A total of 2050 time-expired, 176 fresh, and 400 initial-reactive PLT packs were tested by real-time PCR using broadly reactive 16S primers and a “universal” probe (TaqMan, Invitrogen). PLTs were also tested using a microbial detection system (BacT/ALERT, bioMérieux) under aerobic and anaerobic conditions.

Seven of 2050 (0.34%) time-expired PLTs were found repeat reactive by PCR on the initial nucleic acid extract but none of these was confirmed positive on testing frozen second aliquots. BacT/ALERT testing also failed to confirm any time-expired PLTs positive on repeat testing, although 0.24% were reactive on the first test. Three of the 400 “initial-reactive” PLT packs were found by both PCR and BacT/ALERT to be contaminated (Escherichia coli, Listeria monocytogenes, and Streptococcus vestibularis identified) and 14 additional packs were confirmed positive by BacT/ALERT only. In 13 of these cases the contaminating organisms were identified as anaerobic skin or oral commensals and the remaining pack was contaminated with Streptococcus pneumoniae.

These results demonstrate that the 16S PCR assay is less sensitive than BacT/ALERT and inappropriate for early testing of concentrates. However, rapid PCR assays such as this may be suitable for a strategy of late or prerelease testing.

An immunoglobulin fraction containing IgG, IgM, and IgA was prepared by caprylic acid precipitation of cryoprecipitate-poor plasma. The capacities of the cation exchangers (S HyperCel and CM Ceramic HyperD F) and anion exchangers (HyperCel STAR AX and Q HyperCel) to remove IgA, IgM, and spiked FXI were tested following a design of experiment approach using microplates and chromatographic column scale-up. FXI removal was also evaluated using Mustang S chromatographic membranes. IgG/IgG subclasses, IgA, IgM, and FXI were assessed by enzyme-linked immunosorbent assay, and caprylic acid, by gas chromatography.

Extensive removal of IgA and IgM, but not FXI, was achieved by a two-step chromatographic process combining S HyperCel used in the IgG binding and elution mode and HyperCel STAR AX used in the IgG flow-through mode, providing high IgG and IgG subclass recovery (>85%), high purity (>99.5%), and efficient removal of IgA (<0.5%) and IgM (undetectable). Twenty-six-fold FXI removal was achieved by processing the resulting purified IgG fraction through Mustang S cation-exchanger membranes at pH 6.0 and 12.7 mS/cm. Caprylic acid was removed by S HyperCel.

Combining S HyperCel and HyperCel STAR AX extensively removed IgA and IgM, with good IgG recovery. Mustang® S membranes can be used for dedicated removal of FXI.

A 21-year-old man with sickle cell disease and multiple prior transfusions received two phenotype-matched, compatible RBC units during a brief admission for pain crisis. He developed rapid-onset progressive anemia and hemoglobinuria. Methylprednisolone, erythropoietin, and rituximab were administered. Fifteen days posttransfusion the hemoglobin (Hb) concentration decreased to 3.1 g/dL, with evidence of severe congestive heart failure. No new antibodies were identified. It was felt that his heart failure would not improve without increasing oxygen-carrying capacity. A combination of volume overload, anemia, and hemolysis prompted a novel isovolemic procedure to increase Hb level without removing his own RBCs or causing fluid overload. A cell separator was used operating on the plasma-exchange program, with three cross-match–compatible, washed RBC units as the replacement fluid. After the procedure, there was no evidence of hemolysis. Over the following 6 days, the congestive heart failure resolved, the Hb concentration increased to 7.5 g/dL, and the patient fully recovered. He had a similar event 3 years previously.

Plasma-to-RBC replacement may be beneficial for selected patients with life-threatening anemia. This intervention provides immediate improvement in oxygen-carrying capacity, conserving the patient's own RBCs, while avoiding fluid overload. Although blood transfusion may precipitate further hemolysis, this case report describes successful plasma-to-RBC exchange transfusion with concurrent supportive care to offset hemolysis, including corticosteroid, intravenous immunoglobulin, and rituximab.

A total of 105 CB units were thawed and washed at different time intervals (6, 12, 24, and 36 months). For every thawed CB unit, samples were removed and cell enumeration (total nucleated cells [TNCs], mononuclear cells [MNCs], CD34+, CD133+) was performed. In addition, viability was obtained with the use of flow cytometry, and recoveries were calculated. Also, total absolute colony-forming unit counts were performed and progenitor cell recoveries were studied by clonogenic assays.

Significant differences (p < 0.05) were observed in certain variables (TNCs, MNC numbers, viability) when they were examined in relation with time intervals, while others (CD34+, CD133+) were relatively insensitive (p = NS) to the duration of time interval the CB units were kept in cryostorage condition.

The data presented suggest that cryopreservation of CB units in a mechanical freezer at −150°C may represent an alternative cryostorage condition for CB cryopreservation.

Thirty-six CSP units were thawed, refrigerated, and sampled aseptically at 0, 24, 48, and 120 hours postthaw. Clotting factor activities (Factor [F]V, FVII, FVIII, and fibrinogen) and prothrombin time were measured using an automated coagulation analyzer, and von Willebrand factor (vWF) and ADAMTS13 activities using enzyme-linked immunosorbent assay.

Fibrinogen, FVIII, and vWF activities were unchanged from thaw values after 120 hours of storage; ADAMTS13, FV, and FVII activities were significantly lower than at thaw, but mean reductions were only −2.6, −7.7, and −12%, respectively. Losses were proportionately greater in the first 24 hours of refrigerated storage.

Extending the refrigerated storage of CSP from 1 to 5 days had little impact on product quality. The retention of more than 97% of initial mean ADAMTS13 activity after 5 days of refrigerated storage suggests that the shelf life of thawed refrigerated CSP could be extended without meaningful losses of its likely most important ingredients. CSP postthaw storage could be aligned to that of refrigerated thawed frozen plasma, currently available for transfusion in some jurisdictions for up to 5 days postthaw.

A retrospective cohort study was performed. Age, comorbidities, hemoglobin (Hb), the potential for blood loss, and units of red blood cell (RBC) transfusions were recorded. Preoperative transfusions were evaluated to determine whether they met criteria established by the Mozambican Ministry of Health as well as proposed guidelines based on more restrictive protocols. Avoidable blood transfusions were defined as those preoperative transfusions that were not indicated based on these guidelines. Multivariate logistic regression was used to identify factors that predicted transfusion.

A total of 205 patients (age range, 0.1-86 years) underwent surgery in the main operating room during the 2-week study period. Overall, 35 (17%) patients received 68 transfusions. Of these, 36 transfusions were given preoperatively and 32 were given intraoperatively. Thirty-six percent of preoperative transfusions were avoidable according to national guidelines. Ninety-two percent were avoidable using more restrictive guidelines. The primary predictors of preoperative blood transfusion were lower Hb (odds ratio [OR], 0.390/1 g/dL; p < 0.0001) and the potential for blood loss (OR, 3.73; p = 0.0410).

Adherence to existing Hb thresholds recommended by national blood transfusion guidelines could significantly reduce the number of transfusions and the association risk of transfusion-transmissible infections. Adoption of more restrictive guidelines is recommended to further improve blood transfusion utilization and further reduce the transmission risk of human immunodeficiency virus and hepatitis.

A Markov-based model was constructed to compare the health and financial implications of four alternative antigen-matching strategies for chronically transfused sickle cell patients. The strategies varied by the group of patients receiving matched blood (all patients prophylactically or only patients with a history of alloimmunization [history-based]), and by the extent of antigen matching (limited to C, E, and K, or extended to 11 antigens). Direct medical costs and alloimmunization events were assessed over 10- and 20-year periods, for a hypothetical cohort of initially transfusion-naive patients and for a dynamic population.

Within a hypothetical cohort of initially transfusion-naive patients, implementing prophylactic limited matching for all chronically transfused patients instead of history-based limited matching is expected to cost an additional $765.56 million over 10 years, but result in 2072 fewer alloimmunization events. Within the same cohort, implementing prospective extensive matching is expected to cost $1.86 billion more than history-based extensive matching, but result in 2424 fewer alloimmunization events. Averting a single alloimmunization event using prospective matching would cost $369,482 to $769,284. Among a dynamic population over 10 years, prospective limited matching is expected to cost $358.34 million more than history-based limited matching.

While prospective matching for all transfused patients would reduce alloimmunization, this strategy requires considerable expenditure.

AABB's Biovigilance Tissue Working Group conducted a Web-based survey, distributed to 1069 hospital institutional members in June 2011. Human tissue types used, departmental responsibilities, and views of AABB involvement were queried.

Of the 336 (31%) total respondents, 84% use allogeneic and/or autologous human tissue. Sixty-one percent have stored tissue on consignment. As in 2005, the department of surgery most often had responsibility for tissue use, followed by the blood bank or transfusion service (BBTS). Overall, the BBTS had a smaller role in oversight of autologous tissue acquisition in 2011 versus 2005, but no change in level of responsibility for storage or issue of tissues. Hospitals reported the BBTS and combined blood and tissue services (CBTS) added responsibilities for storing and monitoring eye tissue and heart valves (p < 0.05) since 2005. The BBTS/CBTS increased their degree of responsibility for reporting suspected postimplant infection and other adverse reactions for musculoskeletal allografts (p < 0.01), eye tissue (p < 0.005), and eye tissue recipients recall notification (p < 0.05). The BBTS/CBTS have more responsibility than any other department for stem cell and cord blood management.

In this survey, AABB institutional members reported that BBTS are more involved than previously in the regulatory aspects of human tissue oversight and remain involved in many operational aspects of hospital tissue management.

Errors were prospectively identified from 2005 to 2010. Error data were coded on a secure online database called the Transfusion Error Surveillance System. Errors were defined as any deviation from established standard operating procedures. Errors were identified by clinical and laboratory staff. Denominator data for volume of activity were used to calculate rates.

A total of 15,134 errors were reported with a median number of 215 errors per month (range, 85-334). Overall, 9083 (60%) errors occurred on the transfusion service and 6051 (40%) on the clinical services. In total, 23 errors resulted in patient harm: 21 of these errors occurred on the clinical services and two in the transfusion service. Of the 23 harm events, 21 involved inappropriate use of blood. Errors with no harm were 657 times more common than events that caused harm. The most common high-severity clinical errors were sample labeling (37.5%) and inappropriate ordering of blood (28.8%). The most common high-severity error in the transfusion service was sample accepted despite not meeting acceptance criteria (18.3%). The cost of product and component loss due to errors was $593,337.

Errors occurred at every point in the transfusion process, with the greatest potential risk of patient harm resulting from inappropriate ordering of blood products and errors in sample labeling.

We hypothesized that after we established a neonatal intensive care unit (NICU) transfusion management program in 2009, a decrease in early (first week after birth) RBC transfusion rate and a decrease in the incidence of severe IVH occurred concomitantly.

During a 9-year period 2716 VLBW neonates were admitted to our NICUs. In 2004, 58% of VLBW neonates received one or more RBC transfusions during the first week. After a transfusion compliance program was established in 2009, this rate declined, reaching 25% by 2012. In parallel, the severe IVH rate also declined, from 17% in 2004 to 8% in 2012 (R² = 0.73). IVH occurred in 27% of those who received a RBC transfusion during the first week versus less than 2% of those with no early transfusion (p < 0.001). The decrease in IVH rate occurred exclusively among neonates born in an Intermountain Healthcare perinatal center and not among those initially cared for in an “outside” hospital and subsequently transported to an Intermountain NICU.

It remains unclear whether transfusing VLBW neonates during the first days after birth is a proximate cause of IVH. However, the present report is consistent with previous studies showing that successful efforts to reduce early RBC transfusions is associated with a decrease in the incidence of severe IVH.

All hospitals in the Canadian province of Ontario with transfusion medicine services were invited to participate in a 5-day audit of FP utilization. FP dose, indication, and clinical patient data were collected for each transfusion request. Indications for FP transfusions were independently adjudicated as appropriate, inappropriate, or indeterminate based on predefined criteria.

Seventy-six (49%) of 155 invited hospitals participated in the audit, which included 573 requests for 2012 units of FP. A total of 559 transfusions (1909 units) were administered. Of 573 requests, 164 (28.6%) were deemed inappropriate most often because: 1) they were administered to patients with an international normalized ratio below 1.5 or 2) they were administered in absence of bleeding or emergency surgery. The most frequent indications for FP transfusions were before surgery and warfarin reversal. Overall, patients admitted to the clinical areas of surgery, internal medicine, and the emergency department represented the largest users of FP, but this varied by hospital type (community vs. academic). The most frequently requested doses of FP were 2 and 4 units.

This point-prevalence hospital audit revealed that transfusion of FP is frequently inappropriate. Focusing on reducing the two most common reasons for inappropriate FP transfusions could lead to a significant improvement in FP utilization.

Human PLTs (hPLTs) in plasma with or without single or multiple Mirasol PRT treatments were assessed in vitro by aggregation and percentage of P-selectin expression. In vivo studies included PLT recovery in SCID mice and assessment of ALI in a two-event mouse model in which the sensitizing event was lipopolysaccharide injection and the second event was infusion of Mirasol-treated hPLTs.

A single-dose Mirasol treatment (5 J/cm²) did not induce any change in aggregation in response to adenosine 5′-diphosphate (ADP) while a five-times-repeat Mirasol treatment (5×) increased aggregation response to low concentration of ADP. Mirasol PLTs (1×-5×) had increased percentage of P-selectin–positive PLTs after treatment and decreased aggregation with TRAP as the agonist. In vivo recovery in SCID mice was reduced extensively with Mirasol treatments (1× and 5×). In the two-event model of ALI, only the 5× Mirasol PLTs accumulated in the lung and this was not accompanied by changes in lung histology or increases in MIP-2 levels in bronchoalveolar lavage fluid.

Mirasol PRT treatment induced PLT activation and reduced in vivo recovery in a SCID mouse model of transfusion. In our two-event mouse model of ALI, the 5× Mirasol hPLTs accumulated in the lung, but did not cause signs of ALI. The 1× Mirasol treatment did not lead to PLT lung accumulation or ALI in this model.

We performed an extensive search for clinical ABC studies, focusing on acute normovolemic hemodilution (ANH), intraoperative blood salvage (IBS), or preoperative autologous deposit (PAD). Only ABC studies providing a minimum set of clinical variables were included. Using a clinically validated mathematical model, we then calculated maximal allowable blood loss (efficacy) and increase in red blood cell (RBC) mass (effectiveness) to rank the three techniques.

We identified 21 clinical ABC studies, including 3926 patients, as suitable for our model. Our model shows that IBS with high recovery rates is the most efficacious and effective ABC measure. PAD will reveal nearly similar efficacy and effectiveness, only if sufficient time for RBC regeneration has passed and if 4 PAD units or more are available. Our model further demonstrates that ANH as well as IBS with low recovery rates are the least efficacious and effective alternatives.

IBS appears to be the most efficacious and effective ABC measure. PAD can only reduce the need for allogeneic blood transfusions under certain circumstances. ANH does not appear to play an important role in ABC.

A retrospective analysis was conducted into the percentage of outdated PLT components during the 3 years before and after the adoption of PLT PI technology in our institution. The PLT transfusion dose for both pre-PI and post-PI periods was similar. A retrospective analysis to study clinical safety and component utilization was also performed in the Balearic Islands University Hospital.

As a result of PI implementation in our institution, the PLT production cost increased by 85.5%. However, due to the extension of PLT storage time, the percentage of outdated PLT units substantially decreased (−83.9%) and, consequently, the cost associated with outdated units (−69.8%). This decrease represented a 13.7% reduction of the initial cost increase which, together with the saving in blood transportation (0.1%), led to a saving of 13.8% over the initial cost. Therefore, the initial 85.5% increase in the cost of PLT production was markedly reduced to 71.7%. The mean number of PLT concentrates per patient was similar during both periods.

The extension of PLT storage time can substantially contribute to reducing the financial impact of PI by decreasing the percentage of outdated PLTs while improving blood safety. Since the adoption of PI, there have been no documented cases of PLT transfusion–related sepsis in our region.

Blood was inoculated with 10⁴ or 10⁵ parasites/mL and riboflavin treated with or without ultraviolet (UV) irradiation (40-160 J/mL red blood cells [mL_RBCs]). Parasite genome integrity was assessed by quantitative amplification inhibition assays, and P. falciparum viability was monitored in vitro.

Riboflavin alone did not affect parasite genome integrity or parasite viability. Application of UV after riboflavin treatment disrupted parasite genome integrity, reducing polymerase-dependent amplification by up to 2 logs (99%). At 80 J/mL_RBCs, riboflavin plus irradiation prevented recovery of viable parasites in vitro for 2 weeks, whereas untreated controls typically recovered to approximately 2% parasitemia after 4 days of in vitro culture. Exposure of blood to 160 J/mL_RBCs was not associated with significant hemolysis.

Riboflavin plus irradiation treatment of whole blood damages parasite genomes and drastically reduces P. falciparum viability in vitro. In the absence of suitable malaria screening assays, parasite inactivation should be investigated for prevention of transfusion-transmitted malaria in highly endemic areas.

Plasma pools collected from 31,200 consecutive Polish donors typed as D– were tested by real-time polymerase chain reaction (PCR) for the presence of RHD-specific markers located in Intron 4 and Exons 7 and 10. RHD+ individuals were characterized by PCR or cDNA sequencing and serology.

Plasma cross-pool strategy revealed 63 RHD+ donors harboring RHD*01N.03 (n = 17), RHD*15 (n = 12), RHD*11 (n = 7), RHD*DEL8 (n = 3), RHD*01W.2 (n = 3), RHD-CE(10) (n = 3), RHD*01W.3, RHD*01W.9, RHD*01N.05, RHD*01N.07, RHD*01N.23, and RHD(IVS1-29G>C) and two novel alleles, RHD*(767C>G) (n = 3) and RHD*(1029C>A). Among 47 cases available for serology, 27 were shown to express the D antigen

1) Plasma cross-pool strategy is a reliable and cost-effective tool for RHD screening. 2) Only 0.2% of D– Polish donors carry some fragments of the RHD gene; all of them were C or E+. 3) Almost 60% of the detected RHD alleles may be potentially immunogenic when transfused to a D– recipient.

In a regional health care system consisting of 11 hospitals using a common CPOE, data on the number of plasma orders and alerts that were generated were collected over a 4-month period before prescribers were required to select an indication for plasma. When adaptive ordering was implemented prescribers had to choose from prepopulated indications for plasma: INR of 1.6 or greater with bleeding, INR of 1.6 or greater before an invasive procedure, therapeutic exchange, massive transfusion, and other. Regardless of the antecedent INR the alert did not trigger if massive transfusion or plasmapheresis was selected. Information on prescribers and recipients was collected during this 5-month period.

In the 4-month period before the adaptive alerts were implemented, 42.9% of the plasma orders generated an alert; in the 5-month period thereafter the alert rate was significantly lower at 27.9% (p < 0.0001). The percentage of heeded alerts increased during the adaptive alert period (24.3% vs. 17.1%, respectively, p = 0.004). A significant percentage (45%) of other plasma orders were for periprocedure or bleeding patients whose antecedent INR was less than 1.6. There were significant differences in prescriber specialties among those who ordered plasma using the other indication compared to all plasma orders.

Electronic interventions improve compliance with plasma guidelines but as implemented are not sufficient to completely curtail non–evidence-based ordering.

In the study we 1) compared the effect of two-step cryopreservation and controlled-rate freezing method on the postthaw quality of CB (Study A) and 2) evaluated the postthaw quality of HSC fractions isolated from CB with various methods and frozen with controlled-rate freezing method (Study B). The same cryoprotectant mixture was used for 20 CB units (Study A) and 122 CB units (Study B).

In Study A, 13.79 × 10⁸ and 13.29 × 10⁸ initial white blood cell (WBC) counts decreased to 6.38 × 10⁸ and 6.02 × 10⁸ after thaw for the two methods, respectively. The mononuclear cell (MNC) counts decreased from 5.90 × 10⁸ to 3.71 × 10⁸ and from 5.64 × 10⁸ to 3.47 × 10⁸ dependent on the method. MNC viability decreased from 99.0% to 97.4% for the former and from 98.5% to 97.2% for the latter method. The differences were insignificant. In Study B, postthaw WBC recovery in HSC fractions was 74.4% to 103.5%, MNC recovery 106.4% to 118.5%, CD34+ cell recovery 102.5% to 150.2%, and MNC viability 94.1% to 97.4%.

Neither the cryopreservation procedure nor the freezing of isolated HSCs affected product quality, which may indicate that various freezing methods can be used for cell banking provided the they follow recommendations of good manufacturing practice and Directive 2004/33/EC.

We recently generated transgenic mice with RBC-specific expression of the human KEL glycoprotein, specifically the KEL2 or KEL1 antigens. Herein, we investigate recipient alloimmune responses to transfused RBCs in this system.

Transfusion of RBCs from KEL2 donors into wild-type recipients (lacking the human KEL protein but expressing the murine KEL ortholog) resulted in dose-dependent anti-KEL glycoprotein immunoglobulin (Ig)M and IgG antibody responses, enhanced by recipient inflammation with poly(I:C). Boostable responses were evident upon repeat transfusion, with morbid-appearing alloimmunized recipients experiencing rapid clearance of transfused KEL2 but not control RBCs. Although KEL1 RBCs were also immunogenic after transfusion into wild-type recipients, transfusion of KEL1 RBCs into KEL2 recipients or vice versa failed to lead to detectable anti-KEL1 or anti-KEL2 responses.

This murine model, with reproducible and clinically significant KEL glycoprotein alloantibody responses, provides a platform for future mechanistic studies of RBC alloantibody induction and consequences. Long-term translational goals of these studies include improving transfusion safety for at-risk patients.

System qualification, culture media quality verification, and validation of CB processing by-product (CB-BP) sample as surrogate to final product for sterility testing were followed by studies evaluating method sensitivity, specificity, reproducibility, ruggedness or robustness, and bacteriostatic or fungistatic effect of CB-BP sample. CB-BP cultures and control samples were formulated using BACTEC Plus Aerobic/F, Plus Anaerobic/F, and Myco F/Lytic media. Samples were seeded with selected test organisms (n = 13 at 10-100 colony-forming units [CFUs] per vial) and cultured for 14 days (bacterial) and 30 days (fungal).

Under testing conditions, no stasis effect of test sample on microbial growth and no false-positive or false-negative results were reported. Although a 7-day culture was sufficient to detect all validation test organisms seeded at ≤26 CFUs/vial, growth in actual product sterility testing practice may require a 10- to 14-day culture. Assay reproducibility was uncertain at very low bioburden (<10 CFUs/vial). Growth time to detection neither varied between different media lots nor prolonged in culture vials with loading delay (6-8 hr at room temperature).

BACTEC-FX culture and detection system and BACTEC media formulae have high detection capability and can be effectively validated for sterility testing of CB products.

From one institution, blood utilization data for 134,456 patients, 23,559 of whom were transfused with RBCs, were analyzed. Hb triggers and targets for transfused patients were plotted and graphically compared to the trigger and target ranges from previously published randomized clinical trials.

