The effects of polyvinyl chloride and polyolefin blood bags on red blood cells stored in a new additive solution.

BACKGROUND AND OBJECTIVES: Red blood cells (RBCs) must be stored in polyvinyl chloride (PVC) bags plasticized with di-2-ethylhexyl phthalate or a similar plasticizer to achieve their full storage life with conventional storage solutions. Improved storage solutions might remove this requirement and allow blood storage in other plastics. Experimental Additive Solution-61 (EAS-61), which maintains RBCs for 9 weeks with reduced haemolysis and satisfactory 51Cr 24-h recovery, is an appropriate candidate improved RBC storage solution. MATERIALS AND METHODS: Twenty-four units of packed RBCs were pooled in groups of four units, each pool was realiquoted into four units and stored, six pooled units per arm, in one of the following: 100 ml of EAS-61 in PVC; 200 ml of EAS-61 in PVC; 100 ml of EAS-61 in polyolefin (PO); and 200 ml of EAS-61 in PO. Haemolysis, RBC morphology indices, RBC ATP concentrations, and other measures of RBC metabolism and function were measured weekly. RESULTS: RBC haemolysis exceeded 1% by 7 weeks in PO bags containing 100 ml or 200 ml of EAS-61. In PVC bags, haemolysis was less than 1% at 11 weeks. RBC ATP concentrations were 1 mol/g of haemoglobin (Hb) higher at 2 weeks in the PVC-stored units. CONCLUSIONS: RBCs stored in PVC had markedly less haemolysis and higher RBC ATP concentrations than those stored in PO. Haemolysis would limit RBC storage in PO bags to a duration of 6 weeks, even with EAS-61.     

Menopausal status affects the susceptibility of stored RBCs to mechanical stress

The mechanical fragility index (MFI) is an in vitro measure of sublethal injury to RBCs. In our previous experiments, we demonstrated that an increase in sublethal injury (increasing MFI) was a component of the RBC storage lesion, and that the MFI was significantly higher amongst the RBC units from male donors compared to pre-menopausal female donors during storage. It was hypothesized that hormonal or menstrual factors contributed to this difference. In this study, we found that RBC units donated by post-menopausal women demonstrated an MFI that was significantly higher than those donated by pre-menopausal women throughout storage.     

Deterioration of red blood cell mechanical properties is reduced in anaerobic storage

BackgroundHypothermic storage of red blood cells (RBCs) results in progressive deterioration of the rheological properties of the cells, which may reduce the efficacy of RBC transfusions. Recent studies have suggested that storing RBC units under anaerobic conditions may reduce this storage-induced deterioration.ResultsThe bulk perfusion rates for anaerobically stored RBC were significantly higher than for conventionally stored RBCs over the entire duration of storage for all devices (up to 10% on day 42; up to 14% on day 63). Capillary perfusion rates suggested that anaerobically stored RBC units contained significantly fewer non-deformable RBC capable of transiently plugging microfluidic device capillaries. The number of plugging events caused by these non-deformable RBC increased over the 63 days of hypothermic storage by nearly 16- to 21-fold for conventionally stored units, and by only about 3- to 6-fold for anaerobically stored units.DiscussionThe perfusion measurements suggest that anaerobically stored RBC retain a greater ability to perfuse networks of artificial capillaries compared to conventionally (aerobically) stored RBC. It is likely that anaerobic storage confers this positive effect on the bulk mechanical properties of stored RBC by significantly reducing the number of non-deformable cells present in the overall population of relatively well-preserved RBC.     

