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IMPORTANCE: Gamma irradiation of leukoreduced red blood cells (RBCs) prevents transfusion-associated graft-vs-host disease but also exacerbates storage lesion formation in RBCs. It is unknown whether freshly irradiated RBCs are more efficacious than irradiated and stored RBCs in preterm infants with high transfusion requirements. OBJECTIVE: To examine whether transfusion of freshly irradiated vs irradiated and stored RBC components improves cerebral oxygen delivery in preterm infants with anemia. DESIGN, SETTING, AND PARTICIPANTS: This single-center, double-blinded, proof-of-concept randomized clinical trial was conducted at the neonatal intensive care unit of Wellington Regional Hospital in Wellington, New Zealand, between December 1, 2017, and November 30, 2018. Participants were preterm infants (

Background Laparoscopic Heller myotomy (LHM) and esophageal balloon dilation (BD) are the two mainstays of achalasia treatment—this study examines the outcomes when they are performed simultaneously without fundoplication. Methods All patients undergoing LHM&BD were reviewed for demographic and procedural data, and to see if additional procedures for achalasia had been performed. Patients were surveyed using the Eckardt score and the GERD quality-of-life score (GERD-HRQL) to assess the durability of repair. Results From 2013-2020, 66 patients underwent LHM&BD. There were no esophageal perforations and a median LOS of 1 day. Seven patients have required additional operations or procedures at median 4-years follow up. 31 patients (47%) responded to the survey. The average Eckardt score was 2.9 (goalConclusions LHM&BD allows for a safe, durable repair of achalasia. Reflux symptoms are manageable with PPI without fundoplication and the re-intervention rate similar to published values.

BACKGROUND Gamma irradiation of red blood cells (RBCs) is well recognized to exacerbate storage lesion formation, but the effect of storage after irradiation on in vivo oxygen delivery capacity of transfused RBCs is currently not known. STUDY DESIGN AND METHODS In 24 preterm infants with anemia receiving nonurgent transfusion of irradiated RBCs, we examined cerebral regional tissue oxygenation (crSO2 ) and time spent with peripheral arterial saturation (SpO2 ) less than 88%. Physiologic data were obtained immediately before, immediately after, and 5 days after transfusion. RESULTS We observed linear negative moderate correlations between time since irradiation and the magnitude of change in crSO2 (r = -0.60; 95% CI, -0.81 to -0.27; p = 0.0018) and time spent with SpO2 of less than 88% (r = -0.42; 95% CI, -0.71 to 0.003; p = 0.04) immediately after transfusion. In infants (n = 9) who received fresher RBCs (irradiated

BACKGROUND Hepatitis E virus (HEV) Genotype 3 (G3) infection is a zoonosis that may be transmitted during the acute phase by transfusion. The aim of this study was to determine the incidence of HEV and seroprevalence among Irish blood donors. STUDY DESIGN AND METHODS Anonymized samples from 1076 donations collected in 2012 were tested for HEV immunoglobulin (Ig)G using the Wantai enzyme-linked immunosorbent assay. A total of 24,985 anonymized donations collected between December 2013 and June 2014 were individually tested for HEV RNA using the Procleix HEV assay; reactive donations were confirmed by an in-house real-time polymerase chain reaction (PCR) test. RESULTS Seroprevalence for anti-IgG was 5.3% (95% confidence interval [CI], 4.0%-6.8%), ranging from 1.1% in the 18- to 29-years age group to 33.3% in males over 60 years. HEV RNA screening of 24,985 samples yielded five PCR-confirmed donations (1:4997, 0.02%; 95% CI, 0.0065%-0.0467%), only one of which was serologically reactive (HEV IgM reactive only). Viral loads ranged from 10 to 44,550 IU/mL. Genotype analysis on three samples identified HEV G3 virus. Four of the five viremic donations were from donors in the 18- to 29-years age group (p = 0.01). CONCLUSION Seroprevalence for anti-HEV IgG was low compared to some European countries, but 1 in 5000 donations was viremic. Viremia was predominantly in younger Irish donors. After Department of Health approval the Irish Blood Transfusion Service implemented individual blood donation HEV RNA screening initially for a 3-year period from January 2016.

We performed a prospective analysis of iron status in plateletpheresis donors, using whole blood donors as a control group, to assess the haematinic effects of regular anti-coagulated extracorporeal circulation and platelet collection.Ferritin levels were measured in samples from 31 regular male plateletpheresis donors and from 14 first time male whole blood donors, immediately before and immediately after donation, and immediately before the next donation. An additional 33 regular male plateletpheresis donors and 17 first time male whole blood donors had serum ferritin levels checked predonation.Male plateletpheresis donors had a statistically significant fall in serum ferritin after donation (P = 0.005)*. In addition, male platelet donors had significantly lower serum ferritin levels than first time male blood donors: ferritin20 µg/L was found in 6/64 (9%) of regular platelet donors and 1/31 (3%) of first time blood donors (P 0.001)*.Our studies support the value of serum ferritin measurement in apheresis donor management.

