Search results for: Swati Rajput

Authors: Swati Srivastava, Mattia Lauriola, Yuval Gilad, Adi Kimchi, Yosef Yarden

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The glucocorticoid receptor (GR) has been proposed to play important, but incompletely understood roles in cancer. Glucocorticoids (GCs) are widely used as co-medication of various carcinomas, due to their ability to reduce the toxicity of chemotherapy. Furthermore, GR antagonism has proven to be a strategy to treat triple negative breast cancer and castration-resistant prostate cancer. These observations suggest differential GR involvement in cancer subtypes. The goal of our study has been to elaborate the current understanding of GR signaling in tumor progression and metastasis. Our study involves two cellular models, non-tumorigenic breast epithelial cells (MCF10A) and Ewing sarcoma cells (CHLA9). In our breast cell model, the results indicated that the GR agonist dexamethasone inhibits EGF-induced mammary cell migration, and this effect was blocked when cells were stimulated with a GR antagonist, namely RU486. Microarray analysis for gene expression revealed that the mechanism underlying inhibition involves dexamenthasone-mediated repression of well-known activators of EGFR signaling, alongside with enhancement of several EGFR’s negative feedback loops. Because GR mainly acts primarily through composite response elements (GREs), or via a tethering mechanism, our next aim has been to find the transcription factors (TFs) which can interact with GR in MCF10A cells.The TF-binding motif overrepresented at the promoter of dexamethasone-regulated genes was predicted by using bioinformatics. To validate the prediction, we performed high-throughput Protein Complementation Assays (PCA). For this, we utilized the Gaussia Luciferase PCA strategy, which enabled analysis of protein-protein interactions between GR and predicted TFs of mammary cells. A library comprising both nuclear receptors (estrogen receptor, mineralocorticoid receptor, GR) and TFs was fused to fragments of GLuc, namely GLuc(1)-X, X-GLuc(1), and X-GLuc(2), where GLuc(1) and GLuc(2) correspond to the N-terminal and C-terminal fragments of the luciferase gene.The resulting library was screened, in human embryonic kidney 293T (HEK293T) cells, for all possible interactions between nuclear receptors and TFs. By screening all of the combinations between TFs and nuclear receptors, we identified several positive interactions, which were strengthened in response to dexamethasone and abolished in response to RU486. Furthermore, the interactions between GR and the candidate TFs were validated by co-immunoprecipitation in MCF10A and in CHLA9 cells. Currently, the roles played by the uncovered interactions are being evaluated in various cellular processes, such as cellular proliferation, migration, and invasion. In conclusion, our assay provides an unbiased network analysis between nuclear receptors and other TFs, which can lead to important insights into transcriptional regulation by nuclear receptors in various diseases, in this case of cancer.

Keywords: epidermal growth factor, glucocorticoid receptor, protein complementation assay, transcription factor

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Authors: Swati Mishra

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In recent years, there has been an explosion in Gold NanoParticle (GNP) research, with a rapid increase in publications in diverse fields, including imaging, bioengineering, and molecular biology. GNPs exhibit unique physicochemical properties, including surface plasmon resonance (SPR) and bind amine and thiol groups, allowing surface modification and use in biomedical applications. Nanoparticle functionalization is the subject of intense research at present, with rapid progress being made towards developing biocompatible, multi-functional particles. In the present study, the photochemical method has been done to functionalize various-shaped GNPs like nanostars by the molecules like ninhydrin. Ninhydrin is bactericidal, virucidal, fungicidal, antigen-antibody reactive, and used in fingerprint technology in forensics. The GNPs functionalized with ninhydrin efficiently will bind to the amino acids on the target protein, which is of eminent importance during the pandemic, especially where long-term treatments of COVID- 19 bring many side effects of the drugs. The photochemical method is adopted as it provides low thermal load, selective reactivity, selective activation, and controlled radiation in time, space, and energy. The GNPs exhibit their characteristic spectrum, but a distinctly blue or redshift in the peak will be observed after UV irradiation, ensuring efficient ninhydrin binding. Now, the bound ninhydrin in the GNP carrier, upon chemically reacting with any amino acid, will lead to the formation of Rhumann purple. A common method of GNP production includes citrate reduction of Au [III] derivatives such as aurochloric acid (HAuCl4) in water to Au [0] through a one-step synthesis of size-tunable GNPs. The following reagents are prepared to validate the approach. Reagent A solution 1 is0.0175 grams ninhydrin in 5 ml Millipore water Reagent B 30 µl of HAuCl₄.3H₂O in 3 ml of solution 1 Reagent C 1 µl of gold nanostars in 3 ml of solution 1 Reagent D 6 µl of cetrimonium bromide (CTAB) in 3 ml of solution1 ReagentE 1 µl of gold nanostars in 3 ml of ethanol ReagentF 30 µl of HAuCl₄.₃H₂O in 3 ml of ethanol ReagentG 30 µl of HAuCl₄.₃H₂O in 3 ml of solution 2 ReagentH solution 2 is0.0087 grams ninhydrin in 5 ml Millipore water ReagentI 30 µl of HAuCl₄.₃H₂O in 3 ml of water The reagents were irradiated at 254 nm for 15 minutes, followed by their UV Visible spectroscopy. The wavelength was selected based on the one reported for excitation of a similar molecule Pthalimide. It was observed that the solution B and G deviate around 600 nm, while C peaks distinctively at 567.25 nm and 983.9 nm. Though it is tough to say about the chemical reaction happening, butATR-FTIR of reagents will ensure that ninhydrin is not forming Rhumann purple in the absence of amino acids. Therefore, these experiments, we achieved the functionalization of gold nanostars with ninhydrin corroborated by the deviation in the spectrum obtained in a mixture of GNPs and ninhydrin irradiated with UV light. It prepares them as a carrier molecule totake up amino acids for targeted delivery or germicidal action.

