The role of EBNA binding proteins in cell transformation

Epstein - Barr virus (EBV) infects m ajority of the human population and maintains sub - cl inical infection . However , under certain conditions it is associated with several B - cell malignancies , such as Burkitt lymphoma, Hodgkin’s lymphoma etc. Moreover , EBV also plays a causative role in a cquired immunodeficiency syndrome ( AIDS ) associated lymph omas and post - transplant lymphoproliferative disease (PTLD). EBV maintains latent infection and expresses a particular set of proteins that are necessary for host cell proliferation. Studying function of EBV latent proteins could help us to understand the mechanisms underlying EBV induced B - cell transformation. EBV transformed B cells, i.e. lymphoblastoid cell lines (LCLs) is a well - established in vitro model system to study the molecular mechanisms of B - cell transformation. In the present work , we have ide ntified vitamin D receptor (VDR) as a binding p artner of EBNA3. We showed that EBNA3 can block the VDR mediated gene transactivation and protect s B - cells from vitamin D3 induced growth arrest/ apoptosis. We have observed that hypoxia inducible factor 1 alp ha (HIF1 ) is stabilized in LCLs at normoxic conditions. HIF1 is not hydroxylated and therefore it is not degraded in LCLs . We have shown that prolylhydroxylases 1 and 2 (PHD1 and 2) that are responsible for hydroxylation of HIF1 form complex es with and respectively Due to t his binding catalytic activity of PHDs is block ed , resulting in inhibition of HIF1 hydroxylation and subsequent degradation. Stabilized HIF1 is transcriptionally active and induces genes that are involved in glycolysi s. Moreover , LCLs have high levels of pyruvate and lactate in contrast to mitogen activated B cells , indicating induction of aerobic glycolysis or Warburg effect. We have shown that mitochondrial ribosomal protein MRP S18 - 2 (S18 - 2), an EBNA6 binding protein , can immortalize rat embryonic fibroblasts (REFs). These immortalized cells express stem cell markers like SSEA1, Sox2, Oct3/4 and have the characteristics of embryonic stem cells. S18 - 2 also immortalize d the adult rat skin fibroblasts (RSFs). Moreover, s ingle clones from immortalized REFs and RSFs resulted in tumors in SCID mice. T h i s t hes i s w or k re v ea l s three d i ffere n t aspects of EB V induced B - cell transformation, i .e . pr o t ec ti o n fro m v it a m i n D 3 i ndu c e d apop t o s i s , m e t abo li c a d ap t a ti o n required for proliferation an d hijacking functions of novel protein MRPS18 - 2 for immortalization

Chiral Knots: An Application to Synthetic Chemistry

The purpose of this paper is to serve as an introduction to knot theory by mathematically defining a knotand equivalent knots in 3-space, specifically in terms of topology. Via the concepts of ambient isotopy and the Kauffman X polynomial refine definition of equivalent knots. The definition of equivalence interms of the Kauffman X Polynomial has a significant application to synthetic chemistry. All of this leads us to the study of chirality, determining whether a molecule can be deformed into its mirror image. Certain molecules, especially enzymes react differently if they are chiral, means they cannot be deformed into their mirror image. Key words: Equivalent knots, ambient isotopy, Kauffman X polynomial, chirality

Study the Sustainability and challenges of farm to fork restaurants in Metropolitan city of Maharashtra

Farm to fork concept is established in metropolis cities. The data was collected from 31 farms to fork restaurants to understand the operation out of 48 restaurants. The concept still not getting the amount of sustainable business in the Indian market. Research suggests that there is good protection to successfully run the concept restaurant as the city market demands. The study analyses the challenges faced by the restaurant owners, where creating consistent supply chain management of raw fresh food, control on food cost affects the business sustainability. The study will make understand the new business owners for farm to fork restaurant start-up in India. They will understand the challenges and how to survive in a current market environment. &nbsp

Flow past a square-section cylinder with a wavy stagnation face

Published versio

Use of Nanohybrid Material for Formulation, Development and Evaluation of Pharmaceutical Dosage Form Containing the Low Solubility Active Pharmaceutical Ingredient