Nine hospital services with the highest transfusion rates were selected for analysis. The service with the highest Hb trigger and target was further analyzed by comparing transfusion thresholds for individual providers. Differences among services and among individual providers for mean Hb transfusion triggers and targets were significant (up to 1.5 g/dL, p < 0.0001). The variation between the 10th and 90th percentiles for both trigger and target was also significant (up to 3 g/dL, p < 0.0001). If a restrictive transfusion strategy were implemented, the need for transfusion would be reduced or eliminated in 10% to 50% of patients, depending on the service and the individual provider.

By using these methods for analyzing and presenting RBC utilization data, opportunities can be identified for blood conservation, and educational efforts can be directed toward the appropriate individual hospital services and providers.

Plasma samples of 107 hepatitis B surface antigen (HBsAg)-negative, HBV ID-NAT (Ultrio) positive-yield samples and 29 HBV DNA–negative, HBsAg-positive samples were used for comparison of NAT options in replicate testing of dilutions. Viral loads and relative sensitivities were determined by probit analysis against the Eurohep standard.

Ultrio Plus detected a significantly (p < 0.00001) higher proportion of replicate assays on HBV NAT yields (77%) than Ultrio ID (62%) and TaqScreen MP6 (47%), whereas Ultrio Plus MP4 and MP8 detected 53 and 41%, respectively. On HBsAg-yield samples missed by Ultrio screening, the reactivity rate increased significantly (p < 0.0001) from 23% in Ultrio to 65% in Ultrio Plus and further to 72% (p = 0.10) in the TaqScreen assay. The overall improvement factor of the analytical sensitivity offered by the target enhancer reagent in the Ultrio Plus assay was 2.5 (2.0-3.1)-fold on the Ultrio yield samples, but 43 (11-350)-fold on the HBsAg yields. In ID-NAT format the analytical sensitivity of TaqScreen relative to Ultrio Plus was 2.0 (1.0-4.2), 0.9 (0.7-1.3), and 1.6 (0.9-3.0) on the Eurohep standard, HBV NAT–, and HBsAg-yield samples respectively.

The clinical sensitivity of the currently available commercial NAT methods is mainly driven by the pool size.

This study is a single case report of a non-HSCT candidate who developed skin aGVHD with mild clinical course after decitabine and G-PBSCs combination. The clinical course, chimerism, and aGVHD pathology studies are detailed.

Compared with conventional aGVHD in allo-HSCT recipients and TA-aGVHD, the presentation of this case followed a self-limited clinical course without marrow aplasia or severe progression. However, the patient eventually died of leukemia without a significant graft-versus-leukemia effect.

First, the results demonstrate the existence of aGVHD in elderly non-HSCT candidates receiving adoptive cellular immune therapy. Second, aGVHD occurring under these conditions is probably a unique entity of aGVHD compared to TA-aGVHD and the conventional pattern in allo-HSCT recipients with respect to clinical course and prognosis.

A total of 1659 male and female donors from the Retrovirus Epidemiology Donor Study-II (REDS-II) Donor Iron Status Evaluation (RISE) study who were either first-time/reactivated (FT/RA; no donations for 2 years) or frequent donors were recruited into a longitudinal study of regular donation of RBCs. Of these, 1002 donors returned 15 to 24 months later for a final assessment. Absent iron stores (AIS) was defined as plasma ferritin level of less than 12 μg/L. Logarithm of the ratio of soluble transferrin receptor to ferritin of at least 2.07 (≥97.5% in FT/RA males) was used to define iron-deficient erythropoiesis (IDE). Receiver operating characteristics analysis was performed to assess selected RBC indices (e.g., percentage of hypochromic mature RBCs, proportion of hypochromic mature RBCs [HYPOm], and hemoglobin [Hb] content of reticulocytes [CHr]) in identifying AIS and IDE.

HYPOm and CHr detected IDE with comparable sensitivity, 72% versus 69%, but differed in specificity: HYPOm 68% and CHr 53%. For detecting AIS, sensitivity was improved to 85% for HYPOm and 81% for CHr but specificity was reduced for both. Venous Hb had high specificity but poor sensitivity for IDE and AIS. A plasma ferritin level of less than 26.7 μg/L was a good surrogate for assessing IDE.

RBC indices correlate with AIS and IDE and are more informative than Hb measurement, but lack sufficient sensitivity and specificity to be used as diagnostic tools in blood donors at risk for iron deficiency.

Ninety-eight adult patients undergoing primary THA or TKA were randomly assigned to receive an intraoperative intravenous loading dose of 1.0 g of TXA followed by another 1.0-g dose 3 hours later (TXA group) or a matching volume 0.9% saline placebo (control group). A postoperatively shed autologous blood recovery system was used in all patients and the minimum reinfusion volume set at 250 mL. Red blood cells were transfused if hemoglobin level was less than 8 or if 8 to 10 g/dL with symptoms of anemia.

The proportion of patients receiving autologous blood reinfusion was significantly lower in the TXA group (5/49) compared to placebo (42/49) with an absolute difference of −75.5% (adjusted relative risk, 0.005), and none of the patients in the TXA group received more than 400 mL retransfused. Median total external blood loss during the first 24 hours was lower in the TXA group, 320 mL (range, 80-930 mL), compared to 970 mL (range, 100-2600 mL) in the placebo group (p < 0.001). There were no significant differences in homologous blood transfusions and hematologic variables between groups. Treatment differences were consistent by size and significance when the analysis was repeated separately in patients undergoing TKA or THA.

Addition of TXA to a restrictive transfusion protocol makes the use of a postoperative blood salvage system in patients undergoing primary hip and knee arthroplasty unnecessary.

The rate of donors with unreported risk was estimated by a telephone survey of 2677 donors who answered “no” to risk questions. The number of T. cruzi antibody–positive donors missed by risk questions was estimated from the product of this rate and the selective testing T. cruzi antibody–positive rate. The 95% confidence interval (CI) was estimated by Monte Carlo simulation. To test the model, 60,132 donors were tested for T. cruzi antibody (26% of donors in selected regions, Phase I). In Winnipeg, Manitoba, the highest-risk region, 26,915 donors were tested (92.5% of donors, Phase II).

In the telephone survey, 21 (0.8%) donors reported risk factors that would have identified them for selective testing. Seven were born in Mexico or Central or South America, five had travel risk, and nine had mother or maternal grandmother risk. The 95% CI for predicted number of T. cruzi antibody–positive donors answering “no” to risk questions was 0.71 to 4.38. In Phase I, one Winnipeg donor confirmed positive but had answered risk questions correctly. No other positive donations were identified.

The estimated risk of T. cruzi–positive donors who answer “no” to risk questions is low and is confirmed by the seroprevalence among these donors.

Four areas were explored: circumstances of visit to hospital, external pressure, experience of donating, and potential repeat donation. After donation and consent, research assistants administered 25 questions and, according to literacy, helped with translation and completion.

Of 513 FDs, three-fourths were males (median age, 27 years). Only 1.3% were unemployed and more than 50% were students or teachers. Ties with hospitalized patient were family (76%), friends (13%), colleagues, or sharing place of worship (10%). Donating blood was the reason for visiting in 16.8% and 20.9% had previously donated blood probably as FDs. In one-third of FDs, the family asked for donation of which 10% was pressured by the unjustified reason that not donating was endangering the patient's life. For two-thirds of FDs, donation was given “because individuals were asked.” Donation was a positive experience for 77% of donors, 62% being interested in predonation testing. Repeating donation was acceptable for 99% of 79% FDs answering.

FDs are active in the population, are willing to donate blood if asked, are submitted to little pressure, do not receive incentives, and accept repeat donation. Except for circumstances of donation, FDs are not different from VNRDs and more directly motivated. They constitute a legitimate and important source to improve the blood supply in SSA.

This was a prospective study of adult trauma patients admitted to a Level I trauma center. Demography, Injury Severity Score (ISS), transfusion therapy, and mortality were registered. Hemostatic resuscitation was based on a massive transfusion protocol encompassing transfusion packages and thromboelastography (TEG)-guided therapy.

A total of 182 patients were included (75% males, median age 43 years, ISS of 17, 92% with blunt trauma). Overall 28-day mortality was 12% with causes of death being exsanguinations (14%), traumatic brain injury (72%, two-thirds expiring within 24 hr), and other (14%). One-fourth, 16 and 15% of the patients, received red blood cells (RBCs), plasma, or platelets (PLTs) within 2 hours from admission and 68, 71, and 75%, respectively, of patients transfused within 24 hours received the respective blood products within the first 2 hours. In patients transfused within 24 hours, the median number of blood products at 2 hours was 5 units of RBCs, 5 units of plasma, and 2 units of PLT concentrates. Nonsurvivors had lower clot strength by kaolin-activated TEG and TEG functional fibrinogen and lower kaolin–tissue factor-activated TEG α-angle and lysis after 30 minutes compared to survivors. None of the TEG variables were independent predictors of massive transfusion or mortality.

Three-fourths of the patients transfused with plasma or PLTs within 24 hours received these in the first 2 hours. Hemorrhage caused 14% of the deaths. We introduced transfusion packages and early TEG-directed hemostatic resuscitation at our hospital 10 years ago and this may have contributed to reducing hemorrhagic trauma deaths.

All donations were screened by US-required serologic HBV tests and for HBV DNA by MP-NAT (Novartis/Gen-Probe). Further testing by individual-donation polymerase chain reaction (ID-PCR) confirmed various classes of MP-NAT–reactive or –nonreactive donations. The hepatitis B surface antigen (HBsAg)-yield method was used to calculate incidence and the incidence–window-period model used to define residual risk.

Of approximately 12.8 million donations screened during 2009 to 2011, a total of 1368 HBV confirmed positives including 941 by MP-NAT were observed (combined 4.32% positive predictive value) of which five were seronegative NAT-yield donations (1:2.6 million) and 25 HBsAg-yield (anti-HBc–nonreactive) donations from which an incidence of 1.62/100,000 person-years (vs. 3.43 during 2006-2008) and residual risk of 1:592,000 to 1:754,000 were calculated. With the addition of MP-NAT, and resulting 8.8-day window-period reduction, residual risks decreased to 1:765,000 to 1:1,006,000. Of the 1368 positives, 99.6% were detected by serology and 68.8% by MP-NAT; ID-PCR detected 427 more infected donors than MP-NAT.

HBV MP-NAT and decreases in HBV incidence (likely vaccine-related) in the United States have reduced residual risks to levels comparable to those of human immunodeficiency virus and hepatitis C virus and raise the question of the continued need for all three HBV markers for blood donation screening. Further reductions in residual risk will require the implementation of more sensitive HBV-NAT methods including ID-NAT.

Prospectively collected data from 45,031 Irish whole blood donors were used. Hb cutoff levels for donation were approximately 0.35 mmol/L lower in Ireland than the Dutch cutoff levels (8.07 mmol/L vs. 8.40 mmol/L in men; 7.45 mmol/L vs. 7.80 mmol/L in women). The predictive performance of the models was assessed with calibration plots, calibration-in-the-large, and the concordance (c)-statistic. The models were updated by revising the strength of the individual predictors in the models.

A total of 613 men (2.4%) and 1624 women (8.4%) were deferred from donation because of a low Hb level. Validation demonstrated underestimation of predicted risks and lower c-statistics for men and women compared to the Dutch cohort. The strength of most predictive factors, particularly previous Hb level, was lower in Irish donors. The updated models showed a c-statistic of 0.83 (95% confidence interval [CI], 0.81-0.84) for men and 0.76 (95% CI, 0.74-0.77) for women.

The performance of Dutch prediction models for Hb deferral was limited when validated in Irish whole blood donors. Updating the models resulted in different predictor effects. This improved mainly the model calibration; the improvement in discrimination was small.

We retrospectively reviewed the PBSC collections of 18 consecutive lymphoma and multiple myeloma patients, who had previously mobilized poorly despite the use of plerixafor and received plerixafor again during remobilization.

During the first mobilization attempt, all 18 recombinant granulocyte–colony-stimulating factor (G-CSF; two) or G-CSF plus chemotherapy–mobilized patients (16) had poor response to plerixafor, with peripheral blood (PB) CD34+ counts ranging from 0 to 7.48 × 10⁶/L after the first dose. They collected only 0.15 × 10⁶ to 1.63 × 10⁶ (median, 0.40 × 10⁶) CD34+ cells/kg after one to four collections. The median average daily yield was 0.24 × 10⁶ CD34+ cells/kg. Remobilization began 1 to 4 weeks later with G-CSF, plerixafor, and with (three) or without (15) cyclophosphamide. The PB CD34+ cell counts after the first dose of plerixafor were 3.04 × 10⁶ to 127.54 × 10⁶/L (median, 14.58 × 10⁶/L). After one to four doses of plerixafor, each patient collected an additional 0.39 × 10⁶ to 14.02 × 10⁶ (median, 1.89 × 10⁶) CD34+ cells/kg, and the median daily average was 0.78 × 10⁶ CD34+ cells/kg. Cumulatively, after two rounds of collections, 15 collected more than 2.0 × 10⁶ CD34+ cells/kg. Thirteen have proceeded to autologous stem cell transplantation (ASCT) and successfully engrafted.

In patients who had responded poorly to the use of plerixafor as a mobilization salvage agent, response to remobilization with plerixafor for the second time was variable, but most (83.3%) patients were able to collect enough PBSCs to proceed to ASCT.

We developed a new pathogen detection system that can detect multiple viruses simultaneously, using originally designed degenerate polymerase chain reaction primers to amplify a wide range of viral genotypes. Amplified samples were identified using a DNA microarray of pathogen-specific probes.

We detected very low copy numbers of multiple subtypes of viruses, such as human hepatitis C virus (HCV), human hepatitis B virus (HBV), human parvovirus B19 (PVB19), and West Nile virus (WNV), using a single plate. We also detected all genotypes of human immunodeficiency virus (HIV) but sensitivity was less than for the other viruses.

We developed a microarray assay using novel primers for detection of a wide range of multiple pathogens and subtypes. Our NAT system was accurate and reliable for detection of HIV, HBV, HCV, PVB19, and WNV, with respect to specificity, sensitivity, and genotype inclusivity. Our system could be customized and extended for emerging pathogens and is suitable as a future NAT system.

For analytical sensitivity assessment the World Health Organization HBV, hepatitis C virus (HCV), and human immunodeficiency virus (HIV) international standards and various genotype dilution panels were used. Individual donations (IDs) from 9980 first-time donors were screened simultaneously by serology and both TMA assay versions.

The 50 and 95% limits of detection (LODs) for HBV using Ultrio Plus were 0.8 (0.6-1.0) and 4.6 (3.2-7.2) IU/mL, respectively, 2.4 (1.4-4.8)-fold more sensitive than Ultrio. The TMA assay versions had comparable LODs for HIV-1 and HCV. The improvement factors on analytical sensitivity panels of HBV Genotypes A to G ranged from 1.3 to 7.3 and 50% LODs (95% confidence interval) reduced from 12.5 (10-15) to 3.8 (3.2-4.4) copies/mL. One Ultrio Plus HBV Genotype D yield sample missed by the Ultrio assay in the donor screening study was detected with ninefold higher sensitivity. The specificities of ID nucleic acid test (ID-NAT) and serologic testing in a similar repeat test algorithm were 100 and 99.41%, respectively.

More efficient target capture chemistry in the new TMA assay version significantly improved sensitivity and diminished variability in detecting HBV strains of various genotypes. We recommend a triplicate ID-NAT repeat test strategy to eliminate discriminatory tests on false-non–repeat-reactive (anti-HBc–nonreactive) donations.

Granulocyte apheresis procedures are performed with standard apheresis equipment, including with the frequently used apheresis system for peripheral blood “stem cell” collection, COBE Spectra MNC (Terumo BCT), using the same tubing set as for MNC collection, but a different software protocol, PMN. An automated apheresis system for granulocyte collection, Spectra Optia IDL (Terumo BCT), became available in October 2011. Since then, 70 granulocyte apheresis procedures have been performed at our site, 35 each with the new and old systems.

Apheresis procedures were well tolerated throughout. The target dose of 1 × 10¹⁰ neutrophils was achieved in all but one collection with Spectra Optia IDL. Spectra Optia IDL collections were approximately 20% more efficient. Products contained more nontarget white blood cells (mononuclear cells), but fewer RBCs and platelets. Although less blood had to be processed with Spectra Optia IDL to achieve the same granulocyte dose, clinically relevant differences between the two apheresis devices were not apparent.

Both apheresis systems are similarly capable of generating granulocyte concentrates.

Pooled data on ABT, PNI, 30-day mortality, and length of hospital stay (LHS) from 2547 patients undergoing elective lower-limb arthroplasty (n = 1186) or hip fracture repair (n = 1361) were compared between patients who received either very-short-term perioperative IV iron (200-600 mg; n = 1538), with or without recombinant human erythropoietin (rHuEPO; 40,000 IU), or standard treatment (n = 1009).

Compared to standard therapy, perioperative IV iron reduced rates of ABT (32.4% vs. 48.8%; p = 0.001), PNI (10.7% vs. 26.9%; p = 0.001), and 30-day mortality (4.8% vs. 9.4%; p = 0.003) and the LHS (11.9 days vs. 13.4 days; p = 0.001) in hip fracture patients. These benefits were observed in both transfused and nontransfused patients. Also in elective arthroplasty, IV iron reduced ABT rates (8.9% vs. 30.1%; p = 0.001) and LHS (8.4 days vs.10.7 days; p = 0.001), without differences in PNI rates (2.8% vs. 3.7%; p = 0.417), and there was no 30-day mortality.

Despite known limitations of pooled observational analyses, these results suggest that very-short-term perioperative administration of IV iron, with or without rHuEPO, in major lower limb orthopedic procedures is associated with reduced ABT rates and LHS, without increasing postoperative morbidity or mortality.

A pool-and-split design of leukoreduced CPD–saline-adenine-glucose-mannitol–packed RBC units was used to generate three groups: untreated controls, sham-treated units, and units treated with a cold (1-6°C) rejuvenation solution on Day 28, 35, or 42 of cold storage. Units were followed until Day 49 of storage and assessed for ATP concentration, morphology, deformability, and other in vitro quality variables including hemolysis, pH, and supernatant potassium levels.

At every treatment time, rejuvenation was associated with a significant increase in intracellular ATP (p < 0.01). On Day 28, rejuvenation was accompanied by a significant decrease in deformability 1 week after treatment (p < 0.01). Rejuvenation on Day 28, but not Day 35 or 42, was also associated with a significant change in morphology (p < 0.01). Of the in vitro quality variables measured, most changed during cold storage, but differences among treatment groups were not observed.

The results of our study demonstrate a cold rejuvenation of RBCs during HS increases intracellular ATP, but that this change does not ameliorate, or exacerbate, the metabolic or biochemical symptoms of the storage lesion.

Literature searches were performed in MEDLINE and the Cochrane Database of Systematic Reviews.

We identified nine case reports of pediatric patients who had experienced cardiac arrest during massive transfusion. Serum potassium concentration was reported in eight of those reports; the mean was 9.2 ± 1.8 mmol/L. Risk factors for TAHCA noted in the case reports included infancy (n = 6); age of red blood cells (RBCs; n = 5); site of transfusion (n = 5); and the presence of comorbidities such as hyperkalemia, hypocalcemia, acidemia, and hypotension (n = 9). We also identified 13 clinical studies that examined potassium levels associated with transfusion. Of those 13, five studied routine transfusion, two were registries, and six examined massive transfusion.

Key points identified from this literature search are as follows: 1) Case reports are skewed toward infants and neonates in particular and 2) the rate of blood transfusion, more so than total volume, cardiac output, and the site of infusion, are key factors in the development of TAHCA. Measures to reduce the risk of TAHCA in young children include anticipating and replacing blood loss before significant hemodynamic compromise occurs, using larger-bore (>23-gauge) peripheral intravenous catheters rather than central venous access, checking and correcting electrolyte abnormalities frequently, and using fresher RBCs for massive transfusion.

Quarantine plasma samples were freeze-dried. The clotting factors fibrinogen, Factor (F)V, FVIII, FXI, von Willebrand factor (vWF), protein S, antithrombin, plasminogen, and plasmin inhibitor were determined after manufacturing and after storage at room temperature and refrigeration. Reported adverse transfusion events were evaluated and compared to that of FFP. Clinical effectiveness was estimated by inquiry among experienced users.

Lyophilization resulted in a loss of coagulation factor activity between 0% and up to 20% to 25% (FVIII, vWF). When stored refrigerated, coagulation factors did not lose more than 10% of their activities. Storage at room temperature for 24 months mainly affected vWF/ristocetin cofactor activity and fibrinogen activity. From 2007 to 2011 more than 230,000 units of FDP were delivered. There were no reports about clinical ineffectiveness. The frequency of transfusion reactions was not different from that of FFP.

Lyophilized plasma showed characteristics similar to FFP. Since FDP requires neither complex logistics nor time-consuming thawing, it allows rapid treatment of coagulopathies.

A multicenter, randomized, nonblinded, crossover paired treatment protocol was performed. Thirty patients with orders for at least two TPE procedures were randomly assigned to the AMICUS or the COBE Spectra (TerumoBCT) for the first treatment. Each patient was crossed over to the other device using the same procedure settings from the first procedure. The primary objective compared efficiency of plasma removal (EPR) with secondary objectives of comparing PLT and hemoglobin (Hb) waste plasma content, coagulation factor and complement activation, fluid balance tracking accuracy, procedure length, and adverse events.

The EPR for the AMICUS (81.9 ± 7.62%) was superior to that of the COBE Spectra (75.2 ± 6.29%; p = 0.00001). The AMICUS also demonstrated statistically higher fluid balance accuracy (99.84%) compared to that of the COBE Spectra (98.83%; p < 0.0001) and a statistically shorter procedure time (103.9 ± 30.8 vs. 110.5 ± 27.1 min, p < 0.001). No significant differences with regard to PLT and Hb content in the waste plasma, change in patient PLT count, or changes in markers of coagulation and complement cascade activation were seen. Frequency and severity of adverse reactions were similar.

The AMICUS separator can effectively perform TPE. The AMICUS demonstrated superior plasma removal efficiency compared to the COBE Spectra with no evidence of significant differences in PLT removal, hemolysis, and coagulation or complement activation.

The aim of this study was to perform DNA investigations to identify silent KEL alleles among 10 KEL:–5 patients and 121 individuals presenting the rare KEL:1,−2 phenotype. Serologic investigations were performed on patients' red blood cells and serum. The KEL gene analysis was done by using a BeadChip assay (HEA Version, 1.2, Immucor), real-time polymerase chain reaction, and/or sequencing of all 19 exons of the KEL gene.

In KEL:–5 patients, two novel KELnull alleles were described: 821G>A being the second described KELnull allele on a KEL*01 backbone and 184Tdel. In the 121 KEL:1,−2 individuals, nine (7.4%) were found to display a discordant KEL:1,−2 phenotype and KEL*01/KEL*02 genotype. Three novel silent KEL*02 alleles were described: 1084C>A, 1708G>A, and IVS11+5g>a.

The number of silent KEL alleles and the notion that KEL null alleles are on a KEL*02 background may evolve in the coming years. Systematic DNA analysis showed that the number of discordant phenotype/genotype results, related to silent KEL*02 alleles was higher than expected in France. These data emphasize that clinical practice based on DNA analysis for blood group antigens requires caution and should improve the performance of the blood group phenotype prediction.

DNA and RNA were extracted from white blood cells and polymerase chain reaction–based assays, cloning, and sequencing were done using standard protocols.