Keywords: gold nanostars, ninhydrin, photochemical method, UV visible specgtroscopy

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Authors: Swati Vohra

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A genuine economic and social transformation requires equal contribution and participation from both men and women; however, achieving this gender parity is a global concern. In the patriarchal societies around the world, women have been silenced, oppressed, and subjugated. Girls and women comprise half of the world’s population. This, however, must not be the lone reason for recognizing and providing equal opportunities to them. Every individual has a right to develop through opportunities without the biases of gender, caste, race, or ethnicity. The world today is confronted by pressing issues of climate change, economic crisis, violence against women and children, escalating conflicts, to name a few. Achieving gender parity is thus an essential component in meeting this wide array of challenges in order to create just, robust and inclusive societies. In 2015, The United Nation enunciated achieving 17 Sustainable Development Goals by 2030, one of which is SGD#5- Gender Equality, that is not merely a stand-alone goal. It is central to the achievement of all 17 SDG’s. Without progress on gender equality, the global community will not only fail to achieve the SDG5, but it will also lose the impetus towards achieving the broad 2030 agenda. This research is based on a hypothesis that aims to connect the targets laid by the UN under SDG#5 - 5.4 (Recognize and value unpaid care and domestic work) and 5.5 (Ensure women participation for leadership at all levels of decision-making). The study evaluates the impact of unpaid household responsibilities on women’s leadership in India. In Indian society, women have experienced a low social status for centuries, which is reflected throughout the Indian history with preference of a male child and common occurrences of female infanticides that are still prevalent in many parts of the country. Insistence on the traditional gender roles builds patriarchal inequalities into the structure of Indian society. It is argued that a burden of unpaid labor on women is placed, which narrows the opportunities and life chances women are given and the choices they are able to make, thereby shutting them from shared participation in public and economic leadership. The study investigates theoretical framework of social construction of gender, unpaid labor, challenges to women leaders and peace theorist perspective as the core components. The methodology used is qualitative research of comprehensive literature, accompanied by the data collected through interviews of representatives of women leaders from various fields within Delhi-National Capital Region (NCR). The women leaders interviewed had the privilege of receiving good education and a conducive family support; however, post marriage and children it was not the case and the social obligations weighed heavy on them. The research concludes by recommending the importance of gender-neutral parenting and education along with government ratified paternal leaves for at least six months and childcare facilities available for both the parents at workplace.

Keywords: gender equality, gender roles, peace studies, sustainable development goals, social construction, unpaid labor, women’s leadership

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Authors: Swati Gautam, Salman Akhtar, S. P. Jaiswar, Amita Jain

