bluuebunny/crossref_metadata_2025 huggingface dataset:
Effective ultraviolet irradiation of platelet concentrates in teflon bags
Authors: Capon S. M. , Sacher R. A. , Deeg H. J.
Published: March 2003

Several plastic materials used in blood storage were evaluated for their ability to transmit ultraviolet B (UVB) light. A plastic bag manufactured from sheets of transparent Teflon efficiently (78–86%) transmitted UVB light and was employed in subsequent functional studies of lymphocytes and platelets exposed to UVB light while contained in these bags. In vitro experiments showed a UVB dose‐dependent abrogation of lymphocyte responder and stimulator functions, with concurrent preservation of platelet aggregation responses. In a phase I pilot study, UVB‐treated platelet concentrates were administered to four bone marrow transplant recipients. Adverse effects attributable to the transfusions were not observed, and patients showed clinically effective transfusion responses. No patient developed lymphocytotoxic HLA or platelet antibodies. These studies suggest that platelets can be effectively irradiated with UVB light in a closed system. However, numerous variables, including container material, volume and composition of contents, steady exposure versus agitation, and exact UV wavelength, must be considered.
Cryopreserved platelets have decreased adhesive capacity
Authors: Owens M. R. , Cimino C. , Donnelly J.
Published: March 2003

The technique of freezing blood platelets has proven very useful in transfusion support of some patients who have become alloimmunized by prior transfusions. Although transfused frozen platelets have an acceptable life span in vivo, functional defects have been found when these cells were tested in vitro. The adhesive properties of frozen platelets were investigated by use of a modified Baumgartner chamber to perform paired perfusion studies of fresh versus frozen platelets or fresh versus 5‐day‐stored platelets from the same whole blood unit. Platelets were either frozen in liquid nitrogen with dimethyl sulfoxide as the cryopreservative or stored under standard blood bank conditions for 5 days. The freeze‐thaw recovery of platelets was 73 +/− 8 percent. Frozen platelets exhibited a significant decrease in platelet adhesion as compared to fresh platelets from the same unit; adhesion of frozen platelets was only 53 percent of that of fresh platelets (p = 0.04). A slight, but insignificant decrease was noted with platelets stored for 5 days (86%, p = 0.197). These findings indicate that frozen‐thawed platelets have a significant defect in adhesive capacity as compared to fresh platelets, and that platelets stored under blood bank conditions for 5 days maintain adhesive capacity well.
Creating a uniform donor medical history questionnaire
Authors: Kolins J. , Silvergleid A. J.
Published: March 2003

Blood Centers of California, Inc. (BCC) is a consortium of 19 independent blood centers that collectively draw approximately 800,000 units of whole blood each year. Over the past several years, a committee of BCC, composed primarily of medical and nursing directors at member institutions, developed a Uniform Donor Medical History Questionnaire (UDMHQ). The UDMHQ, consisting of 42 questions, satisfies the requirements of the United States Food and Drug Administration, the State of California, the American Association of Blood Banks, and the special concerns of the members of BCC. The primary benefit of standardizing the donor medical history was an improvement in its content. Other, unproved benefits include delivering a consistent message to donors and potentially providing some legal protection to participating blood centers.
In vitro characteristics of volume‐reduced platelet concentrate stored in syringes
Authors: Pisciotto P. T. , Snyder E. L. , Napychank P. A. , Hopfer S. M.
Published: March 2003

For convenience, small volumes of platelet concentrate (PC) intended for neonatal patients are often dispensed in syringes. The PC, however, may remain in the syringe for up to several hours before the actual transfusion. As there are few data on the effect of such syringe storage on PCs, the in vitro syringe storage properties of small volumes of 1‐ and 5‐day‐old units, and volume‐reduced units of PC were evaluated. In four separate experiments, PCs were stored in syringes in volumes of 10, 15, or 30 mL for up to 6 hours at 20 to 24 degrees C without agitation. Platelets were evaluated for pH, platelet count, and a variety of biochemical and in vitro functional assays. Results showed that even with the equivalent of a full unit of platelets stored in the syringe for up to 6 hours, the pH did not fall below 6.0. Although there was an increase in lactate production and consumption of glucose, which paralleled the decline in pH, the changes were not greater than those seen in platelets stored up to 5 days in gas‐permeable blood bags. Similar results were seen for PCs stored in syringes for 6 hours at 37 degrees C. All of the pH levels recorded at the end of 6 hours of syringe storage were above the minimum required level of pH 6.0. Data from in vitro platelet assays imply that at any time during their shelf life, PCs can be stored in gas‐impermeable polypropylene syringes for up to 6 hours and can maintain acceptable storage characteristics; in vivo data are needed to confirm these observations.
Progressive platelet activation with storage: evidence for shortened survival of activated platelets after transfusion
Authors: Rinder H. M. , Murphy M. , Mitchell J. G. , Stocks J. , Ault K. A. , Hillman R. S.
Published: March 2003

