A study on crosstalk between lung carcinoma and immune cells

Tumor cells are seen to modulate the phenotype of all major immune cells to express tumor favouring phenotypes. Inflammation associated with tumors, a result of such interaction, is increasingly being believed to play a major role in tumor initiation, progression and even metastasis. This modulation is achieved very early when Monocytes, precursors of Macrophages and DCs, from the circulating pool are recruited towards tumors and selectively differentiated. Monocytes, in particular, are thought to generate a cytokine milieu in the microenvironment favourable to tumor. Such a crosstalk and the pathways involved therein are not well established, especially in human models. Using representative human carcinoma cells of different origin including Lung, Colon and Cervix, we show that factor(s) associated with these cells can activate secretion of tumor-associated cytokines, TNF-α, IL-6, IL-10, IL-12p40 but not IL-12p70 or IL-1β from human monocytes. Comparative murine co-cultures are also able to induce similar responses. Treatment of monocytes with TLR-2 blocking antibody inhibits these inflammatory responses upon encountering cell-associated as well as secretory ligand(s) from tumor cells. Pharmacological inhibition of intracellular MAP kinase pathway in carcinoma cells ablates the TLR-2 agonistic activity of carcinoma cells. However, inhibition of EGFR and Ras, two major oncogenic players, had no such effect. Early inflammatory response tends to enhance the proliferation and invasiveness of tumor cells and concurrently, increase the viability of monocytes. These tumor associated inflammatory responses may well be one of the mechanisms to manipulate effector T-cell response against tumors. These results suggest a previously unrecognized pathway that may regulate inflammatory responses triggered by cancer cells from monocytes. Our findings have important implications for understanding Cancer related Inflammation

Anti-Metastatic Propensity of Biscoumarin Scaffold Synthesized Under Catalyst Free Aqueous Phase Microwave Irradiation

An environmentally benign, catalyst free, aqueous phase, microwave assisted method for the synthesis of biscoumarin derivatives using 4-hydroxycoumarin and different aldehydes is reported. The comparative investigation of the same synthetic methodology under conventional refluxing, domestic and scientific programmable microwave synthesizer has been attempted for the purpose of comparing the reaction performance in terms of % yield, reaction time, reproducibility and reaction control. In addition to the increased yields and attenuated reaction times for biscoumarin synthesis, the difficulty of reaction control and result reproducibility usually encountered with domestic microwave ovens have been addressed through an optimized synthetic methodology using microwave synthesizer. Screening bioactive compounds for their propensity towards inhibition of cancer metastasis is a must step in advancement of cancer chemotherapy. The synthesized biscoumarin scaffold was investigated for inhibitory metastatic activity against human lung carcinoma cell line, A549 through in vitro wound healing and invasion assays. The biscoumarin scaffold was found to possess an effective anti-proliferative activity. It was also found to be efficient in ablating the migration and invasiveness of these cells under in vitro conditions. This work is licensed under a Creative Commons Attribution 4.0 International License

Clonal Identification and Molecular Characterization of Saffron (Crocus sativus L.) Clones

Crocus sativus is a triploid sterile plant characterized by its long red stigmas, which produce and store significant quantities of carotenoid derivatives. Saffron is widely used mainly as herbal medicine or food coloring, and as a flavoring agent. It is cultivated only in few countries around the world. Saffron selections of Kashmir showed heterogeneity for stigma length which may be due to the genetic and environmental factors. Identification of high yielding selections using the existing gene pool of saffron shows promise and potential for improving the productivity of this crop. Thirty one morphologically distinct saffron selections/clones were characterized for identification of variation in stigma characteristics and apocarotenoid content. Molecular characterization was done through SSR, ISSR and RAPD markers and comparative gene expression between diverse selections was done through semi-quantitative and quantitative PCR analysis. In present study heterogeneity among 31 saffron selections was observed with respect to stigma length. Apocarotenoid content was estimated through soxhlet extraction and was correlated with stigma length of saffron selections. Significant variation in stigma length (2.86-4.84 cm) and non-heritable change in stigma number was observed across thirty one selected saffron clones. HPLC analysis also revealed significant variation in crocin (40-45mg/g), safranal (0.17-0.28 mg/g) and picrocrocin content (0.87-1.27 mg/gm) contents between the clones. Stigma size viz-a-viz quality evaluation confirmed that saffron of Kashmir is of intrinsically high quality with respect to colouring, aroma and taste. Variability in stigma characteristics observed in saffron selections under study can thus be utilized for saffron crop improvement

