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1.
PLoS One ; 10(7): e0132831, 2015.
Article in English | MEDLINE | ID: mdl-26181416

ABSTRACT

To address the need to study frozen clinical specimens using next-generation RNA, DNA, chromatin immunoprecipitation (ChIP) sequencing and protein analyses, we developed a biobank work flow to prospectively collect biospecimens from patients with renal cell carcinoma (RCC). We describe our standard operating procedures and work flow to annotate pathologic results and clinical outcomes. We report quality control outcomes and nucleic acid yields of our RCC submissions (N=16) to The Cancer Genome Atlas (TCGA) project, as well as newer discovery platforms, by describing mass spectrometry analysis of albumin oxidation in plasma and 6 ChIP sequencing libraries generated from nephrectomy specimens after histone H3 lysine 36 trimethylation (H3K36me3) immunoprecipitation. From June 1, 2010, through January 1, 2013, we enrolled 328 patients with RCC. Our mean (SD) TCGA RNA integrity numbers (RINs) were 8.1 (0.8) for papillary RCC, with a 12.5% overall rate of sample disqualification for RIN <7. Banked plasma had significantly less albumin oxidation (by mass spectrometry analysis) than plasma kept at 25 °C (P<.001). For ChIP sequencing, the FastQC score for average read quality was at least 30 for 91% to 95% of paired-end reads. In parallel, we analyzed frozen tissue by RNA sequencing; after genome alignment, only 0.2% to 0.4% of total reads failed the default quality check steps of Bowtie2, which was comparable to the disqualification ratio (0.1%) of the 786-O RCC cell line that was prepared under optimal RNA isolation conditions. The overall correlation coefficients for gene expression between Mayo Clinic vs TCGA tissues ranged from 0.75 to 0.82. These data support the generation of high-quality nucleic acids for genomic analyses from banked RCC. Importantly, the protocol does not interfere with routine clinical care. Collections over defined time points during disease treatment further enhance collaborative efforts to integrate genomic information with outcomes.


Subject(s)
Biological Specimen Banks/organization & administration , Carcinoma, Renal Cell/genetics , Gene Expression Regulation, Neoplastic , Histones/genetics , Kidney Neoplasms/genetics , RNA, Neoplasm/genetics , Adult , Aged , Aged, 80 and over , Arizona , Carcinoma, Renal Cell/metabolism , Carcinoma, Renal Cell/pathology , Carcinoma, Renal Cell/surgery , Cell Line, Tumor , Chromatin Immunoprecipitation , Female , Gene Library , Histones/metabolism , Humans , Kidney Neoplasms/metabolism , Kidney Neoplasms/pathology , Kidney Neoplasms/surgery , Male , Methylation , Middle Aged , Oxidation-Reduction , Quality Control , RNA, Neoplasm/chemistry , Sequence Analysis, DNA , Sequence Analysis, RNA , Serum Albumin/chemistry
2.
J Robot Surg ; 7(3): 273-9, 2013 Sep.
Article in English | MEDLINE | ID: mdl-27000923

ABSTRACT

Robot-assisted cystectomy surgery may be advantageous for patients. The purpose of this study was to compare anesthetic management and outcomes in patients undergoing robot-assisted versus open radical cystectomy. In a retrospective review of 256 cystectomy procedures, procedure duration, blood loss, respiratory parameters, recovery room opiate consumption, pain scores and antiemetic use in the recovery room, and hospital length of stay were compared. After exclusions, 96 robot-assisted and 102 open procedures were analyzed. Anesthesia and surgery duration were significantly longer in the robot-assisted group, while the length of hospital stay was significantly shorter in the robot-assisted group: 7.1 ± 5.8 versus 9.8 ± 5.03 days, p = 0.0005. Estimated blood loss was 601.8 ± 491.4 ml in the open group versus 257.7 ± 164.3 ml in the robot-assisted group, p < 0.0001. Recovery room opiate consumption was significantly less in the robot-assisted group: 9.5 ± 8.9 versus 12.6 ± 9.9 mg (morphine equivalents), p = 0.02. The highest recorded respiratory rate was significantly higher in the robot-assisted group, as was the highest recorded peak airway pressure. Among patients with arterial blood gas data, the highest arterial partial pressure of CO2 was significantly greater in the robot-assisted group than in the open surgery group: 42.6 ± 5.6 versus 37.4 ± 4.8 mmHg CO2, p = 0.0001. Surgeons and anesthesia providers can expect robot-assisted radical cystectomy surgery to last longer than traditional open surgery, but to be associated with less pain and blood loss. Positioning and abdominal insufflation for robot-assisted surgery may contribute to ventilation challenges.

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