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The participation of tRNAs in fundamental aspects of biology and disease necessitates an accurate, experimentally confirmed annotation of tRNA genes and curation of tRNA sequences. This has been challenging because RNA secondary structure, nucleotide modifications, and tRNA gene multiplicity complicate sequencing and mapping efforts. To address these issues, we developed hydro-tRNAseq, a method based on partial alkaline RNA hydrolysis that generates fragments amenable for sequencing. To identify transcribed tRNA genes, we further complemented this approach with photoactivatable crosslinking and immunoprecipitation (PAR-CLIP) of SSB/La, a conserved protein involved in pre-tRNA processing. Our results show that approximately half of all predicted tRNA genes are transcribed in human cells. We also report nucleotide modification sites and their order of introduction, and we identify tRNA leaders, trailers, and introns. By using complementary sequencing-based methodologies, we present a human tRNA atlas and determine expression levels of mature and processing intermediates of tRNAs in human cells.
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One of the major decisions both prior to initiating a SISI approach and once inspecting the pelvic pathology is whether or not there is an immature or cancerous component to the pathology, for avoidance of rupture in these cases may be critical. We have found that the majority of cases can be approached with SISI, as long as the radiology is consistent with benign pathology (i.e., simple appearing cyst without solid component, hemorrhagic cyst, torsion, or solid mass consistent with teratoma based on appearance of fat or calcifications) and the hormonal markers are normal. Any indication of malignancy (peritoneal studding, retroperitoneal lymphadenopathy, noncalcified or highly vascularized solid mass, or elevated hormonal markers) should discourage use of the SISI technique. In these situations, multiport or multi-instrument single-incision laparoscopy (e.g., LESS) can be considered, although the standard is open surgery.
Teratomas form an interesting group of neoplasms for which the SISI technique is particularly applicable. Usually mature but occasionally massive, they may be bilateral in 10% of cases [22], whether synchronously or metachronously. We follow these patients for a period of at least 5 years after diagnosis, which allows small, metachronous lesions to be identified early, making ovarian preservation easier. Small ovarian dermoids found during surveillance are easily treated using SISI instrumentation to exteriorize the ovary through the umbilicus for excision. 59ce067264
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