The anti-Ku in Proband 1, which caused hemolytic disease and anemia of the fetus and newborn, was a mixture of immunoglobulin (Ig)G₁ and IgG₂ and gave macrophage indexes ranging from 47.8 to 59.3 (>20 is clinically significant) in a monocyte monolayer assay. The proband, her daughter, and compatible sister had a heterozygous deletion of a G in Exon 18 (Nucleotide c.1972_1975delG) in a KEL*02 allele causing a frameshift. The mechanism for silencing of the other KE*02 allele was undetermined. Proband 2 was heterozygous for a nonsense change (KEL*382C/T; Arg128Stop), a missense change (KEL*244T/C; Cys82Arg), and KEL*578T/C (KEL*01/KEL*02). Direct sequencing of cDNA and cloning showed that the KEL*01 allele had 244C, 382C, 578T and the KEL*02 allele carried 244T, 382T, 578C.

We report a novel single-nucleotide deletion, a novel nonsense allele, and a novel missense allele all resulting in the K_null phenotype. The anti-Ku from Proband 1 was clinically important.

Whole blood samples from 22,904 donations were tested for CMV DNA, and CMV DNA–positive donations were categorized as donations from 1) seronegative donors, 2) newly seropositive donors, and 3) long-term seropositive donors.

Twenty-one donors were reproducibly CMV DNA–positive (0.09%). Frequency of detection and concentration of CMV DNA in whole blood were comparable for seronegative and long-term seropositive donors. Nonreproducibly positive results for CMV DNA in whole blood were more frequent in long-term seropositive donors (0.16% vs. 0.01%, p < 0.01). Only low concentrations of CMV DNA in plasma were detectable in two seronegative donors and one long-term seropositive donor. Highest concentrations of CMV DNA in both whole blood and plasma, however, were found in newly seropositive donors.

Prevalences of window period donations among seronegative donors and reactivations among long-term seropositive donors, as well as the CMV DNA concentration in whole blood and plasma samples from these donors, are comparable. Therefore, blood products from both groups could be used for patients at risk for TT-CMV, while those of newly seropositive donors seem to bear an increased risk.

To analyze the interaction between von Willebrand factor (VWF) and ADAMTS13, we used large-pore isoelectric focusing (IEF) analysis followed by detection with anti-ADAMTS13 monoclonal antibody. FFP, CSP, cryoprecipitate (CP), and purified ADAMTS13 were analyzed for their effects on high shear stress–induced platelet aggregation (H-SIPA).

IEF analysis of normal plasma revealed three groups of ADAMTS13 bands with pI of 4.9 to 5.6, 5.8 to 6.7, and 7.0 or 7.5. Two band groups (pI 4.9-5.6 and 5.8-6.7) were found in plasma of a patient with Type 3 von Willebrand disease, in which VWF is absent, whereas no bands were found in plasma of a patient with congenital ADAMTS13 deficiency. Mixing these plasmas generated the bands at pI 7.0 or 7.5, representing the VWF-ADAMTS13 complex; these bands were absent in CSP. FFP and purified ADAMTS13 down regulated H-SIPA in a dose-dependent manner. However, CP did not inhibit H-SIPA in the initial phase, and the degree of inhibition at the endpoint was almost indistinguishable from those of the other two plasma products.

Both plasma products (FFP and CSP) are effective for PE in TTP patients. However, CSP may be more favorable, because it has lower levels of VWF and almost normal ADAMTS13 activity, but lower levels of ADAMTS13 in complex with larger VWF multimers.

We analyzed supernatants and blood smears of human red blood cell (RBC) units that either were or were not leukoreduced before storage for markers of NETs.

We identified extracellular DNA, which was associated with histones and myeloperoxidase, a marker of neutrophil granules, in supernatants and blood smears of nonleukoreduced RBC units. These markers of NETs were absent in leukoreduced RBC units. Importantly, NETs passed through blood transfusion filters and could therefore potentially be infused into patients.

Our studies indicate that NETs are liberated during storage of nonleukoreduced RBC units. Future studies should address whether NETs in RBC units could potentially contribute to transfusion-associated complications.

Polymerase chain reaction amplification, direct sequencing, restriction fragment length polymorphism assays, hemagglutination, flow cytometry, and protein modeling were performed by standard methods.

Proband 1 (KUCI) and her serologically compatible sister were heterozygous for a nucleotide change in Exon 11 (KEL*1271C/T; Ala424Val). Proband 2 (KANT) was heterozygous for KEL*1283G/T (Arg428Leu) and KEL*1216C/T (Arg406Stop) in Exon 11. Red blood cells (RBCs) from Proband 1 and her sister were not agglutinated by plasma from Proband 2; however, RBCs from Proband 2 were agglutinated by plasma from Proband 1. Probands 3, 4, 5, and 6 had the KEL*1391C>T change associated with the previously reported KETI– phenotype. Proband 5 was also homozygous for KEL*905T>C encoding the K11–K17+ phenotype. Hemagglutination studies revealed an association between KUCI, KANT, KETI, and K11. Protein modeling indicated that whereas Ala424 and Arg428 are clustered, Val302 and Thr464 are not.

Ala424 in the Kell glycoprotein is associated with the high-prevalence Kell antigen, KUCI (ISBT 006032), which is detected by the antibody of Proband 1. Arg428 is associated with the high-prevalence Kell antigen, KANT (ISBT 006033). The association between KUCI, KANT, KETI, and K11 and the results of protein modeling are discussed.

The para-Bombay phenotype of the nine Chinese probands was identified by standard serologic techniques. The coding regions of FUT1 and FUT2 genes were amplified by polymerase chain reaction and then directly sequenced. ABO genotyping was performed by polymerase chain reaction with sequence-specific priming method. The FUT1 and FUT2 genotypes and the distribution in all reported Chinese para-Bombay individuals including our study were also summarized.

Five FUT1 genotypes, h1h3 (n = 3), h1h2 (n = 3), h1h1 (n = 1), h3h3 (n = 1), and h2h3 (n = 1), and three functional FUT2 genotypes, Se³⁵⁷Se³⁵⁷ (n = 4), Se³⁵⁷Se^{357, 716} (n = 4), and Se³⁵⁷Se^{357, 385} (n = 1) described before were identified in nine probands.

The review of the literature shows that a total of 17 FUT1 alleles and four FUT2 alleles (Se³⁵⁷, Se^357,716, Se^{357 385}, Se) have been identified in Chinese para-Bombay individuals. The four FUT1 alleles, h1 (547delAG), h2 (880delTT), h3 (C658T), and h4 (C35T; A980C) are most prevalent, which account for more than 90% of all allele counts and are essential to be involved when developing para-Bombay genotyping kit for Chinese.

In this study, further investigation was performed to reveal the underlying mechanism responsible for reduction of B antigen expression in the exceptional B_m individual. Peptide nucleic acid–clamping polymerase chain reaction was carried out to amplify the B-related allele, followed by sequence determination. Electrophoretic mobility assays and promoter assays were performed to examine whether a nucleotide substitution reduced the binding of a transcription factor and induced loss of function of the element.

Sequence determination revealed one point mutation of the GATA motif in the element. The electrophoretic mobility shift assays showed that the mutation abolished the binding of GATA transcription factors, and the promoter assays demonstrated complete loss of enhancer activity of the element.

These observations suggest that the mutation in the GATA motif of the erythroid-specific regulatory element may diminish the binding of GATA transcription factors and down regulate transcriptional activity of the element on the B allele, leading to reduction of B antigen expression in erythroid lineage cells of the B_m individual.

The failure modes and effects occurring throughout the whole process of blood transfusion were studied. Each failure mode was evaluated using three scores: severity of effect (S), likelihood of occurrence (O), and probability of detection (D). Risk priority numbers (RPNs) were calculated by multiplying the S, O, and D scores. The plan-do-check-act cycle was also used for continuous improvement.

Analysis has showed that failure modes with the highest RPNs, and therefore the greatest risk, were insufficient preoperative assessment of the blood product requirement (RPN, 245), preparation time before infusion of more than 30 minutes (RPN, 240), blood transfusion reaction occurring during the transfusion process (RPN, 224), blood plasma abuse (RPN, 180), and insufficient and/or incorrect clinical information on request form (RPN, 126). After implementation of preventative measures and reassessment, a reduction in RPN was detected with each risk. The failure mode with the second highest RPN, namely, preparation time before infusion of more than 30 minutes, was shown in detail to prove the efficiency of this tool.

FMEA evaluation model is a useful tool in proactively analyzing and reducing the risks associated with the blood transfusion procedure.

Variant D types were characterized using molecular typing, RHD sequencing, extended serologic D antigen investigations, and flow cytometric D antigen quantification.

We discovered three novel weak D types termed weak D Types 45.1, 75, and 76 with RHD nucleotide substitutions coding for amino acid exchanges in predicted intracellular RhD polypeptide stretches; antigen densities of approximately 1.990, 900, and 240 D sites per red blood cell were found, respectively. Adsorption-elution technique–supported D epitope mapping of these three weak D types demonstrated the expression of all tested D epitopes. Initial molecular typing of the three investigated samples by RHD gene exon scanning polymerase chain reaction using sequence-specific priming yielded a negative reaction for A1193 located in RHD Exon 9 and could be explained by specific mutations for weak D Types 45.1 (C818T, G1195A), 75 (G1194C), and 76 (A1215C).

All novel weak D types expressed all tested D epitopes. It is of interest that for weak D Types 45.1, 75, and 76, similar alleles with a maximal divergence of one amino acid only, that is, weak D Types 45, 41, and 68, respectively, have been reported so far.

We set up, in a single-tube format, a qualitative and quantitative assay based on multiplex PCR of short fluorescent fragments (QMPSF) to simultaneously amplify all 10 RHD exons on the one hand and all 10 RHCE exons on the other hand.

The test proved to be useful to rapidly identify hybrid genes in hemizygous RHD samples carrying a hybrid D-CE gene and to resolve unknown genotypes by quantifying individual exons in compound heterozygous samples, but also unexpectedly helped to redefine the RHDΨ haplotype. While validating the test, two novel single-point variants, c.648G>C (p.L216F) and c.1048G>C (p.D350H), were found.

For the first time, a QMPSF-based method is reliable to individually quantify the exons of both RH genes, including hybrid D-CE genes in compound heterozygous samples and may help to investigate samples with unknown RHD and/or RHCE status.

MPX, Ultrio, and Ultrio Plus were used to test 2222 pedigreed, marker-positive samples with varying viral loads, each from a unique US blood donor. NAT-positive, seronegative yield samples (16 HBV, 156 HCV, and 23 HIV) were tested in replicates of three; undiluted; and in 1:6, 1:8, and 1:16 dilutions (MP6, MP8, and MP16), simulating various MP sizes. Seropositive samples (1276 HBV, 488 HCV, and 263 HIV) were tested by ID-NAT in singlet.

MPX-MP6 and Ultrio Plus-MP16 had equivalent HCV sensitivity. Although Ultrio Plus-MP16 for HIV trended toward lesser sensitivity, this was not corroborated in a large substudy of low-viral-load samples in which Ultrio Plus-MP8/MP16 showed 100% reactivity. MPX-ID and Ultrio Plus-ID HBV clinical sensitivity were identical, but MPX-MP6 was significantly more sensitive than Ultrio Plus-MP16; the differential yield projected to one HBV NAT yield per 4.72 million US donations. Ultrio Plus HBV sensitivity did not increase at MP8 versus MP16. Ultrio Plus versus Ultrio sensitivity was significantly increased in HBV-infected donors with early acute, late acute or chronic, and occult infections. No difference in sensitivity was noted for any virus for MPX-MP6 versus Ultrio Plus-ID.

Our data support US donation screening with MPX-MP6 or Ultrio Plus-MP16 since the HBV DNA detection of Ultrio Plus was significantly enhanced (vs. Ultrio) without compromising HIV or HCV RNA detection.

Detailed data were collected on patient demographics, diagnoses, transfusion history, history of known allo- and autoantibodies, and specific serologic tests performed for 6077 consecutive serologic work ups in 3608 antibody-positive patients between 2009 and 2011 at four US academic medical centers. Direct testing costs were also determined at each site for each serologic test performed to calculate total costs per work up and per patient over the duration of the study.

The mean direct cost of serologic testing was $114 per work up and $195 per patient. The mean cost per patient was significantly higher for 12 of 19 diagnostic categories evaluated, including autoimmune hemolytic anemia (mean cost per patient, $1490; p < 0.001), hematologic malignancies ($640, p < 0.001), and transplant recipients ($462, p = 0.019). Patient transfusion and serologic testing characteristics associated with greatest increases in costs included history of a warm autoantibody ($626, p < 0.001) and more than five prior transfusions ($404, p < 0.001).

Antibody-positive patients with complex diagnoses or transfusion histories require significantly more resources and incur greater cost to assess red blood cell antibody status.

We conducted a literature search of MEDLINE, Cochrane Controlled Register of Clinical Trials, EMBASE, and PubMed databases to April 2012.

A total of 788 citations were reviewed and 30 reports were included in the analysis. Most studies did not include technologies currently in use for HLA typing or detection of HLA antibodies as 75% were conducted before the year 2000. None of the studies were adequately powered to detect an effect on mortality or hemorrhage. HLA-matched PLTs did not reduce alloimmunization and refractoriness rates beyond that offered by leukoreduction, and utilization was not consistently improved. HLA-matched PLTs led to better 1-hour posttransfusion count increments and percentage of PLT recovery in refractory patients; however, the effect at 24 hours was inconsistent.

The correlation of the PLT increment with other clinical outcomes and the effect of leukoreduction on HLA-matched PLT transfusion could not be determined. Prospective studies utilizing current technology and examining clinical outcomes are necessary to demonstrate the effectiveness of HLA-matched PLT transfusion.

The KEL1/2 single-nucleotide polymorphism was polymerase chain reaction (PCR) amplified with one adjoining base, and the PCR product was sequenced using a genome analyzer (GAIIx, Illumina); several millions of PCR sequences were analyzed.

The results demonstrated the feasibility of diagnosing the fetal KEL1 or KEL2 blood group from cell-free DNA purified from maternal plasma.

This method requires only one primer pair, and the large amount of sequence information obtained allows well for statistical analysis of the data. This general approach can be integrated into current laboratory practice and has numerous applications. Besides DNA-based predictions of blood group phenotypes, platelet phenotypes, or sickle cell anemia, and the determination of zygosity, various conditions of chimerism could also be examined using this approach. To our knowledge, this is the first report focused on antenatal blood group determination using NGS.

Data on current hospital blood inventories were collected from 11 hospitals and three blood centers in five nations. A general predictive model for the age of RBCs at the time of issue was developed based on considerations of demand for RBCs in the hospital.

Age of RBCs at issue is sensitive to the following factors: ABO group, storage age at the time of receipt by the hospital, the restock interval, inventory reserve, mean demand, and variation in demand.

A simple model, based on hospital demand, may serve as the basis for examining factors affecting the storage age of RBCs in hospital inventories. The model suggests that the age of RBCs at the time of their issue to the patient depends on factors external to the hospital transfusion service. Any substantial change in the expiration date of stored RBCs will need to address the broad variation in demand for RBCs while attempting to balance considerations of availability and blood wastage.

We have developed a new HPA-genotyping method for simultaneous detection of HPA-1 to -16 based on suspension array technology. A total of 216 samples from Chinese Han donors in Xi'an were genotyped using the developed method, and all the samples again were genotyped using polymerase chain reaction (PCR) sequence-based typing (PCR-SBT), which is considered the gold standard.

All 216 samples were successfully genotyped for HPA-1 to -16 using both our method and PCR-SBT. Results showed that the genotype and allele frequencies obtained using our method were fully consistent with those obtained using PCR-SBT.

Our method provides accurate, high-throughput, and simultaneous genotyping of HPA-1 to -16 and will serve as the foundation for large-scale clinical genotyping of HPAs and for the establishment of an HPA-typed PLT donor registry.

Seeking to understand the biochemical mechanisms underlying these observed effects, we analyzed signal transduction in PLT concentrates after riboflavin and ultraviolet light (UV; Mirasol) treatment and subsequent storage focusing on the phosphorylation levels of selected protein kinases.

Among identified candidates, p38MAPK increased fourfold in phosphorylation after PRT. Incubation of PLT concentrates with a p38MAPK-specific inhibitor before PRT significantly improved numerous PLT quality measures. Phosphorylation levels of the p38MAPK substrates AKT, VASP, and HSP27 also decreased with inhibitor treatment. Phospho-HSP27 decrease in the presence of the inhibitor correlated with a reduction in PLT activation determined by surface expression of P-selectin.

These findings support a model of one dominant underlying molecular signaling mechanism that is impacted by the riboflavin and UV (Mirasol) PRT process resulting in alterations in PLT quality. The identification of such a target should assist in the development of strategies to ameliorate this negative aspect of an otherwise beneficial and important safety development for transfusion medicine.

A closed, automated cell processor, the ACP 215 from Haemonetics Corporation, was used to wash RCCs and determine optimal pre- and postwash storage times. Two postwash storage solutions, additive solution (AS)-3 and saline-adenine-glucose-mannitol (SAGM), were compared. The in vitro quality of leukoreduced RCCs, prepared from citrate-phosphate-dextrose–anticoagulated whole blood, was determined postwash and compared to existing guidelines for RCC quality (hemoglobin content, hematocrit, and hemolysis) and predetermined criteria for ATP and supernatant potassium levels. A criterion for visual hemolysis was also applied.

The prewash storage time, postwash storage time, and the postwash resuspension solution all contributed to RCC quality postwash. Levels of hemolysis were greater when washed RCCs were resuspended in SAGM (p = 0.01), while AS-3 proved worse at maintaining ATP levels postwash (p < 0.01). Immediately postwash, all units had supernatant K⁺ levels below the detection limit of the instrument (<1 mmol/L), but these increased to above acceptable levels within 14 days.

Based on all acceptance criteria, a maximum 14-day prewash storage period and 7-day postwash storage period in SAGM preservative was found to be optimal. The longer outdate time postwashing should help lessen challenges in providing components to patients before expiry.

CB buffy coat cells and isolated CD34-positive (CD34^pos) cells from premature and full-term CB and adult blood were tested with several combinations of growth factors while omitting xenogeneic proteins from the culture medium. Cell differentiation was analyzed serially during 21 days using flow cytometry, progenitor assays, and high-performance liquid chromatography.

Expanded CB buffy coat cells resulted in a threefold higher number of erythroblasts than the isolated CD34^pos cells. However, the RBCs contaminating the buffy coat remained present during the culture with uncertain quality. Premature and full-term CB CD34^pos cells had similar fold expansion capacity and erythroid differentiation. With the use of interleukin-3, stem cell factor, and erythropoietin, the fold increases of all CD34^poscell sources were similar: CB 3942 ± 1554, adult peripheral mobilized blood 4702 ± 1826, and bone marrow (BM) 4143 ± 1908. The proportion of CD235a expression indicating erythroblast presence on Day 21 was slightly higher in the adult CD34^pos cell sources: peripheral blood stem cells (96.7 ± 0.8%) and BM (98.9 ± 0.5%) compared to CB (87.7 ± 2.7%; p = 0.002). We were not able to induce further erythroid maturation in vitro.

This explorative study showed that fairly pure autologous erythroid-expanded cell populations could be obtained by a simple culture method, which should be optimized. Future challenges comprise obtaining ex vivo enucleation of RBCs with the use of a minimal manipulating approach, which can add up to autologous RBCs derived from CB in the treatment of anemia of prematurity.

We performed phenotype investigations by serology studies, analyzed the DNA sequence of the ABO gene by direct sequencing or sequencing after cloning, and evaluated promoter activity by reporter assays.

In 62 rare ABO alleles, we identified 29 novel ABO subgroup alleles in 43 apparently unrelated subgroup individuals and their four available pedigrees. Of these alleles, one was a deletion-mutation allele, four were hybrid alleles, and 24 were point-mutation alleles. Most of the point mutations were detected in Exons 6 to 7, while several others were also detected in Exons 1 to 5 or splicing regions. One ABO promoter mutation, −35 to −18 del, was found and verified to reduce promoter activity, as determined by dual luciferase assays. Two mutations, 7G>T and 52C>T, carrying the premature terminal codons E3X and R18X in the 5′-region, were found to be associated with the very weak ABO subgroups “Ael” and “Bel.”

Twenty-nine ABO subgroup alleles were newly linked to different kinds of ABO variations. We provide the first evidence that promoter abnormality is involved in the formation of weak ABO phenotypes. We also described the first naturally occurring ABO alleles with premature terminal codons in the 5′-region that led to Ael and Bel phenotypes.

The authors compared 51 patients undergoing staged bilateral TKA who received TXA (2 g; subjects) with 70 who did not (controls). There were no significant differences between the groups in terms of demographics or preoperative Hb. For each TKA, 1 g of TXA was administered intravenously 15 minutes before incision and 1 g was administered intravenously at tourniquet release. Blood loss, Hb levels, and transfusions were recorded. Statistical analyses were performed using computer software. Significance was set at 0.05.

Subjects had a significantly lower (p < 0.001) mean (±SD) blood loss (373.8 ± 264.6 mL vs. 871.6 ± 457.7 mL), significantly higher (p < 0.005) Hb levels on Postoperative Days 1 and 2, and a significantly lower (p < 0.001) mean (±SD) number of transfused allogenic blood units (0.60 ± 0.84 units vs. 1.53 ± 1.30 units).

TXA reduces blood loss, improves postoperative Hb, and decreases the allogenic blood transfusion requirements for patients undergoing bilateral staged TKA. TXA is an option for patients choosing bilateral staged TKA to decrease the risks associated with blood transfusion or when autologous blood is not available.

This cross-sectional study was conducted between January 2009 and March 2011 at four Brazilian blood centers to identify the population of HIV-negative eligible blood donors that answered face-to-face interviews without disclosing risks, but subsequently disclosed deferrable risk factors by ACASI. Compared to the donor interview, the ACASI contained expanded content on demographics, sexual behavior, and other HIV risk factors questions.

A total of 901 HIV-negative blood donors were interviewed. On the ACASI, 13% of donors (n = 120) declared a risk factor that would have resulted in deferral that was not disclosed during the face-to-face assessment. The main risk factors identified were recent unprotected sex with an unknown or irregular partner (49 donors), sex with a person with exposure to blood or fluids (26 donors), multiple sexual partners (19 donors), and male–male sexual behavior (10 donors). Independent factors associated with the disclosure of any risk factor for HIV were age (≥40 years vs. 18-25 years; adjusted odds ratio [AOR], 0.45; 95% confidence interval [CI], 0.23-0.88) and blood center (Hemope vs. Hemominas; AOR, 2.51; 95% CI, 1.42-4.44).

ACASI elicited increased disclosure of HIV risk factors among blood donors. ACASI may be a valuable modality of interview to be introduced in Brazilian blood banks.

Donations were screened simultaneously for HBV serologic markers and ID-NAT, followed by discriminatory assay and confirmatory test algorithms. Window period (WP) reduction and residual HBV transmission risk were computed using mathematical modeling.

