Comprehensive Characterization of Surface-Bound Proteins and Measurement of Fibrin Fiber Thickness on Extracorporeal Membrane Oxygenation Circuits Collected From Patients.

IF 4 2区 医学 Q1 CRITICAL CARE MEDICINE
Pediatric Critical Care Medicine Pub Date : 2024-11-01 Epub Date: 2024-08-13 DOI:10.1097/PCC.0000000000003591
Tengyi Cai, Samantha J Emery-Corbin, Conor McCafferty, Suelyn Van Den Helm, Natasha Letunica, Chantal Attard, Rebecca Barton, Stephen Horton, Steve Bottrell, Bradley Schultz, Graeme MacLaren, Roberto Chiletti, Derek Best, Amy Johansen, Fiona Newall, Warwick Butt, Yves d'Udekem, Laura F Dagley, Jumana M Yousef, Paul Monagle, Vera Ignjatovic
{"title":"Comprehensive Characterization of Surface-Bound Proteins and Measurement of Fibrin Fiber Thickness on Extracorporeal Membrane Oxygenation Circuits Collected From Patients.","authors":"Tengyi Cai, Samantha J Emery-Corbin, Conor McCafferty, Suelyn Van Den Helm, Natasha Letunica, Chantal Attard, Rebecca Barton, Stephen Horton, Steve Bottrell, Bradley Schultz, Graeme MacLaren, Roberto Chiletti, Derek Best, Amy Johansen, Fiona Newall, Warwick Butt, Yves d'Udekem, Laura F Dagley, Jumana M Yousef, Paul Monagle, Vera Ignjatovic","doi":"10.1097/PCC.0000000000003591","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To characterize surface-bound proteins and to measure the thickness of fibrin fibers bound to extracorporeal membrane oxygenation (ECMO) circuits used in children.</p><p><strong>Design: </strong>Single-center observational prospective study, April to November 2021.</p><p><strong>Setting: </strong>PICU, Royal Children's Hospital, Melbourne, Australia.</p><p><strong>Patients: </strong>Patients aged less than 18 years on venoarterial ECMO and without preexisting disorder.</p><p><strong>Interventions: </strong>None.</p><p><strong>Measurements and main results: </strong>ECMO circuits were collected from six patients. Circuit samples were collected from five different sites, and subsequently processed for proteomic and scanning electron microscopy (SEM) studies. The concentration of proteins bound to ECMO circuit samples was measured using a bicinchoninic acid protein assay, whereas characterization of the bound proteome was performed using data-independent acquisition mass spectrometry. The Reactome Over-representation Pathway Analyses tool was used to identify functional pathways related to bound proteins. For the SEM studies, ECMO circuit samples were prepared and imaged, and the thickness of bound fibrin fibers was measured using the Fiji ImageJ software, version 1.53c ( https://imagej.net/software/fiji/ ). Protein binding to ECMO circuit samples and fibrin networks showed significant intra-circuit and interpatient variation. The median (range) total protein concentration was 19.0 (0-76.9) μg/mL, and the median total number of proteins was 2011 (1435-2777). A total of 933 proteins were commonly bound to ECMO circuit samples from all patients and were functionally involved in 212 pathways, with signal transduction, cell cycle, and metabolism of proteins being the top three pathway categories. The median intra-circuit fibrin fiber thickness was 0.20 (0.15-0.24) μm, whereas the median interpatient fibrin fiber thickness was 0.18 (0.15-0.21) μm.</p><p><strong>Conclusions: </strong>In this report, we have characterized proteins and fiber fibrin thickness bound to ECMO circuits in six children. The techniques and approaches may be useful for investigating interactions between blood, coagulation, and the ECMO circuit and have the potential for circuit design.</p>","PeriodicalId":19760,"journal":{"name":"Pediatric Critical Care Medicine","volume":null,"pages":null},"PeriodicalIF":4.0000,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Pediatric Critical Care Medicine","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1097/PCC.0000000000003591","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/8/13 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"CRITICAL CARE MEDICINE","Score":null,"Total":0}
引用次数: 0

Abstract

Objective: To characterize surface-bound proteins and to measure the thickness of fibrin fibers bound to extracorporeal membrane oxygenation (ECMO) circuits used in children.

Design: Single-center observational prospective study, April to November 2021.

Setting: PICU, Royal Children's Hospital, Melbourne, Australia.

Patients: Patients aged less than 18 years on venoarterial ECMO and without preexisting disorder.

