Mutation analysis of KRAS and BRAF genes in metastatic colorectal cancer: A first large scale study from Iran

Background: The investigation of mutation patterns in oncogenes potentially can make available a reliable mechanism for management and treatment decisions for patients with colorectal cancer (CRC). This study concerns the rate of KRAS and BRAF genes mutations in Iranian metastatic colorectal cancer (mCRC) patients, as well as associations of genotypes with clinicopathological features. Materials and Methods: A total of 1,000 mCRC specimens collected from 2008 to 2012 that referred to the Mehr Hospital and Partolab center, Tehran, Iran enrolled in this cross sectional study. Using HRM, Dxs Therascreen and Pyrosequencing methods, we analyzed the mutational status of KRAS and BRAF genes in these. Results: KRAS mutations were present in 33.6 cases (n=336). Of KRAS mutation positive cases, 85.1 were in codon 12 and 14.9 were in codon 13. The most frequent mutation at KRAS codon 12 was Gly12Asp; BRAF mutations were not found in any mCRC patients (n=242). In addition, we observed a strong correlation of KRAS mutations with some clinicopathological characteristics. Conclusions: KRAS mutations are frequent in mCRCs while presence of BRAF mutations in these patients is rare. Moreover, associations of KRAS genotypes with non-mucinous adenocarcinoma and depth of invasion (pT3) were remarkable

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Isolation of Mesenchymal Stem Cells From Dental Pulp of Exfoliated Human Deciduous Teeth

Objective: The exfoliated human deciduous tooth (SHED) contain multipotent stemcells that identified to be a population of highly proliferative and clonogenic .Thesecells are capable of differentiating into a variety of cell types including neural cells,adipocytes, and odontoblasts.Material and Methods: Normal exfoliated human deciduous incisors collected fromsix- to nine-years-old children. The pulp was separated from the crown and digestedwith collagenase .Single cell solutions were cultivated in α-MEM supplemented withES-FCS. After two to three days, the cells reached confluency and were trypsinizedand cultured for further passages. The passage-4 cells were analyzed with CD34,CD45, CD105, CD166, CD31, CD90 and CD146 markers that indicated these cellshad a mesenchymal stem cell (MSC) identity. We examined the cells for AlkalinePhosphatase activity to investigate the mesenchymal (stromal) nature.Finally, thecells were differentiated into the osteoblastic and adipocytic lineages in differentsubcultures and analysed by RT-PCR and different staining protocols.Results: Viable cells growing out of the explants showed elongated shapes inclusters. These cells showed alkaline phosphatase activity. Flow cytometry resultsrevealed high expression of pluripotent stem cell markers .In some area of theosteoinductive cultures nodule-like structures were observed that showed redmineralizing area upon staining with Alizarin Red.In adipogenic cultures lipid vesiclesappeared after five weeks of induction with Oil Red.Conclusion: This study show that pulp contains cells with high plasticity andproliferation capacity and can be easily isolated without any serious intervention