Introduction: Circulating microparticles (MP) have been described in sickle cell anaemia (SCA); however, their interaction with endothelial markers remains unclear. We investigated the relationship between MP, protein C (PC), free protein S (PS), nitric oxide (NO), endothelin-1 (ET-1) and adrenomedullin (ADM) in a large cohort of paediatric patients.Method: A total of 111 children of African ethnicity with SCA: 51 in steady state; 15 in crises; 30 on hydroxyurea (HU) therapy; 15 on transfusion; 17 controls (HbAA) of similar age/ethnicity. MP were analysed by flow cytometry using: Annexin V (AV), CD61, CD42a, CD62P, CD235a, CD14, CD142 (tissue factor), CD201 (endothelial PC receptor), CD62E, CD36 (TSP-1), CD47 (TSP-1 receptor), CD31 (PECAM), CD144 (VE-cadherin). Protein C, free PS, NO, pro-ADM and C-terminal ET-1 were also measured.Results: Total MP AV was lower in crisis (1.26×106 ml−1; 0.56–2.44×106) and steady state (1.35×106 ml−1; 0.71–3.0×106) compared to transfusion (4.33×106 ml−1; 1.6–9.2×106, p0.9, p

Gamma irradiation of red blood cells (RBCs) is well recognized to exacerbate storage lesion formation, but the effect of storage after irradiation on in vivo oxygen delivery capacity of transfused RBCs is currently not known.In 24 preterm infants with anemia receiving nonurgent transfusion of irradiated RBCs, we examined cerebral regional tissue oxygenation (crSOWe observed linear negative moderate correlations between time since irradiation and the magnitude of change in crSOOur findings indicate that storage after gamma irradiation may have a detrimental effect on the oxygen delivery capacity of RBCs given to anemic preterm infants.

As stem cell technologies move from the developmental to the commercial stage strategies must be developed to monitor culture operations. These will ensure consistency of differentiation programs and maintenance of optimum cell viability during production runs. Due to the sensitivity of stem cells to their environment, and their variability in response to external stimuli, accurate monitoring of in vitro conditions will be crucial for effective large-scale culturing of therapeutic stem cells. Here we describe a simple method to monitor the expansion and maturation of adult human haematopoietic stem/progenitor cells into red blood cells in vitro by measuring the oxygen consumption rate of cultures. Cell cultures followed a characteristic pattern of oxygen consumption that is reflective of in vivo erythroid maturation. This method could be easily developed as an online system to map erythroid differentiation and maturation of cultured cells as effectively as the more time consuming process of flow cytometric analysis of surface marker expression patterns.

Men and women have different mean haemoglobin levels in health in venous blood - women have mean levels approximately 12% lower than men. A similar sex-related difference in haemoglobin levels in adult animals is found in many species of mammals, birds and reptiles, indicating that it is an important physiological phenomenon. It is probably a direct effect of sex hormones, both oestrogen and androgens, on erythropoiesis. However, since there is no difference in erythropoietin levels between the sexes, this effect most likely takes place in the kidney, rather than in the bone marrow. Oestrogens dilate and androgens constrict the renal microvasculature: dilation and vasoconstriction in vessels below 300 μm in diameter respectively increase and decrease the haematocrit in blood in arterioles, capillaries and venules, altering the oxygen delivery per unit red cell mass, and providing a mechanism for varying the red cell mass without compensatory changes in erythropoiesis.

By the time vCJD was first described in 1996, it was already far too late to offset further disaster from transmission of the disease by blood transfusion: almost all the humans who would be infected and infectious were already diseased. Nothing done by the blood transfusion services around that time, with the exception of excluding transfusion recipients as blood donors, would have made any useful contribution to containing the extent of the epidemic. The ability to spread emerging diseases before the problem is manifest or understood is a fixed and unavoidable feature of blood transfusion as it is practiced today. A second fixed property of blood transfusion is that the root cause of disaster is not within the control of the blood transfusion universe. Strategies that have emerged to cope with similar threat in other enterprises that also contain these properties comprise the components of robust design: surveillance, preparedness for action, engagement, herding together, evasion or avoidance, early adoption of potentially useful measures, engineered resilience, defence in depth, damage limitation including modularity and removal of feedback loops, and contingency, redundancy and failure management, and ultimately, individual escape. Early adoption of leucodepletion based on the possibility that it might work rather than any hard evidence was a good example of threat management. Exclusion of previously transfused donors is a robust mechanism for containing any future infection; optimal blood use structures that provide a national transfusion rate as low as possible also constitute an effective threat management strategy.

