Cellular properties of human erythrocytes preserved in saline–adenine–glucose–mannitol in the presence of <scp>L</scp>‐carnitine

Arduini, Arduino; Minetti, Giampaolo; Ciana, Annarita; Seppi, Claudio; Brovelli, A; Profumo, Antonella; Vercellati, Cristina; Zappa, Manuela; Zanella, Alberto; Dottori, Secondo; Bonomini, Mario

doi:10.1002/ajh.20753

Cited by 16 publications

(25 citation statements)

References 36 publications

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“…RBCs were purified from leukocytes and platelets as described [19] and treated with 5 mM diisopropylfluorophosphate as a serine protease inhibitor for 10 min at 25 °C as a 20% haematocrit (Ht) suspension in PBS [20]. RBCs of different densities were separated in polyarabinogalactan density gradients as described [21,22]. RBCs of densities d=1.075, d=1.095 and d=1.110, as representative of, respectively, young, middle-age and old RBCs, were used for analyses of Hb, Ht, SDS-PAGE, and preparation of ghosts.…”

Section: Cell Handlingmentioning

confidence: 99%

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Membrane Remodelling and Vesicle Formation During Ageing of Human Red Blood Cells

Ciana

Achilli

Gaur

et al. 2017

Cell Physiol Biochem

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Background/Aims: A high surface-to-volume ratio and a spectrin membrane-skeleton (MS) confer to the mammalian red blood cells (RBCs) their characteristic deformability, mechanical strength and structural stability. During their 120 days of circulatory life in humans, RBCs decrease in size, while remaining biconcave disks, owing to a coordinated decrease in membrane surface area and cell water. It is generally believed that part of the membrane is lost with the shedding of spectrin-free vesicles of the same type that can be obtained in vitro by different treatments. If this were true, an excess of MS would arise in old RBCs, with respect to the lipid bilayer. Aim of this paper was to investigate this aspect. Methods: Quantification of spectrin by electrophoretic methods was carried out in RBCs of different age. Results: Spectrin decreases, on a per cell basis, with RBC ageing. On the other hand, the membrane raft protein marker flotillin-2, while decreasing in the membrane of old cells, was found to be strongly depleted in the membrane of in vitro-induced vesicles. Conclusion: Part of the membrane-skeleton is probably lost together with part of the lipid bilayer in a balanced way. These findings point to a mechanism for the in vivo release of membrane that is different from that which is known to occur in vitro.

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Section: Cell Handlingmentioning

confidence: 99%

“…Hb was quantified with the Drabkin's reagent according to the manufacturer instructions (Sigma-Aldrich S.r.l., Milan, Italy). Protein 4.1a/4.1b ratio was measured as a cell age parameter in each RBC subpopulation [22]. Ghost membranes were prepared according to Dodge [23] as detailed in [24].…”

Section: Cell Handlingmentioning

confidence: 99%

Membrane Remodelling and Vesicle Formation During Ageing of Human Red Blood Cells

Ciana

Achilli

Gaur

et al. 2017

Cell Physiol Biochem

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“…Other studies have shown that RBC MCHC and MCV change during blood storage and the type and extent of the changes depend on the additive preservation solution that is used to store the RBCs [21,23,24]. Therefore, in our simulations, RBC MCHC was varied from 14 to 22 mM and the volume of RBC was decreased from 90 to 60 mm 3 .…”

Section: Examined Effects Of Rbc Mchc MCV and Permeabilitymentioning

confidence: 99%

“…In the simulation, permeability was varied from P m (5000 mm s obtain an estimate of the maximum value of permeability needed to explain our experimental results. Although some reports show that MCV increased during storage in certain additive solutions [23,24], we used a substantially lower value in simulations of older RBCs in order to maxmize effects of volume changes and thereby obtain the minimum necessary changes in permeability.…”

Section: Effects Of Rbc Mchc and MCV On No Scavengingmentioning

confidence: 99%

“…The mean corpuscular hemoglobin concentration decreases, and the structure of RBC membrane changes significantly during storage [21,22]. In addition, RBC volume varies during blood storage but the type of change depends on which additive preservation solutions is used [23,24]. Changes in RBCs during storage are referred as blood storage lesion.…”

Section: Introductionmentioning

confidence: 99%

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Mechanism of Faster NO Scavenging by Older Stored Red Blood Cells