The use of the mechanical fragility test in evaluating sublethal RBC injury during storage

Background The mechanical fragility index (MFI) is an in vitro measurement of the extent of RBC sublethal injury. Sublethal injury might constitute a component of the RBC storage lesion, thus the MFI was determined serially during routine RBC storage. Methods Leucoreduced AS‐5‐ and SAGM‐preserved RBCs were stored under routine blood bank conditions. The mechanical fragility (MF) of each unit was serially measured during storage. Results For both AS‐5 and SAGM units, male and female RBCs demonstrated statistically significant increases in the MFI during storage. The MFI was significantly lower in AS‐5 units compared to SAGM units throughout storage. Female RBCs had significantly lower MFI vs. male RBCs in both AS‐5 and SAGM units at all times. No significant differences in MFI were observed between ABO groups for both genders for AS‐5 RBCs. Conclusions The MF of RBCs increases during storage. Both gender and preservation solution influenced the MFI; however, the male:female MFI ratios were similar at all time‐points and remained stable, suggesting that gender‐based biological differences exist independent of storage solution. The MF could be a useful test for evaluating the effect of novel interventions intended to mitigate the susceptibility of RBCs to sublethal injury during storage.     

Glucose‐6‐phosphate dehydrogenase deficiency in transfusion medicine: the unknown risks

The hallmark of glucose‐6‐phosphate dehydrogenase (G6PD) deficiency is red blood cell (RBC) destruction in response to oxidative stress. Patients requiring RBC transfusions may simultaneously receive oxidative medications or have concurrent infections, both of which can induce haemolysis in G6PD‐deficient RBCs. Although it is not routine practice to screen healthy blood donors for G6PD deficiency, case reports identified transfusion of G6PD‐deficient RBCs as causing haemolysis and other adverse events. In addition, some patient populations may be more at risk for complications associated with transfusions of G6PD‐deficient RBCs because they receive RBCs from donors who are more likely to have G6PD deficiency. This review discusses G6PD deficiency, its importance in transfusion medicine, changes in the RBC antioxidant system (of which G6PD is essential) during refrigerated storage and mechanisms of haemolysis. In addition, as yet unanswered questions that could be addressed by translational and clinical studies are identified and discussed.     

Effect of gamma irradiation on blood from glucose 6 phosphate dehydrogenase deficient blood donors

Recently, the irradiation of cellular blood components has received increased attention. Normal red blood cells (RBC) when subjected to gamma irradiation show increase in haemolysis and increased storage lesions. Glucose 6 phosphate dehydrogenase (G6PD) deficient blood is lacking in anti-oxidant properties which protect the (RBC) membrane and therefore when subjected to gamma irradiation, it may lead to increased haemolysis. In this study, 500 healthy non-remunerated blood donors were screened for G6PD deficiency. About 350 ml of whole blood was collected from 10 G6PD deficient donors (cases) and 10 units were collected from donors who were not deficient for G6PD (controls). All units were subjected to gamma irradiation of 25 Gy by a self contained gamma cell irradiator (Nordion Canada) on day 0 and then stored at 4°C. Sampling was done from these cases and controls on day 0, 7, 14 and 21.

Estimation of free plasma hemoglobin (Hb), free plasma potassium (K) and lactate dehydrogenase (LDH) was performed. It was found that there were no significant changes between the cases and controls in any of the parameters assessed, although the values for all the parameters were higher in case group at the end of storage period, these differences were not statistically significant.     

The need to label red blood cell units with their haemoglobin content: a single centre study on haemoglobin variations due to donor-related factors

Background

Red blood cell (RBC) transfusions are given as “number of units” without considering the haemoglobin (Hb) content of these units. Donor factors influencing Hb level in whole blood donors and, ultimately, in RBC units have not been studied.

Materials and methods

Donor data for a period of 1.5 years were retrospectively analysed and the effects of age, gender and weight on the Hb level of the donors were determined. The correlation between donor’s Hb concentration with total Hb in the RBC unit was analysed. Additionally, actual Hb content of 125 RBC units was determined. The total Hb content of these RBC units was also mathematically calculated based on the blood donors’ Hb. The ability of this mathematically calculated Hb to predict actual Hb content per RBC unit was then analysed.

Results

The mean Hb level in female donors was 1.79 g/dL lower than in the male donors (p<0.001). Increasing age was associated with a lower mean Hb in the donors (p<0.01), while a higher body weight correlated weakly (r=0.06) but significantly with increased mean Hb (p<0.01). Logistic regression analysis showed that in blood donors, female gender had a stronger influence on lowering the mean Hb than either older age or lower weight. A variation of nearly 100% (42.3–80.8 g Hb per unit) was seen in the total Hb content of the RBC units tested. Mathematically calculated Hb content correlated well (r=0.6; p<0.01) with the actual Hb content of the RBC units.