Hepatitis E virus (HEV) Genotype 3 (G3) infection is a zoonosis that may be transmitted during the acute phase by transfusion. The aim of this study was to determine the incidence of HEV and seroprevalence among Irish blood donors.Anonymized samples from 1076 donations collected in 2012 were tested for HEV immunoglobulin (Ig)G using the Wantai enzyme-linked immunosorbent assay. A total of 24,985 anonymized donations collected between December 2013 and June 2014 were individually tested for HEV RNA using the Procleix HEV assay; reactive donations were confirmed by an in-house real-time polymerase chain reaction (PCR) test.Seroprevalence for anti-IgG was 5.3% (95% confidence interval [CI], 4.0%-6.8%), ranging from 1.1% in the 18- to 29-years age group to 33.3% in males over 60 years. HEV RNA screening of 24,985 samples yielded five PCR-confirmed donations (1:4997, 0.02%; 95% CI, 0.0065%-0.0467%), only one of which was serologically reactive (HEV IgM reactive only). Viral loads ranged from 10 to 44,550 IU/mL. Genotype analysis on three samples identified HEV G3 virus. Four of the five viremic donations were from donors in the 18- to 29-years age group (p = 0.01).Seroprevalence for anti-HEV IgG was low compared to some European countries, but 1 in 5000 donations was viremic. Viremia was predominantly in younger Irish donors. After Department of Health approval the Irish Blood Transfusion Service implemented individual blood donation HEV RNA screening initially for a 3-year period from January 2016.

BACKGROUND: Blood transfusion in rural sub-Saharan Africa presents special challenges. Transfusions are primarily given for emergencies—life-threatening blood loss or anemia; blood is usually collected from family or replacement donors; and facilities to store an adequate reserve in a hospital bank are constrained. We report the everyday and organizational practices in a medium-sized district hospital in Northern Ghana. STUDY DESIGN AND METHODS: Information and data on blood transfusion practices at West Gonja Hospital, Damongo, were available from the laboratory reports, from day books and workbooks, and from direct observation in the following four areas: blood collection and blood donors; blood donation testing; blood storage and logistics; and clinical transfusion practice, adverse events, and follow-up. RESULTS: The hospital serves a rural community of 86,000. In 2009, a total of 719 units of whole blood were collected, a rate of 8.36 units per 1000 population. All donors were family or replacement donors. Positivity rates for infectious disease markers were 7.5% (64/853) for hepatitis B surface antigen, 6.1% (50/819) for hepatitis C virus, 3.9% (33/846) for human immunodeficiency virus, and 4.7% (22/468) for syphilis. Supply of laboratory materials was sometimes problematic, especially for temperature-critical materials. Difficulties in sample labeling, storage of blood and laboratory supplies, and disposal of waste were also incurred by operational, material, and financial constraints. Follow-up for outcomes of transfusion is not currently feasible. CONCLUSIONS: The operational, demographic, and financial environment pertaining in a rural hospital in Northern Ghana differs substantially from that in which much of current blood transfusion practice and technology evolved. Considerable effort and innovation will be needed to address successfully the challenges posed.

Bacteria in transfused platelets can cause serious morbidity and, rarely, death. Most contaminating bacteria enter the blood at the time of venepuncture. While many of these contaminants fail to grow in the platelet unit, storage of platelets at 20–24 °C facilitates growth of some organisms, and the cumulative risk of severe sepsis increases with the storage age of platelet components. Several methods have been developed or adapted to attempt to detect contaminating bacteria with high sensitivity and specificity, but the perfect test has yet to be found. Testing early in the platelet component’s shelf life, even using exquisitely sensitive culture-based tests, is compromised by major problems of sample error – there may be too few bacteria present at this stage to ensure that any practical sample volume contains even one of them. Culture techniques are too slow to be useful as a release test. On the other hand, available rapid tests are too insensitive to use early in the shelf life, and have yet to show convincingly that they are sensitive enough for testing close to the time of transfusion. Nevertheless testing for bacteria in platelet components represents a significant advance in blood transfusion safety, and prevents the transfusion of many hundreds of bacterially-contaminated platelet units each year.

The cultivation of erythroid cells at large scale would have to be performed in suitable bioreactors which would most likely employ some mode of agitation to ensure optimal mass and gas transfer and prevent culture inhomogeneity. The effect of low agitation at 15-20 rpm on ex vivo erythropoiesis of PB CD34+ derived cultures was investigated and found to have significant impact on erythroid development. Agitated cultures showed a reduced lag phase and increased cell expansion during the early stages of culture. Additionally, agitation accelerated erythroid differentiation as seen by the loss of early development markers, acquisition of late erythroid markers and premature cell cycle arrest, although not yielding higher fractions of terminally differentiated cells in comparison to stationary culture. However, agitation at 20 rpm led to significantly increased loss of cell viability after day 15 in culture, an effect that could be reduced by decreasing the agitation rate to 15 rpm. On the one hand these results imply that agitation may improve cell yields and reduce expensive cytokine-dependent early culture stages but on the other hand it might introduce the risk of increased cell death in large scale culture.