Liu

Janes

et al. 2013

Free Radical Biology and Medicine

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a b s t r a c tThe blood storage lesion involves morphological and biochemical changes of red blood cells (RBCs) that occur during storage. These include conversion of the biconcave disc morphology to a spherical one, decreased mean corpuscular hemoglobin concentration, varied mean corpuscular volume, reduced integrity of the erythrocyte membrane with formation of microparticles, and increased cell-free hemoglobin. We studied the extent that older stored red blood cells scavenge nitric oxide (NO) faster than fresher stored red blood cells. Using electron paramagnetic resonance spectroscopy and stoppedflow absorption spectroscopy to measure the rate of NO uptake and reaction with hemoglobin in red cells, we found that older stored red blood cells scavenge NO about 1.8 times faster than fresher ones. Based on these experimental data, we simulated NO scavenging by fresher or older stored red blood cells with a biconcave or spherical geometry, respectively, in order to explore the mechanism of NO scavenging related to changes that occur during blood storage. We found that red blood cells with a spherical geometry scavenges NO about 2 times slower than ones with a biconcave geometry, and a smaller RBC hemoglobin concentration or volume increases NO scavenging by red blood cells. Our simulations demonstrate that even the most extreme possible changes in mean corpuscular hemoglobin concentration and mean corpuscular volume that favor increased NO scavenging are insufficient to account for what is observed experimentally. Therefore, RBC membrane permeability must increase during storage and we find that the permeability is likely to increase between 5 and 70 fold. Simulations using a two-dimensional blood vessel show that even a 5-fold increase in membrane permeability to NO can reduce NO bioavailability at the smooth muscle. Background: Transfusion of older stored blood may be harmful. Results: Older stored red blood cells scavenge nitric oxide more than fresher cells. Conclusion: As stored red blood cells age, structural and biochemical changes occur that lead to faster scavenging. Significance: Increased nitric oxide scavenging by red blood cells as a function of storage age contributes to deleterious effects upon transfusion.

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Mechanism of faster NO scavenging by older stored red blood cells

Liu

Janes

et al. 2014

Redox Biology

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The blood storage lesion involves morphological and biochemical changes of red blood cells (RBCs) that occur during storage. These include conversion of the biconcave disc morphology to a spherical one, decreased mean corpuscular hemoglobin concentration, varied mean corpuscular volume, reduced integrity of the erythrocyte membrane with formation of microparticles, and increased cell-free hemoglobin. We studied the extent that older stored red blood cells scavenge nitric oxide (NO) faster than fresher stored red blood cells. Using electron paramagnetic resonance spectroscopy and stopped-flow absorption spectroscopy to measure the rate of NO uptake and reaction with hemoglobin in red cells, we found that older stored red blood cells scavenge NO about 1.8 times faster than fresher ones. Based on these experimental data, we simulated NO scavenging by fresher or older stored red blood cells with a biconcave or spherical geometry, respectively, in order to explore the mechanism of NO scavenging related to changes that occur during blood storage. We found that red blood cells with a spherical geometry scavenges NO about 2 times slower than ones with a biconcave geometry, and a smaller RBC hemoglobin concentration or volume increases NO scavenging by red blood cells. Our simulations demonstrate that even the most extreme possible changes in mean corpuscular hemoglobin concentration and mean corpuscular volume that favor increased NO scavenging are insufficient to account for what is observed experimentally. Therefore, RBC membrane permeability must increase during storage and we find that the permeability is likely to increase between 5 and 70 fold. Simulations using a two-dimensional blood vessel show that even a 5-fold increase in membrane permeability to NO can reduce NO bioavailability at the smooth muscle.BackgroundTransfusion of older stored blood may be harmful.ResultsOlder stored red blood cells scavenge nitric oxide more than fresher cells.ConclusionAs stored red blood cells age, structural and biochemical changes occur that lead to faster scavenging.SignificanceIncreased nitric oxide scavenging by red blood cells as a function of storage age contributes to deleterious effects upon transfusion.

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Cellular properties of human erythrocytes preserved in saline–adenine–glucose–mannitol in the presence of L‐carnitine

Cited by 16 publications

References 36 publications

Membrane Remodelling and Vesicle Formation During Ageing of Human Red Blood Cells

Membrane Remodelling and Vesicle Formation During Ageing of Human Red Blood Cells

Mechanism of Faster NO Scavenging by Older Stored Red Blood Cells

Mechanism of faster NO scavenging by older stored red blood cells

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