Discussion

We demonstrated the effect of gender, age and weight on Hb levels in whole blood donors. Dissimilarities in the donor Hb caused nearly 100% variations in the Hb content of the RBC units. It would, therefore, be prudent to label RBC units with their total Hb content. This total Hb content can be predicted fairly accurately from the donor’s pre-donation Hb level.     

Effect of gamma irradiation on blood from glucose 6 phosphate dehydrogenase deficient blood donors

Recently, the irradiation of cellular blood components has received increased attention. Normal red blood cells (RBC) when subjected to gamma irradiation show increase in haemolysis and increased storage lesions. Glucose 6 phosphate dehydrogenase (G6PD) deficient blood is lacking in anti-oxidant properties which protect the RBC membrane and therefore when subjected to gamma irradiation, it may lead to increased haemolysis. In this study, 500 healthy non-remunerated blood donors were screened for G6PD deficiency. About 350 ml of whole blood was collected from 10 G6PD deficient donors (cases) and 10 units were collected from donors who were not deficient for G6PD (controls). All units were subjected to gamma irradiation of 25 Gy by a self contained gamma cell irradiator (Nordion Canada) on day 0 and then stored at 4 degrees C. Sampling was done from these cases and controls on day 0, 7, 14 and 21. Estimation of free plasma hemoglobin (Hb), free plasma potassium (K) and lactate dehydrogenase (LDH) was performed. It was found that there were no significant changes between the cases and controls in any of the parameters assessed, although the values for all the parameters were higher in case group at the end of storage period, these differences were not statistically significant.     

Transitory interruption of recommended storage conditions does not cause significant changes in in vitro parameters of leucocyte‐depleted red blood cells

Background and Objectives Conventional quality control studies of the shelf life of RBC units do not consider cold chain interruptions that occur during cross‐matching or between the release of RBCs from the blood bank and their return from the ward. These interruptions may, however, lead to a considerable loss of quality. On the other hand, differences in the quality of RBCs may derive from the different manufacturing processes employed in various blood centres. Materials and Methods One day after the expiry date of the RBC unit, we analysed complete blood count, blood gas, potassium, LDH, hydroxybutyrate dehydrogenase, glucose, lactate, total and free haemoglobin (Hb) and ATP and compared the results with regard to the frequency of storage interruptions and to two manufacturers of these RBCs. Results We could not find any correlations between the frequency of interruptions (0–11) and these parameters in any of the data sets. However, we found significant differences when comparing the two suppliers. RBCs of manufacturer A (‘A’, inline filtration of whole blood) contained 25% more Hb than those of manufacturer B (‘B’, inline filtration after buffy coat reduction). Sixteen percentage of ‘A’‐RBC, but none of ‘B’‐RBC, exceeded a haemolysis of 0·8%. Conclusions Transitory interruptions of cold chain do not measurably impair the quality of RBCs. The effect on storage of RBCs in the blood bank is not as significant a factor as the differences that exist between RBC manufacturing procedures.     

Influence of irradiation on in vitro red-blood-cell (RBC) storage variables of leucoreduced RBCs in additive solution PAGGS-M