HBV NAT-yield rates for both WP and occult HBV infection (OBI) increased significantly from 1:34,471 to 1:17,362 (p = 0.036) and from 1:5120 to 1:2450 (p < 0.0001), despite a 1.2- and 1.6-fold decrease in hepatitis B surface antigen (HBsAg) incidence and prevalence rates respectively. After adjusting for this bias, the WP and OBI NAT-yield improvement factors were 2.3 and 3.4, respectively, higher than a less than 1.5-fold increase estimated from analytical sensitivity studies on HBV Genotype A and C standards. The current WP transmission risk with Ultrio Plus screening was estimated at 1:55,000 compared to 1:22,000 with HBsAg testing.

The observed greater than twofold enhanced WP NAT yield with the Ultrio Plus assay can be explained by greater than 10-fold increased analytical sensitivity in detecting the HBV Genotype B and C strains in Hong Kong. Direct comparison studies of the two assay versions on dilutions of HBV NAT-yield samples are required to confirm this hypothesis.

Twenty beagles were challenged with 1-hour similar infusions of (200 μmol/L) metHb (n = 5), oxyHb (n = 5), albumin (n = 5), or saline (n = 5). Measurements were taken over 3 hours.

Infusions of the two pure Hb species resulted in increases in mean arterial blood pressure (MAP), systemic vascular resistance index, and NO consumption capacity of plasma (all p < 0.05) with the effects of oxyHb being greater than that from metHb (MAP; increase 0 to 3 hr; 27 ± 6% vs. 7 ± 2%, respectively; all p < 0.05). The significant vasoconstrictive response of metHb (vs. albumin and saline controls) was related to in vivo autoreduction of metHb to oxyHb, and the vasoactive Hb species that significantly correlated with MAP was always oxyHb, either from direct infusion or after in vivo reduction from metHb. Clearance of total Hb from plasma was faster after metHb than oxyHb infusion (p < 0.0001).

These findings indicate that greater NO consumption capacity makes oxyHb more vasoactive than metHb. Additionally, metHb is reduced to oxyHb after infusion and cleared faster or is less stable than oxyHb. Although we found no direct evidence that metHb itself is involved in acute vascular effects, in aggregate, these studies suggest that metHb is not inert and its mechanism of vasoconstriction is due to its delayed conversion to oxyHb by plasma-reducing agents.

In the current study we compare the efficacy of lenograstim and filgrastim in terms of number of circulating CD34+ cells/μL during the fifth day of G-CSF administration, the number of days of apheresis required to obtain the target cell dose, the median of CD34+ cells collected on the first day of apheresis, or the median number of total CD34+ cells collected at the end of the procedure, in a series of 146 healthy donors undergoing hematopoietic stem cell mobilization for allogeneic transplantation.

We observed that, using a comparable dose for the two products, no significant differences were observed between the two groups.

In conclusion, the current retrospective study shows that lenograstim and filgrastim are similar in terms of efficacy for the mobilization of hematopoietic stem cells in healthy donors.

AS-1 RBC units stored up to 42 days were sampled at selected storage times. Samples were added to aortic rings ex vivo, a system where NO-mediated vasodilation could be experimentally controlled.

RBC units showed storage-dependent changes in plasma hemoglobin (Hb), RBC 2,3-diphosphoglycerate acid, and RBC adenosine triphosphate conforming to expected profiles. When freshly collected (Day 0) blood was added to rat aortic rings, methacholine (MCh) stimulated substantial NO-mediated vasodilation. In contrast, MCh produced no vasodilation in the presence of blood stored for 42 days. Surprisingly, the vasoinhibitory effects of stored RBCs were almost totally mediated by RBCs themselves: removal of the supernatant did not attenuate the inhibitory effects, while addition of supernatant alone to the aortic rings only minimally inhibited MCh-stimulated relaxation. Stored RBCs did not inhibit vasodilation by a direct NO donor, demonstrating that the RBC-mediated vasoinhibitory mechanism did not work by NO scavenging.

These studies have revealed a previously unrecognized vasoinhibitory activity of stored RBCs, which is more potent than the described effects of free Hb and works through a different mechanism that does not involve NO scavenging but may function by reducing endothelial NO production. Through this novel mechanism, transfusion of small volumes of stored blood may be able to disrupt physiologic vasodilatory responses and thereby possibly cause adverse clinical outcomes.

HBsAg-negative samples from 65,800 voluntary blood donors were tested with the cobas TaqScreen MPX test in pools of 6 on the Roche cobas s 201 blood screening platform. Samples positive for HBV DNA and negative for HBsAg were quantitated with the Roche COBAS AmpliPrep/COBAS TaqMan HBV test. In addition, serologic tests for HBsAg, hepatitis B surface antibody, anti-hepatitis B core antigen (anti-HBc), anti-hepatitis B e antigen (anti-HBe), and hepatitis B e antigen (HBe) were done using the Roche electrochemiluminescence immunoassay.

A total of 80 nucleic acid amplification technology (NAT) test-reactive pools were identified and 59 pools (74%) resolved to a reactive sample. All samples were HBV DNA reactive and the viral load in each sample was quantitated. The viral loads of the samples ranged from less than 20 to 34,600 IU/mL; 13 samples (22%) had viral loads of more than 20 IU/mL, 27 samples (45.8%) had viral loads of less than 20 IU/mL, and 19 samples (32.2%) had undetectable viral loads. Of the 59 NAT-reactive samples, 40 (67.8%) were anti-HBc positive. Fifteen of the 59 samples could not be confirmed as NAT reactive either by an alternative NAT test or by serology.

The HBV NAT yield in blood donors in Qingdao is 0.06% (38/65,800). This study confirmed the value of NAT for interdicting HBV-positive donations and preventing transfusion-transmitted HBV infections.

The aim of this study was to determine the CTL2 transcript pattern in human peripheral blood cells and tissues and its capacity to bind HNA-3a antibodies. RNA was isolated from human whole blood, isolated neutrophils, mononuclear blood cells, leukoreduced platelets, human lung, liver, and colon. After reverse transcription, the single-stranded cDNA was amplified using primer combinations specific for the respective transcript. Plasmids containing the entire CTL2 coding cDNA of the transcript variant TV1 or TV2 served as controls. HEK293T cells expressing both variants were used to determine the binding of HNA-3a antibodies.

The shorter TV2 transcript was demonstrated in each RNA sample derived from human peripheral blood tested so far, as well as in human lung and liver, whereas the longer TV1 transcript was only detected in human lung and colon. TV1 and TV2 had the same binding capacity to HNA-3a antibodies.

The expression of TV1 and TV2 is tissue and cell specific, with peripheral blood cells expressing only TV2. This does not affect binding of HNA-3a antibodies. Whether the unequal expression might be relevant in the pathogenesis of TRALI remains to be investigated.

Oxygenation patterns among four VLBW infants who developed transfusion-related NEC (TR-NEC) were compared to four VLBW infants with similar gestational age who were transfused but did not develop NEC (non-NEC). Cerebral and mesenteric patterns were recorded before, during, and 48 hours after RBC transfusion using near-infrared spectroscopy (NIRS) technology. Percentage change from mean baseline regional oxygen saturation values and cerebrosplanchnic oxygenation ratios were analyzed.

All TR-NEC infants (24-29 weeks’ gestation; 705-1080 g) demonstrated greater variation in mesenteric oxygenation patterns surrounding transfusions than non-NEC infants (27.6-30 weeks’ gestation; 980-1210 g). TR-NEC infants received larger mean volumes of total blood (27.75 ± 8.77 mL/kg) than non-NEC infants (15.25 ± 0.5 mL/kg).

Wide fluctuation and decreases in mesenteric oxygenation patterns are more pronounced in TR-NEC infants, especially before TR-NEC onset, compared to non-NEC infants. Greater total volume of infused blood was associated with TR-NEC in preterm infants. Using NIRS, larger prospective studies are needed to further evaluate potential risk factors for NEC in this high-risk population.

A two-center retrospective case-referent study was performed where case patients and control subjects were sampled from all consecutive patients who had received their first and subsequent RBC transfusions in one of the two centers only. Cases were defined as patients who developed a first RBC alloantibody. Control subjects were patients without detectable RBC alloantibodies, who were matched to the case patients regarding number of RBC transfusions. Binary logistic regression analysis was used to examine the association between storage time of RBCS and the occurrence of alloimmunization.

A total of 144 cases and 286 controls were selected for our study, who had received a total 5478 RBC units. Comparing patients receiving units stored shorter than a certain number of days versus older units (with various storage periods up to 4 weeks) did not reveal an association or a trend between alloimmunization risk and storage time categories.

Our findings suggest that storage times of transfused RBCs between 1 and 4 weeks do not affect the risk of alloimmunization.

The accumulation of choline-containing phospholipids (LPC, sphingomyelin [SM], and phosphatidylcholine [PC]), LPA, and ATX during the storage of autologous blood and the changes caused by leukoreduction were investigated. A total of 26 orthopedic patients were enrolled. Autologous blood was collected as whole blood and, after leukoreduction, preserved refrigerated until use. Prestorage leukoreduced (LR) and non-LR autologous blood samples were analyzed. The time-dependent changes and the effect of the filtration were compared.

A time-dependent and significant increase in the levels of LPA was observed in both non-LR and LR samples. The concentration of LPA was significantly reduced in LR compared to non-LR samples. The concentration of LPC was higher in LR compared to non-LR samples. The levels of PC, SM, and ATX were not affected by either the storage period or the leukoreduction.

Leukoreduction of autologous whole blood effectively reduced the accumulation of LPA. On the other hand, prestorage leukoreduction resulted in an increased concentration of LPC, without significantly affecting ATX. Further studies are necessary to confirm the role of LPA in the pathogenesis of adverse effects of blood transfusion, especially TRALI.

Microbiologic culture findings on consecutive HPCs and other cell preparations at a single institution derived from peripheral blood, marrow, cord blood, and mesenchymal stromal cells during all phases of manipulation were retrospectively examined from 2005 through 2011. Results were classified as confirmed positive, false positive, and indeterminate.

During the 6-year surveillance period, 365 patients underwent 912 procedures involving HPC or other cell-based transfusion. True positive microbial contamination was found in five of 663 (0.8%) peripheral blood and two of 34 (5.9%) marrow preparations (p = 0.04), while no contamination was found in 118 preparations from other sources. True-positive microbial contaminants included coagulase-negative staphylococci in autologous HPC products derived from peripheral blood from two patients with asymptomatic central venous catheter infections at time of apheresis and Propionibacterium acnes in one apheresis and two marrow products. Organism loads were low in all cases (≤500 colony-forming units/mL), and no adverse sequelae occurred in four patients that received contaminated products.

The incidence of microbial contamination of progenitor cell products in our institution over a 6-year period was low (0.8% overall), with contaminants originating from infected central venous catheters or from skin flora. All contaminants were bacterial species of low virulence, present in low titers and, if transfused, did not result in adverse reactions.

RHD nucleotide sequences were determined from genomic DNA. D epitope patterns were established with commercial monoclonal anti-D panels.

DIV comprises several variants of the D antigen with distinct serology, molecular structures, evolutionary origins, and ethnic prevalences. The DIV phenotype is determined by 350H shared by all, but not limited to, DIV variants which are further divided into DIVa and DIVb. The DIVa phenotype is expressed by DIV Type 1.0 harboring 350H and the dispersed amino acids 62F, 137V, and 152T. The DIVb phenotype is expressed by DIV Type 3 to Type 5 representing RHD-CE-D hybrids. Four of the six postulated DIV variants were encountered among 23 DIV samples analyzed. Of 12 DIV carriers with anti-D, 10 were female and seven likely immunized by pregnancy. Two DIV-related alleles are newly described: DWN, which differs from DIV Type 4 by 350D and epitope pattern. DNT carries 152T, known to cause a large D antigen density.

DIV alleles arose from at least two independent evolutionary events. DIV Type 1.0 with DIVa phenotype belongs to the oldest extant human RHD alleles. DIV Type 2 to Type 5 with DIVb phenotype arose from more recent gene conversions. Anti-D immunization, especially dreaded in pregnancies, will be avoided not only in carriers of DVI but also in carriers of other D variants like DIV, if our proposed D typing strategy is adopted.

This is a retrospective analysis using data on donation testing for TTIs (2005-2010) from the national blood service donor database and data on postdonation interviews with TTI-positive donors (2008-2010) from a risk factor database incorporating responses to standardized interview questions. The study measured the prevalence and incidence of TTIs in Australia and assessed their time trends. Multivariate analysis of time trends was conducted using Poisson regression models.

Overall, the prevalence and incidence of TTIs in 2005 to 2010 remained low and steady. The prevalence of hepatitis C virus decreased (rate ratio [RR], 0.93; 95% confidence interval [CI], 0.89-0.97) and the prevalence of active syphilis increased (RR, 1.51; 95% CI, 1.15-1.99) significantly during the study period. Prevalence of TTIs among Australian blood donors was substantially lower than that in the general population and no unique risk factors were identified in test-positive blood donors when compared with the general population.

Both the prevalence and the incidence of TTIs in Australian blood donors remained low, with a steady or declining trend for most infections except active syphilis. The lower prevalence of TTIs in blood donors compared with the general population reflects the effectiveness of donor education and donor selection measures in Australia.

SAGM RBCs, prepared from WB after overnight RTH (n = 10), were compared to SAGM RBCs prepared using an apheresis device (Alyx, n = 10). As a control, SAGM RBCs were also prepared within 2 hours of WB collection (2-hr WB, n = 10). All RBCs were stored at 4°C for 42 days with weekly assay of in vitro variables, cytokines and/or chemokines, and neutrophil activation after incubation with RBC supernatant.

RTH WB RBCs exhibited decreased levels of 2,3-diphosphoglycerate acid (2.3 μmol/g hemoglobin [Hb] ± 2.1 vs. 13.7 ± 1.3 μmol/g Hb) and morphology (160 ± 10 vs. 192 ± 5) on Day 1 and increased hemolysis (0.45 ± 0.21% vs. 0.31 ± 0.09%) and microparticles (6.1 ± 2.8/10³ RBCs vs. 3.9 ± 1.1/10³ RBCs) on Day 42 compared to apheresis RBCs. Gro-α and ENA-78 cytokine levels were significantly higher in RTH WB than Alyx RBCs during storage. CD11b expression was highest in neutrophils exposed to supernatant from RTH WB RBCs (p < 0.05).

RBC preparation method has a meaningful effect on the RBC storage lesion, which should be taken into account in addition to length of storage.

A total of 54,267 whole blood (WB) donors who donated between January 1 and March 31, 2008, at the five blood centers in China were followed for 2.5 years. Logistic regression was conducted to identify factors associated with their return behavior. A recurrent-event Cox proportional-hazard model was used to evaluate the overall effect of demographic variables and return behavior among first-time donors.

Donors with previous donation history were more likely to return and the number of previous returns was positively associated with future return (odds ratios, 3.31, 4.82, and 8.16 for one, two to three, and more than three times compared to none). Thirty-four percent of donors (first-time donor, 21%; repeat donor, 54%) made at least one return donation, with 14% returning in the first 9 months. The multivariable logistic regression model for all WB donors and the Cox proportional hazard model for first-time donors showed consistent predictors for return: female sex, older age (≥25 years), larger volume (300 or 400 mL), and donating in satellite collection site.

Encouraging first-time donors to make multiple donations is important for keeping adequate blood supply. The finding that first-time and repeat donors shared the same predictors for return indicates that retention strategies on repeat donors may be effective on first-time donors. Studies on motivators and barriers to return are needed, so that successful retention strategies can be tailored.

We retrospectively analyzed 29 patients who underwent TA from 2007 to 2011. Fifteen of the patients underwent TA procedure with VR and 14 of the patients underwent TA procedure without VR.

WBC reduction was significantly higher in patients who underwent TA with VR (p < 0.001). Early mortality rate was significantly lower in leukemia patients who underwent TA with VR than in patients who underwent TA without VR (p < 0.01); early mortality rates were 6.7% for 7-day and 13.8% for 100-day survivals. The mortality rates in the TA without VR group, however, were 42.9 and 71.4% for 7- and 100-day survivals, respectively.

Decreased early mortality rate in TA with VR group may be associated with prompt reduction of WBCs achieved with TA with VR and may also be associated with removal of the cytokines related to leukostasis. TA with VR would give more time for induction chemotherapy and increased overall survival rate.

In this case series, we describe ITP patients from our practice who achieved durable responses to the TRAs romiplostim and eltrombopag. Patients were classified as having a definite TRA-induced remission if PLT counts increased above 100 × 10⁹/L after TRA treatment and remained above 100 × 10⁹/L even after the medication was discontinued; or a possible TRA-induced remission if PLT counts increased above 100 × 10⁹/L, remained elevated for at least 3 months after the medication was discontinued, but a subsequent relapse occurred or the effect of other disease-modifying therapies could not be excluded.

Of 31 patients with chronic ITP treated with TRAs in our practice, nine patients achieved a PLT count response with either romiplostim (n = 6) or eltrombopag (n = 3) that was maintained even after the medications were discontinued. Three patients met criteria for a definite TRA-induced remission, each after exposure to romiplostim. Patients had ITP for a median of 7.8 years and had failed a median of four prior therapies including eight patients who had a splenectomy. We documented a progressive decline in anti-glycoprotein IIbIIIa PLT autoantibodies in one patient while on treatment.

Some patients with ITP can achieve sustained PLT count responses after the use of TRAs. This observation raises the possibility that these agents may restore immune tolerance to PLT antigens in some patients and supports the practice of down titrating the dose.

A 65-year-old woman with liver and renal insufficiency was transferred to our hospital for liver transplant evaluation. Two days after a 5-day course of ceftriaxone, her PLT count declined from a stable baseline of approximately 70 × 10⁹/L to a value of 3 × 10⁹/L, with coincident onset of mucocutaneous purpura. Her PLT count remained in the 1 × 10⁹ to 6 × 10⁹/L range until her death 13 days later, despite intravenous immune globulin, steroids, and PLT transfusions. The persistently low PLT count impeded central catheter placement for hemodialysis and possible therapeutic plasmapheresis. A strong ceftriaxone-dependent, PLT-reactive antibody was identified in a sample drawn 7 days after ceftriaxone was last administered, and ceftriaxone remained detectable in her serum for at least 8 days after the last dose.

A ceftriaxone-dependent, PLT-reactive antibody was responsible for the persistent thrombocytopenia in this patient. Although DIT is generally expected to improve within a few days of drug discontinuation, impaired drug clearance can significantly alter the outcome. This case highlights the importance of altered drug metabolism and clearance in critically ill patients, especially those with combined hepatic and renal dysfunction. DIT should be strongly suspected in patients with acute thrombocytopenia, and all treatment options to reduce serum drug levels should be seriously considered.

In a prospective survey we established the ethical acceptability of collection, research, and therapy with UCB HSCs versus hESCs among health care professionals, pregnant women, patients undergoing in vitro fertilization therapy, parents, and HSC donors and recipients in Switzerland.

There was overall agreement about an ethical justification for the collection of UCB for research and therapy in the majority of participants (82%). In contrast, research and therapy with hESCs was acceptable only by a minority (38% of all responders). The collection of hESCs solely created for HSC collection purposes met overall with the lowest approval rates. Hematologists displayed among the participants the highest acceptance rates for the use of hESCs with 55% for collection, 63% for research, and 73% for therapy.

This is the first study assessing the perception of hESCs for research and therapy in comparison with UCB HSCs in different target groups that are exposed directly, indirectly, or not at all to stem cell–based medicine. Our study shows that the debate over the legitimacy of embryo-destructive transplantation medicine is far from over as particularly hESC research continues to present an ethical problem to an overwhelming majority among laypersons and even among health care professionals.

We studied the effect of donor and recipient sex on outcome of 9038 single-donor PLT transfusions.

Using standard corrected count increment or percent PLT recovery (PPR) calculations, male patients showed inferior recovery rates, irrespective of donor sex. Using an adjusted PPR, which takes into account differences in blood volume between males and females, neither donor nor recipient sex played any role in PLT recovery after transfusion. Similarly, the time to next PLT transfusion was unaffected by both donor and recipient sex. In a subgroup analysis of patients with graft-versus-host disease after allogeneic HSCT, male recipients of a female allograft—which may carry anti-H-Y T cells and antibodies—had significantly lower time to next PLT transfusion. However, this occurred after both male donor and female donor PLT transfusions, arguing against an involvement of alloreactivity against H-Y antigens on PLTs.

This large analysis found no evidence that donor-recipient sex matching influences the outcome of PLT transfusion.

During the first 6 months of the program, 26,415 donors were screened using a single random plasma glucose (RPG) level. All donors were asked to eat before donation. Low-, moderate-, and high-risk groups were formed based on RPG levels (<140, 140-200, and >200 mg/dL). Contact with a telephone questionnaire was made with 139 of 178 (78%) of the persons in the high-risk group with 33 new cases of diabetes diagnosed by the donor's physician and 26 donors indicating that they were not diagnosed with diabetes. Sex- and age-matched donors in the low- and moderate-risk groups were contacted and administered the same questionnaire.

The three risk groups were similar, except for body mass index (28.1 ± 5.4 kg/m² vs. 29.9 ± 5.5 kg/m² vs. 32.7 ± 5.6 kg/m², p < 0.001). The discriminative effectiveness of screening was evaluated by the area under the receiver operating characteristics (AROC) curve. The AROC curve was 0.950 (95% confidence interval, 0.920-0.979) for the identification of diabetes. Using a RPG cutoff of 200 mg/dL, sensitivity was 100%, specificity was 82%, and positive predictive value was 56%. Cost analyses showed that the mean cost to screen, per donor, was less than $1. Cost per case identified was estimated to be less than $500 for a RPG cutoff of 200 mg/dL.

Screening during the blood donation process appears to be accurate, convenient, and inexpensive.

This retrospective study analyzed 38,436 whole blood donations made by young (aged 16 to 18 years) first-time donors. The effect of collection volume on vasovagal reaction was compared among different weight subgroups for both sexes by chi-square test.

For females in all weight subgroups and two of the male lower-weight subgroups, the reduction percentages ranged from 35% to 58% (p < 0.05). It was also noted that, among the females, a higher weight was associated with a higher percent reduction in the reaction rate.

With reduced collection volume, this study detected large and significant reduction in reaction rates among all females, as well as lower-weight males.

BALB/cJ mice were transfused with either untreated or riboflavin and UVB–treated C57Bl/6J platelet-rich plasma (PRP) containing WBCs. Circulating alloantibody and allospecific splenocyte cytokine responses were measured.

Pathogen reduction of allogeneic WBC-enriched PRP using riboflavin and UVB light before transfusion prevented alloimmunization, with a loss of both alloantibody generation and priming of secondary cytokine responses ex vivo. When mice given treated transfusions were subsequently given untreated transfusions, they produced normal levels of alloantibodies but had reduced secondary cytokine responses ex vivo. This immune modulation was antigen specific and was dependent on the presence of WBCs in the treated product.

UVB plus riboflavin treatment of WBC-enriched PRP effectively blocks alloimmunization and modulates immune responses to subsequent exposures.

This retrospective cohort study included patients undergoing low-to-moderate–risk cardiac surgery with cardiopulmonary bypass without (control; n = 531) or with cell salvage (n = 433; Autolog, Medtronic). Study endpoints, including 24-hour blood loss and RBC requirements, were evaluated using adjusted logistic regression.

