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Utility of total cell-free DNA levels for surgical damage evaluation in patients with urological surgeries.

ABSTRACT: The evaluation of surgical damage is challenging because of the lack of specific biomarkers. Total cell-free DNA (cfDNA) levels have been reported to increase with external trauma and may be a biomarker for tissue damage. To investigate the utility of perioperative total cfDNA levels in evaluating surgical damage in urological surgeries. This multicenter, prospective, observational study included 196 patients scheduled for urological surgeries between September 2020 and July 2021. The primary outcome was the change in total cfDNA levels before and after urological surgery. The secondary outcome was the effect of surgical type on total cfDNA ratio before and after urological surgery. The postoperative median total cfDNA level of the 196 patients was significantly increased 2.5-fold compared to the preoperative level (185.2 ng/mL vs. 406.7 ng/mL, P < 0.001). The median total cfDNA before/after ratio was greater than four-fold for kidney transplantation, open cystectomy, and open adrenalectomy. The ratio was less than two-fold for laparoscopic adrenalectomy and robot-assisted radical prostatectomy. Major surgery showed a significant postoperative increase in total cfDNA levels, while minor surgery did not. Total cfDNA levels increased 2.5-fold after urological surgery and it can be used as an acute-phase biomarker for surgical damage.

SUBMITTER: Konishi S

PROVIDER: S-EPMC8585863 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Simple and low-cost recombinant enzyme-free seamless DNA cloning methods have recently become available. In vivo Escherichia coli cloning (iVEC) can directly transform a mixture of insert and vector DNA fragments into E. coli, which are ligated by endogenous homologous recombination activity in the cells. Seamless ligation cloning extract (SLiCE) cloning uses the endogenous recombination activity of E. coli cellular extracts in vitro to ligate insert and vector DNA fragments. An evaluation of the efficiency and utility of these methods is important in deciding the adoption of a seamless cloning method as a useful tool. In this study, both seamless cloning methods incorporated inserting DNA fragments into linearized DNA vectors through short (15-39 bp) end homology regions. However, colony formation was 30-60-fold higher with SLiCE cloning in end homology regions between 15 and 29 bp than with the iVEC method using DH5? competent cells. E. coli AQ3625 strains, which harbor a sbcA gene mutation that activates the RecE homologous recombination pathway, can be used to efficiently ligate insert and vector DNA fragments with short-end homology regions in vivo. Using AQ3625 competent cells in the iVEC method improved the rate of colony formation, but the efficiency and accuracy of SLiCE cloning were still higher. In addition, the efficiency of seamless cloning methods depends on the intrinsic competency of E. coli cells. The competency of chemically competent AQ3625 cells was lower than that of competent DH5? cells, in all cases of chemically competent cell preparations using the three different methods. Moreover, SLiCE cloning permits the use of both homemade and commercially available competent cells because it can use general E. coli recA- strains such as DH5? as host cells for transformation. Therefore, between the two methods, SLiCE cloning provides both higher efficiency and better utility than the iVEC method for seamless DNA plasmid engineering.

Project description:BackgroundWe evaluated the utility and quality of surgical videos posted on the main retinal YouTube channels by surgeons at different career stages and assessed how well the steps of the vitrectomy videos conformed to the parameters in the Casey Eye Institute Vitrectomy Indices Tool for Skills assessment (CEIVITS) scale.MethodsForty-five videos were included from nine retinal YouTube channels posted from 2011 to 2021. For each surgeon, 10 videos were randomized and the utility, quality, and educational content were assessed. For each video, the surgeons also assessed how the validated CEIVITS items were presented in the videos. The surgeons were divided based on years of experience: fellows (0-3 years), young surgeons (4-10 years), and senior surgeons (more than 10 years).ResultsThe video image quality was rated as good in 63.52% of evaluations, moderate in 30.37%, and poor in 6.11%. The quality assessment of the videos among the groups did not differ. The fellows rated the use of the videos as educational tools higher (3.99) than the young (3.87) and senior surgeons (3.47) (p < 0.0002, Kruskal-Wallis test); 34.76% of the fellows reported learning something new from the videos compared with 19.17% of the senior surgeons (p < 0.05). The CEIVITS scale item that was seen more frequently was related to core vitrectomies (72.29%) and the least represented was about checking infusion lines (80.17%).ConclusionsVitreoretinal surgical videos are useful educational tools during all stages of surgeons' careers, and the evaluation of the quality of the images did not differ significantly among the groups, however, surgeons with expertise shorter than 10 years report significantly greater use of videos than experienced surgeons. Videos posted to the public domain on different social media, most often YouTube, are widespread and unregulated for providing complimentary surgical education. Retinal societies should formulate guidelines and improve the educational value of the surgical videos posted on the Internet. Trial Registration The Federal University of São Paulo institution's Research Ethics Committee reviewed and approved this study protocol (Approval Number, 4.726.589).

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Utility of total cell-free DNA levels for surgical damage evaluation in patients with urological surgeries.

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