Background The effect of gamma irradiation on leucoreduced red‐blood‐cells (RBCs) stored in an additive solution (AS) containing phosphate, adenine, glucose, guanosine, saline and mannitol (PAGGS‐M) has not yet been studied, and there are different recommendations about storage time of leucoreduced RBCs after irradiation. Study Design and Methods We studied 63 leucoreduced RBC units. All RBCs were stored in AS PAGGS‐M and leucoreduced on the collection day. Twenty‐one components were irradiated on Day +14 with 30 Gy and 22 served as non‐irradiated controls. Samples were drawn and analysed from these 43 units on Day +7, +14, +21, +28, +35, +42 and +49 from the collection day. From 20 units, no samples were taken earlier than on Day +49. Of these, 10 components had been irradiated on Day +14 with 30 Gy and 10 served as non‐irradiated controls. Results Gamma irradiation induced an enhanced in vitro haemolysis rate in the irradiated components. One of the irradiated units showed a haemolysis rate over the recommended limit of 0·8% on Day +42 and four on Day +49. The leakage of potassium ions from irradiated RBCs started to increase faster than that of unirradiated RBCs from the day of irradiation. Lactate dehydrogenase levels increased faster in irradiated units 3 weeks after irradiation. We showed that taking samples weekly does not affect the final result. Conclusions Our findings show that the European recommendations should not be changed in regard to the limitation of the storageability after irradiation of leucoreduced RBCs. The damage after irradiation and storage cannot be prevented by using the high‐quality AS PAGGS‐M.     

Causes of microcytic anaemia and evaluation of conventional laboratory parameters in the differentiation of erythrocytic microcytosis in blood donors candidates*

ABSTRACT

Context and Objective: Microcytic anaemia results from defective synthesis of haemoglobin in the erythroid precursors, causing a reduction in its mean corpuscular volume (MCV). The most common causes of microcytosis, without the increase in HbA₂ levels, are iron deficiency anaemia (IDA) and α-thalassemia. The aim of this study was to identify the causes of microcytic anaemia and evaluate the haematological parameters from blood donors deemed ineligible (due to the low haematocrit level) that would differentiate the IDA and α-thal, whether isolated or in association.

Methods: Genomic DNA was submitted to the polymerase chain reaction multiplex for the diagnosis of the most common allele deletions of α-thal and erythrogram and in order to verify haematological parameters. Iron deficiency (ID) was determined through the measurement of serum ferritin.

Results: Of the 204 samples, 82 (40.2%) were identified with ID, 24 (17.8%) with α-thal and 10 (4.9%) with ID associated with α-thal. In the α-thal with ID group haemoglobin (Hb), MCV, mean corpuscular Hb concentration (MCHC) and mean corpuscular Hb (MCH) values were significantly lower compared to the isolated α-thal. In the group with ID Hb, MCV, MCHC and MCH values were significantly lower compared to those with isolated α-thal. The α-thal with ID group, showed Hb, MCV, MCHC and MCH significantly reduced when compared to those with IDA.

Conclusions: This study showed that the values of haematological parameters, especially haematocrit, Hb, MCV, MCH, MCHC and red blood cell distribution width (RDW), are lower in patients with IDA, especially when associated with α-thal and therefore it may be useful to discriminate between the different types of microcytic anaemia.     

Dose‐dependent effect of stored‐blood transfusions on recipient red blood cell indices,deformability and density

We prospectively studied the dose‐dependent effect of transfused stored red blood cells (RBCs) on recipient RBC indices, deformability and cell density in 10 patients administered stored RBCs for blood transfusion during general surgery. There were dose‐dependent decreases in mean corpuscular volume and increases in mean corpuscular haemoglobin concentration after completion of 4‐ and 6‐unit stored RBC transfusions. The amount of dense populations increased proportionately with the amount of stored RBCs transfused. The maximal deformability index value was significantly and dose‐dependently decreased, suggesting that hemodynamic blood flow, especially the microcirculation may be impaired in patients who receive large amounts of stored RBCs.     

Beta thalassemia minor is a beneficial determinant of red blood cell storage lesion

