All Data

In recent years, not only mRNA (messenger RNA) but also other small non-coding RNA have focused on molecular diagnosis and therapy in oncology fields. Especially in human medicine, many studies elucidate the ability and function of many microRNAs, which are small non-coding RNAs. However, there are still not many studies in the veterinary field. In my PhD study, I focused on the non-coding small RNA in canine oncology fields. In the first chapter, I studied the dysregulated micro RNA in canine oral melanoma. At first, I performed the microarray-based miRNA profiling of canine malignant melanoma (CMM) tissue obtained from the oral cavity. Then, I also confirmed the differentially expressed microRNA by quantitative reverse transcription-PCR (qRT-PCR). An analysis of the microarray data revealed 17 dysregulated miRNAs; 5 were up-regulated, and 12 were downregulated. qRT-PCR analysis was performed for 2 up-regulated (miR-204 and miR-383), 3 down-regulated (miR-122, miR-143, and miR-205) and 6 additional oncogenic miRNAs (oncomiRs; miR-16, miR-21, miR-29b, miR-92a, miR-125b and miR-222). The expression levels of seven of the miRNAs, miR16, miR-21, miR-29b, miR-122, miR-125b, miR-204, and miR-383 were significantly up-regulated, while the expression of miR-205 was down- 2 regulated in CMM tissues compared with normal oral tissues. The microarray and qRT-PCR analyses validated the up-regulation of two potential oncomiRs, miR-204 and miR-383. I also constructed a protein interaction network and a miRNA–target regulatory interaction network using STRING and Cytoscape. In the proposed network, was a target for miR-383, and were targets for miR-204, and was a target for both. The miR-383 and miR-204 were potential oncomiRs that may be involved in regulating melanoma development by evading DNA repair and apoptosis. In my second chapter, I focused on non-coding RNA other than microRNA, and I compared canine hepatocellular carcinomas (HCC) and hepatocellular adenomas (HCA). I elucidated the differential expression of Y RNA-derived fragments because Y RNA-derived fragments have yet to be investigated in canine HCC and HCA. I used qRT-PCR to determine Y RNA expression in clinical tissues, plasma, and plasma extracellular vesicles, and two HCC cell lines (95-1044 and AZACH). Y RNA was significantly decreased in tissue, plasma, and plasma extracellular vesicles for canine HCC versus canine HCA and healthy controls. Y RNA was decreased in 95-1044 and AZACH cells versus normal liver tissue and 3 in AZACH versus 95-1044 cells. In plasma samples, Y RNA levels were decreased in HCC versus HCA and Healthy controls and increased in HCA versus Healthy controls. Receiver operating characteristic analysis showed that Y RNA could be a promising biomarker for distinguishing HCC from HCA and healthy controls. Overall, the dysregulated expression of Y RNA can distinguish canine HCC from HCA. However, further research is necessary to elucidate the underlying Y RNA-related molecular mechanisms in hepatocellular neoplastic diseases. To the best of my knowledge, this is the first report on the relative expression of Y RNA in canine HCC and HCA. In conclusion, I have demonstrated the up-regulation of potential oncomiRs, miR-16, miR-21, miR-29b, miR-122, miR-125b, miR-204 and miR383 in CMM tissues. In particular, the strong up-regulation of miR-383 in CMM tissues compared with normal oral tissues identified by microarray screening was confirmed by qRT-PCR. I conclude that miR-383 and miR-204 may promote melanoma development by regulating the DNA repair/checkpoint and apoptosis. Then, I also demonstrated the Y RNA dysregulation in the cHCC. Especially to my knowledge, this is the first report on Y RNA in canine tumors. Interestingly, this ncRNA has distinctive characteristics and differentiates malignant tumors (HCC) from benign 4 tumors (HCA). The expression pattern of Y RNA is consistent across clinical samples and cell lines. Thus, Y RNA has promising potential for differentiating HCC from HCA. Further research is required to fully elucidate the role of Y RNA in the development and progression of canine HCC and HCA.

In Japan, China, and Singapore, several studies have reported increased incidences of peripheral venous catheter-related bloodstream infection by Bacillus cereus during the summer. Therefore, we hypothesized that bed bathing with a B. cereus-contaminated “clean” towels increases B. cereus contact with the catheter and increases the odds of contaminating the peripheral parenteral nutrition (PPN). We found that 1) professionally laundered “clean” towels used in hospitals have B. cereus (3.3×10^4 colony forming units (CFUs) / 25cm^2), 2) B. cereus is transferable onto the forearms of volunteers by wiping with the towels (n=9), and 3) B. cereus remain detectable (80∼660 CFUs /50cm^2) on the forearms of volunteers even with subsequent efforts of disinfection using alcohol wipes. We further confirmed that B. cereus grow robustly (10^2 CFUs /mL to more than 10^6 CFUs /mL) within 24hours at 30°C in PPN. Altogether we find that bed bathing with a towel contaminated with B. cereus leads to spore attachments to the skin, and that B. cereus can proliferate at an accelerated rate at 30°C compared to 20°C in PPN. We therefore highly recommend ensuring the use of sterile bed bath towels prior to PPN administration with catheter in patients requiring bed bathing.