Platelets are known to become activated during storage, but it is unclear whether such activation affects recovery or survival after platelet concentrate (PC) transfusion. With the use of flow cytometry to determine the percentage of platelets expressing the alpha‐granule membrane protein 140 (GMP‐140), a known adhesive ligand appearing on the platelet surface after activation, several studies were conducted. These investigations evaluated 1) the occurrence of significant platelet activation over time in PCs (n = 46) stored under standard blood bank conditions; 2) the correlation between platelet activation and platelet recovery in normal subjects after PC storage (n = 12), as assessed by the recovery of Indium‐labeled platelets; and 3) the recovery of activated and unactivated platelets in thrombocytopenic cancer patients transfused with standard PCs (n = 11). It was determined 1) that an increasing duration of storage of PC was associated with increasing platelet activation as measured by the percentage of platelets expressing GMP‐140, progressing from a mean of 4 +/− 2 percent (SD) on the day of collection to a mean of 25 +/− 8 percent by 5 days of storage: 2) that, in normal subjects, posttransfusion recovery of autologous platelets stored for 2 to 4 days and then labeled with In111 was inversely correlated with the percentage of activated platelets in the transfused PC (r = ‐0.55, p = 0.05); and 3) that, when thrombocytopenic patients were transfused with standard PCs, the recovery of the activated platelets in the transfused PCs averaged only 38 +/− 15 percent of the number predicted by the absolute platelet increment. It can be concluded that significant platelet activation occurs with standard platelet storage over 5 days and that activated platelets that express GMP‐140 are preferentially cleared from the circulation after transfusion.
A multicenter study on the efficiency of white cell reduction by filtration of red cells
Authors: Masse M. , Andreu G. , Angue M. , Babault C. , Beaujean F. , Bidet M.L. , Boudart D. , Calot J.P. , Cotte C. , Follea G. , Gérota Jean , Hau Françhise , Hurel Catherine , Legrand Edwige , Marchesseau Brigitte , Nasr Olivier , Robert Françoise , Royer Didier , Schooneman François , Tardivel René , Vidal‐Obert Madeleine
Published: March 2003

To evaluate accurately the current performance of filtration, the French Produits Sanguins Labiles study group, composed of 21 transfusion teams, conducted a large‐scale 6‐month study involving over 1400 filtrations and 3000 controls. Some 745 standard red cell concentrates (RBC concentrates) and 690 concentrates previously white cell (WBC)‐reduced by removal of buffy coat (BC‐poor RBC concentrates) were filtered using six commercially available filters: at least 170 results were collected per filter, spread among a minimum of three teams. Prefiltration controls show that the removal (manual and automated) of the buffy coat results in an initial WBC reduction of approximately 63 percent, along with a hemoglobin loss of 4 g (7%). After filtration, residual WBCs were counted in the Nageotte manual counting chamber. The reliability of this counting method, which is simple and adapted to low WBC concentrations, was characterized in this study by a 25‐percent coefficient of variation (CV) for a concentration of 2.5 WBCs per microL (i.e, 0.6 × 10(6) WBCs/filtered unit). The analysis of the results shows that, for five of six filters (1 filter was excluded), the postfiltration median value of residual WBCs was 1.1 × 10(6) in filtered RBC concentrates (n = 590), whereas it was 0.34 × 10(6) in filtered BC‐poor RBC concentrates (n = 581). The difference is significant (p < 10(−8), Wilcoxon test). Hemoglobin loss due to filtration varies according to the filter, from 5.7 ±/− 2.2 to 17.3 ±/− 2.5 g. The study data confirm that the combination of prefiltration BC removal plus filtration results in a reduced number of residual WBCs (99% of filtered BC‐poor RBC con‐ centrates contained <5 times 105 WBCs, as compared to only 81% of filtered RBC concentrates). However, these combined methods result in a significantly higher overall hemoglobin loss than filtration alone (10 g overall hem lobin loss for filtration alone vs. 15 for combination of BC removal plus filtration).
Unaltered lymphocyte subsets in hepatitis C virus‐seropositive blood donors
Authors: Prince H.E. , Fang C.T.
Published: March 2003