Blood donor genetics and lifestyle affect the quality of red blood cell (RBC) storage. Heterozygotes for beta thalassemia (bThal⁺) constitute a non-negligible proportion of blood donors in the Mediterranean and other geographical areas. The unique hematological profile of bThal⁺ could affect the capacity of enduring storage stress, however, the storability of bThal⁺ RBC is largely unknown. In this study, RBC from 18 bThal⁺ donors were stored in the cold and profiled for primary (hemolysis) and secondary (phosphatidylserine exposure, potassium leakage, oxidative stress) quality measures, and metabolomics, versus sex- and age-matched controls. The bThal⁺ units exhibited better levels of storage hemolysis and susceptibility to lysis following osmotic, oxidative and mechanical insults. Moreover, bThal⁺ RBC had a lower percentage of surface removal signaling, reactive oxygen species and oxidative defects to membrane components at late stages of storage. Lower potassium accumulation and higher uratedependent antioxidant capacity were noted in the bThal⁺ supernatant. Full metabolomics analyses revealed alterations in purine and arginine pathways at baseline, along with activation of the pentose phosphate pathway and glycolysis upstream to pyruvate kinase in bThal⁺ RBC. Upon storage, substantial changes were observed in arginine, purine and vitamin B6 metabolism, as well as in the hexosamine pathway. A high degree of glutamate generation in bThal⁺ RBC was accompanied by low levels of purine oxidation products (IMP, hypoxanthine, allantoin). The bThal mutations impact the metabolism and the susceptibility to hemolysis of stored RBC, suggesting good post-transfusion recovery. However, hemoglobin increment and other clinical outcomes of bThal⁺ RBC transfusion deserve elucidation by future studies.     

Hypoxia and hypocapnia storage of γ-irradiated red cell concentrates

Backgroundγ-irradiation is used to treat red blood cell (RBC) concentrates (RCCs) transfused to immunosuppressed patients. This treatment damages RBCs and increases storage lesions. Several studies have shown the beneficial effect of reducing O₂ content during RBC storage. The present research work investigated the effect of γ-irradiation on RCCs stored under normal and hypoxia/hypocapnia conditions.Materials and methodsO₂ concentration (measured as oxyhaemoglobin fraction, sO₂) and ABO-matched RCCs from whole blood donations, leukoreduced and prepared in phosphate, adenine, glucose, guanosine, saline and mannitol (PAGGSM) were pooled and split in two identical RCCs within 24 h post donation. One bag (Hx) was submitted to O₂ and CO₂ adsorption for 3 h on an orbital shaker at 22±2 °C and then transferred to a storage bag impermeable to gas. The other bag (Ctrl) was left as it was. The two bags were then stored at 4 °C. γ-irradiation (25 Gy) was applied at day 2 or 14, and the RCCs were stored until day 43. Different parameters (metabolites, haemolysis, morphology) were measured.ResultsStarting sO₂ values were 63.7±18.4% (n=12) in Ctrl and 20.8±9.8% (n=12) in Hx bags, and reached 90.8±9.1% and 6.6±5.9% at day 43, respectively. As expected, an increase in glycolysis rate was observed after deoxygenation. Extracellular potassium concentrations were identical and reached around 70 mM at expiry with an irradiation-dependent kinetic release. No difference in haemolysis was observed after irradiation on day 2 in either group (<0.40%, p>0.9999). When irradiated at day 14, haemolysis was lower (p=0.033) in RCCs under hypoxia at the end of storage (day 28, 0.67±0.16%) compared to control (1.06±0.33%). Percentages of spherocytes were lower under hypoxia.DiscussionThe storage under hypoxia provided equivalent storage when RCCs were irradiated at day 2 and was advantageous when irradiated at day 14. In summary, O₂-depletion of RCCs enable a better storage of RBCs, particularly when late irradiation is applied.     

Gender difference in rheologic properties of blood and risk of cardiovascular diseases

According to official statistical data there is a significant difference between pre-menopausal women and age-matched men in morbidity and mortality from cardiac diseases and especially from myocardial infarction. There are several speculations regarding the nature of this phenomenon which have both supporting and refuting evidence. Our hypothesis was that due to regular physiologic bleeding, rheological properties of blood of pre-menopausal women are superior to those of men, and place such women at a lower risk of cardiovascular diseases than men in any age group. We believe that this difference in hemorheological properties is due to the reduced concentration of red blood cells (RBCs) and due to greater population of younger and less population of older RBCs in female blood. We studied mechanical properties of blood from 47 pre-menopausal women and 50 age-matched men. Compared to female blood, male blood had higher viscosity and RBC aggregation and lower RBC deformability. Oxygen Delivery Index, calculated as a ratio of hematocrit to blood viscosity, was found to be significantly lower in male blood. Decreased oxygen delivery along with increased RBC aggregation and decreased RBC deformability may contribute to the higher risk for the development of cardiovascular diseases. Regular blood donation may reduce hematocrit and blood viscosity, improve rheological properties of blood, and increase oxygen delivery in men.     