This study investigated whether filtration of leucodepleted red cells in SAG-M through the P-CAPT filter in order to prevent the potential risk of vCJD infection associated with prion transmission through transfusion has any deleterious effect on red-cell quality. Bottom-and-top SAG-M leucodepleted red-cell concentrates (24 units) were prion-reduction filtered on the day following collection, with half of the units undergoing irradiation on day 14. A control group (12 units) was not prion filtered. Units were sampled at 7-day intervals up to day 35 and tested using standard measures of red-cell quality as well as prothrombin content (to examine prion filter efficacy). Haemoglobin loss per unit was approximately 9 g and in some cases levels were below standard specification (40 g). Haemolysis increased significantly after filtration [0.01 (0.00-0.05) vs. 0.23 (0.07-0.52, p

Microparticles (MP) derived from vascular endothelium or circulating blood cells circulate in the peripheral blood. They originate from blebbing and shedding from cell membrane surfaces in physiological and pathological conditions and are present in low concentrations in normal plasma. Increased levels are generated by a number of mechanisms including platelet activation, direct vascular endothelial damage, thrombin activity on the cell surface, C5b-9 activation, and PF4-heparin-antibody interaction. Several techniques are currently used to study the generation and nature of circulating microparticles. In particular, the genesis and role of microparticles, derived from platelets, endothelial cells and monocytes, in sepsis (especially meningococcal-induced), heparin-induced thrombocytopenia (HIT), thrombotic thrombocytopenic purpura (TTP), aplastic anaemia, paroxysmal nocturnal haemoglobinuria (PNH) and sickle cell disease (SCD) have been well studied, and provide important insights into the underlying diseases. A defect in the ability to form microparticles leads to the severe bleeding disorder of Scott syndrome, which in turn provides a revealing insight into the physiology of coagulation. In addition the complex role of microparticles in vascular and cardiovascular diseases is an area of immense interest, that promises to yield important advances into diagnosis and therapy.

The study was conducted to assess the efficacy and tolerance of solvent-detergent (SD) plasma in neonates, in obstetric and gynaecological patients and in patients with liver disease in three large hospitals in Dublin during the period 1 April 2002 (when SD plasma was introduced) through 31 October 2003. Forty-one neonates received 67 transfusions of SD plasma at a mean dose +/- standard deviation of 18.4 +/- 13.2 mL kg(-1). Thirty-one (75.6%) patients had coagulopathy without haemorrhage (either disseminated intravascular coagulopathy or other coagulopathy); a further eight (19.5%) had clinical haemorrhage (excluding intraventricular haemorrhage), which complicated a coagulopathy. Thirty-eight obstetric and gynaecological patients received 57 SD plasma transfusions at a mean dose of 15.3 +/- 7.7 mL kg(-1). Thirty-six women (94.7%) had haemorrhage with mean blood loss per patient of 3345.8 +/- 2738.1 mL. Fifteen children with liver disease received 33 SD plasma transfusions at a mean volume of 38.0 +/- 41.5 mL kg(-1) body weight. Seventeen adult patients with severe end-stage liver disease were transfused with SD plasma either following liver transplantation or prior to other invasive procedures, at a mean dose of 10.2 +/- 3.4 mL kg(-1). There were statistically significant decreases in mean activated partial thromboplastin time and prothrombin time in neonates, in obstetric and gynaecological patients and in patients with liver disease. Complete correction of coagulation parameters was not achieved in any case in the liver transplant group. No adverse reactions were observed for SD plasma infusion. Use of SD plasma in critically ill neonates, in women with obstetric and gynaecological emergencies and in patients with liver disease appears safe, and improves laboratory indices of coagulopathy.

Objective Fetomaternal mismatch for human platelet antigen (HPA)-1a accounts for approximately 85% of cases of neonatal alloimmune thrombocytopenia. The purpose of the study was to determine the prevalence of the HPA-1a negative platelet phenotype in a cohort of pregnant women in Ireland, the rate of alloimmunisation to HPA-1a in HPA-1 mismatched pregnancies and the associated incidence of neonatal alloimmune thrombocytopenia. Design A prospective case–control study. Setting The antenatal clinics of a large maternity teaching hospital. Population or sample Pregnant women, regardless of parity, presenting at the antenatal clinics. Methods An enzyme-linked immunosorbent assay (ELISA) designed for simultaneous HPA-1a typing and antibody detection was used. Further analysis for HPA-1a alloantibodies was performed using commercial ELISA's (GTI PakPlus and Pak1) and the monoclonal antibody immobilisation of platelet antigens assay. Confirmation of serological typing was by the polymerase chain reaction technique using sequence-specific primers (PCR-SSP). Main outcome measures The presence of the HPA-1a negative phenotype and its association with the development of maternal anti-HPA-1a and infant thrombocytopenia. Results Eighty-four of 4090 consecutive women enrolled in the study tested positive for HPA-1a in the screening ELISA. Confirmatory genotyping was performed on 67 women (representing 80% of the cohort), and 54 women (representing 3272 non-selected pregnancies), were homozygous for the HPA-1b allele (1.7%). Three of 34 (9%) women who delivered HPA-1a positive babies had detectable anti-HPA-1a and all three babies had neonatal alloimmune thrombocytopenia, for an overall incidence of 1:1100 non-selected pregnancies. Conclusions The observed prevalence of 1.7% for the HPA-1a negative platelet phenotype is as expected from studies in other countries. While we have demonstrated the practicability of antenatal HPA-1a screening, further research is warranted to investigate maternal parameters predictive of severe fetal thrombocytopenia in HPA-1a alloimmunised pregnancies.