Anti-Metastatic Propensity of Biscoumarin Scaffold Synthesized Under Catalyst Free Aqueous Phase Microwave Irradiation

An environmentally benign, catalyst free, aqueous phase, microwave assisted method for the synthesis of biscoumarin derivatives using 4-hydroxycoumarin and different aldehydes is reported. The comparative investigation of the same synthetic methodology under conventional refluxing, domestic and scientific programmable microwave synthesizer has been attempted for the purpose of comparing the reaction performance in terms of % yield, reaction time, reproducibility and reaction control. In addition to the increased yields and attenuated reaction times for biscoumarin synthesis, the difficulty of reaction control and result reproducibility usually encountered with domestic microwave ovens have been addressed through an optimized synthetic methodology using microwave synthesizer. Screening bioactive compounds for their propensity towards inhibition of cancer metastasis is a must step in advancement of cancer chemotherapy. The synthesized biscoumarin scaffold was investigated for inhibitory metastatic activity against human lung carcinoma cell line, A549 through in vitro wound healing and invasion assays. The biscoumarin scaffold was found to possess an effective anti-proliferative activity. It was also found to be efficient in ablating the migration and invasiveness of these cells under in vitro conditions. This work is licensed under a Creative Commons Attribution 4.0 International License

Molecular Characterization of Saffron-Potential Candidates for Crop Improvement

In this study, thirty one (31) morphologically distinct selections of saffron crop were used for molecular characterization. Molecular characterization was done through SSR, ISSR and RAPD markers. RAPD and ISSR markers showed significant variation; however, SSR markers did not reveal any variation between the selected clones. The Jaccard’s similarity coefficient ranged from 0.94 to 1.00 with an average of 0.98 among all 31 selections used. Minimum similarity value (0.94) was observed between CITH-S-107 and PAM-S-116 selections. The study provides sufficient knowledge to identify clones with better stigma characteristics for further crop improvement programs

Bioactive supra decorated thiazolidine-4-carboxylic acid derivatives attenuate cellular oxidative stress by enhancing catalase activity

International audienc

Atomic Insight into the Altered O⁶-Methylguanine-DNA Methyltransferase Protein Architecture in Gastric Cancer

<div><p>O⁶-methylguanine-DNA methyltransferase (MGMT) is one of the major DNA repair protein that counteracts the alkalyting agent-induced DNA damage by replacing O⁶-methylguanine (mutagenic lesion) back to guanine, eventually suppressing the mismatch errors and double strand crosslinks. Exonic alterations in the form of nucleotide polymorphism may result in altered protein structure that in turn can lead to the loss of function. In the present study, we focused on the population feared for high exposure to alkylating agents owing to their typical and specialized dietary habits. To this end, gastric cancer patients pooled out from the population were selected for the mutational screening of a specific error prone region of MGMT gene. We found that nearly 40% of the studied neoplastic samples harbored missense mutation at codon¹⁵¹ resulting into Serine to Isoleucine variation. This variation resulted in bringing about the structural disorder, subsequently ensuing into a major stoichiometric variance in recognition domain, substrate binding and selectivity loop of the active site of the MGMT protein, as observed under virtual microscope of molecular dynamics simulation (MDS). The atomic insight into MGMT protein by computational approach showed a significant change in the intra molecular hydrogen bond pattern, thus leading to the observed structural anomalies. To further examine the mutational implications on regulatory plugs of MGMT that holds the protein in a DNA-Binding position, a MDS based analysis was carried out on, all known physically interacting amino acids essentially clustered into groups based on their position and function. The results generated by physical-functional clustering of protein indicated that the identified mutation in the vicinity of the active site of MGMT protein causes the local and global destabilization of a protein by either eliminating the stabilizing salt bridges in cluster C3, C4, and C5 or by locally destabilizing the “protein stabilizing hing” mapped on C3-C4 cluster, preceding the active site.</p></div

Three dimensional time dependent Phi/Psi distribution showing the change in the Gibbs free energy by the mutation in the Clusters 3 and 4 (black = wt, Red = Mu).

<p>Three dimensional time dependent Phi/Psi distribution showing the change in the Gibbs free energy by the mutation in the Clusters 3 and 4 (black = wt, Red = Mu).</p

Illustrative representation of drastic conformational variability that a mutant structure (arrow specifies the site of an mutated residue) undergoes when compared wild type protein structure.

<p>The snapshots were retrieved at every 5 ns interval along the 30 ns simulation.</p