Hematological and Molecular Characterization of a Novel Hb A2 Variant with Homozygous α-Thalassemia-2 in a Southern Thai Individual

We report here the hematological and molecular features of a novel δ-globin chain variant found in a Southern Thai woman. Her complete blood count was as follows: red blood cell (RBC) count 5.90?×?10¹²/L, hemoglobin concentration (Hb) 12.6?g/dL, packed cell volume (PCV) 0.41?L/L, mean corpuscular volume (MCV) 69.5 fL, mean corpuscular Hb (MCH) 21.4?pg, mean corpuscular Hb concentration (MCHC) 30.7?g/dL and RBC distribution width (RDW) 13.1%. The blood smear demonstrated microcytic hypochromic RBCs suggestive of thalassemia trait. Hemoglobin analysis identified Hb A₂?+?Hb A₂-Kiriwong (2.4%) and Hb F (0.1%) on high performance liquid chromatography (HPLC). To characterize the α-thalassemia (α-thal) genotype, common α-thal-1 and α-thal-2 alleles were characterized by multiplex gap-polymerase chain reaction (gap-PCR). The results revealed homozygous α-thal-2 (–α^3.7/–α^3.7) in this case. DNA sequencing showed the presence of a novel δ-globin gene mutation [δ77(EF1)His→Arg; HBD: c.233A>G] that we named Hb A₂-Kiriwong for the village from where the proband lived. In summary, the presence of microcytic hypochromic RBCs in this case was likely the result of the homozygous –α^3.7 (rightward) deletion and was not affected by this Hb A₂ variant.     

Storage of whole blood overnight in different blood bags preceding preparation of blood components: in vitro effects on red blood cells

Background

Routines for the storage of whole blood (WB) overnight for the preparation of blood components on the following day are of increasing interest primarily for logistic reasons. The present study focuses on in vitro effects during storage for 6 weeks on red blood cells (RBC) prepared in different blood containers after being held overnight.

Study design and methods

Five different blood collection systems were used with either inline leucocyte reduction red cell filters for the preparation of RBC, buffy coat (BC) and plasma or WB filters for the preparation of RBC and plasma. A new container with an integrated WB filter removing leucocytes but not platelets was also included for the preparation of leucocyte-reduced RBC, BC and plasma units. Standard CPD solution (63 or 70 mL) and SAG-M solution (100 or 110 mL) were used for the collection of either 450 or 500 mL blood. All WB units were stored at room temperature, either overnight for 18–24 hours (test groups, n=104) or for up to 8 hours (reference groups, n=20). In addition, five test units were stored overnight under refrigeration.

Results

In test groups (overnight storage at room temperature) we found significantly lower levels of extracellular potassium, 2,3-DPG and pH (up to day 28). During storage, higher levels of ATP (Terumo, CaridianBCT until day 35, Fresenius until day 14, Fenwal throughout storage) were seen in test groups than in reference groups. When WB was stored overnight at 2–6°C before WB filtration, the levels of ATP and haemolysis were higher than in the corresponding reference.

Conclusion

Significant differences in in vitro parameters were observed between RBC prepared within 8 hours and 18–24 hours after blood collection. The results were consistent irrespective of the blood container used. New alkaline solutions may decrease the differences.     