Baseline characteristics were similar between groups. The cell saver group received 568 ± 267 mL of autologous blood. Median number of allogeneic RBC transfusions was higher in the control group (2 [1-5]) compared with the cell salvage group (1 [0-3]; p < 0.001). There were no clinically relevant differences in postoperative coagulation test results between groups. The relative risk (RR) for postoperative RBC transfusion was reduced to 0.76 (95% confidence interval [CI], 0.70-0.83; p < 0.0001) in the cell salvage group. Moreover, patients in the cell salvage group had a lower chance for myocardial infarction (RR, 0.26; 95% CI, 0.08-0.91; p = 0.035), whereas the cell salvage group was associated with a higher probability for intensive care discharge within 24 hours after surgery (RR, 1.08; 95% CI, 1.02-1.14; p = 0.009).

The use of cell salvage throughout the entire procedure reduces postoperative blood loss and allogeneic RBC transfusion. These findings advocate implementation of cell salvage in all patients undergoing on-pump cardiac surgery, irrespective of anticipated surgery-related blood loss.

Blood samples from consenting donors in southeastern CT were collected from mid-August through early October 2009 and tested by IFA for immunoglobulin G antibodies and real-time polymerase chain reaction (PCR) for B. microti DNA. IFA specificity was determined using blood donor samples collected in northwestern Vermont (VT), an area nonendemic for Babesia.

Of 1002 CT donors, 25 (2.5%) were IFA positive and three (0.3%) were real-time PCR positive. Among the three real-time PCR–positive donors, two were also IFA positive, while one was IFA negative and may represent a window period infection. The two IFA- and real-time PCR–positive donors appeared to subsequently clear infection. The other real-time PCR–positive donor did not provide follow-up samples. Of 1015 VT donors tested by IFA, only one (0.1%) was positive, but may have acquired infection during travel to an endemic area.

We prospectively identified several real-time PCR–positive blood donors, including an IFA-negative real-time PCR–positive donor, in an area highly endemic for B. microti. These results suggest the need to include nucleic acid testing in planned mitigation strategies for B. microti.

Viral loads were assessed by bDNA and RT-PCR assays and if below 100 copies (cps)/mL, by Ultrio limiting dilution probit analysis. The probability of virus transmission by WP and EC donations was estimated for different levels of the 50% minimum infectious dose (ID₅₀) using Poisson distribution statistics.

The equal distribution of WP donations plotted by log HIV-RNA levels indicated a random appearance of donors in the ramp-up phase. The HIV p24 Ag assay detected 45% of WP samples and the cutoff crossing point was estimated at 8140 (bDNA)/22,710 (RT-PCR) cps/mL. On replicate retesting of 40 HIV p24 Ag–negative ID-NAT WP-yield samples Ultrio minipool (MP)8, Ultrio-Plus MP8, and TaqScreen MP6 detected 79, 81, and 78%, respectively. Modeling with an estimated ID₅₀ of 31.6 virions/RBC indicated that 15% of p24 Ag–negative ID-NAT WP-yield donations would have transmitted HIV if MP6-8 NAT had been used. Only 2% of RBC transfusions from ECs are estimated to be infectious with a worst-case ID₅₀ estimate of 316 virions.

Our analysis of viremia and infectivity of WP and EC donations enables comparison of the efficacy of NAT options in preventing HIV transmission risk.

We performed a retrospective cohort study of neonates who received, in the first month after birth, between January 1, 2007, and December 31, 2008, at least one PLT transfusion. Seventy-four neonates who received 197 volume-reduced PLTs transfusions, 68 neonates who received 105 PASII PLT transfusions, and eight neonates who received eight plasma PLT transfusions were analyzed. Early (within 8 hr after transfusion) and follow-up count increments (16-24 hr after transfusion) were evaluated for 191 and 81 volume-reduced PLTs, 77 and 56 PASII PLTs, and six and five plasma PLT transfusions, respectively, using a random-effects model.

Volume-reduced PLTs were transfused at twice the dose in one-fifth the volume of PASII and plasma PLTs. The early posttransfusion count increment was higher for volume-reduced PLTs at 111 × 10⁹/L (95% confidence interval [CI], 86-135) compared to PASII PLTs at 62 × 10⁹/L (95% CI, 40-84; p = 0.000) and plasma PLTs at 47 × 10⁹/L (95% CI, 14-79). The follow-up count increment was also higher for volume-reduced PLTs at 60 × 10⁹/L (95% CI, 19-100) compared to PASII PLTs at 38 × 10⁹/L (95% CI, −0.2 to 77; p = 0.082) and plasma PLTs at 4 × 10⁹/L (95% CI, −38 to 46).

Neonates who received twice the PLT dose by volume-reduced PLTs had twice as high early and follow-up count increment showing similar efficacy of products.

Questions about previous blood donation and interest in future donation were added to the National HIV Behavioral Surveillance survey questionnaire, which periodically gathers risk behavior information from MSM in San Francisco.

Overall, 77.3% of 475 MSM respondents expressed interest in donating. By lower-risk criteria, 10.1% had no sexual contact in the past 6 months (2.3% in the past 12 months) and 1.9% had only lower-risk sexual contact in the past 6 months (1.5% in the past 12 months). Of the 23.4% who answered yes to having donated in the past, at least 25.2% did not comply with the current deferral of no male–male sex since 1977.

The majority of MSM are interested in donating blood. Depending on how the policy would be changed (i.e., either a temporary or a behavior-based deferral criterion), substantial numbers of MSM would be eligible.

Samples from 1019 whole blood donors were tested for anti-HEV immunoglobulin (Ig)G by enzyme-linked immunosorbent assay and Western blot. The incidence of HEV and presence of HEV RNA in donors who seroconverted were determined by testing archive samples and recipients of viremic donations were traced. Anti-HEV IgM and alanine transaminase (ALT) testing were also performed to assess the value of such measures in the prevention of TT-HEV infections.

A total of 69 of 1019 donors tested positive for anti-HEV IgG (6.8% seroprevalence), and seroconversion for anti-HEV IgG occurred in seven of 69 donors within 2 years (incidence, 0.35%/year). Three of seven (42.8%) seroconverting donors provided an archive sample in which HEV RNA was detectable. One recipient of these donations was traceable; anti-HEV IgG, IgM, and HEV RNA testing were negative 41 days after transfusion. Neither ALT levels nor anti-HEV IgM detection correlated with the presence of HEV RNA.

The seroprevalence of HEV was 6.8%, and the annual incidence 0.35%. HEV RNA was detectable in several seroconverting donors, without evidence for HEV transmission in the only traceable recipient. Since neither ALT nor anti-HEV IgM testing correlate with the presence of HEV RNA, HEV nucleic acid testing currently provides the only method for the prevention of TT-HEV infection. However, before implementation, more data about clinical relevance of TT-HEV infections and infectious dose of HEV are required.

Blood hemoglobin (Hb) was measured upon admission, at venipuncture, and immediately after collecting 400 mL of whole blood from 736 donors. Shifted fluid volume was calculated using a formula that included Hb levels and estimated total blood volume.

By the end of blood collection, 188 ± 80 and 211 ± 82 mL of fluid, which is equivalent to almost half of the total amount of withdrawn blood, had entered the intravascular space in male and female donors, respectively. The difference between the sexes was significant despite the lower body weight and circulating blood volume of the female donors. Body weight increased, whereas age decreased the volume of shifted fluid in female donors.

Blood loss after donation is quickly compensated by an interstitial fluid shift into the intravascular space and may not be the only direct cause of VVR in the setting of a whole blood donation of 400 mL.

Sixty patients undergoing posterior spine surgery were enrolled in a prospective, randomized trial comparing intraoperative blood loss using unipolar cautery alone or with use of a bipolar tissue sealant device. Hb concentration and fluid volume were measured from all surgical sponges, suction canisters, and the cell salvage device. Using the volume and concentration of Hb from each solution allowed calculation of Hb mass, which was converted into volume of blood lost and compared with estimates of blood loss documented by the anesthesia team. A single-sample t test of no difference was used to compare estimated with measured blood loss.

Mean estimated blood loss exceeded measured blood loss by 246 mL (860 mL vs. 614 mL, p < 0.0001).

Estimated blood loss exceeded measured blood loss by 40% on average. The likely etiology of this discrepancy relates to the inability to visually determine Hb concentration of sanguineous solutions in suction canisters and surgical sponges. Ramifications of excessive bleeding estimates include unnecessary transfusion and overadministration of intravenous fluids, both of which may have deleterious effects.

Hemagglutination of RBCs from two individuals known to express a single copy of functional ABCG2 were compared to RBCs from eight unrelated, previously characterized, Jr(a+^W/–) donors. Standard polymerase chain reaction–based methods were used to characterize ABCG2 alleles.

Two monoclonal anti-Jr^a clones agglutinated RBCs from the eight Jr(a+^W/–) study subjects. Two of these subjects were homozygous for a missense ABCG2 change (c.1858A; Asp620Asn). Two were heterozygous for two missense changes; one was c.1858G>A and c.421C>A (Asp620Asn; Gln141Lys), and the other was c.1714A>C and c.421C>A (Ser572Arg; Gln141Lys). The remaining four subjects were heterozygous for c.421C>A (Gln141Lys), and for one of four null alleles.

We have identified three ABCG2 alleles that are newly associated with weakened Jr^a expression. One of these is novel, the missense allele c.1714A>C (Ser572Arg) and two that have been previously described c.421C>A (rs2231142; Gln141Lys) and c.1858G>A (rs34783571; Asp620Asn). In addition, we found a novel, presumed null allele, c.1017_1019delCTC (Ser340del).

Four aerobic bacteria (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mitis, and Bacillus cereus), five anaerobic bacteria (Fusobacterium necrophorum, Clostridium perfringens, Bacteroides fragilis, Prevotella loescheii, and Propionibacterium acnes), and one fungus (Candida albicans) were spiked into apheresis HPC products at concentrations of 10, 10², 10³, and 10⁴ colony-forming units (CFUs)/mL. Aerobic and anaerobic bottles were incubated until positive or for up to 5 days. DNA was simultaneously extracted for polymerase chain reaction amplification of 16S ribosomal RNA (rRNA) gene.

All aerobic bacteria grew in both bottles at all concentrations tested within 24 hours, and the time to positivity (TTP) was significantly shorter with aerobic bottles. C. albicans grew in the aerobic media at all concentrations within 30 hours. Anaerobes grew in the anaerobic bottle at all concentrations within 5 days. No bacteria were detected by using 16S rRNA gene amplification at 10⁴ CFUs/mL.

Compared to culture, 16S rRNA gene amplification of HPCs does not improve sensitivity or turnaround time for HPC sterility testing. The VersaTREK system is a reliable tool for detecting microbial contamination of apheresis HPC products with a limit of detection of less than or equal to 10 CFUs/mL. Inclusion of both the aerobic and the anaerobic culture bottles achieves the shortest TTP for all species tested.

We fractionated UCMCs by physicochemical charge-based methods. Avoiding Miltenyi particles and HESPAN yielded 30 x 10⁶ to 54 x 10⁶ UCMCs/20 mL UCB by Ficoll-Isopaque for serial depletion fractionation, using nylon wool column (NWC) or direct rosetting with sheep red blood cells (SRBCs) without incubation in the cold.

CD34+ cell yields (approx. 5.12%) were 39 times greater than 0.13% (Korean study, 11,098 UCB units) and 10 to 20 times greater than 0.25% to 0.3% harvested by anti-CD34 Miltenyi particles. SRBC depletion of most high-specific-gravity T cells achieved considerable enrichment of CD34+ and BY55+ cells. Using NWC achieved 2.5-fold enrichment of CD34+ cells and twofold enrichment of BY55+ cells. Direct SRBC rosetting provided better or higher enrichment of CD34+ cells. Overall CD34+ cell yield in low-density fraction was more than twice after direct rosetting (38% vs. 16%) in contrast to separation by NWC followed by SRBC rosetting. CD3+ cell yields (by three CD markers) were approximately 8.83%, far below approximately 30 x 10⁷/kg considered acceptable to avoid graft-versus-host disease. Natural killer cell yields (CD16+/CD56+ and BY55+/ CD160+) are in perfect agreement.

Achieving approximately 5% CD34+ cell yields from single UCB donations, a major advance, holds great promise for CD34+ cell therapy of adults and larger children, and cheaper cultured RBC manufacture.

Patients with GVHD and LTR receiving ECP with either UVAR XTS or CELLEX (Therakos) were prospectively recruited for this study. A complete cell count with differential was performed on preprocedural peripheral blood and samples from the collected buffy coats. Correlation analysis and linear regression were performed between cell counts in peripheral blood and buffy coat. Collection efficiency was compared between UVAR XTS and CELLEX.

In all 52 patients, lymphocyte counts in buffy coat and peripheral blood showed strong correlation (r values were 0.85 and 0.983 for UVAR XTS and CELLEX, respectively; p < 0.001) with slopes of 2 and 5.1 for UVAR XTS and CELLEX, respectively (p < 0.001). The quantitative relationship remained robust in patients stratified by diagnoses. Monocytes also showed consistent correlation and linearity, but not neutrophils or combined white blood cells, red blood cells, or platelets. CELLEX enriched approximately twice as many lymphocytes and monocytes than UVAR XTS per procedure (p < 0.001).

The preprocedural peripheral lymphocyte count can predict the number of lymphocytes within the buffy coat collected during ECP, which may justify the use of peripheral lymphocyte count as a surrogate for the cell dose treated per procedure. Peripheral monocyte counts may serve as an alternative. CELLEX is more efficient in collecting lymphocytes and monocytes than UVAR XTS under conditions tested.

Patient serum was studied for antibodies that recognize protamine, heparin-protamine complexes, and platelets (PLTs) treated with protamine using flow cytometry, enzyme-linked immunosorbent assay, and serotonin release from labeled PLTs.

A high-titer immunoglobulin G antibody was detected in patient serum that recognizes protamine in a complex with heparin or PLT surface glycosaminoglycans (GAGs) and activates PLTs treated with protamine at concentrations achieved in vivo after protamine infusion. The antibody is distinctly different from those found in patients with heparin-induced thrombocytopenia on the basis of its failure to recognize heparin in a complex with PLT factor 4 (PF4) and to release serotonin from labeled PLTs in the absence of protamine.

Findings made suggest that the patient's antibody is specific for conformational changes induced in protamine when it reacts with heparin or a PLT surface GAG. Development of severe thrombocytopenia after treatment of this patient with protamine defines a previously undescribed mechanism of drug-induced immune thrombocytopenia. Patients given protamine who produce this type of antibody may be at risk of experiencing thrombocytopenia if given the drug a second time while antibody is still present.

RBCs were harvested from FVB.HOD mice that express an RBC-specific model transgene (HOD) and stored for 14 days with either ascorbic acid in saline or saline alone. Twenty-four-hour posttransfusion recovery of RBCs was tracked by flow cytometry. Alloimmunization was monitored by flow cytometry crossmatch. Cytokines were monitored by multiplex bead arrays.

RBCs stored under standard conditions had decreased 24-hour posttransfusion recovery and increased induction of both alloantibodies and interleukin (IL)-6 and monocyte chemoattractant protein (MCP)-1 secretion in the mouse recipients. Addition of ascorbic acid from 3.6 to 10.8 mmol/L resulted in a significant decrease in microparticle formation, an improved RBC 24-hour posttransfusion recovery (p < 0.01), and a decrease in recipient alloimmunization (p = 0.0001). Induction of MCP-1 and IL-6 secretion was not decreased by ascorbic acid.

These data indicate that the addition of ascorbic acid solution to RBCs during storage has a beneficial effect on recovery and immunogenicity of RBCs, but not cytokine induction. The addition of ascorbic acid (or other antioxidants) to human RBCs may have beneficial effects.

A standard lookback was conducted on untested cellular blood components from donors later confirmed to be HTLV positive, for the 4 to 5 years before 2002, and on the last tested negative donation from donors who had seroconverted.

A total of 437 red blood cell and platelet components were included and an outcome was reported for 84% of these. Just over half of identified recipients were dead at the time of lookback; blood samples for testing were obtained from 77% of identified living recipients. HTLV infection was confirmed in seven recipients, but one was discounted as not transfusion transmitted.

Although numbers are small, our results provide evidence of the efficacy of leukoreduction in reducing the likelihood of HTLV transmission through transfusion of cellular blood components. The HTLV-positive rate in recipients of leukoreduced components was 3.7%, a reduction of 93% compared with nonleukoreduced components. Importantly, the one infected recipient of a leukoreduced component had existing risk factors for HTLV infection. HTLV lookback was much less efficient in identifying infected recipients than was hepatitis virus C lookback.

We encapsulated the human cord blood mononuclear cells (CBMCs) in alginate 3D static and rotating culture systems, compared the cell number amplification, the proportion of CD34+ cells, and the colony-forming capacity of these systems to those of the conventional two-dimensional (2D) system. The amplified cells were transplanted into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice to confirm the hematopoiesis reconstruction capacity of the cells.

The increase in the cell number and the proportion of CD34+ cells in the CBMCs was more effective in these 3D alginate culture systems than in the conventional 2D culture system under the same conditions (p < 0.05). The stem cell maintenance capability was confirmed by flow cytometry and colony-forming assay ex vivo and NOD/SCID mice xenogeneic transplantation model in vivo.

Our results demonstrated that these 3D alginate culture systems are an efficient way to amplify cord blood HSCs for extended periods without having them lose their self-renewal capacity in vivo. These novel 3D alginate culture systems are promising for the amplification of UCB-derived HSCs for clinical application in the future.

Blood was drawn from 45 human immunodeficiency virus–positive men either with Kaposi's sarcoma (KS; n = 21) or without KS (n = 24) and subject to leukoreduction filtration. HHV-8 viral load was measured in plasma and in blood before and after filtration.

Twelve subjects, all with KS, had detectable HHV-8 viremia before filtration with viral loads of 10² to 10⁵ copies/mL (mean, 3 × 10⁴ copies/mL). After filtration, seven of 12 subjects no longer had detectable HHV-8 in their blood, and five of 12 subjects had detectable HHV-8 that was 90% reduced on average from prefiltration levels. The presence of HHV-8 in the blood after filtration was strongly associated with prefiltration viral loads greater than 1000 copies/mL and the presence of cell-free virus in plasma. None of the subjects without KS had detectable levels of HHV-8 virus in blood before or after filtration.

Cell-associated HHV-8 appeared to be effectively removed by leukoreduction filtration. Cell-free HHV-8 was present in 42% of subjects as 1% to 20% of the total virus which was not removed by filtration.

A 31-year-old US Army soldier tested positive for HTLV-I 44 days after receipt of emergency blood transfusions for severe improvised explosive device blast injuries. One donor's unit tested HTLV-I positive on the DoD-mandated retrospective testing. Both the donor and the recipient tested reactive with enzyme immunoassay and supplemental confirmation by HTLV-I Western blot. The donor and recipient reported no major risk factors for HTLV-I. Phylogenetic analysis of HTLV-I sequences indicated Cosmopolitan subtype, Subgroup B infections. Comparison of long terminal repeat and env sequences revealed molecular genetic linkage of the viruses from the donor and recipient.

This case is the first report of transfusion transmission of HTLV-I in the US military during combat operations. The emergency fresh whole blood policy enabled both the donor and the recipient to be notified of their HTLV-I infection. While difficult in combat, predonation screening of potential emergency blood donors with Food and Drug Administration–mandated infectious disease testing as stated by the DoD Health Affairs policy should be the goal of every facility engaged with emergency blood collection in theater.

Fifty blood donor and patient samples with discrepant results of D phenotyping were subjected to routine serology to define the D phenotype including monoclonal anti-D immunoglobulin M and indirect antiglobulin test. Commercially available panels of monoclonal anti-D were used for identification of partial D and weak D phenotypes. Genomic DNA was evaluated using allele-specific amplification polymerase chain reaction with sequence-specific primers to define weak D type.

Molecular typing confirmed most of the serology results; three samples that were not clear-cut serologically were identified by molecular typing, two samples as weak D Type 4.2 (DAR), and one sample as weak D Type 4.0. Another two samples identified by serologic panel as weak D were unresolved by molecular typing. A sample with partial D Type II by serology revealed a Weak D Type 4.0 by molecular typing. Results interestingly showed the high frequency of weak D Type 4.2 (DAR) in Egypt.

RHD molecular typing can solve discrepancies during routine testing due to partial and weak D phenotypes for better transfusion outcome.

The pH of 13,693 PCs produced for transfusion was monitored daily using Blood Storage, Inc.'s pH sterile, automated fluoroscopic evaluation technology. Upon indication of compromised quality or expiration, PCs were returned and in vitro tests were performed.

A total of 998 PCs were returned, of which 962 outdated, 26 had a positive BacT/ALERT reaction, seven had aggregates, one was without swirl, one had low pH, and one had high pH. BacT/ALERT was faster in identifying bacterial contamination than pH measurements. The pH at the end of the storage period was significantly lower than at the beginning. In vitro tests indicated that while the PC quality was acceptable upon expiration, it rapidly declined after expiration.

In this setting where the vast majority of PCs were of good quality and within acceptable pH limits, daily, noninvasive routine pH measurement has limited added value in identifying quality-compromised PCs.

Frozen Babesia-spiked whole blood was microcentrifuged, and the supernatant transferred and microcentrifuged again to concentrate the parasite. The DNA was extracted and amplified using real-time polymerase chain reaction (PCR) using Babesia-specific primers. The assay was employed in three series of experiments: 1) a validation and optimization spiking experiment, 2) a blinded serial dilution probit analysis to determine the limit of detection, and 3) evaluation of two blinded panels of clinical samples from possible babesiosis cases.

At a decreasing inoculum of 445, 44.5, and 4.45 copies/mL, the assay had positive rates of 100, 97.5, and 81%, respectively. The blinded probit analysis demonstrated a detection rate of 95 and 50% at 12.92 and 1.52 parasites/2 mL of whole blood, respectively. Evaluation of clinical samples showed 13 of 21 samples to be positive, with a range of 85 to 4.8 million parasites/mL. There were no positives detected among 48 healthy donors

We have developed a highly sensitive and specific, quantitative real-time PCR-based assay for detection of B. microti that could have a useful role in blood screening. It can also be employed broadly to understand Babesia epidemiology, disease pathogenesis, and host immunology.

A Markov-based decision tree was constructed to compare ATR rates and associated costs expected from current practice and from alternative strategies of using APs stored in PAS. The potential use of pretransfusion medication was also incorporated. Using a hospital perspective and including direct medical expenses only (US$2012), Monte Carlo microsimulations were run to evaluate outcomes under a base-case analysis. One-way and probabilistic sensitivity analyses were used to assess outcome uncertainty.

Under base-case variables, using APs stored in PAS for all patients as an initial transfusion protocol is expected to avert ATRs and associated costs, compared to current practice. Using PAS for all patients along with pretransfusion medication would be cost-saving only when the additional cost of PAS is below $9.14. If PAS storage could eliminate pretransfusion medication use, it is expected to result in cost savings when the additional unit cost of PAS is under $11.90. At a PAS cost of $15, averting one ATR would cost $701.95. Using PAS storage only in response to recurring mild ATRs is associated with cost savings under all costs of PAS evaluated.

Using PAS storage for all AP transfusions to prevent ATRs may be financially and clinically beneficial, compared to current practice.