BACKGROUND: Citrate infusion during apheresis procedures can cause lowering of the plasma ionized calcium leading to prolongation of the QT interval and hypotension. At the myocardial level, QTc measurement is a sensitive marker of subphysiologic calcium values caused by citrate toxicity. STUDY DESIGN AND METHODS: Seventy-six regular platelet donors were recruited into this study. QTc intervals were measured continuously for 3 minutes at four different points during the apheresis procedures. The blood pressure was recorded every 5 minutes throughout the procedure. RESULTS: The baseline QTc value for men was 406.5 ± 13.1 milliseconds1/2 and for women was 417.6 ± 13.3 milliseconds1/2 (p = 0.0016). QTc prolongation occurred in all procedures in all donors. Maximum recorded values were significantly higher in women than in men (450.8 ± 20.8 vs. 424.8 ± 14.3 ms1/2; p = 0.0025). All QTc values returned to baseline by 15 minutes after the procedure. Changes in blood pressure were minimal and no donor experienced severe symptoms. CONCLUSIONS: QTc prolongation always occurs during plateletpheresis. This prolongation is greater in women than in men. This study indicates that it may be advisable to assess donors for family or personal history of syncope and family history of sudden death to exclude those at increased risk of arrhythmias because of asymptomatic carriage of a long-QT gene. In addition baseline QTc measurement is a simple noninvasive procedure that could be applied to further studies with a view to enhancing donor safety in apheresis.

An understanding of the metabolic profile of cell proliferation and differentiation should support the optimization of culture conditions for hematopoietic stem and progenitor cell (HSPC) proliferation, differentiation, and maturation into red blood cells. We have evaluated the key metabolic parameters during each phase of HSPC culture for red blood cell production in serum-supplemented (SS) and serum-free (SF) conditions. A simultaneous decrease in growth rate, total protein content, cell size, and the percentage of cells in the S/G2 phase of cell cycle, as well as an increase in the percentage of cells with a CD71(-)/GpA(+) surface marker profile, indicates HSPC differentiation into red blood cells. Compared with proliferating HSPCs, differentiating HSPCs showed significantly lower glucose and glutamine consumption rates, lactate and ammonia production rates, and amino acid consumption and production rates in both SS and SF conditions. Furthermore, extracellular acidification was associated with late proliferation phase, suggesting a reduced cellular metabolic rate during the transition from proliferation to differentiation. Under both SS and SF conditions, cells demonstrated a high metabolic rate with a mixed metabolism of both glycolysis and oxidative phosphorylation (OXPHOS) in early and late proliferation, an increased dependence on OXPHOS activity during differentiation, and a shift to glycolytic metabolism only during maturation phase. These changes indicate that cell metabolism may have an important impact on the ability of HSPCs to proliferate and differentiate into red blood cells.

Glycocalicin (GC) is the soluble portion of platelet membrane protein GP1b, and may be cleaved from the platelet surface during platelet activation. Previous study has indicated that plasma glycocalicin/platelet (GC/plt) levels are elevated in patients presenting with acute stroke. The present study was undertaken to determine the GC/plt levels in patients being treated for transient ischaemic episodes, to assess whether the elevated GC/plt level in acute stroke is due to a detectable, constitutive premorbid state of platelet activation. In sixteen consecutive patients attending a vascular surgery clinic, GC levels were measured on a citrated plasma sample, and corrected for circulating platelet count. Since 15 of 16 patients were taking aspirin when seen at clinic, a control study was undertaken to assess the effect of aspirin on sequential plasma GC/plt levels measured over 10 days--5 pre and post daily aspirin for 5 days, 4 acting as non-aspirinated controls. Plasma GC/plt levels in normal plasma were 21.6 +/- 8.0 fg; mean +/- SD. In the 16 patients the GC/plt levels were 13.1 fg/plt; SD 5.4, range 2.9-24.3. All platelet counts were in the normal range in all patients involved. While a masking effect due to aspirin cannot be completely ruled out, these studies indicate that plasma GC/plt level is not useful as a predictor of acute stroke in the premorbid population.

Objectives:To determine whether topical fibrin sealant reduced suture line bleeding during carotid endarterectomy with polytetrafluoroethylene (PTFE) patch closure.Design:Prospective randomised non-blinded control trial.Setting:Regional vascular surgery unit.Materials:Seventeen patients undergoing carotid endarterectomy were randomised either to receive fibrin sealant as a topical haemostatic agent at the arteriotomy suture line or to act as control.Outcome measures:Time taken to achieve haemostasis at the suture line. Intraoperative blood loss. Total operative time.Results:The median time to achieve haemostasis was 5.5 min (range 4–31 min) in the treatment group and 19 min (range 10–47 min) in the control group. This difference was statistically significant p < 0.005 by Mann-Whitney test. There was no statistical difference in total operative time. Operative blood loss was lower in the treatment group (median 420ml, range 300–500ml) than in the control group (median 550ml, range 350–1200ml) but this difference was not statistically significant. One patient in the control group suffered a perioperative thrombo-embolic event.Conclusion:Fibrin sealant is an effective topical haemostatic agent for arteriotomy suture lines involving PTFE material.