Evolution of adverse changes in stored RBCs

Recent studies have underscored questions about the balance of risk and benefit of RBC transfusion. A better understanding of the nature and timing of molecular and functional changes in stored RBCs may provide strategies to improve the balance of benefit and risk of RBC transfusion. We analyzed changes occurring during RBC storage focusing on RBC deformability, RBC-dependent vasoregulatory function, and S-nitrosohemoglobin (SNO-Hb), through which hemoglobin (Hb) O(2) desaturation is coupled to regional increases in blood flow in vivo (hypoxic vasodilation). Five hundred ml of blood from each of 15 healthy volunteers was processed into leukofiltered, additive solution 3-exposed RBCs and stored at 1-6 degrees C according to AABB standards. Blood was subjected to 26 assays at 0, 3, 8, 24 and 96 h, and at 1, 2, 3, 4, and 6 weeks. RBC SNO-Hb decreased rapidly (1.2 x 10(-4) at 3 h vs. 6.5 x 10(-4) (fresh) mol S-nitrosothiol (SNO)/mol Hb tetramer (P = 0.032, mercuric-displaced photolysis-chemiluminescence assay), and remained low over the 42-day period. The decline was corroborated by using the carbon monoxide-saturated copper-cysteine assay [3.0 x 10(-5) at 3 h vs. 9.0 x 10(-5) (fresh) mol SNO/mol Hb]. In parallel, vasodilation by stored RBCs was significantly depressed. RBC deformability assayed at a physiological shear stress decreased gradually over the 42-day period (P < 0.001). Time courses vary for several storage-induced defects that might account for recent observations linking blood transfusion with adverse outcomes. Of clinical concern is that SNO levels, and their physiological correlate, RBC-dependent vasodilation, become depressed soon after collection, suggesting that even "fresh" blood may have developed adverse biological characteristics.     

Oxidative stress in erythrocytes of banked ABO blood

Objective: To understand the responses of A, B, and O blood groups to oxidative stress (OS) induced through storage.

Methods: A, B, and O blood units were obtained from the blood bank at KIMS Hospital, Bangalore, and stored for 35 days at 4°C in citrate phosphate dextrose adenine-1 solution. Every fifth day, hemoglobin (Hb) was assessed in whole blood and erythrocytes were isolated from each group. OS markers such as (i) antioxidant enzymes [superoxide dismutase and catalase] and superoxides were assessed in hemolysate; (ii) lipid peroxidation product – malondialdehyde (MDA) and protein oxidation products [protein carbonyls, advanced oxidation protein products (AOPP), and protein sulfhydryls] were assessed in membrane ghosts.

Results: Antioxidant enzymes and Hb were similar in all groups. Superoxides increased in blood group O. MDA and AOPP differed between the groups, where levels in blood group O were lower than blood groups A and B. Sulfhydryls were maintained throughout storage.

Discussion: The antioxidant defense in A, B, and O groups were similar as evident from our results of Hb, antioxidant enzymes and sulfhydryls. However, the response of blood group O diverged from that of A and B, substantiated by the results of MDA, AOPP, and superoxides. Thus blood group O endured oxidative insult more efficiently than A and B. This study forms the basis for future studies on erythrocyte membrane and exploring blood group O as a potential candidate for prolonging storage.     

Influence of late irradiation on the in vitro RBC storage variables of leucoreduced RBCs in SAGM additive solution

Background There exists only few data on in vitro and in vivo effects of gamma irradiation of leucoreduced red blood cells (RBCs). Reported studies reflect the effects of early irradiation and subsequent storage. The effects of irradiation on RBCs shortly before the end of their shelf‐life have not been examined. Study Design and Methods We studied 160 RBC units that were stored in the additive solution saline–adenine–glucose–mannitol and leucoreduced on the collection day. Forty components were irradiated on day +14 with 30 Gy, 40 on day +28, 40 on day +35, and 40 served as nonirradiated controls. In vitro evaluation of all units was performed on days +3, +7, +14, +21, +28, +35, and +42 from the collection day. Results Gamma irradiation induced leakage of potassium ions and lactate dehydrogenase and enhanced in vitro haemolysis rate in the irradiated components, which started to increase faster than that of nonirradiated RBCs from the day of irradiation, i.e. from day +14 in units that were irradiated on day +14, from day +28 in units that were irradiated on day +28, and from day +35 in units that were irradiated on day +35. Conclusions This study presents data on the in vitro quality of leucoreduced RBCs that have been irradiated on days +14, +28, or +35 after collection. Our findings support the proposal that the current limitation of the age of RBCs on the day of gamma irradiation may be replaced by staged limitations depending on the time of irradiation.