Lymphocyte subsets were evaluated by dual‐color flow cytometry in whole blood specimens from 35 blood donors who were seropositive on enzyme‐linked immunosorbent assay (ELISA) for hepatitis C virus (HCV) and whose sera reacted in a four‐antigen recombinant immunoblot assay (RIBA) (referred to as the HCV+R group), 15 donors who were seropositive on ELISA for HCV with indeterminate or negative RIBA results (the HCV+I/N group), and 25 HCV‐seronegative controls (HCV ‐ group). The cell subsets assessed included natural killer cells, B cells, T cells, CD4 and CD8 subsets of T cells, and T‐cell subsets defined by the coexpression of markers that appear (HLA‐DR, CD25, CD38) or disappear (CD45RA) after activation. A one‐way analysis of variance revealed no significant differences among the three study groups. These findings show that, unlike cytomegalovirus‐ and human immunodeficiency virus‐positive individuals, HCV‐positive individuals do not exhibit lymphocyte alterations indicative of the immune activation caused by chronic viral infection.
Prevention of transfusion‐associated cytomegalovirus infection in neonatal patients by the removal of white cells from blood
Authors: Eisenfeld L. , Silver H. , McLaughlin J. , Klevjer‐Anderson P. , Mayo D. , Anderson J. , Herson V. , Krause P. , Savidakis J. , Lazar A. , Rosenkrantz T. , Pisciotto P.
Published: March 2003

The usual methods employed to reduce the risk of transfusion‐associated cytomegalovirus (TA CMV) disease have been to transfuse blood or cellular blood components that are CMV antibody‐negative or to administer deglycerolized frozen red cells. To determine if the reduction of white cells (WBCs) in blood by filtration will also eliminate TA CMV disease in a high‐risk population, 48 surviving very low birth weight (less than 1250 g) neonatal infants born to CMV‐ seronegative mothers at three participating institutions in the Hartford, Connecticut area and receiving at least one CMV‐seropositive blood transfusion were studied. The incidence of TA CMV disease in 26 neonatal patients who received blood prepared by a modified spin‐cool‐ filter technique and in 22 neonatal patients who received blood filtered through a WBC‐reduction filter was compared with the incidence of transfusion‐associated disease in similar populations reported in other studies. The CMV antibody prevalence of the blood donor population was found to be 37 percent. At the time of discharge of the individual neonatal infants in the population studied, and/or 2 to 6 months later, 47 of the 48 who had undergone transfusion had CMV antibody‐negative serologic tests and/or urine culture. The other infant transiently seroconverted because of passive transfer of the antibody. None of the 48 neonatal infants had clinical evidence of CMV infection. This study indicates that WBC reduction of donor blood can reduce and perhaps prevent TA CMV disease in high‐risk neonatal patients.
Ultraviolet‐B irradiation of platelets: a preliminary trial of efficacy
Authors: Sherman L. , Menitove J. , Kagen L.R. , Davisson W. , Lin A. , Aster R.H. , Buchholz D.H.
Published: March 2003

Prior studies established that ultraviolet‐B light (UVB) irradiation of platelet concentrates (PCs) at appropriate doses can eliminate the mixed lymphocyte culture‐stimulating and ‐responding capacity of lymphocytes in the PCs without adversely affecting in vitro platelet function. The in vivo recovery and survival and in vitro characteristics of UVB‐irradiated platelets were investigated in paired studies. PCs were stored for 1 day and then exposed to UVB. Platelet recovery, survival, and function were comparable to those of nonirradiated platelets. Recovery and survival of platelets stored for 5 days before UVB exposure were decreased relative to controls, although they were considered clinically acceptable. Paired transfusion studies were also performed in seven thrombocytopenic patients by using platelets obtained by apheresis. Comparable posttransfusion platelet increments and bleeding time corrections were obtained with both irradiated and control (nonirradiated) platelets. It can be concluded that platelets survive and function relatively normally in vivo after UVB irradiation sufficient to abolish lymphocyte reactivity in mixed lymphocyte culture. Long‐term studies of UVB‐irradiated PCs are needed to assess their potential in reducing recipient alloimmunization.
The efficacy of subcutaneous recombinant human erythropoietin in the correction of phlebotomy‐induced anemia in autologous blood donors
Authors: Biesma D.H. , Kraaijenhagen R.J. , Marx J.J. , van de Wiel A
Published: March 2003