Jean-Pierre Allain and colleagues argue that, while unintended, the foreign aid provided for blood transfusion services in sub-Saharan Africa has resulted in serious negative outcomes, which requires reflection and rethinking.

Blood transfusion in rural sub-Saharan Africa presents special challenges. Transfusions are primarily given for emergencies--life-threatening blood loss or anemia; blood is usually collected from family or replacement donors; and facilities to store an adequate reserve in a hospital bank are constrained. We report the everyday and organizational practices in a medium-sized district hospital in Northern Ghana.Information and data on blood transfusion practices at West Gonja Hospital, Damongo, were available from the laboratory reports, from day books and workbooks, and from direct observation in the following four areas: blood collection and blood donors; blood donation testing; blood storage and logistics; and clinical transfusion practice, adverse events, and follow-up.The hospital serves a rural community of 86,000. In 2009, a total of 719 units of whole blood were collected, a rate of 8.36 units per 1000 population. All donors were family or replacement donors. Positivity rates for infectious disease markers were 7.5% (64/853) for hepatitis B surface antigen, 6.1% (50/819) for hepatitis C virus, 3.9% (33/846) for human immunodeficiency virus, and 4.7% (22/468) for syphilis. Supply of laboratory materials was sometimes problematic, especially for temperature-critical materials. Difficulties in sample labeling, storage of blood and laboratory supplies, and disposal of waste were also incurred by operational, material, and financial constraints. Follow-up for outcomes of transfusion is not currently feasible.The operational, demographic, and financial environment pertaining in a rural hospital in Northern Ghana differs substantially from that in which much of current blood transfusion practice and technology evolved. Considerable effort and innovation will be needed to address successfully the challenges posed.

A complex relationship exists between venous hemoglobin level and thrombosis risk. The hemoglobin content of capillary blood varies in healthy individuals in relation to that of venous blood with sex, age, season of the year, and venous hemoglobin level. (Murphy WG, et al. Blood 2010;116:2861-2). To study further the dynamics of this relationship, venous and capillary (finger pulp blood) hemoglobin levels were compared in 462 frequent blood donors with hereditary hemochromatosis. A linear mixed effects model was fitted to compare capillary hemoglobin with the difference between the venous and capillary hemoglobin. Random slopes and intercepts for each donor were modelled to account for inter-individual variability. A linear mixed effects model was fitted to hemoglobin levels over cumulative time since first donation for each person in separate analyses for capillary and venous hemoglobin levels. Random intercepts and random slopes were modelled within individuals. For each change of 1g/dL in capillary hemoglobin, the difference between venous and capillary hemoglobin changed by -0.694 g/dL, significantly different from zero (p After venesection capillary hemoglobin rose by a mean of 0.496 mg/dL/day, (p These data indicate that healthy individuals preserve venous hemoglobin at the expense of capillary hemoglobin as the red cell mass rises and falls, and that the capillary space buffers the venous hemoglobin level from changes in red cell mass. Since this space occupies > 10% of the intravascular volume (Chaplin H, et al. JCI 1953:32;1309-16) we estimate that 27% or more of a decrease or increase in red cell mass is accommodated by changes in hemoglobin level of capillary blood relative to the venous hemoglobin level. The mechanisms underlying these dynamics, such as NO-induced vasodilation by deoxyhemoglobin and modulation of the Fåhraeus effect, remain to be elucidated; however their implications for the relationship between hemoglobin level and thrombosis may be considerable.

The continuous production of large numbers of red blood cells (RBCs) ex vivo is a challenging task due to process economics and complex culture conditions. In any serial passaging process, the culture conditions and operation mode are important criteria for achieving high proliferation with optimal passage lengths. The optimal inoculation concentration for serial passaging is a factor that affects both the kinetics and the total expansion performance. As part of our attempt to develop a scalable, economical and reproducible system for production of RBCs we used mathematical expressions to define the growth curves of peripheral blood derived erythroid progenitors over the course of their expansion process. We used a Gompertz function to evaluate the specific growth rate for the optimisation of inoculation concentration and passage lengths to achieve optimal expansion. This led to values of 3 × 10 5 cells/ml as the optimum inoculation concentration and 36 h as the optimum passage length. Also the variations in growth curves confirmed the altered growth kinetics of erythroid progenitors during sequential passaging in expansion process. Cost analysis suggested a 60-h passage length at every passage, resulting in a 42.9% process-cost reduction. However, this has increased the process duration in achieving the similar expansion factor. This methodology for optimising the expansion process of peripheral derived erythroid progenitors based on optimum culture conditions could provide us with a direction and an improved performance for scale-up applications.