In oral absorption of a drug, the drug first dissolves and then is absorbed by diffusion through gastrointestinal membranes. The gastrointestinal environment is aqueous in nature and it is well-known that one-third of the drug population is water insoluble. Hence, there is a need for enhancement of the solubility and dissolution of such drugs. In this work, enhancement of the solubility and dissolution of the practically insoluble drug Rosuvastatin Calcium was achieved by formation of nanohybrids using microwave-induced diffusion (MIND), which ultimately leads to bioavailability enhancement. nanohybrids were formed by using natural carriers such as acacia,guar gum and PVP k-30 with the help of microwaves. Selection of carriers was based on their surfactant and wetting properties. Solubility studies were carried out to establish the solubility-enhancing property of the nanohybrids. To support solubility analysis results, dissolution studies (i.e. powder dissolution and in-vitro dissolution) were carried out. The nanohybrids were characterized by Fourier transform infrared spectroscopy, differential scanning calorimetry, X-ray diffraction studies, scanning electron microscopy and transmission electron microscopy. It was found that as the concentration of polymer in the composite increased the solubility and dissolution of rosuvastatin calcium were enhanced. The optimised ratio (drug : polymer) for all the composites was found to be 1:4. The novelty of this work is the green and cost-effective way of forming drug nanocomposites with the help of microwave, which can be scaled up to an industrial level. The method gives an immaculate means of solubilisation by generating drug dispersion at the micro and nanoscale level in natural biodegradable stabilising media. Hence, this study demonstrates the use of nanohybrids in solubility and dissolution enhancement. Keywords- BCS Class II, Nanohybrids, Solubilization, Enhancement of solubility, Optimized drug dissolution and drug release

Formulation and Optimization of Sustained Release Floating Matrix Tablets of Baclofen

This investigation concerns the development of sustained released floating matrix tablets of Baclofen for improving its bioavailability by prolonging gastric residence time. Floating matrix tablets (FMT) of Baclofen were prepared using 32 optimization designs.Carbopol 934p, HPMC K4M and gas generating agents such as sodium bicarbonate and citric acid were used in formulation. The fabricated tablets were evaluated for their physical characteristics such as hardness, drug content, buoyancy, swelling properties and in vitro release studies in 0.1N HCl. The tablets without gas generating agents and HPMC K4M did not float at all. Tablets with gas generating agents and HPMC K4M and Carbopol 934p floated up to 12 h without complete erosion and showed slower drug release. HPMC K4M and Carbopol 934P maintain the integrity of the FMT and sustaining the drug release. The increase in the concentration of HPMC K4M and Carbopol 934p in FMT from 75 mg to 125 mg and 20 mg to 60mg respectively resulted in decrease in release rate of drug. The possibility of drug polymer interaction was determined by differential scanning calorimetric (DSC) and Fourier transform infrared (FTIR) spectrometer, and confirmed no interaction between drug and polymers. The mechanism of drug release is mainly described by diffusion and swelling mechanism of polymers

A RECENT REVIEW ON NASAL MICROEMULSION FOR TREATMENT OF CNS DISORDER

Nasal route is found to be valuable for targeting drugs to CNS via a different mechanism. The advantages, disadvantages, various aspects of nasal anatomy and physiology, mechanism of drug transport from nose brain, drug selection criteria to cross BBB/Blood-CSF barrier are discussed briefly. Nowadays many drugs have better systemic bioavailability through nasal route as compared to oral administration. In addition, intranasal drug delivery enables dose reduction, rapid attainment of therapeutic blood levels, quicker onset of pharmacological activity, and fewer side effects. There are various approaches in delivering a therapeutic substance to the target site. One such approach is using microemulsion as a carrier for the drug. The main purpose of this study is the use of microemulsion technology in drug targeting to the brain along with mechanism of the nose to brain transport, formulation and formation of the microemulsion and its characterization

Identifying Reliable Diagnostic/Predictive Biomarkers for Rheumatoid Arthritis

Introduction and objective: Elevated C-reactive protein is usually a good indicator of rheumatoid arthritis (RA); however, there are limitations that compromise its specificity and therefore there is an urgent need to identify more reliable diagnostic biomarkers to detect early stages of RA. In addition, identifying the correct therapeutic biomarker for the treatment of RA using methotrexate (MTX) would greatly increase the benefits experienced by the patients. Materials and methods: Primary normal synoviocytes human fibroblast-like synoviocytes (HFLS) and its phenotype rheumatic HFLS-RA cells were chosen for this study. The HFLS-RA–untreated and MTX-treated cells were subjected to microarray analysis. Results: Microarray data identified 74 differentially expressed genes. These genes were mapped against an RA inflammatory pathway, shortlisting 10 candidate genes. Gene expression profiling of the 10 genes were studied. Fold change (FC) was calculated to determine the differential expression of the samples. Discussion: The transcription profiles of the 10 candidate genes were highly induced in HFLS-RA cells compared with HFLS cells. However, on treating the HFLS-RA cells with MTX, the transcription profiles of these genes were highly downregulated. The most significant expression FC difference between HFLS and HFLS-RA (treated and untreated) was observed with HSPA6, MMP1, MMP13, and TNFSF10 genes. Conclusions: The data from this study suggest the use of HSPA6, MMP1, MMP13, and TNFSF10 gene expression profiles as potential diagnostic biomarkers. In addition, these gene profiles can help in predicting the therapeutic efficacy of MTX

Can curcumin improve the methotrexate based treatment for rheumatoid arthritis?