The efficacy of subcutaneous recombinant human erythropoietin (rhEPO) (500 U/kg; administered twice a week during the 3 weeks before surgery) in the recovery of preoperative hemoglobin concentrations within a 3‐ week period was studied in 40 patients, each of whom donated 2 units (900 mL) of blood for their own use before total hip replacement surgery. Twenty autologous blood donors received rhEPO (EPO group) and 20 were not treated (control group). The initial hemoglobin concentration (14.0 ± 1.0 g/dL [140 ± 10 g/L]) was completely recovered before surgery (14.0 ± 1.6 g/dL [140 ± 16 g/L]) in the EPO group, while a decrease from 13.8 ± 1.1 to 12.2 ± 1.3 g per dL (138 ± 11 to 122 ± 13 g/L) was observed in the control group. The preoperative reticulocyte count showed more than sixfold increase in the EPO group, whereas a twofold to threefold increase was found in the control group. Serum ferritin concentration fell to 42 ± 29 micrograms per L in the EPO group and to 54 ± 35 micrograms per L in the control group. The postoperative serum erythropoietin concentration in the EPO group was significantly lower than that in the control group, but it did not differ from the pretreatment value and was attended by a higher hemoglobin concentration after surgery. Only transient flu‐like symptoms were mentioned by patients who were treated with rhEPO. Changes in blood pressure or platelet count or other adverse events were not observed. This study demonstrates that subcutaneous rhEPO is safe and effective for the complete correction of the loss of 2 units of blood within a 3‐week period.
Red cell alterations associated with virucidal methylene blue phototreatment
Authors: Wagner S.J. , Storry J.R. , Mallory D.A. , Stromberg R.R. , Benade L.E. , Friedman L.I.
Published: March 2003

Several red cell storage properties were evaluated following phototreatment with methylene blue (MB) under conditions that inactivated > or = 6 log10 of added vesicular stomatitis virus. Red cell 2,3 DPG levels were similar to untreated controls throughout conventional 42‐day storage at 4 degrees C. Plasma hemoglobin levels were elevated approximately twofold in MB‐phototreated samples, and morphology scores were 5 percent lower after 42‐day storage. ATP levels declined 30 percent in phototreated samples and in a control sample containing MB and not exposed to light. Lipid peroxidation was not observed in treated or control cells, nor were differences observed in sodium dodecyl sulfate‐polyacrylamide gel electrophoresis of ghost membranes derived from phototreated and control samples. Phototreated cells exhibited enhanced ion permeability; sodium and potassium levels approached equilibrium with the suspending medium within 4 to 7 days after treatment. Direct agglutination tests using rabbit anti‐human IgG or rabbit anti‐human serum albumin on MB‐phototreated cells indicated that serum proteins had absorbed to the surface of treated red cells. Plasma depletion by washing red cells prior to phototreatment did not prevent protein binding upon subsequent addition of untreated autologous or group AB plasma. To a much smaller extent, phototreatment of plasma resulted in IgG association with untreated red cells. The addition of glutathione to red cell suspensions prevented IgG binding to phototreated red cells but did not prevent enhanced ion permeability. Taken together, these data suggest that the red cell surface is altered by virucidal MB phototreatment of vesicular stomatitis virus.
Use of erythropoietin to increase the volume of autologous blood donated by orthopedic patients
Authors: Mercuriali F. , Zanella A. , Barosi G. , Inghilleri G. , Biffi E. , Vinci A. , Colotti M.T.
Published: March 2003

For patients who donate blood for autologous use and undergo major orthopedic surgery, low basal hematocrit (Hct) is the major cause of allogeneic blood exposure. To determine whether recombinant human erythropoietin (rHuEPO) could increase autologous blood procurement and reduce allogeneic blood exposure, a prospective randomized study was conducted in 50 women undergoing total hip replacement who had basal Hct < 40 percent (0.40). Patients were randomly placed in three groups: those receiving placebo, those receiving 300 U of rHuEPO per kg, and those receiving 600 U of rHuEPO per kg every 3 to 4 days for 21 days. Oral iron (125‐270 mg/day) was given; in the last 24 patients, 100 mg of iron saccharate was administered intravenously at each donation. At each visit, 350 mL of blood was collected if Hct was > or = 34 percent (0.34). Patients receiving rHuEPO donated a greater amount of blood for autologous use than did patients in the placebo group (4.5 +/− 1.1 vs. 2.8 +/− 0.6 units; p < 0.05) and received a significantly lower amount of allogeneic blood (1.2 +/− 1.4 vs. 0.4 +/− 0.8 units; p < 0.05). No difference between the effects of the two doses of rHuEPO was observed. Iron support was a critical factor in the efficacy of treatment. No untoward effects were observed. The rHuEPO emerged as a safe and effective treatment, with adequate iron support, by which to increase preoperative deposit of autologous blood and to reduce exposure to allogeneic blood for patients with low basal Hct.
A pattern of 5‐1‐1 and c100‐3 only on hepatitis C virus (HCV) recombinant immunoblot assay does not reflect HCV infection in blood donors
Authors: Busch M.P. , Tobler L. , Quan S. , Wilber J.C. , Johnson P. , Polito A. , Steane E. , Zola A. , Bahl C. , Nelles M. , Lee S.R.
Published: March 2003

