ABSTRACT
Antimicrobial (AM) resistance is a growing threat in human and veterinary medicine, spreading across species and perceived as One-Health issue. Prudent use of AM products is essential in mitigating this risk in both human and veterinary medicine. Farm veterinarians, responsible for prescribing appropriate AM and offering advice to farmers on their proper usage, are regarded as key players in the livestock industry. An online survey of farm veterinarians (n = 1,531) was conducted to assess their educational experience, beliefs, current status of prescription, practical behavior, and self-efficacy regarding the prudent use of antimicrobials (PUA). The data from 170 respondents were analyzed using descriptive statistics, regression and mediation analysis. Participants chose AM based on their experiences, academic resources, and peer veterinarians. Approximately 77% did not routinely conduct antibiotic-susceptibility-test. Most participants believed in the importance of PUA and the role of veterinarians to reduce AM resistance, but they reported a lack of awareness among farmers and insufficient national support interfere with their practical behavior with regard to PUA. Half of the participants had PUA-education, and 78.6% reported that education had a positive impact on their behavior with PUA. Self-efficacy partly mediates the relationship between belief and behavior for PUA, which accounted for 23.77% of the total effect that beliefs have on behavior for PUA. The findings of the study show that enhancing veterinarians' abilities to practice and their self-efficacy through education tailored to the current status of farms can contribute to the reduction of AMs in the livestock sector.
ABSTRACT
PURPOSE: Breast cancer has a high prevalence in Korea. To achieve personalized therapy for breast cancer, long-term follow-up specimens are needed for next-generation sequencing (NGS) and multigene analysis. Formalin-fixed paraffin-embedded (FFPE) samples are easier to store than fresh frozen (FF) samples. The objective of this study was to optimize RNA extraction from FFPE blocks for NGS. METHODS: RNA quality from FF and FFPE tissues (n=5), expected RNA amount per unit area, the relationship between archiving time and quantity/quality of FFPE-extracted RNA (n=14), differences in quantitative real-time polymerase chain reaction (qRT-PCR) and NGS results, and comparisons of both techniques with tissue processing at different institutions (n=96) were determined in this study. RESULTS: The quality of RNA did not show any statistically significant difference between paired FF and FFPE specimens (p=0.49). Analysis of tumor cellularity gave an expected RNA amount of 33.25 ng/mm2. Archiving time affected RNA quality, showing a negative correlation with RNA integrity number and a positive correlation with threshold cycle. However, RNA from samples as old as 10 years showed a 100% success rate in qRT-PCR using short primers, showing that the effect of archiving time can be overcome by proper experiment design. NGS showed a higher success rate than qRT-PCR. Specimens from institution B (n=46), which were often stored in a refrigerator for more than 6 hours and fixed without slicing, showed lower success rates and worse results than specimens from the other institutes. CONCLUSION: Archived FFPE tissues can be used to extract RNA for NGS if they are properly processed before fixation. The expected amount of RNA per unit size calculated in this study will be useful for other researchers.