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经Mirasol病原体灭活技术处理后血小板线粒体完整性的评估。

Evaluation of platelet mitochondria integrity after treatment with Mirasol pathogen reduction technology.

作者信息

Li Junzhi, Lockerbie Owen, de Korte Dirk, Rice Joe, McLean Robyn, Goodrich Raymond P

机构信息

Navigant Biotechnologies, Inc., Lakewood, Colorado 80215, USA.

出版信息

Transfusion. 2005 Jun;45(6):920-6. doi: 10.1111/j.1537-2995.2005.04381.x.

Abstract

BACKGROUND

Previous studies showed that Mirasol (Navigant Biotechnologies, Inc.) pathogen reduction technology (PRT) treatment resulted in an increase in platelet (PLT) glucose consumption and lactate production rates and decrease in pH in media during PLT storage. Increased glycolytic flux could result from damage to mitochondria and/or increased ATP consumption.

STUDY DESIGN AND METHODS

PLT concentrates were collected by standard automated blood component collection system (Trima, Gambro BCT) procedure on Day 0 and treated with Mirasol PRT treatment on Day 1. PLT mitochondrial transmembrane potential was evaluated by staining PLTs with JC-1 followed by flow cytometry analysis. Mitochondrial enzymatic activity was measured by the MTT assay. ATP content and pH were also quantified. The values for these measurements were compared among control, untreated, and pathogen reduction technology (PRT)-treated PLTs during PLT storage for up to 7 days.

RESULTS

No significant changes were found in pH, JC-1 signal, MTT activity, and ATP content of the PLTs immediately after PRT treatment. The treated PLTs exhibited a moderate but significantly accelerated decrease in pH and lower ATP content after 7-day storage when compared to control PLTs. Neither the JC-1 assay nor the MTT assay, however, showed a significant difference between control and treated PLTs during PLT storage.

CONCLUSIONS

There is no evidence from these studies that Mirasol PRT treatment alters PLT mitochondrial structural and functional integrity immediately after treatment and during PLT storage. An increased demand for ATP may be the driving force for observed increases in both the glycolytic flux and the oxidative metabolism observed in treated PLTs.

摘要

背景

先前的研究表明,Mirasol(Navigant生物技术公司)病原体灭活技术(PRT)处理会导致血小板(PLT)在储存期间葡萄糖消耗增加、乳酸生成速率加快以及培养基pH值降低。糖酵解通量增加可能是由于线粒体受损和/或ATP消耗增加所致。

研究设计与方法

血小板浓缩物于第0天通过标准自动血液成分采集系统(Trima,甘布罗BCT)程序采集,并于第1天用Mirasol PRT处理。用JC-1对血小板进行染色,随后进行流式细胞术分析,以评估血小板线粒体跨膜电位。通过MTT法测定线粒体酶活性。还对ATP含量和pH值进行了定量分析。在血小板储存长达7天的过程中,比较了对照、未处理和经病原体灭活技术(PRT)处理的血小板的这些测量值。

结果

PRT处理后立即检测血小板的pH值、JC-1信号、MTT活性和ATP含量,未发现显著变化。与对照血小板相比,处理后的血小板在储存7天后pH值呈中度但显著加速下降,ATP含量降低。然而,在血小板储存期间,JC-1检测和MTT检测均未显示对照血小板和处理后血小板之间存在显著差异。

结论

这些研究没有证据表明Mirasol PRT处理会在处理后及血小板储存期间改变血小板线粒体的结构和功能完整性。ATP需求增加可能是导致处理后血小板中糖酵解通量和氧化代谢增加的驱动力。

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