Current criteria for a reactive (positive) interpretation on hepatitis C virus (HCV) recombinant immunoblot assay (RIBA) require > or = 1+ reactivity to at least two of the four HCV antigens present in the assay. Given that 5‐1‐1 is a subcomponent of c100‐3, there is concern that donor samples reacting only with these two antigens (and not with c22‐3 or c33c) could be incorrectly classified as positive on the basis of limited reactivity to only one HCV gene product. It is determined that 0.23 to 0.44 percent of HCV enzyme immunoassay‐repeatably reactive donor sera demonstrate a pattern of 5‐1‐1 and c100‐3 only on RIBA. Evaluation of six such donor sera using peptide enzyme immunoassays spanning the c100‐3 antigen showed highly restricted reactivity to the 5‐1‐1 N‐terminal region of c100‐3, in contrast to broad 5‐1‐1 and c100‐ 3 C‐terminal peptide reactivity observed in the majority of donor sera with other positive RIBA patterns. HCV polymerase chain reaction and follow‐up serologic evaluations of four of these donors indicated the absence of viremia or evolving seroconversion in all cases. It is concluded that, in the blood donor setting, a pattern of only 5‐1‐1 and c100‐3 reactivity is typically not indicative of HCV infection. To avoid overinterpretation, it is recommended that RIBA grading criteria be revised to require reactivity to two or more HCV‐encoded gene products.
The effect of solvent/detergent‐treated plasma on stored platelet concentrates
Authors: Tocci L. J. , Napychank P. , Cable R. G. , Snyder E. L.
Published: March 2003

The treatment of fresh‐frozen plasma (FFP) with a solvent/detergent (S/D) solution to inactivate contaminating viruses has been shown to be effective in reducing virus transmission while maintaining the hemostatic properties of the plasma. FFP is treated with tri(n‐ butyl)phosphate (solvent) and Triton X‐100 (detergent) and then purified; the in vivo effect of the residual S/D has been reported to be minimal. In clinical transfusion practice, ABO‐incompatible, HLA‐ matched, single‐donor platelets may have to be resuspended in ABO‐ compatible plasma. The use of S/D‐treated plasma for this purpose would remove the added risk of transfusion‐transmitted diseases due to the use of another blood component. As there are no data on the use of S/D‐ treated plasma as a platelet‐resuspending medium, the potential toxicity of the residual solvent and detergent on the in vitro function and integrity of platelets stored in S/D‐treated plasma for 5 days was studied. A repeated‐measures analysis of variance was used for statistical analysis. Results showed that, as compared to controls (non‐ S/D‐treated plasma), platelets resuspended in S/D‐treated plasma maintained their functional properties, including morphology score and osmotic recovery, for up to 5 days of storage (p > 0.05, NS). No significant changes were seen among S/D‐treated plasma and control groups for platelet count, lactate dehydrogenase discharge, beta‐ thromboglobulin release, glucose utilization, or generation of lactate. Measurement of pO2 and pCO2 values showed some differences between S/D‐ treated plasma and control groups that were significant, but not clinically significant. The pH values for all four groups ranged from 7.1 to 7.4 on Day 5. As compared to plasma controls, platelets resuspended in S/D‐treated plasma and stored for up to 5 days maintain acceptable in vitro storage characteristics for use in clinical transfusion practice.
Analysis of the repertoire of human B‐lymphocytes specific for type A and type B blood group terminal trisaccharide epitopes
Authors: Conger J.D. , Chan M.M. , DePalma L.
Published: March 2003