We describe an investigation to determine possible common sources of infection among donors or factors associated with PLT collection or storage and to determine whether human transfusion-associated listeriosis cases had been reported. We also reviewed all isolates with PLTs as a source sent to the CDC National Listeria Reference Laboratory between November 1, 2005, and December 31, 2011.

Each PLT donor–associated isolate had a distinct pulsed-field gel electrophoresis pattern combination. Other than these four cases, no other cases of Lm-contaminated PLTs were identified by the American Red Cross or by CDC during 2005. However, two additional cases of Lm isolated from donated PLTs were detected, one in 2008 and one in 2011.

Although the source of contamination for these PLT units is unclear, and a source common to all units was not identified, this investigation underscores the value of screening for bacterial contaminants of PLTs.

Recipients of previous donations from OBI donors were investigated through lookback (systematic retrieval of recipients) or traceback (triggered by clinical cases). Serologic and genomic studies were undertaken on consenting donors and recipients. Multiple variables potentially affecting infectivity were examined.

A total of 45 of 105 (42.9%) donor-recipients pairs carried antibodies to HBV core (anti-HBc) as evidence of previous HBV infection. Subtracting 15% of anti-HBc population background, the adjusted transmission rate was 28%. Anti-HBc prevalence increased to 28 of 44 (63.8%) in unvaccinated recipients receiving anti-HBs–negative OBI blood products. In contrast, four of 26 (15.4%) recipients of anti-HBs–positive products were anti-HBc positive. Transmission with anti-HBs–negative products depended on volume of plasma transfused (85%-100% with 200 mL of fresh frozen plasma [FFP], 51% with 50 mL in platelet concentrates [PCs], and 24% with 20 mL in red blood cells [RBCs], p < 0.0001 FFP vs. RBCs). The 50% minimum infectious dose of OBI HBV DNA was estimated at 1049 (117-3441) copies. Donor and recipient strains sequence homology of at least 99% confirmed transfusion-transmitted infection in 10 cases and excluded it in one case.

Blood products from donors with OBI carry a high risk of HBV transmission by transfusion. This risk is dependent on presence of anti-HBs and viral dose. This may justify safety measures such as anti-HBc and HBV nucleic acid test screening depending on epidemiology.

A 34-year-old female patient developed massive hemolysis during infusion of 50 mL of iomeprol. Serologic studies were performed using standard techniques.

Before hemolysis, the patient's serum was weakly positive with e+ red blood cells (RBCs; autoanti-e) and the direct antiglobulin test (DAT) was negative. After hemolysis, the patient's serum samples became significantly reactive with e– RBCs in the presence of iomeprol but not in the presence of two other similar CM. The DAT became strongly positive only with anti-C3d.

Initially, an allergic reaction was suggested, and as the hemolysis became obvious, a toxic hemolysis was suspected. However, serologic reexamination revealed an iomeprol-dependent antibody. IHA related to CM has yet only been described in one patient in 1991. The hemolysis in that patient was related to Isopaque, an older ionic CM. Here, we describe an additional patient and recommend that CM should be considered as a rare risk in the development of IHA.

Apheresis PLTs from pairs of ABO-identical male donors were pooled and divided into four volumes. One volume was stored without filtration, whereas the other three were filtered with different devices. PDMPs, LDMPs, and RDMPs were measured by flow cytometry during 2 weeks of controlled-temperature (22°C) agitated storage.

On average, PDMPs doubled over 5 days of storage, followed by a much steeper increase by which PDMPs on Day 14 were nearly 20 times higher than on Day 0. LDMP and RDMP counts were relatively stable over 14 days. Significant differences among filtered and nonfiltered products did not emerge.

Although the conditions of this study showed no favorable or unfavorable effects of three different filters on microparticle formation, surveillance and investigation of unanticipated outcomes in other experimental and clinical circumstances should continue.

Neutrophil-reactive antibodies were detected by agglutination, microscopic immunofluorescence, and monoclonal antibody (MoAb)-specific immobilization of neutrophil antigens (MAIGA) assay. For epitope mapping of FcγRIIIb-reactive antibodies, recombinant chimeric variants of FcγRIIIb were used in the MAIGA assay. Genotyping of FCGR3B was performed by allele-specific polymerase chain reaction.

Both mothers were typed FCGR3B*01+, *02–, *03+. Antibody screening revealed the presence of alloantibodies reactive with FcγRIIIb encoded by FCGR3B*02, but not with FcγRIIIb encoded by FCGR3B*03. MAIGA with recombinant, partly chimeric FcγRIIIb variants demonstrated that the antigen recognized by maternal antibodies is characterized by two amino acids, Ala78 and Asp82. Among the FCGR3B alleles, the sequence Ala78---Asn82 is exclusively encoded by FCGR3B *02.

A previously unrecognized second antigen, HNA-1d, is present on FcγRIIIb encoded by FCGR3B*02. This antigen is characterized by the sequence Ala78---Asn82. It appears that only individuals carrying the HNA-1c phenotype can form anti-HNA-1d alloantibodies. The HNA-1 system now consists of four antigens encoded by three alleles.

A retrospective study was conducted to examine the demographic and clinical outcome of this group of donors. All confirmed culture-positive cases under the BST were retrieved and those donors with S. bovis bacteremia were contacted for follow-up. Viable culture samples were sent for detailed microbiologic analysis.

From 1998 to 2012, a total of 16 donors were found to have S. bovis bacteremia, giving an estimated prevalence of 1 in 110,800 donations. They consisted of nine men and seven women with median age of 43.5 years. Eight donors had undergone colonoscopy with colonic carcinoma detected in two and benign adenoma in four. Five of the 16 isolates could be retrieved for 16S DNA sequencing. Four were identified as S. gallolyticus ssp. pasteurianus and one as S. gallolyticus ssp. gallolyticus. The two patients with colonic carcinoma had one each of subspecies pasteurianus and gallolyticus.

The findings highlight a close association of S. bovis and colonic carcinoma. We recommend prompt donor follow-up if S. bovis bacteremia is detected. Besides, all confirmed S. bovis should be fully characterized by molecular technique.

DMSO (5% final concentration) was added to buffy coat–derived PLTs, followed by centrifugation to concentrate and freezing at −80°C. Cryopreserved PLTs (n = 12 per group) were thawed at 37°C, reconstituted in a unit of thawed frozen plasma, SSP+, or PAS-G. In vitro PLT quality was examined before freezing, immediately after thawing, and 6 and 24 hours after thawing.

After thawing and reconstitution, PLTs in plasma and PAS-G displayed similar recovery (69 and 73%, respectively), while PLT recovery in SSP+ was lower (62%). All PLTs maintained an acceptable pH and metabolic activity during postthaw storage. Frozen PLTs were activated, although the extent differed depending on the reconstitution solution, with PLTs in PAS-G retaining better aggregation responses than PLTs in plasma or SSP+.

Thawing cryopreserved PLTs in PAS-G, without plasma supplementation, resulted in PLTs with similar recovery and in vitro quality indicators as those suspended in plasma. Importantly, using PAS-G enables the PLTs to be ready for use significantly faster than when having to thaw frozen plasma, which may be beneficial in trauma situations.

The deployed qualification of CBBs is based on retrospective and prospective review of the CBB.

Forty-one CBBs were evaluated retrospectively: seven CBBs were disqualified based on failed quality control (QC) results. Eight CBBs did not meet the criteria for retrospective qualification because fewer than 3 cord blood units were received and the CBB was not accredited. As of March 2012, three US and one non-US CBBs have been qualified prospectively. One CBB withdrew from the qualification process after successful completion of the comprehensive survey and subsequent failure of the provided QC unit to pass the minimum criteria. One CBB failed the prospective qualification process based on processing methods that were revealed during the paper portion of the evaluation.

A CBB qualification process is necessary for a transplant center to manage the qualification of the large number of CBBs needed to support a umbilical cord blood transplantation program. A transplant center that has utilized cord blood for a number of years before implementation of a qualification process should use a retrospective qualification process along with a prospective process.

Existing testing records of apheresis PLTs screened by PGD from July 2008 to April 2012 were reviewed. All PLT units were initially screened by routine postcollection culture methods. Secondary screening using PGD was performed for indated PLTs on PLT storage Day 4 and for outdated PLTs on Day 8.

A total of 8535 apheresis PLTs were available in inventory during the study period. Of these, 5030 (58.9%) were dispensed and transfused before PGD testing and 3505 (41.1%) underwent PGD testing on Day 4. Twenty-five units tested on Day 4 were PGD initial reactive (0.71%). All were confirmed to be false positive by repeat PGD testing in triplicate (n = 20) or by confirmatory culture (n = 5). An additional 364 units that were PGD nonreactive on Day 4 were approved for transfusion on Day 6 or Day 7 due to urgent clinical need. A total of 371 outdated units underwent repeat PGD testing before discard on Day 8; all were nonreactive.

Secondary PGD testing of culture-screened apheresis PLTs results in low yield in a medium-sized transfusion service. Use of PGD testing on Day 4 may allow for extension of the apheresis PLT shelf life to Day 7 for hospitals that face supply constraints.

Whole blood donors who were permanently disqualified because of a false-reactive test between 1990 and 2007 in the province of Quebec were compared to donors who remained eligible, matched for baseline characteristics. The incidence of CHD after entry into the study was determined through hospitalization and death records. We compared eligible and disqualified donors using an “intention-to-treat” framework.

Overall, 12,357 donors who were permanently disqualified were followed for 124,123 person-years of observation, plus 50,889 donors who remained eligible (516,823 person-years). On average, donors who remained eligible made 0.36 donation/year during follow-up and had an incidence of hospitalizations or deaths attributable to CHD of 3.60/1000 person-years, compared to 3.52 among permanently disqualified donors (rate ratio, 1.02; 95% confidence interval, 0.92-1.13).

Donors who remained eligible did not have a lower risk of CHD, compared to donors who were permanently disqualified due to a false-reactive TD marker. Because of the quasi-random nature of false-reactive screening tests, this natural experiment has a level of validity approaching that of a randomized trial evaluating the effect of regular blood donation on CHD risk. These results do not support the iron hypothesis.

In this study, microvesiculation and changes in the composition of the RBC membrane were investigated throughout 49 days of storage and were correlated with in vitro assays examining membrane quality. Leukoreduced RBC units produced using the buffy coat method were collected and stored at 1 to 6°C and were tested weekly for hemolysis, osmotic fragility, deformability, ATP, hematologic indices, and morphology. Microvesiculation was assessed using multicolor flow cytometry. High-performance liquid chromatography and mass spectrometry were used to determine the composition and quantity of phospholipids (PLs) and cholesterol (C) on Days 2 and 43.

The assessment of RBCs throughout storage revealed significant increases in percent hemolysis, while significant decreases in ATP concentrations, and the mean corpuscular hemoglobin concentration were observed. Flow cytometry analysis revealed a significant increase in the mean number of microparticles per microliter during storage. Throughout storage, significant decreases were identified in the amount of PLs and total lipids within the RBC membrane. No significant change in the amount of C in the RBC membrane was identified.

Significant changes to the RBC membrane occur during storage. The length of storage will influence RMP generation, osmotic fragility, hemolysis, and changes in deformability. These changes in RBC in vitro quality may contribute to transfusion reactions and negative posttransfusion outcomes.

Plasma and platelet (PLT) concentrates were inoculated with either P. falciparum– or P. yoelii–infected red blood cells (RBCs). Aliquots from each unit were collected after inoculation, after addition of riboflavin, and after treatment. In vitro P. falciparum growth was assessed using thin blood films of duplicate samples at 24, 48, 72, and 96 hours. P. yoelii parasitemia was followed in mice for 14 days postinoculation.

In the in vitro studies, the mean P. falciparum parasitemia increased 12- to 19-fold in pretreatment samples, both before and after addition of riboflavin, after 96-hour culture. Few parasites were observed in Mirasol-treated units at 24 hours; those that were observed were degenerating. Through the remainder of the 96-hour culture period, cultures of treated samples were negative. In the in vivo study, mouse plasma containing P. yoelii–infected RBCs had a mean starting titer of 4.6 log mouse infectious dose 50%/mL. No infectious parasite was detected in treated samples.

Treatment with riboflavin and UV light was effective at reducing viable P. falciparum in both PLT and plasma products by at least 3.2 logs. Additionally, an at least 4.4-log reduction was observed with P. yoelii.

Two experiments were performed: in the first one, volume reduction of the UCB units was carried out before analysis. In the second one, analysis was carried out with no previous manipulation. Samples were stored at room temperature and one aliquot was taken daily for analysis. We examined CD34+ cell, B-cell precursor, mature B and T lymphocyte, monocyte, granulocyte, and mesenchymal stem cell (MSCs) concentrations.

Thirty-six UCB units were analyzed. CD34+ cells and mature T lymphocytes increased (viability 99%). Mature B lymphocytes and MSCs decreased, maintaining viability. Granulocytes decreased with loss of viability. Monocytes and immature B lymphocytes remained stable. Clonogenic assays showed a decrease in colony-forming unit (CFU) number in UCB units stored for 96 hours.

UCB manipulation did not influence cell viability. All cell subsets remained viable until 96 hours after collection. CD34+ cells and T lymphocytes increased, probably due to the loss of other subsets. CFU growth during the period analyzed and confirmed stem cell functionality, despite the decrease at 96 hours. Results demonstrated that UCB units could probably be processed up to 96 hours after collection.

In this comparative, block-randomized, open-label, active-controlled, crossover Phase I trial, 60 healthy adult volunteers received single transfusions of 1200 mL of parent product (in Period 1) and of the LG plasma product (in Period 2) or vice versa. In both periods, plasmapheresis (600 mL) preceded the transfusion. Blood samples were drawn before and after apheresis and 15 minutes, 2 hours, 24 hours, and 7 days after end of plasma transfusion, to assess recovery, safety, and tolerability. The primary efficacy endpoints were the changes in coagulation factors and hemostatic variables compared to baseline; their relative recovery was computed in the per-protocol analysis (n = 43). Safety and tolerability were assessed (n = 60).

Variations in coagulation factors and hemostatic variables over time were similar between the two treatments and within normal range; 90% confidence intervals for the derived recovery data were within predefined limits of equivalence. Both products were well tolerated. The advanced manufacturing process also significantly increased plasmin inhibitor concentrations after transfusion in vivo.

The LG plasma product was bioequivalent to its predecessor with respect to recovery of clotting factors and demonstrated comparable safety and tolerability in healthy volunteers. Both products compensated well for the loss of clotting factors after apheresis (NCT01063595).

D– women participating in the routine antenatal RHD screening program in the capital region of Denmark were retrospectively studied. We used a standard dilution curve to quantify the amounts of cell-free fetal DNA (cffDNA) and divided women into groups according to cffDNA levels. PAPP-A was measured at 11 to 14 weeks. Information about pregnancy outcome and complications was obtained from the National Fetal Medicine Database, medical charts, and discharge letters.

The odds ratio (OR) of developing severe preeclampsia given a cffDNA level above the 90th percentile compared to cffDNA below the 90th percentile was 8.1 (95% confidence interval [CI], 2.6-25.5). The OR of developing mild preeclampsia given a cffDNA level below the 5th percentile compared to cffDNA levels above the 5th percentile was 3.6 (95% CI, 1.1-11.7). PAPP-A levels below the 5th percentile were associated with mild preeclampsia, but adding it to the analysis did not increase the detection rate (DR).

Women with cffDNA levels below the 5th percentile and above the 90th percentile quantified at 25 weeks' gestation are at increased risk of developing preeclampsia. Adding PAPP-A levels to the analysis did not increase the DR of preeclampsia.

We have prospectively compared by fluorescence-activated cell sorting analyses the cellular composition of three blood sources on the one hand—peripheral blood (PB; n = 10) versus granulocyte–colony-stimulating factor (G-CSF)-mobilized PB (GCSF-PB, n = 10) versus cord blood (CB, n = 10)—and of three graft sources on the other hand—unmanipulated bone marrow (uBM, n = 5) versus leukapheresis product (LP, n = 10) versus thawed CB graft (n = 7).

All median absolute numbers of cell subsets were found significantly higher in GCSF-PB and LP, except for monocytoid dendritic cells (mDCs) in CB and uBM. The most impressive results were the median quantities of memory T and B lymphocytes but also of plasmacytoid DCs (pDCs) contained in LP compared to thawed CB graft, with ratios of 375, 318, and 247, respectively. The proportions of naive and CD4+/CD8− T cells, transitional B cells, and CD5+ and naive B lymphocytes were found significantly higher in CB samples while the proportions of mDCs and pDCs were found significantly lower.

Our study shows strong differences in terms of quantitative and qualitative cellular composition between several blood or graft sources, possibly explaining the differences observed in terms of outcomes after transplant.

Cancer risk after blood transfusion was evaluated in a US population-based case–control study using 552,951 elderly cases identified from cancer registries and 100,000 frequency-matched controls. Transfusions received 0 to 12, 13 to 30, and 31 to 48 months before cancer diagnosis or selection date were identified using Medicare claims. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using logistic regression models. A Bonferroni correction adjusted for multiple testing.

Transfusions received 0 to 12 months before cancer diagnosis and/or selection were associated with significantly elevated risk of cancer overall (OR, 2.05; 95% CI, 1.95-2.16) and cancer of the stomach; cancer of the colon; cancer of the liver, kidney, renal pelvis, and/or ureter; lymphoma; myeloma; and leukemia. No significant associations for cancer overall were observed for the two earlier intervals. No site was associated with transfusions received 13 to 30 or 31 to 48 months before diagnosis and/or selection. Nonetheless, overall cancer risk increased with the number of transfused periods (p-trend < 0.0001).

Risk of overall cancer and specific sites was elevated 0 to 12 months after blood transfusion and associated with multiple transfusions, possibly due to reverse causation, that is, incipient cancers or cancer precursors causing anemia.

PCL was prepared by S/D treatment, oil extraction, and hydrophobic interaction chromatography. The content of TGF-β in PCL was determined by enzyme-linked immunosorbent assay. Cultured CD4+ T cells were used to investigate the effects of PCL on expression of transcription factor forkhead box P3 (Foxp3), the inhibition of T-cell proliferation, and cytokine production. The regulatory function of PCL-converted CD4+ T cells was analyzed by suppressive assay. The BALB/c mice were given PCL-converted CD4+ T cells before ovalbumin (OVA) sensitization and challenge using an asthma model. Inflammatory parameters, such as the level of immunoglobulin E (IgE), airway hyperresponsiveness (AHR), bronchial lavage fluid eosinophils, and cytokines were assayed. Recombinant human (rHu) TGF-β1 was used as control.

PCL significantly enhanced the development of CD4+Foxp3+-induced regulatory T cells (iTregs). Converted iTregs produced neither Th1 nor Th2 cytokines and inhibited normal T-cell proliferation. PCL- and rHuTGF-β-converted CD4+ T cells prevented OVA-induced asthma. PCL- and rHuTGF-β-modified T cells both significantly reduced expression levels of OVA-specific IgE and significantly inhibited the development of AHR, airway eosinophilia, and Th2 responses in mice.

S/D-treated PCL promotes Foxp3+ iTregs and exerts immunosuppressive and anti-inflammatory properties. This finding may help to understand the clinical properties of platelet lysates.

Path analysis was used to examine the simultaneous relationships among syncopal reactions, donation anxiety, needle pain, donor satisfaction, and donation intention in predicting repeat donation. Participants included 421 first- and second-time donors recruited for a study comparing the effects of predonation water loading with and without the use of applied muscle tension during donation (52% female, 60.8% first-time donor, mean age 20.3 years). For this longitudinal follow-up study, donor database records were accessed 2 years after the index donation to assess repeat donation.

Results of a series of path analyses demonstrated the influential role of donor anxiety in shaping donor retention (final model χ² = 35.75, root mean square error of approximation 0.03, comparative fit index 0.98, weighted root mean square residual 0.74). First, anxiety exerted a direct negative influence on donation intention, the proximal and sole direct predictor of repeat donation. Second, anxiety increased the likelihood of donor-reported needle pain, adversely affecting donation satisfaction and, subsequently, donation intention. Finally, anxiety was associated with donor ratings of syncopal reactions through its impact on needle pain, which also contributed to decreased donation intention.

These results provide novel evidence that donation anxiety plays a central role in shaping future donation behavior. Individual differences in anxiety must be considered when developing and testing strategies to enhance blood donor retention.

A suspected case of FMAIT was investigated for PLT-specific antibodies using a panel of both HPA-typed and paternal PLTs. HPA typing was performed by polymerase chain reaction with sequence-specific primers and further DNA analysis was performed using direct sequencing of the coding regions of the ITGA2B and ITGB3 genes.

Maternal antibodies reactive only with paternal PLTs were localized to glycoprotein (GP)IIb/IIIa using the monoclonal antibody immobilization of PLT antibody assay. A single-nucleotide polymorphism was detected in Exon 23 of ITGA2B in the father and affected child, which predicted a V740L substitution in the mature protein. Recombinant V740L mutated GPIIb expressed in HEK293 cells was specifically recognized by maternal antibodies. The polymorphism was not detected either in the mother or in a cohort of 100 donors.

The V740L polymorphism defines a new low-frequency antigen implicated in two cases of FMAIT in a single family. Low-frequency HPAs are clinically important and their elucidation requires both crossmatch studies and gene sequencing in cases where there is strong clinical evidence of FMAIT but initial laboratory investigations do not support the diagnosis.

Adult patients with GVHD or bronchiolitis obliterans syndrome (BOS) after lung transplant undergoing ECP at our center were alternatively assigned, within the same ECP cycle (composed by two procedures each), to MNC collection with either device. Patients' characteristics, procedure, and product-related variables were compared.

Thirty-nine patients (24 with GVHD and 15 with BOS) underwent a total of 126 ECP procedures, with good compliance to both devices. Product volume and platelet (PLT) and red blood cell contamination were significantly lower with the Spectra Optia. MNC collection efficiency (CE), purity, and PLT loss were similar between the two devices, while white blood cells CE was in favor of the COBE Spectra.

The Spectra Optia device proved to be a good option for MNC collection in the difficult ECP setting, since it ensures high-quality MNC collection, while at the same time saving personnel's time, guaranteeing optimal monitoring and care to this frail patient population.

Samples from 93 nontransfused males were tested using five different lots of a multiplex bead-based anti-HLA detection kit. A subset of 17 samples was tested on 5 days using a single lot. An additional 96 samples from donations with varied anti-HLA levels were tested using kits from two different lots. Results were reported as a normalized background (NBG) ratio.

For the 93 nontransfused donors, NBG values generated using the reference lot were significantly higher than those obtained with three of the four comparator lots. However, for the 96 samples with low-, moderate-, and higher-level anti-HLA, Class I (CL-I) values were 1.4 times lower and Class II (CL-II) values were 1.2 times lower using the reference versus comparator lot. For CL-I antibodies the between-lot standard deviation (SD) was 1.36 (95% confidence interval [CI], 1.19-1.60), while the between-day SD was 1.27 (95% CI, 1.08-1.52). Similarly, for CL-II antibodies the between-lot SD was 0.81 (95% CI, 0.70-0.95), while the between-day SD was 0.50 (95% CI, 0.43-0.60).

There is interlot variability in the tested HLA detection assay as well as significant bias between lots. It may be reasonable to develop a new cutoff when a new lot is obtained.