Interventions: None.

Measurements and main results: ECMO circuits were collected from six patients. Circuit samples were collected from five different sites, and subsequently processed for proteomic and scanning electron microscopy (SEM) studies. The concentration of proteins bound to ECMO circuit samples was measured using a bicinchoninic acid protein assay, whereas characterization of the bound proteome was performed using data-independent acquisition mass spectrometry. The Reactome Over-representation Pathway Analyses tool was used to identify functional pathways related to bound proteins. For the SEM studies, ECMO circuit samples were prepared and imaged, and the thickness of bound fibrin fibers was measured using the Fiji ImageJ software, version 1.53c ( https://imagej.net/software/fiji/ ). Protein binding to ECMO circuit samples and fibrin networks showed significant intra-circuit and interpatient variation. The median (range) total protein concentration was 19.0 (0-76.9) μg/mL, and the median total number of proteins was 2011 (1435-2777). A total of 933 proteins were commonly bound to ECMO circuit samples from all patients and were functionally involved in 212 pathways, with signal transduction, cell cycle, and metabolism of proteins being the top three pathway categories. The median intra-circuit fibrin fiber thickness was 0.20 (0.15-0.24) μm, whereas the median interpatient fibrin fiber thickness was 0.18 (0.15-0.21) μm.

Conclusions: In this report, we have characterized proteins and fiber fibrin thickness bound to ECMO circuits in six children. The techniques and approaches may be useful for investigating interactions between blood, coagulation, and the ECMO circuit and have the potential for circuit design.

体外膜氧合回路表面结合蛋白的综合表征和患者体内纤维蛋白纤维厚度的测量。
目的:确定表面结合蛋白的特征,并测量用于儿童的体外膜肺氧合(ECMO)回路的纤维蛋白纤维厚度:描述体外膜氧合(ECMO)回路表面结合蛋白的特征,并测量结合在儿童体外膜氧合回路上的纤维蛋白纤维的厚度:单中心前瞻性观察研究,2021 年 4 月至 11 月:地点:澳大利亚墨尔本皇家儿童医院 PICU:干预措施:无:测量和主要结果收集了六名患者的 ECMO 循环样本。从五个不同部位采集回路样本,随后进行蛋白质组学和扫描电子显微镜(SEM)研究。使用双喹啉酸蛋白质测定法测量了 ECMO 循环样本中结合的蛋白质浓度,而结合的蛋白质组的特征描述则使用数据独立采集质谱法进行。Reactome 过度呈现通路分析工具用于确定与结合蛋白相关的功能通路。在扫描电子显微镜研究中,制备了 ECMO 电路样本并进行了成像,使用 Fiji ImageJ 软件 1.53c 版(https://imagej.net/software/fiji/)测量了结合纤维蛋白纤维的厚度。蛋白质与 ECMO 循环样本和纤维蛋白网络的结合在循环内和患者间存在显著差异。蛋白质总浓度的中位数(范围)为 19.0 (0-76.9) μg/mL,蛋白质总数的中位数为 2011 (1435-2777)。所有患者的 ECMO 循环样本中通常结合有 933 种蛋白质,它们在功能上参与了 212 种通路,其中信号转导、细胞周期和蛋白质代谢是前三大通路类别。回路内纤维蛋白纤维厚度的中位数为 0.20 (0.15-0.24) μm,而患者间纤维蛋白纤维厚度的中位数为 0.18 (0.15-0.21) μm:在这份报告中,我们描述了六名患儿 ECMO 循环中结合的蛋白质和纤维蛋白厚度。这些技术和方法可用于研究血液、凝血和 ECMO 电路之间的相互作用,并有可能用于电路设计。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Pediatric Critical Care Medicine
Pediatric Critical Care Medicine 医学-危重病医学
CiteScore
7.40
自引率
14.60%
发文量
991
审稿时长
3-8 weeks
期刊介绍: Pediatric Critical Care Medicine is written for the entire critical care team: pediatricians, neonatologists, respiratory therapists, nurses, and others who deal with pediatric patients who are critically ill or injured. International in scope, with editorial board members and contributors from around the world, the Journal includes a full range of scientific content, including clinical articles, scientific investigations, solicited reviews, and abstracts from pediatric critical care meetings. Additionally, the Journal includes abstracts of selected articles published in Chinese, French, Italian, Japanese, Portuguese, and Spanish translations - making news of advances in the field available to pediatric and neonatal intensive care practitioners worldwide.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信