in the pediatric intraocular malignancy, Retinoblastoma (RB). In the present study, we aim to understand the multiple stromal cellular subtypes and tumor stromal interactions expressed in RB tumors. Materials and Methods: Immunohistochemistry for stromal cell markers CD31, CD68, alpha-smooth muscle (α-SMA), vimentin and glial fibrillary acidic protein (GFAP) was performed on formalin fixed paraffin embedded tissues sections of RB (n=12). The differential expression of stromal target molecules; fibroblast activation protein (FAP), tenascin-C (TNC), osteopontin (SPP1), bone marrow stromal antigen 2 (BST2), stromal derived factor 2 and 4 (SDF2 and SDF4) in primary RB tumors (n=20) and normal retina (n=5) was studied by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. The differential expression was correlated with the histopathological features of RB. The interaction between RB cell lines (Weri-Rb-1, NCC-RbC-51) and Bone marrow stromal cells (BMSC) was also studied using direct co-culture and indirect co-culture methods. The functional effect of the co-culture methods on the RB cells was evaluated by invasion and proliferation assays. Global gene expression was studied by using Affymetrix 3’ IVT microarray. Pathway prediction was performed using KEGG and the key molecules were validated using qRT-PCR. Results: The immunohistochemistry revealed the presence of several stromal cell types such as endothelial cells (CD31+;Vim+/-); macrophages (CD68+;Vim+/-); Fibroblasts (Vim+; CD31-;CD68- );myofibroblasts (α-SMA+/ Vim+) and invading retinal astrocytes/ differentiated retinal glia (GFAP+; Vim+). A characteristic distribution of these stromal cell types was observed in the tumor microenvironment, with endothelial cells predominantly seen in blood vessels and macrophages near actively proliferating tumor or necrotic areas. Retinal astrocytes and glia were predominant near the optic nerve regions in invasive tumors with sparse distribution in tumor foci. Fibroblasts were widely distributed with rare evidence of myofibroblasts in the tumor. Both gene and protein expression revealed statistically significant (P<0.05) up-regulation of FAP, TNC and BST2 in primary RB tumors compared to the normal retina. Co-culture of BMSC with RB cells promoted invasion and proliferation of RB cells in direct and indirect contact methods respectively. Direct co-culture of RB cell lines with BMSC resulted in gene expression changes in ECM-receptor interaction, focal adhesion, IL-8 and TGF-β signaling pathways associated with cancer. In contrast, various metabolic pathways such a glucose, fructose and amino acid metabolism were significantly altered under the indirect co-culture condition. Conclusion: The study suggests that the close interaction between RB cells and the stroma might be involved in RB tumor invasion and progression which is likely to be mediated by ECM-receptor interactions and secretory factors. Targeting the tumor stroma would be an attractive option for redesigning treatment strategies for RB.

Keywords: gene expression profiles, retinoblastoma, stromal cells, tumor microenvironment

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