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Background: FGTB is now a major global health problem mostly in developing countries including India. In humans, Mycobacterium Tuberculosis (M.tb) is a causating agent of infection. High index of suspicion is required for early diagnosis due to asymptomatic presentation of FGTB disease. In macrophages Toll Like Receptor-2 (TLR-2) is one which mediated host’s immune response to M.tb. The expression of TLR-2 on macrophages is important to determine the fate of innate immune responses to M.tb. TLR-2 have two work. First its high expression on macrophages worsen the outer of infection and another side, it maintains M.tb to its dormant stage avoids activation of M.tb from latent phase. Single Nucleotide Polymorphism (SNP) of TLR-2 gene plays an important role in susceptibility to TB among different populations and subsequently, in the development of infertility. Methodology: This Case-Control study was done in the Department of Obs and Gynae and Department of Microbiology at King George’s Medical University, U.P, Lucknow, India. Total 300 subjects (150 Cases and 150 Controls) were enrolled in the study. All subjects were enrolled only after fulfilling the given inclusion and exclusion criteria. Inclusion criteria: Age 20-35 years, menstrual-irregularities, positive on Acid-Fast Bacilli (AFB), TB-PCR, (LJ/MGIT) culture in Endometrial Aspiration (EA). Exclusion criteria: Koch’s active, on ATT, PCOS, and Endometriosis fibroid women, positive on Gonococal and Chlamydia. Blood samples were collected in EDTA tubes from cases and healthy control women (HCW) and genomic DNA extraction was carried out by salting-out method. Genotyping of TLR2 genetic variants (Arg753Gln and Arg677Trp) were performed by using single amplification refractory mutation system (ARMS) PCR technique. PCR products were analyzed by electrophoresis on 1.2% agarose gel and visualized by gel-doc. Statistical analysis of the data was performed using the SPSS 16.3 software and computing odds ratio (OR) with 95% CI. Linkage Disequiliribium (LD) analysis was done by SNP stats online software. Results: In TLR-2 (Arg753Gln) polymorphism significant risk of FGTB observed with GG homozygous mutant genotype (OR=13, CI=0.71-237.7, p=0.05), AG heterozygous mutant genotype (OR=13.7, CI=0.76-248.06, p=0.03) however, G allele (OR=1.09, CI=0.78-1.52, p=0.67) individually was not associated with FGTB. In TLR-2 (Arg677Trp) polymorphism a significant risk of FGTB observed with TT homozygous mutant genotype (OR= 0.020, CI=0.001-0.341, p < 0.001), CT heterozygous mutant genotype (OR=0.53, CI=0.33-0.86, p=0.014) and T allele (OR=0.463, CI=0.32-0.66, p < 0.001). TT mutant genotype was only found in FGTB cases and frequency of CT heterozygous more in control group as compared to FGTB group. So, CT genotype worked as protective mutation for FGTB susceptibility group. In haplotype analysis of TLR-2 genetic variants, four possible combinations, i.e. (G-T, A-C, G-C, and A-T) were obtained. The frequency of haplotype A-C was significantly higher in FGTB cases (0.32). Control group did not show A-C haplotype and only found in FGTB cases. Conclusion: In conclusion, study showed a significant association with both genetic variants of TLR-2 of FGTB disease. Moreover, the presence of specific associated genotype/alleles suggest the possibility of disease severity and clinical approach aimed to prevent extensive damage by disease and also helpful for early detection of disease.

Keywords: ARMS, EDTA, FGTB, TLR

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Authors: Swati Bhomia, C. M. Kishtawal, Neeru Jaiswal

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Traditionally, monsoon forecasts have encountered many difficulties that stem from numerous issues such as lack of adequate upper air observations, mesoscale nature of convection, proper resolution, radiative interactions, planetary boundary layer physics, mesoscale air-sea fluxes, representation of orography, etc. Uncertainties in any of these areas lead to large systematic errors. Global circulation models (GCMs), which are developed independently at different institutes, each of which carries somewhat different representation of the above processes, can be combined to reduce the collective local biases in space, time, and for different variables from different models. This is the basic concept behind the multi-model superensemble and comprises of a training and a forecast phase. The training phase learns from the recent past performances of models and is used to determine statistical weights from a least square minimization via a simple multiple regression. These weights are then used in the forecast phase. The superensemble forecasts carry the highest skill compared to simple ensemble mean, bias corrected ensemble mean and the best model out of the participating member models. This approach is a powerful post-processing method for the estimation of weather forecast parameters reducing the direct model output errors. Although it can be applied successfully to the continuous parameters like temperature, humidity, wind speed, mean sea level pressure etc., in this paper, this approach is applied to rainfall, a parameter quite difficult to handle with standard post-processing methods, due to its high temporal and spatial variability. The present study aims at the development of advanced superensemble schemes comprising of 1-5 day daily precipitation forecasts from five state-of-the-art global circulation models (GCMs), i.e., European Centre for Medium Range Weather Forecasts (Europe), National Center for Environmental Prediction (USA), China Meteorological Administration (China), Canadian Meteorological Centre (Canada) and U.K. Meteorological Office (U.K.) obtained from THORPEX Interactive Grand Global Ensemble (TIGGE), which is one of the most complete data set available. The novel approaches include the dynamical model selection approach in which the selection of the superior models from the participating member models at each grid and for each forecast step in the training period is carried out. Multi-model superensemble based on the training using similar conditions is also discussed in the present study, which is based on the assumption that training with the similar type of conditions may provide the better forecasts in spite of the sequential training which is being used in the conventional multi-model ensemble (MME) approaches. Further, a variety of methods that incorporate a 'neighborhood' around each grid point which is available in literature to allow for spatial error or uncertainty, have also been experimented with the above mentioned approaches. The comparison of these schemes with respect to the observations verifies that the newly developed approaches provide more unified and skillful prediction of the summer monsoon (viz. June to September) rainfall compared to the conventional multi-model approach and the member models.

Keywords: multi-model superensemble, dynamical model selection, similarity criteria, neighborhood technique, rainfall prediction

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