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This report describes an improved protocol to generate stranded, barcoded RNA-seq libraries to capture the whole transcriptome. By optimizing the use of duplex specific nuclease (DSN) to remove ribosomal RNA reads from stranded barcoded libraries, we demonstrate improved efficiency of multiplexed next generation sequencing (NGS). This approach detects expression profiles of all RNA types, including miRNA (microRNA), piRNA (Piwi-interacting RNA), snoRNA (small nucleolar RNA), lincRNA (long non-coding RNA), mtRNA (mitochondrial RNA) and mRNA (messenger RNA) without the use of gel electrophoresis. The improved protocol generates high quality data that can be used to identify differential expression in known and novel coding and non-coding transcripts, splice variants, mitochondrial genes and SNPs (single nucleotide polymorphisms).

Metastatic castrate resistant prostate cancer (PCa) remains an incurable entity. In the era of immunotherapy, the complex PCa microenvironment poses a unique challenge to the successful application of this class of agents. However, in the last decade, a tremendous effort has been made to explore this field of therapeutics. In this review, the physiology of the cancer immunity cycle is highlighted in the context of the prostate tumor microenvironment, and the current evidence for use of various classes of immunotherapy agents including vaccines (dendritic cell based, viral vector based and DNA/mRNA based), immune checkpoint inhibitors, Chimeric antigen receptor T cell therapy, antibody-mediated radioimmunotherapy, antibody drug conjugates, and bispecific antibodies, is consolidated. Finally, the future directions for combinatorial approaches to combat PCa are discussed.

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amount of time added to the OR due to the use of the X-ray (41-50 min, p[0.8). However, there was significant discordance between surgeons' and nurses' perceptions of the frequency of lost surgical sharps (p[0.025), the frequency of miscounted surgical sharps (p[0.003), and the frequency of lost surgical sharps not recovered (p[0.003). Percentages are listed in Table 1. When performing Kruskal-Wallis ANOVA test to include anesthesiologists, improvement in perception are seen in all categories.CONCLUSIONS: These data provide initial evidence that OR staff may perceive the frequency of near-miss and retained surgical sharps differently. This hints at a lack of communication regarding these sentinel events. Patient safety can be negatively impacted if certain members of the OR team are not aware of lost, miscounted or retained surgical sharps. Further research is needed to evaluate the impact this may have on patient safety, OR culture and the effective reporting of RSI events.