No abstract

In this study, we investigated the antibacterial and antioxidant activities of Crocus sativus L. Kashmirianus c.v. extracts (callus and stigmas). Profuse callus was obtained on MS medium enriched with BAP (20 µM) + NAA(15 µM) under in vitro conditions from corm slices. Four pathogenic bacterial strains (Staphylococcus aureus CD0001, Escherichia coli CD0006, Pseudomonas aeruginosa CD0023 and Shigella flexneri CD0033) were used for determining the antibacterial activity of extracts. The antioxidant activity was determined by DPPH assay, DNA protection assay, FTC method, TBA assay and lipid peroxidation assay. The methanol stigma extract of saffron was found to be more effective in inhibiting all the pathogenic strains. The stigma extract also showed significant radical scavenging or chelation capacities in four of the methods; however, callus extract exhibited maximum inhibition of peroxy-radicals in lipid peroxidation assay. The protocol for callus production is described. It was concluded that as well as the specific parts of plants displaying diverse pharmacological activities, callus produced under in vitro conditions will assist in enhancing the production of secondary metabolites, which will reduce the pressure on natural saffron.Keywords: antioxidant activity; callus; in vitro culture; MIC; methanol; saffron; stigma Abbreviations: BAP = 6-benzyl adenine purine BHT = butylated hydroxyl toluene FTC = ferric thiocyanate MIC = minimum inhibitory concentration NAA = naphthalene acetic acid TBA = thiobarbituric acid.

Several lines of evidence indicate that Fibronectin Extra Domain A (EDA) promotes metastatic capacity of tumor cells by engaging cell surface α9β1 integrins. This interaction mediated by the C-C loop of EDA activates pro-oncogenic signaling pathways leading to epithelial to mesenchymal transition (EMT) of tumor cells, thus signifying its importance in control of metastatic progression. In this context the present study was designed to explore the active compounds from selected ethno-medicinal plants of western Himalayan region for targeting EDA of Fibronectin in lung carcinoma cells. Structure based informatics for drug designing and screening was employed to generate a lead compound(s) feed that were conformationally and energetically viable. Out of 120 compounds selected, Irigenin showed best binding-affinity with C-C loop of EDA. Irigenin specifically targeted α9β1 and α4β1 integrin binding sites on EDA comprising LEU46, PHE47, PRO48, GLU58, LEU59 and GLN60 in its C-C loop as evaluated by energy decomposition per residue of Irigenin–EDA complex. In-vitro cell motility assays complemented with EDA knock-in and knockdown assays distinctively demonstrated that Irigenin prevents metastatic capacity of lung cancer cells by selectively blocking EDA. The results presented thus project Irigenin as a lead compound to overcome Fibronectin EDA induced metastatic progression in lung carcinoma cells.

Proteomic analysis using multiplex affinity reagents is perhaps the most reliable strategy to capture differentially expressed proteins that are slightly or immensely modified. In addition to expressional variation, it is comprehensively evident that the immunogenicity of a protein can be a deciding factor for instigating an inflammation afflicted-carcinogenesis. Considering both these factors, a simple and systematic strategy was designed to capture the immunogenic cancer biomarkers from sera of colorectal cancer patients. The affinity reagent, in the form of an antibody repertoire against the secretome of the HT29 cell line was used to grade the sera samples on the basis of the degree of immuno-reactivity and to capture differentially expressed antigens from the patient sera. Following affinity based 2DE-MALDI-TOF; the proteins were identified as (1) soluble vimentin; and (2) TGF-beta-inhibited membrane-associated protein (PP16B), in colon cancer sera and (3) keratin, type II cytoskeletal protein in rectal cancer sera. Pathway reconstruction and protein-protein networking of identified proteins predicted only Vimentin to be physically and genetically engaged in close proximity with the most established colorectal cancer associated tumorigenic pathways. Furthermore, our findings suggest that a possible surface stoichiometric shift in the structure of protein could be due to mutations in the coding sequence of Vimentin that may elicit its enhanced secretion possibly due to protein-hyperphosphorylation. Of the three proteins identified, only Vimentin showed higher expression in sera of colon cancer patients alone. Thus, it could be argued that vimentin might help in predicting individuals at higher risk of developing colon cancers. Our data are therefore suggestive of using vimentin as an antigen for tumor vaccination in an autologous set-up for colon cancers.