Microfabricated platforms to quantitatively investigate cellular behavior under the influence of chemical gradients

Understanding cellular behavior in a quantitative manner will provide valuable information to reveal the mechanism of diseases and immune defense and for the discovery of new drugs and new treatment strategies. Today one of the biggest limitations is the traditional methods and tools that we use to investigate cellular behavior. Since conventional techniques are not adequate enough to be selective, specific and precise, rare cells such as metastatic or drug-resistant ones or events such as the onset symptoms of tumors or infections are masked by the majority of cells or events in the population. As a consequence, our diagnosis or treatment approaches might not target the right cells at the right time in the right microenvironment. Therefore, missed and delayed diagnosis or the wrong treatment strategies emerge. To overcome these limitations, microfabricated tools in conjunction with live-cell imaging provide quantitative, accurate, reproducible and repeatable information at single-cell resolution related to the behavior of rare cells in their natural microenvironments. Moreover, these modern methods reduce assay time, sample consumption and waste production, while improving sensitivity, throughput and precision. Herein, we present an integrated microfabricated microfluidic platform for cell culture and drug exposure investigations

Prevalence of K-Ras mutations in hepatocellular carcinoma: A Turkish Oncology Group pilot study

İstanbul Bilim Üniversitesi, Tıp Fakültesi.Hepatocellular carcinoma (HCC) is the fifth most common male-predominant type of cancer worldwide. There is no effective treatment regimen available for advanced‑stage disease and chemotherapy is generally ineffective in these patients. The number of studies on the prevalence of K‑Ras mutations in HCC patients is currently limited. A total of 58 patients from 6 comprehensive cancer centers in 4 metropolitan cities of Turkey were enrolled in this study. Each center committed to enroll approximately 10 random patients whose formalin‑fixed paraffin‑embedded tumor tissues were available for K‑Ras, exon 2 genotyping. Two methods were applied based on the availability of adequate amounts of tumor DNA. In the first method, the samples were processed using TheraScreen. The genomic DNA was further used to detect the 7 most frequent somatic mutations (35G>A; 35G>C; 35G>T; 34G>A; 34G>C; 34G>T and 38G>A) in codons 12 and 13 in exon 2 of the K‑Ras oncogene by quantitative polymerase chain reaction (PCR). In the second method, the genomic DNA was amplified by PCR using primers specific for K‑Ras exon 2 with the GML SeqFinder Sequencing System's KRAS kit. The identified DNA sequence alterations were confirmed by sequencing both DNA strands in two independent experiments with forward and reverse primers. A total of 40 samples had adequate tumor tissue for the mutation analysis. A total of 33 (82.5%) of the investigated samples harbored no mutations in exon 2. All the mutations were identified via a direct sequencing technique, whereas none were identified by TheraScreen. In conclusion, in our patients, HCC exhibited a remarkably low (<20%) K‑Ras mutation rate. Patients harboring K‑Ras wild‑type tumors may be good candidates for treatment with epidermal growth factor inhibitors, such as cetuximab