Establishment of Canine Transitional Cell Carcinoma Cell Lines Harboring BRAF V595E Mutation as a Therapeutic Target

Transitional cell carcinoma (TCC) is the most common malignant tumor of the canine urinary tract and tends to have a poor prognosis due to its invasive potential. Recent studies have reported that up to 80% of canine urothelial carcinoma has the BRAF V595E mutation, which is homologous to the human V600E mutation. Activating the BRAF mutation is an actionable target for developing effective therapeutic agents inhibiting the BRAF/mitogen-activated protein kinase (MAPK) pathway in canine cancer as well as human cancer. We established novel canine TCC cell lines from two tumor tissues and one metastatic lymph node of canine TCC patients harboring the BRAF V595E mutation. Tumor tissues highly expressed the BRAF mutant and phosphorylated extracellular signal-related kinases (ERK)1/2 proteins. The derived cell lines demonstrated activated MAPK pathways. We also evaluated the cell lines for sensitivity to BRAF inhibitors. Sorafenib, a multiple kinase inhibitor targeting RAF/vascular endothelial growth factor receptor (VEGFR), successfully inhibited the BRAF/MAPK pathway and induced apoptosis. The established canine TCC cell lines responded with greater sensitivity to sorafenib than to vemurafenib, which is known as a specific BRAF inhibitor in human cancer. Our results demonstrated that canine TCC cells showed different responses compared to human cancer with the BRAF V600E mutation. These cell lines would be valuable research materials to develop therapeutic strategies for canine TCC patients

Seasonal Changes in Antibiotic Resistance Genes in Rivers and Reservoirs in South Korea

The fate of antibiotic resistance genes (ARGs) in aquatic environments, especially in rivers and reservoirs, is receiving growing attention in South Korea because reservoirs are an important source of drinking water in this country. Seasonal changes in the abundance of 11 ARGs and a mobile genetic element (int1) in two reservoirs in South Korea, located near drinking water treatment plants in Cheonan and Cheongju cities, were monitored for 6 mo. In these drinking water sources, total ARG concentrations reached 2.5 ?? 107 copies mL‒1, which is one order of magnitude higher than in influents of some wastewater treatment plants in South Korea. During the sampling periods in August, October, and November 2016 and January 2017, sulfonamides (sul1), ??-lactam antibiotics (blaTEM), and tetracycline (tetA) resistance genes were the most abundant genes at the two sites. The ARG abundance consistently increased in January relative to 16S ribosomal ribonucleic acid (rRNA) counts. General stress responses to oxidative stress and other environmental factors associated with the cold season could be significant drivers of ARG horizontal gene transfer in the environment. Accordingly, removal of ARGs as a key step in water treatment warrants more attention