Predicting storage‐dependent damage to red blood cells using nitrite oxidation kinetics, peroxiredoxin‐2 oxidation, and hemoglobin and free heme measurements

Oh, Joo–Yeun; Stapley, Ryan; Harper, Victoria M.; Marques, Marisa B.; Patel, Rakesh P.

doi:10.1111/trf.13248

Cited by 16 publications

(21 citation statements)

References 34 publications

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“…We used a spectral deconvolution approach to measure heme-containing species, which we recently showed avoids artifacts associated with widely used and commercially available assays that do not distinguish between free heme and hemoglobin [ 52 ]. Consistent with our previous reports, levels of hemoglobin and free heme were in the hundreds of micromolar range in stored RBC supernatants [ 25 , 52 ]. These levels led to an increase in circulating free hemoglobin, free heme, and NTBI after resuscitation, indicated by these species being elevated after resuscitating mice with stored RBCs.…”

Section: Discussionsupporting

confidence: 92%

“…Another limitation of these measures is that we sampled segments attached to the bags transfused due to constraints limiting the ability to sample the bags directly. Previous studies have shown that segment levels of free hemoglobin are higher than those of their paired bags, whereas the opposite relationship has been reported for free heme [ 24 , 25 ], suggesting that the relative amount of free heme administered is in fact be higher than shown.…”

Section: Discussionmentioning

confidence: 86%

“…There are numerous components in stored RBCs that have the potential to cause organ injury that include, but are not limited to, free heme, free hemoglobin, iron, and microvesicles [ 25 , 52 ]. Consistent with our previous studies [ 25 , 36 ], free oxyHb, metHb, and heme levels were increased after 14 d of RBC storage, being 930 ± 125 μM, 40.3 ± 8.8 μM, and 168 ± 44.7 μM, respectively (mean ± SEM, n = 7). Fig 2 shows that free hemoglobin and free heme were higher 4 h after resuscitation with stored RBCs compared to fresh RBCs ( n = 3–7 as indicated); NTBI level was also higher, but this difference did not reach statistical significance ( p = 0.07).…”

Section: Resultsmentioning

confidence: 99%

“…Our previous studies have shown that the ability of transfused RBCs to improve microcirculatory function and tissue oxygenation in trauma patients progressively diminishes with RBC storage age [ 22 ]. Storage of RBCs is accompanied by biochemical and morphological changes referred to as “storage lesion.” Key amongst these changes are hemolysis and release of free and microvesicle-associated hemoglobin, free heme, and iron [ 23 – 25 ]. Transfusion of these molecular species increases oxidative stress, inhibits nitric oxide bioavailability, results in a pro-inflammatory state, and provides substrate (low molecular weight iron) for microbial growth [ 17 , 18 , 26 – 39 ].…”

Section: Introductionmentioning

confidence: 99%

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Role of heme in lung bacterial infection after trauma hemorrhage and stored red blood cell transfusion: A preclinical experimental study

Wagener

et al. 2018

PLoS Med

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BackgroundTrauma is the leading cause of death and disability in patients aged 1–46 y. Severely injured patients experience considerable blood loss and hemorrhagic shock requiring treatment with massive transfusion of red blood cells (RBCs). Preclinical and retrospective human studies in trauma patients have suggested that poorer therapeutic efficacy, increased severity of organ injury, and increased bacterial infection are associated with transfusion of large volumes of stored RBCs, although the mechanisms are not fully understood.Methods and findingsWe developed a murine model of trauma hemorrhage (TH) followed by resuscitation with plasma and leukoreduced RBCs (in a 1:1 ratio) that were banked for 0 (fresh) or 14 (stored) days. Two days later, lungs were infected with Pseudomonas aeruginosa K-strain (PAK). Resuscitation with stored RBCs significantly increased the severity of lung injury caused by P. aeruginosa, as demonstrated by higher mortality (median survival 35 h for fresh RBC group and 8 h for stored RBC group; p < 0.001), increased pulmonary edema (mean [95% CI] 106.4 μl [88.5–124.3] for fresh RBCs and 192.5 μl [140.9–244.0] for stored RBCs; p = 0.003), and higher bacterial numbers in the lung (mean [95% CI] 1.2 × 107 [−1.0 × 107 to 2.5 × 107] for fresh RBCs and 3.6 × 107 [2.5 × 107 to 4.7 × 107] for stored RBCs; p = 0.014). The mechanism underlying this increased infection susceptibility and severity was free-heme-dependent, as recombinant hemopexin or pharmacological inhibition or genetic deletion of toll-like receptor 4 (TLR4) during TH and resuscitation completely prevented P. aeruginosa–induced mortality after stored RBC transfusion (p < 0.001 for all groups relative to stored RBC group). Evidence from studies transfusing fresh and stored RBCs mixed with stored and fresh RBC supernatants, respectively, indicated that heme arising both during storage and from RBC hemolysis post-resuscitation plays a role in increased mortality after PAK (p < 0.001). Heme also increased endothelial permeability and inhibited macrophage-dependent phagocytosis in cultured cells. Stored RBCs also increased circulating high mobility group box 1 (HMGB1; mean [95% CI] 15.4 ng/ml [6.7–24.0] for fresh RBCs and 50.3 ng/ml [12.3–88.2] for stored RBCs), and anti-HMGB1 blocking antibody protected against PAK-induced mortality in vivo (p = 0.001) and restored macrophage-dependent phagocytosis of P. aeruginosa in vitro. Finally, we showed that TH patients, admitted to the University of Alabama at Birmingham ER between 1 January 2015 and 30 April 2016 (n = 50), received high micromolar–millimolar levels of heme proportional to the number of units transfused, sufficient to overwhelm endogenous hemopexin levels early after TH and resuscitation. Limitations of the study include lack of assessment of temporal changes in different products of hemolysis after resuscitation and the small sample size precluding testing of associations between heme levels and adverse outcomes in resuscitated TH patients.ConclusionsWe provide evidence that ...