Between 1988 and 1993, 17 (3%) out of a total 654 patients underwent reoperation for control of haemorrhage following repair of abdominal aortic aneurysm in a vascular surgery unit. The first operation was performed for rupture in 12 cases and electively in five. The incidence of reoperation for postoperative bleeding was 1.7% following elective operation and 3.3% following emergency operation. Case-controls, matched for sex and primary operation, were identified. The mortality rate in those requiring reoperation was 58% compared with 23% in the control group (p = 0.037). Seven patients suffered progressive deterioration and died in the early postoperative period. Of the remaining ten patients, four suffered unexpected serious complications; two a fatal cerebro-vascular accident (CVA), one a fatal myocardial infarction (MI) and the fourth a non-fatal CVA. The patients requiring reoperation had greater blood loss (p0.05), greater transfusion requirements and lower core temperatures (p0.05) at the end of their first operation than the control group. All except one of the patients who bled had evidence of coagulopathy and had lower platelet counts than the control group both before and after the first operation. At reoperation there were multiple minor bleeding points in 11 patients, no active bleeding points in two patients and a discrete bleeding point in four patients. In conclusion, re-operation for control of postoperative haemorrhage is an uncommon complication which is strongly associated with coagulopathy, may predispose to "rebound" postoperative thrombotic episodes, and carries a poor prognosis.

Background and Objectives Variant Creutzfeldt-Jakob (vCJD) is a fatal transfusion transmissible prion infection. No test for vCJD in the donor population is currently available. Therefore, prion removal by filtration of red cell concentrate (RCC) is an attractive option for prevention. Materials and Methods Twenty patients were recruited with ethical permission, to receive clinically necessary transfusion containing one unit of pfRCC. Follow-up at 24 hours, 6 weeks and 6 months was undertaken. A second pfRCC was administered to 6 patients with similar follow up. pfRCC were prepared using the CE marked P-Capt device by the IBTS. Results In 20 transfused patients undergoing one exposure to a prion filtered unit, no attributable adverse events were noted. A subset of these (n = 6) underwent re-exposure to a further filtered unit without incident. Conclusions This phase 1/11 clinical study provides encouraging data on safety of prion filtration which can be used to plan more extensive studies on the use of filtered blood in adults and children.

Thrombotic thrombocytopenic purpura (TTP) is characterized by thrombocytopenia and disseminated platelet thrombi throughout the microvasculature. Studies by our group have demonstrated calcium- dependent proteolytic activity (calpain) that is no longer detectable in the serum of patients with acute TTP after their recovery. The purpose of this study was to investigate if the protease activity of TTP was detectable in plasma and, therefore, not an in vitro phenomenon secondary to the formation of serum. Additionally, we looked for evidence of membrane association of the active protease in the patients' samples, which would explain the persistence of its activity in the presence of plasma inhibitors. Acute TTP samples, both serum and plasma, were collected from 10 patients with TTP. Calpain was measured using bioassays for enzyme activity and also by detection of the protein using immunoblotting with an anticalpain monoclonal antibody (MoAb). In all instances, calpain could be detected both functionally and antigenically in the acute TTP sera and plasma. No calpain activity could be detected in any of the controls, although antigenic calpain was detectable in one sample from a patient who had undergone cardiopulmonary bypass surgery. To investigate whether the calpain was associated with microparticles in the plasma, the TTP plasma samples were ultrafiltered and ultracentrifuged. Activity was not lost by passage across a 0.2-micron filter but was detectable only in the pellet following ultracentrifugation. Membrane association of the calpain in the microparticles also was demonstrated using solubilization with Triton X-100. Immunoprecipitation studies demonstrated that the calpain activity could be removed by MoAbs against platelet membrane glycoproteins (IX and IIb/IIa) but not by a MoAb against red blood cell membrane glycophorin. These studies indicate that active calpain is associated with platelet microparticles in plasma from patients with TTP.

Background EU law requires a haemoglobin of ≥ 12·5 g/dl for women or ≥ 13·5 g/dl for men at the time of donation. As capillary and venous haemoglobin values may differ in the same subject, we examined whether a capillary haemoglobin level of 12·0 g/dl for women or 13·0 g/dl for men, is equivalent to a venous haemoglobin level of ≥ 12·5 g/dl and ≥ 13·5 g/dl, respectively, to avoid unnecessary loss of blood donations. Methods Over a continuous 42-month period, 36 258 paired capillary and venous samples were taken from 25 762 females and 10 496 males, when the capillary haemoglobin was

The potential of peripheral blood derived CD34+ cells for ex vivo erythropoiesis was investigated in a stroma-free culture system using a novel strategy of daily passaging. By expanding PB-derived CD34+ cells up to 1.5 x 10(6)-fold this method achieved expansion factors previously only reported for CD34+ cells derived from more potent stem cell sources such as cord blood, bone marrow and mobilized peripheral blood. Analysis of cell surface markers showed differentiation of immature CD34+ cells to populations with 80% CD71-/GpA+ cells and up to 45% enucleated cells, indicating a significant amount of terminal maturation. Cell crowdedness was found to have decisive effects on in vitro erythropoiesis. Cell density per surface area rather than cell concentration per media volume determined cell expansion during exponential growth where more crowded cells showed reduced overall expansion. In late stage erythropoiesis, however, when cells no longer proliferating, increased cell density was seen to enhance cell viability. These results indicate that peripheral blood derived haematopoietic stem cells can be an alternative to cells sourced from bone marrow, cord blood or leukapheresis in terms of expansion potential. This provides distinct advantages in terms of availability for studies of conditions for scale-up and maturation, and may have particular clinical applications in the future.