Rheumatoid arthritis (RA) is an autoimmune disorder characterised by its varied and unpredictable origin, eventual destruction of cartilage and bone and the perpetual length of treatment involved. Even though significant developments have taken place over the past couple of decades in the treatment, the efficacy of drugs for RA is a major concern due to the side effects involved. Methotrexate (MTX) is currently used as first line treatment due to its ability to modify the rheumatic conditions so that disease progression can be prevented. However, at the same time prolonged exposure to MTX can lead to severe side effects such as lung fibrosis and hepatotoxicity. Recent research has focused on developing an alternative to MTX with similar efficacy but with reduced adverse effects. One such promising option is curcumin, an active compound extracted from Indian spice Turmeric. Various studies have indicated the synergistic properties exhibited by curcumin and its ability to modulate the underlying inflammatory pathways involved in RA. However, no previous studies have been reported with regards to using a combination of MTX and curcumin for the treatment of RA. A novel RP-HPLC stability indicating method was developed in order to establish the compatibility of the two compounds. The method was developed using Waters Reverse Phase (XBridgeTM Shield RP18 4.6x250 mm, 5 µM) column. A gradient system, consisting of two mobile phases with acetonitrile concentrations of 35% and 60% respectively, was designed to optimise the separation of the two compounds while taking into consideration the difference in hydrophobicity. The wavelengths for detection were 305 nm and 430 nm for MTX and curcumin, respectively. The retention time for MTX and curcumin was 4.8 ± 0.10 min and 12.3 ± 0.10 min, respectively. The Page 4 of 192 total run time of analysis was 25 minutes. The developed method was validated for parameters such as accuracy, precision, linearity, limit of detection and lower limit of quantification. The system was tested through intraday and interday repeatability and reproducibility. The method was used to analyse the MTX and curcumin under different stress conditions such as pH, UV radiation, temperature and humidity to establish their compatibility. The degradation products are successfully separated and therefore, the method can be effectively used as stability indicating method. Gene expression profiling was performed using HFLS-RA cells treated with MTX and curcumin separately and concurrently. The DNA microarray data identified 53 genes that were downregulated and 21 genes that were upregulated in all the treated samples using both stringent and non-stringent filtering. The total of 13 genes were selected based on the fold change obtained in the microarray data and their potential as therapeutic biomarkers based on previous research. The gene validation using qRTPCR confirmed the higher efficacy of curcumin in the inhibition of the proinflammatory genes such as ANGPTL7, CD248, CH25H, COL14A1, CXCL12, CYTL1, IFITM1 and IL7. Curcumin was found to also increase the expression levels of genes associated with anti-inflammatory roles, namely BCAR4, CD274, HSPA6, OTP and RELT. The increased gene expression in samples treated with both MTX and curcumin confirmed the possible synergistic activity which is encouraging, taking in to account the fact that these compounds are compatible according to the stability studies carried out and thus could be used in combination for the treatment of RA. This could improve the current treatment by reducing the severity of side-effects attributed to MTX while maintaining the efficacy of the treatment due to the ability of curcumin to modulate specific therapeutic biomarkers involved in the RA pathogenesis

Micro-RNA: biomarker for the treatment of cancer

MicroRNAs (miRNAs) are presently experiencing a renewed height of attention not simplest as diagnostics, however specifically as noticeably promising novel objectives or gear for medical therapy in several different malignant diseases. MicroRNAs have emerged as key publish-transcriptional regulators of gene expression, involved in diverse physiological and pathological approaches. It changed into located that several miRNAs are without delay worried in human cancers, along with lung, breast, mind, liver, colon most cancers and leukemia. Biomarkers have many capacity packages in oncology, together with danger assessment, screening, differential analysis, dedication of analysis, prediction of reaction to treatment, and tracking of progression of disease. In this evaluation, we summarize the present day know-how and ideas concerning the involvement of microRNAs in cancer, which have emerged from the take a look at of cellular tradition and animal model systems, which includes the regulation of key cancer-related pathways, which include mobile cycle control and the DNA damage response. Importantly, microRNA molecules are already getting into the health facility as diagnostic and prognostic biomarkers for patient stratification and also as therapeutic targets and marketers. Keywords: MiRNA, Biomarkers, cancer