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 86%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Role of heme in lung bacterial infection after trauma hemorrhage and stored red blood cell transfusion: A preclinical experimental study

Wagener

et al. 2018

PLoS Med

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“…The strengths of our study include the consecutive and unselected sample of medical ICU patients, plasma sampling before and after transfusion, sampling from the RBC storage bag rather than the bag segment appendage, and a patient sample size adequate for multivariable analysis. Our study has several limitations.…”

Section: Discussionmentioning

confidence: 99%

Total plasma heme concentration increases after red blood cell transfusion and predicts mortality in critically ill medical patients

et al. 2019

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BACKGROUND: Relationships between red blood cell (RBC) transfusion, circulating cell-free heme, and clinical outcomes in critically ill transfusion recipients are incompletely understood. The goal of this study was to determine whether total plasma heme increases after RBC transfusion and predicts mortality in critically ill patients. STUDY DESIGN AND METHODS:This was a prospective cohort study of 111 consecutive medical intensive care patients requiring RBC transfusion. Cellfree heme was measured in RBC units before transfusion and in the patients' plasma before and after transfusion.RESULTS: Total plasma heme levels increased in response to transfusion, from a median (interquartile range [IQR]) of 35 (26-76) μmol/L to 47 (35-73) μmol/L (p < 0.001). Posttransfusion total plasma heme was higher in nonsurvivors (54 [35-136] μmol/L) versus survivors (44 [31-65] μmol/L, p = 0.03). Posttransfusion total plasma heme predicted hospital mortality (odds ratio [95% confidence interval] per quartile increase in posttransfusion plasma heme, 1.76 [1.17-2.66]; p = 0.007). Posttransfusion total plasma heme was not correlated with RBC unit storage duration and weakly correlated with RBC unit cell-free heme concentration. CONCLUSIONS:Total plasma heme concentration increases in critically ill patients after RBC transfusion and is independently associated with mortality. This transfusion-associated increase in total plasma heme is not fully explained by RBC unit storage age or cell-free heme content. Additional studies are warranted to define mechanisms of transfusion-related plasma heme accumulation and test prevention strategies.R ed blood cell (RBC) transfusion is associated with increased risk of death in critically ill patients. 1 This risk may be explained by storage-related changes in banked RBCs. Such changes include depletion of adenosine triphosphate, nitric oxide, and 2,3diphosphoglycerate, excess cellular stiffness and fragility, and low-grade hemolysis. [2][3][4][5] The resulting cell-free hemoglobin (Hb; free in solution and contained in RBC microparticles) 3,4,6-8 can undergo oxidation and release cellfree heme. 9,10 These mediators may cause toxicity when transfused because cell-free Hb promotes circulatory dysfunction by scavenging nitric oxide 4,7,11,12 and cell-free ABBREVIATIONS: ICU = intensive care unit; IQR = interquartile range; MICU = medical intensive care unit; NTBI = non-transferrinbound iron; SOFA = sequential (sepsis-related) organ failure assessment. From the PIETROPAOLI ET AL.Fig. 5. Relationship between posttransfusion total plasma heme concentration and the pretransfusion plasma haptoglobin concentration. Box plots show the median (horizontal line) and 25th and 75th percentiles (lower and upper limits of the box).The dots represent outliers beyond the whiskers that designate the 10th and 90th percentiles. Comparison was made with Kruskal-Wallis test.

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Resonance Raman spectroscopy of hemoglobin in red blood cells

Wood

Kochan

Marzec

2020

Vibrational Spectroscopy in Protein Research

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Predicting storage‐dependent damage to red blood cells using nitrite oxidation kinetics, peroxiredoxin‐2 oxidation, and hemoglobin and free heme measurements

Cited by 16 publications

References 34 publications

Role of heme in lung bacterial infection after trauma hemorrhage and stored red blood cell transfusion: A preclinical experimental study

Role of heme in lung bacterial infection after trauma hemorrhage and stored red blood cell transfusion: A preclinical experimental study

Total plasma heme concentration increases after red blood cell transfusion and predicts mortality in critically ill medical patients

Resonance Raman spectroscopy of hemoglobin in red blood cells

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