This preliminary study evaluated a brief intervention, based on the theory of planned behavior, designed to encourage high school students to practice testicular self-examination (TSE). Ninety-nine male ninth and 10th grade students were exposed to a message challenging beliefs about outcomes of performing TSE as suggested by the theory of planned behavior, a message providing information about TSE and testicular cancer, or a control message, after which they completed a questionnaire operationalizing components of the theoretical model. Four weeks later, students reported their practice of TSE. As predicted, students exposed to the theory-based message reported a stronger intention to perform TSE and, at follow-up, were more likely to report having performed the exam than study participants exposed to other messages. Data provide partial support for the utility of the theory of planned behavior as a framework for constructing health-promoting messages. Future research recommendations are offered.

Transfusion of blood and blood products has been associated with transmission of infectious agents. However, it is probable that blood products are currently very safe and that pooled virus-inactivated products from remunerated donors are now safer than untreated single voluntary donor components. Although the transmission events of the past and the present are reasonably well understood, reliance on a linear approach to predict safety in the future is open to criticism. Indeed, it was not possible to predict the extent or consequences of the AIDS epidemic or of hepatitis C transmission. Moreover, although variant Creutzfeldt-Jakob disease (vCJD) may not be transmitted to any large extent by transfusion of manufactured blood products, this will be due more to good fortune than good judgement – this agent could have escaped the screening, testing and eradication methods on which current confidence in blood product safety depends. Similarly, the emergence of a highly resistant non-enveloped virus, or even of another previously unrecognised disease-causing agent, could result in new threats from transfusion of blood components and products. The ecology of blood transfusion is exquisitely sensitive to variations in starting conditions, a situation typical of a chaotic rather than a linear system. Seemingly trivial events, often apparently unrelated to blood transfusion, have had enormous consequences in this field. Whatever the events that introduced simian immunodeficiency virus to humans or scrapie to cattle, they were a long way from those involved in the manufacture of blood products. In such a setting, reliance on methods that deal effectively with known threats (such as encapsulated viruses and bacteria) without adequate investigation and management of the intrinsic sensitivity to unpredictable events, leaves open the possibility of further infections emerging in the future. It is this reality that will ultimately result in the eradication of the transfusion of donor-derived blood and blood products in the developed world. In addition, all infections with a long disease-free incubation period in the host that can be transmitted in blood will eventually be over-expressed in groups that are exposed to blood either recreationally or professionally. As in the past, this could have occurred before testing or decontamination processes have been developed for emerging pathogens. Failure to be able to rely on completely risk-free donors, in both the voluntary and non-voluntary sides of the blood industry, continues to offer the potential for the transmission of infectious diseases in the future.

BACKGROUND: Most bacteria present in blood components are normal skin flora, particularly Staphylococcus epidermidis and other coagulase-negative staphylococci. Growth patterns of these bacteria and the effects of different methods of component preparation may depend on variations in behavior between different isolates of the same species. STUDY DESIGN AND METHODS: Whole-blood units were inoculated with 19 different coagulase-negative staphylococcus (CNS) isolates at 1 to 10 and 10 to 100 CFUs per mL. After overnight holding at 22°C, the units were processed into components. The components were cultured before inoculation and during processing, including before and after WBC reduction. RESULTS: At low inoculum levels, CNS was detected in 15 (79%) of 19 whole-blood units and in 12 (63%) of 19 RBCs after separation; after filtration, bacteria were detected in 3 (16%) of 19 (p = 0.0069). For platelet concentrates, 6 (32%) of 19 grew bacteria before filtration and 1 of 18 after filtration (difference not statistically significant). Three (16%) of 19 plasmas were positive before and after freezing. At high inoculum levels, 16 (89%) of 18 whole-blood samples and RBCs were positive before filtration; 6 (33%) of 18 RBCs were positive after filtration (p = 0.0002); 8 (44%) of 18 platelets were positive before filtration; 3 (17%) of 18 were positive after filtration (difference not statistically significant), and 7 (37%) of 18 plasma samples were positive before and after freezing. CONCLUSION: The growth characteristics of CNS in blood components vary with differences either in the subtype of bacteria or in the donor blood. Filtration reduces but does not eradicate contamination of RBCs and platelets by CNS. Plasma may act as a reservoir for CNS infection.

To the editor: Mean red cell mass and hematocrit levels are higher in men than women. How or why this is the case has never been explained. If decreased erythropoiesis due to iron loss or blood loss in females or increased erythropoiesis due to androgens in males were causative, there would be a