In 2009 to 2011, a total of 77 HIV-positive and 649 HIV-negative consented donors who screened nonreactive for hepatitis B virus, hepatitis C virus, syphilis, and alanine aminotransferase in four Retrovirus Epidemiology Donor Study-II Chinese regions received and completed a questionnaire by mail regarding their recent and past medical procedures, drug use, and sexual behaviors, etc. Exploratory and confirmatory factor analyses grouped questions into three risk factors. Multivariable logistic regression analysis examined the relationship between risk factors and HIV status adjusting for center, age, sex, and education.

The three risk factors were test-seeking tendency, medical-related risks, and behavioral risks. In multivariable logistic regression analysis, greater test-seeking tendency and behavioral risks were associated with HIV infection, with the adjusted odds ratios (ORs) being 2.2 (95% confidence interval [CI], 1.2-4.1) and 3.8 (95% CI, 1.8-7.9), respectively, but medical risks were not (OR, 1.2; 95% CI, 0.6-2.2). In comparison to less high school education, high school and more education was associated with lower risks for HIV infection, with the ORs being 0.35 (95% CI, 0.17-0.70) and 0.17 (95% CI, 0.09-0.33), respectively.

Test-seeking tendency and high-risk sexual behaviors are important predictors of HIV infection in Chinese blood donors, suggesting that the health history inquiry used in donor selection process needs improvement to defer high-risk donors more effectively.

During a 39-month period, 1236 donors deferred for fingerstick hemoglobin (Hb) level of less than 12.5 g/dL and 400 nondeferred “control” donors underwent health screening and laboratory testing (complete blood count, ferritin, iron, transferrin). Pica and RLS were assessed by direct questioning. Deferred donors and iron-deficient control donors were given 325 mg of ferrous sulfate daily for 60 days. Reassessments were performed and additional iron tablets dispensed at subsequent visits.

Pica was reported in 11% of donors with iron depletion or deficiency, compared with 4% of iron-replete donors (p < 0.0001). Pagophagia (ice pica) was most common and often of extraordinary intensity. Female sex, younger age, and lower mean cell volume and transferrin saturation values were strongly associated with pica. Donors with pica given iron reported a marked reduction in the desire to consume the nonnutritive substance by Days 5 to 8 of therapy, with disappearance of symptoms by Days 10 to 14. RLS was reported in 16% of subjects with iron depletion or deficiency compared with 11% of iron-replete donors (p = 0.012). Iron replacement generally resulted in improvement of RLS symptoms; however, at least 4 to 6 weeks of iron therapy was necessary.

The presence of pica is associated with a high probability of iron depletion or deficiency in blood donors; however, RLS lacks a strong correlation in this population. Screening questions for pagophagia may be useful in the ascertainment of iron deficiency in donors and may identify those who would benefit from oral iron.

Control, gamma-irradiated, and Mirasol-treated apheresis PCs were assayed on Days 1 and 5 of storage by means of gel-based analytical approaches (two-dimensional gel electrophoresis) and mass spectrometry–based identification of significant (p < 0.05 analysis of variance) differential proteins. Supernatants were then assayed for metabolism and oxidative stress-related metabolites through multiple reaction monitoring mass spectrometry.

Only a handful of modifications could be observed in the PLT proteome profiles in response to the Mirasol treatment, which included proteins involved in oxidative stress responses, PLT metabolism, and activation. Results confirmed increased metabolic rate and oxidative stress in the supernatants of treated PLTs (both gamma irradiated and Mirasol treated).

From this investigation, it emerges that, from a proteomics standpoint, gamma irradiation results in the acceleration of PLT storage lesions and the Mirasol treatment only moderately exacerbates these phenomena.

Two Italian scientific societies, the Italian Society of Hemapheresis and Cell Manipulation (SIdEM) and the Italian Group for Bone Marrow Transplantation (GITMO), joined to develop and disseminate recommendations on appropriate application of ECP treatment in patients with GVHD. Accordingly, SIdEM and GITMO named an expert panel that first selected 16 questions that were considered relevant for clinical practice: the questions were subsequently addressed through a revision of the available literature and in consensus meetings. The whole group discussed the proposed recommendations according to the nominal group technique.

The above-described approach in turn allowed the panel to agree on 47 practice recommendations. SIdEM and GITMO will disseminate such recommendations to the national transplant centers.

In conclusion, SIdEM and GITMO have made a scientific effort to provide a useful tool to physicians involved in the field, thus supporting daily clinical practice, as well as strategic decisions in the setting of ECP treatment of GVHD.

Data were collected via three methods: a review of study publications, study case report forms, and a questionnaire sent to the authors. Authors of randomized controlled trials of PLT transfusion that used bleeding as an outcome measure were identified from the searches reported by two recent systematic reviews. Twenty-four authors were contacted, and 13 agreed to participate. Data submitted were reviewed and summarized.

More recent studies with trained bleeding assessors, detailed documentation, and expanded grading systems have reported higher overall levels of bleeding. The World Health Organization grading system was widely used to grade bleeding, but there was no consistency in the bleeding grade definitions. For example, bleeding classified as Grade 2 in some studies (spreading petechiae) was classified as Grade 1 in other studies.

This study has highlighted differences in the method of recording and grading bleeding, which may account for some of the variation in reported bleeding rates. To ensure that differences between studies can be attributed to trial interventions or types of participant included, this study group is developing consensus bleeding definitions, a standardized approach to record and grade bleeding, and guidance notes to educate and train bleeding assessors.

This study was a retrospective cohort analysis of a population data set that captures diagnostic and procedure data on all hospitalizations from more than 300 hospitals within the Australian state of Victoria from 1998 to 2008.

There were 3149 incident cases of MDS. The age-standardized incidence rate was higher than reported from local cancer registries (for 2007 9.6 per 100,000 [95% confidence interval {CI}, 9.2-10.0] vs. 4.8). Median age was 79 years, 56.3% were males, and 34.6% were TD-MDS. Overall number of hospitalizations with transfusion increased over the study period, but not median transfusion episodes per patient. TD-MDS was associated with new diagnoses of congestive heart failure (CHF; incident rate ratio [IRR], 1.92; 95% CI, 1.41-2.60), but not diabetes (IRR, 1.29; 95% CI, 0.54-3.04) or liver disease (IRR, 1.91; 95% CI, 0.63-5.78). TD-MDS was associated with bacterial (IRR, 1.75; 95% CI, 1.37-2.24) and fungal infections (IRR, 3.13; 95% CI, 1.70-5.75) and leukemia (relative risk [RR], 1.42; 95% CI, 1.07-1.88) and sepsis as cause of death (RR, 1.23; 95% CI, 1.03-1.47) but not CHF (RR, 0.97; 95% CI, 0.71-1.32).

There was a higher incidence of MDS compared with that reported by cancer registries. Overall hospitalizations increased over the study period with no change in transfusion episodes per patient. There were more incident cases of CHF and infections in TD-MDS; however, CHF was not a more frequent cause of death.

We conducted a single-institution, retrospective cohort study to measure blood utilization in 89 consecutive burn patients with 15% to 65% total body surface area (TBSA) burn within 60 days of injury. We also evaluated the relationship of blood product utilization with clinical variables including anticoagulant usage and mortality.

We determined that: 1) the predictors for increased red blood cells (RBCs) and plasma transfusions were high TBSA burn and the use of argatroban anticoagulation (for suspected heparin-induced thrombocytopenia [HIT]); 2) TBSA burn and patient age were independent predictors of mortality, but not RBC or plasma transfusion; and 3) the incidence of symptomatic venous thromboembolic events is not uncommon (11.2%), although HIT is rare (1.1%).

Despite concerns about adverse correlation between increased number of transfusions and mortality in other clinical settings, we did not find this association in our study. However, we demonstrated that the type and intensity of anticoagulation carries substantial risk for increased RBC as well as plasma usage.

New donors (n = 4861) were invited to fill out a questionnaire before their first donation (response, 61%). The questionnaire assessed variables from the Theory of Planned Behavior (intention, self-efficacy, attitude, and norms), conscientiousness, and donation anxiety. Three years later it was determined who became whole blood or plasma donor. Multivariable linear regression analyses for intention were fitted separately for whole blood and plasma donors. A logistic regression analysis was executed to estimate the effect of intention at the beginning of a donor career on becoming a plasma donor.

Plasma donors had a higher intention, self-efficacy, attitude, and conscientiousness and a lower anxiety than whole blood donors. In plasma and whole blood donors, both self-efficacy and cognitive attitude were positively related to intention but with different strength (plasma, β = 0.47 and β = 0.30; whole blood, β = 0.57 and β = 0.17). Having a high level of intention increased the odds of becoming a plasma donor (odds ratio, 1.33; 95% confidence interval, 1.12-1.59).

Motivational differences already exist between future whole blood and plasma donors before their first donation. Although a feeling of self-efficacy is necessary for all new donors, more favorable cognitions are important for future plasma donors. Recruitment strategies for plasma donors should focus on attracting the more self-confident donors by highlighting the usefulness of plasma donation.

This prospective, randomized, crossover study compared Mirasol-treated (MIR) and standard reference (REF) PLT transfusions. PLT counts and TEG measurements were taken at pretransfusion and 1- and 24-hour-posttransfusion time points. The primary outcome measure was the pretransfusion to 1-hour-posttransfusion change in maximum amplitude (ΔMA_1hr). Secondary endpoints included ΔMA among other time points, relative MA, and the PLT count–MA correlation.

Of 16 enrolled patients, one withdrew before study treatment and three did not require two transfusions, leaving 12 patients in the efficacy analyses (seven MIR-REF, five REF-MIR). ΔMA_1hr (mean ± SD) was 10.60 ± 6.47 mm for MIR and 14.33 ± 5.38 mm for REF (p = 0.20, n = 10). ΔMA_24hr was 9.49 ± 7.94 for MIR and 7.13 ± 3.08 for REF (p = 0.38, n = 9); ΔMA_24hr-1hr was −1.11 ± 2.95 for MIR and −7.20 ± 4.81 for REF (p = 0.016, n = 8). MA values for MIR and REF correlated with the log of PLT count (r_MIR = 0.6901, r_REF = 0.7399).

TEG is sensitive to changes in hemostatic function resulting from a single PLT transfusion. MIR and REF provided similar increments in hemostatic function in the immediate posttransfusion period and at 24 hours. A significant difference detected for ΔMA_24hr-1hr suggests different PLT clearance mechanisms. The relationship of these variables to clinically meaningful outcomes, for example, bleeding events or transfusion requirements, has yet to be determined.

Sequential sera from pregnant women with expected FNAIT were assessed for HPA-1a alloantibodies using SPR. Group I (n = 6) was treated with intravenous immunoglobulin (IVIG) and steroids beginning at 19 weeks of gestation (w.g.), and Group II (n = 4) received intrauterine PLT transfusions (IUT) beginning at 22 w.g. Maternal alloantibodies were quantified using an HPA-1a monoclonal antibody (MoAb) as a standard. Antibody avidity was determined as the ratio of B₇₀₀ (end of the dissociation phase) to B₃₅₀ (end of the association phase); the area under the curve (AUC) was calculated to determine overall antibody binding.

After 22 w.g., alloantibody characteristics remained stable in both groups, while there was a steep decrease in B₇₀₀ and B₃₅₀ values between 16 and 22 w.g. (assessed only in Group I), indicating a decrease in anti-HPA-1a alloantibody concentrations. Interestingly, the AUCs of the last maternal sample before elective delivery appeared to be correlated with fetal and neonatal PLT counts (p = 0.014 and 0.017, respectively).

SPR provides quantitative information on HPA-1a alloantibody characteristics in addition to monoclonal antibody–specific immobilization of platelet antigens. SPR results can be calibrated using a MoAb standard and should be further assessed for a potential correlation with fetal PLT count.

Here, we evaluated the K562 erythroleukemic cell line to study the cellular effects of a murine anti-GPC. Cell proliferation was evaluated after treatment with anti-GPC. Flow cytometry was used to evaluate exofacial phosphatidylserine (PS) expression and cell viability (propidium iodide binding). Cell morphology was evaluated under light microscopy with cytospin preparations stained with May-Grünwald Giemsa.

Anti-GPC dramatically inhibited K562 proliferation and increased PS expression, consistent with cytoplasmic blebbing, suggesting evidence of apoptosis. Z-VAD-FMK, an inhibitor of classical apoptosis, was unable to reverse the suppressive effect of anti-GPC. However, hemin was able to attenuate growth suppression.

Together, the data suggest that anti-GPC suppresses erythroid proliferation through the induction of nonclassical apoptosis.

Samples from D– – or Rh_null individuals and their family members, from families for whom Rh phenotype and/or serologic data were unexplained by inheritance of conventional RH alleles, were analyzed. Genomic DNA and transcripts were tested by sequencing analysis.

The first silent allele was a RHCE*cE allele carrying an intronic IVS3+5G>A mutation. The second was a RHCE*ce allele carrying an intronic IVS7-2A>G mutation, whereas the third was a silent RHCE*ce allele carrying a 5-bp deletion (Nucleotides 679-683) in Exon 5.

In addition to hybrid alleles and nucleotide deletion, intronic mutations may be associated with the nonexpression of RhCE antigens. Regarding the RH system, silent alleles may not be investigated among D– – or Rh_null individuals only. Rh phenotype and/or serologic data unexplained by inheritance of conventional RH alleles should lead to molecular investigations.

We report the successful treatment of AVWS with anti-CD20 monoclonal antibody therapy (375 mg/m² rituximab as four weekly doses followed by 375 mg/m² every 90 days) in a patient with concurrent monoclonal B-cell lymphocytosis allowing for the early discontinuation of blood product support after only 2 g/kg IVIG achieved acute hemostasis control.

This is the first documentation of the successful long-term management of AVWS without prolonged blood product or IVIG support. This result contrasts sharply to previously reported rituximab strategies that were deemed ineffective in AVWS.

A maintenance regimen of rituximab may be an effective long-term management strategy for AVWS associated with lymphoproliferative disorders, which may minimize the use of blood products and IVIG.

Plasma Protein Therapeutics Association member companies collected more than 200 prion removal studies on plasma protein manufacturing steps, including precipitation, adsorption, chromatography, and filtration, as well as combined steps. The studies used a range of model spiking agents and bench-scale process replicas. The results were grouped based on key manufacturing variables to identify factors impacting removal. The log reduction values of a group are presented for comparison.

Overall prion removal capacities evaluated by independent groups were in good agreement. The removal capacity evaluated using biochemical assays was consistent with prion infectivity removal measured by animal bioassays. Similar reduction values were observed for a given step using various spiking agents, except highly purified prion protein in some circumstances. Comparison between combined and single-step studies revealed complementary or overlapping removal mechanisms. Steps with high removal capacities represent the conditions where the physiochemical differences between prions and therapeutic proteins are most significant.

The results support the intrinsic ability of certain plasma protein manufacturing steps to remove prions in case of an unlikely contamination, providing a safeguard to products.

In the Leukocyte Antibody Prevalence Study (LAPS), 7920 volunteer blood donors were screened for anti-HLA and surveyed about prior pregnancies and transfusions. A secondary analysis of the LAPS database was performed.

D– women not more than 40 years old (presumed to have received antenatal with or without postpartum RhIG in all pregnancies) had a significantly lower HLA sensitization rate than D+ women (relative risk, 0.58; 95% confidence interval [CI], 0.40-0.83). When stratified by deliveries (one, two, three, or four or more), D– women not older than 40 were HLA sensitized less often than D+ women in every case. In contrast, a clear relationship between D type and HLA sensitization was not seen in older previously pregnant women whose childbearing years are presumed to have preceded the use of routine RhIG prophylaxis. In a multivariable logistic regression model, D– women not more than 40 years old remained significantly less likely to be HLA sensitized compared with D+ women after adjusting for parity, time from last pregnancy, lost pregnancies, and transfusions (odds ratio [OR], 0.55; 95% CI, 0.34-0.88).

Consistent with a nonspecific immunosuppressive effect of RhIG, younger previously pregnant D– women were less likely than previously pregnant D+ women to be HLA sensitized.

A retrospective cohort study was performed between 2008 and 2011. Pediatric patients with hematologic disorders, solid tumors, primary immunodeficiency disorders, or inherited metabolic disorders were transfused with M-sol-R-PCs between 2010 and 2011; the transfusions of P-PCs administered between 2008 and 2011 were compared in terms of frequency and severity of ATRs, corrected count increment (CCI), and occurrence of bleeding. Data were collected for 6 consecutive months on a per-patient basis.

Data obtained during 2008 to 2011 showed that of the 78 patients receiving 515 P-PC transfusions, 14 (17.9%) had 17 ATRs (3.3%); 14 and three ATRs were of Grades 1 and 2, respectively. In 2010 to 2011, 49 patients received 620 transfusions of M-sol-R-PCs, and two patients (4.1%) had Grade 1 ATRs (0.3%). Thus, the frequency of ATRs per bag and per patient differed significantly between the two transfusions. No steroid agents were used for the prevention or treatment of ATRs in the M-sol-R-PC group. The CCI (24 hr) for M-sol-R-PCs did not differ from that for P-PCs.

M-sol-R-PCs were found to be effective in preventing ATRs without loss of transfusion efficiency in children; however, its efficacy should be further evaluated in prospective clinical trials.

A 34-week neonate born to a Hispanic mother with anti-Ge3 developed late-onset hemolysis with hyperbilirubinemia and was successfully treated with transfusions from her mother. Relevant clinical findings and laboratory results for this case are summarized and compared to three other previously reported cases; all babies were born from a mother of Hispanic ethnicity.

Hemolytic disease of the fetus and new born associated with anti-Ge3 is rare but should be considered when working up a broadly reactive RBC antibody screen in women of Hispanic ethnicity. Early identification of pregnant women with anti-Ge3 is recommended for prenatal transfusion planning and close monitoring of the newborn infant for evidence of late-onset anemia.

Blood samples from 120 patients with autoimmune hemolytic anemia (AIHA) and IgG-coated red blood cells (RBCs) were studied by serologic methods. Some autoantibodies were investigated by immunochemical methods.

Autoantibodies against the third external loop of Band 3 have a distinctive pattern of reactivity in that they fail to react after RBC treatment with α-chymotrypsin and pronase, whereas papain, ficin, and trypsin have no effect. Eleven (9%) patients had pure anti-Band 3 autoantibodies. Autoanti-Band 3 antibodies were associated with other specificities in 66 (55%) patients. Immunoprecipitation and rare RBCs from a Wu+ homozygote, known to have an unusual pattern of reactivity after protease treatment, were used to confirm the Band 3 specificity. Treatment with sodium hypochlorite, believed to oxidize the Met residue at Position 559 in the third loop, showed that these autoantibodies were heterogeneous. Most antibodies reacted optimally at 37°C, but two patients had incomplete cold IgG autoantibodies. Unlike autoantibodies to Rh proteins, warm autoanti-Band 3 activate complement and are almost totally bound to autologous RBCs.

We describe the first cases of warm IgG autoantibodies specific for the third loop of Band 3. This external loop also appears as a major target in patients with warm antibody AIHA.

A Markov model has been developed to determine lifetime cost and quality-adjusted life-years (QALYs) of patients. To estimate the annual cost of each method, a cross-sectional study was conducted among two groups of patients who received DFO and DFX (n = 100 and n = 45, respectively). Also a time trade-off method was used to estimate the utility of two strategies. Finally a one-way and probabilistic sensitivity analysis was conducted to examine the strength of the results.

Our base-case analysis showed that estimated total lifetime costs per patient for DFX and DFO were 47,029 international dollar ($Int) and $Int143,522, respectively, while the estimated total discounted QALYs per person were 12.28 and 7.76, respectively. Calculated incremental cost-effectiveness ratio showed that DSX is a dominant therapy and its estimated lifetime net monetary benefit was $Int273,528.

We conclude that the use of DFX instead of DFO represents a cost-effective use of resources for treatment of iron overload in patients with β-thalassemia from Iran's society perspective.

This retrospective observational study was designed as an interrupted time series (1999-2010). The intervention was the introduction of an EPO protocol in THA patients in 2003. Patients were classified according to preoperative hemoglobin (Hb) level: 10 to 13 g/dL (eligible patients for EPO) and more than 13 g/dL. The primary outcome was the percentage of patients receiving an ABT. Segmented regression analysis was used to estimate changes in outcome after the intervention.

A total of 4568 THA patients were included. The absolute reductions in ABTs after the intervention were 17% (95% confidence interval [CI], 6%-29%) for the total study population and 25% (95% CI, 11%-39%) and 8% (95% CI, −5% to 21%) for the Hb groups 10 to 13 and more than 13 g/dL, respectively. In the postintervention period, 46% of the eligible patients (Hb level, 10-13 g/dL) actually received EPO. The transfusion rate in the EPO group was lower compared to the non-EPO group: 14 and 50%, respectively (p < 0.01).

Introduction of a preoperative EPO protocol reduced the transfusion rate in THA patients in daily clinical practice. The reduction must be seen as part of a multifaceted blood management program, in which increased awareness of blood transfusion contributes simultaneously and substantially to the reduction in transfusion rate.

We compared cryopreservation efficiency of the ex vivo expanded CB cells using the standard protocol (freezing solution human serum albumin (HSA)-dimethyl sulfoxide [DMSO]) with the newly designed protocol based on an enriched freezing solution (HP01-DMSO) with respect to the viability index, number of CD34+ and total cells, and recovery of CPs (colony-forming units) and HSCs (NOG/Scid/gamma–null mice engraftment).

Cryopreservation and thawing of expanded CB cells using the “standard” procedure (HSA-DMSO) reduced recovery of the CPs (40%) and HSCs (drastically decreasing engraftment capacity). HP01-based protocol resulted in improvement of preservation of both CPs (>60%) and HSCs (nonaltered engraftment capacities).

Functional maintenance of the expanded graft by cryopreservation is feasible in conditions compatible with human cell therapy requirements.

We performed an anonymous survey, mailed to 40,000 donors in 2008, including questions about symptoms, height, weight, sex, and donation status. Reaction rates were compared to those recorded in our database. Possible risk factors were assessed for various reactions.

The response rate was 45.5%. A total of 32% of first-time and 14% of repeat donors reported having any adverse symptom, most frequently bruising (84.9 per 1000 donors) or feeling faint or weak (66.2 per 1000). Faint reactions were two to eight times higher than reported in our database, although direct comparison was difficult. Younger age, female sex, and first-time donation status were risk factors for systemic and arm symptoms. In females, low estimated blood volume (EBV) was a risk factor for systemic symptoms. Only 51% of donors who consulted an outside physician also called Canadian Blood Services. A total of 10% of first-time donors with reactions found adverse effects information inadequate.

This study allowed us to collect more information about adverse reactions, including minor symptoms and delayed reactions. Based on our findings of the risk factors and frequency of adverse reactions, we are implementing more stringent EBV criteria for younger donors and providing more detailed information to donors about possible adverse effects and their management.

A prototype of the Reveos system was used to process WB. RBCs were resuspended in SAGM, leukoreduced, and assayed for in vitro quality variables during a 42-day storage period at 2 to 6°C. Twenty-four-hour in vivo recovery was determined on Day 42. Plasma was assayed for cellular contamination and activation variables. IPUs were pooled with SSP+ additive solution for in vitro quality assessments during a 7-day storage period at room temperature.

Reveos-produced RBCs and plasma units met the predefined requirements. RBC recovery was superior to control units. On Day 42, hemolysis was below 0.8% and in vivo recovery was above 75% for all RBCs. Cellular contamination was lower for Reveos-produced plasma. PLT yield was higher with overnight-stored WB. PLT quality was well maintained during storage with no significant differences between the two groups.

Blood components prepared with the Reveos from fresh or overnight-held WB meet quality criteria without any relevant difference between the two groups. The Reveos system has the potential to increase efficacy and standardization of blood component preparation.

A total of 430 blood samples with D phenotype ambiguity were recruited for this study. The three most frequent weak D alleles (i.e., weak D, Type 1; weak D, Type 2; and weak D, Type 3), which altogether account for 60% to 90% of the atypical RHD alleles in the Caucasian population, were first identified by Tm-shift genotyping. The remaining unidentified samples were then subjected to a single-tube multiplex polymerase chain reaction (PCR) amplification of all 10 RHD exons followed by direct sequencing.

Optimal conditions for efficient and reliable identification of the three most common weak D variants by Tm-shift genotyping were established. All 10 RHD exons were successfully amplified in a single-multiplex PCR procedure. Employment of the two-step analysis identified RHD variants in 91.6% of the 430 studied samples. Two of the nine previously undescribed variants, c.335G>T and c.939G>A, were found to cause aberrant mRNA splicing by means of a splicing minigene assay.

The new two-step analysis proved to be much easier and cheaper than the DHPLC method and therefore is convenient to be used as a routine, medium-throughput approach for RHD genotyping.

This study was performed to establish the impact on coagulation of RMPs isolated from blood units. Using calibrated automated thrombography, we investigated whether RMPs affect thrombin generation (TG) in plasma.

We found that RMPs were not only able to increase TG in plasma in the presence of a low exogenous tissue factor (TF) concentration, but also to initiate TG in plasma in absence of exogenous TF. TG induced by RMPs in the absence of exogenous TF was neither affected by the presence of blocking anti-TF nor by the absence of Factor (F)VII. It was significantly reduced in plasma deficient in FVIII or F IX and abolished in FII-, FV-, FX-, or FXI-deficient plasma. TG was also totally abolished when anti-XI 01A6 was added in the sample. Finally, neither Western blotting, flow cytometry, nor immunogold labeling allowed the detection of traces of TF antigen. In addition, RMPs did not comprise polyphosphate, an important modulator of coagulation.

Taken together, our data show that RMPs have FXI-dependent procoagulant properties and are able to initiate and propagate TG. The anionic surface of RMPs might be the site of FXI-mediated TG amplification and intrinsic tenase and prothrombinase complex assembly.

We conducted a single-center retrospective study of 2102 patients who had CABG in Ontario to determine whether anemia at hospital discharge was associated with increased 30-day hospital readmissions, readmission secondary to cardiac disease, and 30-day mortality using administrative data.

Of the 2102 patients, 224 patients (11%) were readmitted within 30 days of hospital discharge. Infection was the leading cause of readmissions (24%), followed by heart failure (13%), pulmonary disease (7%), and hemorrhagic disease (7%). Overall, 2.6% of patients were readmitted because of cardiac disease. Of patients discharged, 48% were discharged with a hemoglobin (Hb) level between 8 and 10 g/dL and 42% between 10 and 12 g/dL. Predischarge Hb concentration was not a significant independent predictor of 30-day readmission to the hospital due to all causes, readmission to the hospital due to cardiac causes, or 30-day mortality. A higher comorbidity score, adjusted odds ratio (OR) of 2.1 (95% confidence interval [CI], 1.3-3.6), leg and sternal wound infections OR of 1.9 (95% CI, 1.2-3.0), and postoperative renal failure OR of 1.4 (95% CI, 1.2-2.0) were associated with increased 30-day readmission rates.

The predischarge Hb concentration after CABG was not associated with 30-day readmissions.

Viable cell counts by flow cytometry and colony-forming unit (CFU) recoveries were assessed on cord blood (CB), mobilized peripheral blood stem cell (PBSC), and bone marrow (BM) samples over 72 hours using two different storage conditions, refrigerated (4-8°C) or room temperature (19-22°C). To determine the effects of delayed freezing on progenitor recoveries, paired samples were evaluated before and after cryopreservation.

All samples maintained at refrigerated temperatures resulted in higher recoveries than those at room temperature in all variables assessed. Specifically, when assessing for CFU yields after thawing, the impact of time on BM resulted in a significant loss as soon as 24 hours (n = 10, 36.4 ± 28.0%, p = 0.003). This decrease was also observed for PBSCs and CB but at 48 hours of fresh storage (PBSCs n = 11, 32.7 ± 26.2%, p = 0.006; CB n = 10, 39.6 ± 26.4%, p = 0.001).

Our data suggest that HPC products are better maintained at refrigerated temperatures before cryopreservation. Delaying cryopreservation should be minimized to avoid significant losses in cell potency.

This cross-sectional assessment included the 40 highest-volume facilities collecting and transfusing blood nationally identified in a previous survey. At each facility, study representatives completed a standardized instrument assessing staff performance of transfusion-related activities and performed rapid testing for human immunodeficiency virus, syphilis, and hepatitis B and C with rapid diagnostic tests on clinically discarded specimens. Reactive samples received confirmatory testing. Descriptive statistics were generated, with differences analyzed using chi-square or Fisher's exact tests.

Between November 2010 and May 2011, a total of 332 blood donor collection procedures were observed. Only 52.4% of observed encounters correctly screened and deferred donors by international criteria. Public and private facilities demonstrated glove use, proper sharps disposal, and patient counseling and relayed screening test results in less than 75% of observed events, significantly less likely than military facilities (p < 0.01). Of 1612 specimens assessed, confirmed cases of hepatitis B (n = 6), hepatitis C (n = 1), and syphilis (n = 3) were detected among units already prescreened and accepted for transfusion.

Lapses in proper donor screening contributed to the presence of confirmed-positive units available for transfusion, as detected in this study. Steps must be taken to ensure standardization of testing kits requirements, documentation, and mandatory training and continuing education for blood bank staff with regard to counseling, drawing, processing, and transfusion of blood products.

Blood donors deferred for malaria risk (travel, residence, or previous infection) provided blood samples and completed a questionnaire. Plasma was tested for Plasmodium antibodies by enzyme immunoassay (EIA); repeat-reactive (RR) samples were considered positive and tested by real-time polymerase chain reaction (PCR). Accepted donors provided background testing data.

During 2005 to 2011, a total of 5610 malaria-deferred donors were tested by EIA, including 5412 travel deferrals. Overall, 88 (1.6%) were EIA RR; none were PCR positive. Forty-nine (55.7%) RR donors previously had malaria irrespective of deferral category, including 34 deferred for travel. Among 1121 travelers to Mexico, 90% visited Quintana Roo (no or very low risk), but just 2.2% visited Oaxaca/Chiapas (moderate or high risk). Only two Mexican travelers tested RR; both previously had malaria not acquired in Mexico.

Travel to Mexico represents a large percentage of US donors deferred for malaria risk; however, these donors primarily visit no- or very-low-risk areas. No malaria cases acquired in Mexico were identified thereby supporting previous risk estimates. Consideration should be given to allowing blood donations from US donors who travel to Quintana Roo and other low-risk areas in Mexico. A more effective approach to preventing TTM would be to defer all donors with a history of malaria, even if remote.

Epidemiologic and serologic data were collected retrospectively for 344 chronically HBV-infected blood donors identified from July 2005 to June 2010. Additional laboratory testing was carried out to determine the HBV genotype, viral load, and prevalence of clinically significant mutations and to detect hepatitis delta virus (HDV) coinfection.

Five HBV genotypes (A-E) were found, Genotypes D (45%), A (20%), and E (20%) were the most prevalent. A strong association was seen between genotype and donor ethnicity (p < 0.001) and between genotype and place of residence (p = 0.006). Clinically significant mutations were observed across hepatitis B surface antigen (17%), basal core promoter (25%) and precore (78%) regions. An antiviral resistance profile was identified in one donor. Evidence of HDV coinfection was found in 2% of donors.

The data show the diversity of HBV in asymptomatic chronic infections detected in blood donors in England and North Wales and demonstrates the presence of mutations which may impact on disease. The global nature of these infections and the inability to identify chronically infected donors before donation highlights the importance of using screening assays capable of detecting a broad range of genotypes and mutations. Furthermore, the integration of the virologic and demographic data allows us to more accurately construct a profile of our chronically HBV-infected blood donors.

Blood samples were obtained from Shanghai Blood Center. D– samples typed in routine laboratories were tested using a monoclonal immunoglobulin M reagent, an indirect antiglobulin test, and a commercial panel of monoclonal anti-D (Diagast D-Screen). RHD nucleotide sequencing that included adjacent flanking intron regions was performed. The RHD zygosity was determined. Bioinformatics analysis was performed.

Four different RHD alleles were identified among six blood samples, namely, RHD IVS3+3G>C, RHD IVS6-14del3, RHD IVS4+5G>A, and RHD IVS4+5G>T. The serologic tests were consistent with weak D phenotypes. The bioinformatics results indirectly suggest that these sequence changes affect splicing.

This study is the first to describe weak D types caused by intronic variations near the splice sites in the RHD gene, which is supported by the genotyping results combined with serologic profiles and bioinformatics analysis. The identification of the four novel RHD alleles represents a new significant addition to the molecular bases that underlie weak D, which would deepen our understanding of the mechanism of the low expression of the RhD antigen. These nucleotide changes are predicted to have regulatory effects on RHD splicing.

Six units of PLTs in plasma and 6 units of riboflavin and ultraviolet (Mirasol, TerumoBCT)-treated PLTs in plasma were sampled on Days 2, 6, 8, and 10 after donation. PLT concentration, Annexin 5A staining, ThromboLUX (LightIntegra) thrombelastography, and P-selectin expression, both in unstimulated PLTs and in response to concentration series of adenosine diphosphate, collagen-related peptide, and thrombin receptor–activating peptide (TRAP), were measured.

For PLTs in plasma Annexin 5A expression increased by 0.60% (95% confidence interval [CI], 0.40%-0.80%) and P-selectin expression increased by 1.2% (95% CI, 0.80%-1.6%) per day. Responsiveness to TRAP simultaneously decreased by 1.3% (95% CI, 0.80%-1.8%) per day. After Mirasol treatment ThromboLUX scores decreased 3.3 points (95% CI, 0.2-6.4 points) from 22 to 19 points, Annexin 5A expression increased by 4.8% (95% CI, 3.3%-6.2%), and P-selectin expression increased by 13% (95% CI, 10%-16%), all averaged over the entire storage period. Responsiveness to TRAP simultaneously decreased by 19% (95% CI, 17%-21%).

Our results suggest flow cytometric measurement of agonist-induced P-selectin expression can measure PLT quality decline over the entire range encountered during 10-day storage of both standard PLTs and Mirasol-treated PLTs in plasma.

Data on CHIKV viremic profiles were obtained from a case-control study carried out in a chikungunya-affected area during the 2009 epidemic in Songkhla, Thailand. CHIKV-infected individuals were classified based on a combination of the patient's history and clinical and laboratory findings.

There were 134 laboratory-proven CHIKV-infected cases, of whom 122 (91.0%) were symptomatic and 12 (9.0%) were asymptomatic. The viremic levels in the symptomatic infected individuals peaked on the first 3 days and lasted up to 8 days as defined by viral isolates. CHIKV genomic products were detected as late as Day 17 of illness. The viral loads observed in the symptomatic individuals (median, 5.6 × 10⁵ plaque-forming units per milliliter [pfu/mL]; range, 1.3 × 10¹-2.9 × 10⁸ pfu/mL) were higher than but not significantly different from those observed in the viremic asymptomatic individuals (median, 3.4 × 10³ pfu/mL; range, 8.4 × 10¹-2.9 × 10⁵ pfu/mL [p = 0.22, Wilcoxon test]).

CHIKV infection is highly symptomatic and is associated with high-titred viremia. The viremic levels in asymptomatic CHIKV-infected individuals were in the range known to be capable of transmitting the disease to experimental animals. Asymptomatic CHIKV viremia individuals could be potential disseminators of transfusion-associated chikungunya.

Hematologic patients were assigned to receive PR-PLTs or gamma-irradiated conventional PCs, both prepared in PLT additive solution (PAS). One-hour CCI (primary endpoint), 24-hour CCI, time to next PLT transfusion, and transfusion requirement of red blood cells and plasma were analyzed.

Forty-four patients assigned to PR-PLTs received 220 PR-PLTs and 136 conventional PCs; 72 controls received 517 conventional PCs. No differences between patient groups were observed for mean (±standard deviation [SD]) 1-hour CCI (11.4 [±4.9] for PR-PLT vs. 11.0 [±4.9] for controls), mean (±SD) 24-hour CCI (6.1 [±4.4] for PR-PLTs vs. 6.2 [±4.8] for controls), and for the other evaluated outcomes. No differences between PC types were observed for mean (±SD) 1-hour CCI (10.6 [±6.7] for PR-PLTs vs. 9.9 [±6.2] for conventional PCs) and mean 24 hour-CCI (3.3 [±3.9] for PR-PLTs vs. 4.2 [±5] for conventional PCs). Thirty-five percent of PR-PLTs and 38% of conventional PCs (p = 0.63) were associated with 1-hour CCIs of less than 7.5. Inadequate 24-hour CCIs were observed for 72% of PR-PLTs and 64% of conventional PCs (p = 0.002).

Transfusion efficacy of single-donor apheresis PCs in PAS treated with amotosalen PR versus gamma irradiation is comparable.

Adult patients undergoing isolated coronary artery bypass grafting (CABG) surgery were randomly assigned to receive processed (n = 99) or unprocessed (control; n = 98) residual CPB volume in this single-center randomized controlled trial. The intensive care unit team, patients, and assessors were blinded to treatment assignment and a transfusion protocol was followed. Surgeons were permitted to use retrograde autologous priming to minimize crystalloid pump prime.

The processed study bag was of a smaller volume (280 [0, 550] mL vs. 590 [215, 726] mL; p < 0.01) but a higher hematocrit (29% [0%, 34%] vs. 23% [20%, 25%]; p < 0.01) than control. The rate of transfusion with homologous blood was 39% in both groups (p = 0.92). There was no difference in the volume transfused (processed 323 ± 585 mL vs. control 276 ± 520 mL; p = 0.56). There was also no difference in the proportion of patients transfused with any blood product (processed 44% vs. control 45%; p = 0.95) or in the volume of chest tube output (processed 600 [500, 940] mL vs. control 670 [490, 932] mL; p = 0.62).

Ultrafiltration of residual CPB volume in adults undergoing isolated CABG surgery does not reduce the need for transfusion or bleeding.

This study included 4598 Dutch whole blood donors. Information on ZPP levels measured at the previous visit was added to the existing prediction models to estimate the risk of Hb deferral. Models were compared using the following measures: concordance (c)-statistic, continuous net reclassification improvement (NRI), and clinical net benefit (NB).

Seventy-six male donors (2.9%) and 69 female donors (3.5%) were deferred because of a low Hb level. Previous ZPP level was associated with risk of Hb deferral (odds ratio for interquartile range of previous ZPP level, men 2.0 [95% confidence interval {CI}, 1.7-2.3]; women 2.2 [95% CI, 1.9-2.4]) in a multivariable risk model. Addition of ZPP into the models resulted in an increase of the c-statistic from 0.93 to 0.94 for men and from 0.80 to 0.85 for women. The added value of ZPP was confirmed by measures of clinical usefulness. NRI for men was 0.42, and for women, 0.56. At relevant threshold probabilities between 10 and 15%, NB was higher for models considering ZPP.

This study shows that ZPP measurements obtained at the previous visit may have added value in the risk prediction of Hb deferral in whole blood donors.

The study population comprised 5280 Dutch whole blood donors, who passed the Hb criteria for donation. During donor screening, Hb levels were measured in capillary samples (finger prick), and venous blood samples were taken for measurements of ZPP and other iron variables. These variables included ferritin, transferrin saturation, soluble transferrin receptor (sTfR), hepcidin, red blood cell mean corpuscular volume (MCV), and mean cell Hb (MCH).

With a ZPP cutoff level of at least 100 μmol/mol heme, subclinical iron deficiency was present in 6.9% of male donors and in 9.8% of female donors. Based on other iron variables, iron deficiency was also observed. Prevalence rates ranged from 4.8% (based on transferrin saturation) to 27.4% (based on hepcidin concentration) in men and from 5.6% (based on sTfR concentration) to 24.7% (based on hepcidin concentration) in women.

Results from this study showed that subclinical iron deficiency is prevalent among blood donors that meet the Hb criteria for blood donation, based on ZPP levels and on other iron variables. This finding needs attention because these donors are at increased risk of developing iron deficiency affecting Hb formation and other cellular processes.

Two assays based on cell-free fetal DNA extracted from maternal blood samples and on real-time polymerase chain reaction (QPCR) were developed: an allele-specific QPCR specifically targeting the polymorphic sequence in HPA-1 and the study of the variation in the high-resolution melting curve of amplicons containing the polymorphic region.

All results from the 49 samples obtained from 29 pregnant women were consistent with expectations. Six women were compatible with their fetuses (three HPA-1aa women and three HPA-1bb women), 41 HPA-1bb women were incompatible with their fetuses, as were two HPA-1aa women.

Two fetal PLT genotyping assays on maternal blood samples proved to be reliable as of 15 weeks of gestation, thereby avoiding invasive techniques such as amniocentesis.

We used near-infrared time-resolved spectroscopy to measure cerebral hemoglobin oxygen saturation (ScO₂) and cerebral blood volume (CBV) before and after transfusion in 19 infants (24 measurements) with anemia of prematurity. The median gestational age was 27 weeks 0 days, median birth weight was 751 g, and median postconceptual age at transfusion was 30 weeks 4 days.

bHb levels before and after transfusion (mean ± SD) were 9.3 ± 1.4 and 13.7 ± 1.3 g/dL, respectively. After transfusion, CBV significantly decreased from 2.63 ± 0.60 to 2.13 ± 0.26 mL/100 g of brain, and ScO₂ significantly increased from 72.8 ± 4.3% to 74.7 ± 4.2%.

After transfusion, CBV changes were significantly greater with low compared to high pretransfusion Hb levels. This reflected the physiologic response to severe anemia in premature infants, which is to increase CBV and decrease ScO₂. Therefore, CBV and ScO₂ may be useful markers for determining the need for transfusion in very-low-birth-weight infants.

Six randomized clinical trials of pathogen-inactivated PLT products are reviewed and associated methodologic issues are discussed.

The variation in the trial designs, outcomes, and methods of analysis suggest the need to harmonize the way trials of pathogen-reduced PLT products are conducted to facilitate comparisons between studies and the synthesis of results. Recommendations are made with this goal in mind and to increase the rigor and relevance of findings from future trials.

Future randomized trials of pathogen-reduced PLT products should be based on a clearly stated hypothesis driven by an important research question, a design that is optimal for the research question, outcomes that relate to the research question, clearly defined observation periods, and statistical analyses that lead to valid tests of these hypotheses and associated estimates of treatment effect.

We measured blood group H and A antigens on PLTs from 100 normal volunteers using fluorescent-conjugated reagents and flow cytometry. Individuals were also genotyped at the ABO locus using a commercially available genotyping system.

Expression of A and H antigen varied widely on PLTs from different individuals. Among group A and AB persons, H antigen expression was significantly greater than A antigen (p < 0.0001). The ratio of H-to-A antigen expression varied more than 100-fold in a predictable fashion according to genotype with values lowest in A¹/A¹ < A¹/O < A²/A² < A²/O. H and A antigen expression was unaffected by secretor status. The proportion of PLTs with high A expression also varied directly according to genotype.

Blood group A and H antigen expression on PLTs varies in a predictable fashion according to genotype. Flow cytometry and genotyping identify individuals who strongly express A antigens, a finding that may be relevant to clinical PLT transfusion across ABO barriers.

Since February 2009, platelets were not produced from at-risk donors. Since May 2010, at-risk donors were tested for Trypanosoma cruzi antibodies. Donors testing positive were interviewed about risk factors, and lookback studies were initiated.

There were 7255 at-risk donors of 421,979 donors screened (1.72%). Risk factors were born in Latin America (50.6%), mother or maternal grandmother born in Latin America (28%), and 6 months or more travel history or residence in Latin America (19%). Sixteen (16) at-risk donors had T. cruzi repeat-reactive test results of whom 13 confirmed positive. Eleven of 13 were born in Latin America (nine in Paraguay and two in Argentina), and the other two were born in Canada but had short-term travel history and mothers who had been born in Latin America. Ten of the donors spoke German as their first language (all of those born in Paraguay and one born in Canada). There were 148 previous donations (176 components transfused) evaluated by lookback, of which 28% of recipients could be tested. None were positive.

Selective testing has mitigated a small risk to the blood supply with very few false-positive results. Most positive donors were born in a risk country, with a concentration of German-speaking immigrants from Paraguay. Residency or travel alone were not clear risk factors.

Donor samples that were positive by CLEIA-B19V screening were tested for B19V DNA. The sensitivity of CLEIA-B19V was tested using samples of all three genotypes and B19V DNA–positive donations. B19V DNA–positive donations and pooled plasma were quantitatively assayed for B19V DNA. B19V DNA–positive donations were phylogenetically analyzed by polymerase chain reaction direct sequencing.

The sensitivity of CLEIA-B19V was inferred to be approximately 6.3 log IU/mL with the genotype samples and 6.4 log IU/mL with B19V DNA–positive donor samples. Of 417 CLEIA-B19V–positive samples from 1,035,560 donations in Hokkaido, Japan, 101 were positive for B19V DNA. The 198 strains of B19V DNA–positive donations in Hokkaido over the past 15 years clustered exclusively with Genotype 1. After introduction of CLEIA-B19V, the viral load for B19V DNA in all 772 pooled plasma for fractionation from donors in nationwide Japan did not exceed 4 log IU/mL.

CLEIA-B19V can detect all three genotypes of B19V (viral load >6.3 log IU/mL) and limit the viral load (<4 log IU/mL) in pooled plasma, and thus such screening has further reduced the risk of transfusion-transmitted B19V infection. These results show that CLEIA-B19V screening at the JRC Blood Centers can be an alternative approach to comply with recommendations regarding B19V in the United States and Europe.

We tested 2150 blood donors in MN for the presence of antibodies against B. microti using an immunofluorescent assay (IFA). Donors identified as positive (≥64) were also tested by real-time polymerase chain reaction (PCR) for the presence of parasite DNA. Seropositive donors were contacted by phone and asked questions regarding tick exposure. Donors positive by IFA were indefinitely deferred from donating blood.

A total of 2150 donations were tested between October 2010 and November 2011. Forty-two donors (2.0%) were positive by IFA and one was also PCR positive. All positive donors reported extended outdoor activities, 12 recalled finding ticks on their body, and six had flu-like symptoms since their last blood draw.

This study provides new data about B. microti seroprevalence in MN blood donors. Possibly because the targeted collection areas were mostly expected to be endemic for the parasite, the observed seroprevalence levels were higher than expected, although the geographic distribution of positive donors did not completely overlap with the distribution of reported clinical cases in MN.