Naturally occurring serum antibodies specific for the A and B blood group isoantigens are of great importance in medicine. By using A‐type terminal trisaccharide (ATS) or B‐type terminal trisaccharide (BTS) coupled to albumin as coating antigens, an enzyme‐linked immunosorbent assay capable of detecting all ATS/BTS‐binding antibodies was performed. The combination of this enzyme‐linked immunosorbent assay with limiting‐dilution methodology, using a polyclonal B‐lymphocyte activator, permitted a monoclonal analysis of the human antibody repertoire that is specific for ATS and BTS in persons of different blood types. Most (78%) positive supernatants from type O cultures were monospecific for either ATS or BTS, and these were present at roughly equivalent frequencies. Nine supernatants with reactivity toward both ATS and BTS were tested by red cell adsorption; six had properties expected for true dually reactive monoclonal antibodies: adsorption with either A1 or B red cells eliminated both anti‐ATS and anti‐BTS activity. This finding accords with a monoclonal origin for anti‐A,B. The analysis of cultures of peripheral blood lymphocytes from type A and B donors unexpectedly showed significant numbers of clones with apparent autospecificity. However, none of the anti‐ATS‐positive supernatants from type A cultures or anti‐BTS‐positive supernatants from type B cultures were adsorbable with A1 or B red cells, respectively. Consideration of only true (adsorbable) positives indicates that the type A and B anti‐trisaccharide repertoires differ significantly from the type O repertoire, probably as a result of the action of normal self‐tolerance mechanisms.
Safety and efficacy of preoperative donation of blood for autologous use by patients with end‐stage heart or lung disease who are awaiting organ transplantation
Authors: Goldfinger D. , Capon S. , Czer L. , Leibfreid J. , Trento A. , Ross D. , Waters P. , Klapper E. , Pepkowitz S.
Published: March 2003

Many patients are, perhaps inappropriately, denied the benefits of autologous blood transfusion, because they are thought to be too ill to donate blood safely. The safety and efficacy of autologous blood donation by selected patients with end‐stage heart or lung disease who are awaiting organ transplantation were studied to determine if even these critically ill patients could be suitable candidates for autologous blood donation. Seventy‐two adults awaiting heart or lung transplantation were evaluated for autologous blood donation in a hospital‐based blood collection facility. Phlebotomy was performed if the patient met the required medical eligibility protocol, and if he or she consented to participate. Units of blood were separated into packed red cells and plasma and stored in a frozen state. Of 48 heart transplant candidates, 31 (65%) were each able to donate 1 to 8 units of blood. The median number of exposures to allogeneic components was 1 for patients who donated and 7 for nondonors (p = 0.0141). Among patients who donated, 54 percent required allogeneic components, as compared to 88 percent of nondonors (p = 0.0968). Of 24 lung transplant candidates, 15 (63%) made 1 to 6 donations each. The median number of exposures to allogeneic components was 0 for donors and 2 for nondonors (p = 0.1871), but only 45 percent of donors required allogeneic components, as compared to 100 percent of nondonors (p = 0.0418). No serious complications during or following phlebotomy were observed. It is concluded that autologous blood donation by patients with end‐stage heart or lung disease may be safe. However, to delineate fully which patients can participate, additional stuchs of safety are required, including more objective measurements of postdonation hemodynamics in these and other “high‐risk' patients. But, because avoidance of exposure to allogeneic components or significant reduction in total exposures has been possible for many patients, it is imperative that further studies of safety and effectiveness be undertaken. The out‐ come of this current study sug ests that the benefits of preoperative autologous blood donation can be extended safely to man patients previously considered too sick to participate.
Storage of pools of six and eight platelet concentrates
Authors: Moroff G. , Holme S. , Dabay M.H. , Sawyer S. , Heaton W.A. , Law P. , Alsop P.
Published: March 2003

Platelet concentrates (PCs) prepared from units of whole blood are routinely stored singly at 20 to 24 degrees C and pooled prior to transfusion. Studies have been conducted to evaluate the in vitro properties of pools of six (n = 19) and eight (n = 17) ABO‐identical PCs after storage, with comparative studies involving single units (n = 33). The pools were prepared using the sterile connecting device. One‐ day‐old and 3‐day‐old PCs were pooled and stored for a total of 5 days in a container system consisting of two 1000‐mL polyolefin containers. The pooled platelet suspension was divided approximately equally between the two containers. The platelet count was reduced by less than 5 percent during storage of the pools, which is similar to the reduction found with storage of control units of single PCs. The volume loss due to pooling was 9.6 +/− 1.9 percent (mean +/− 1 SD). The pH of the PC pools was approximately 7.0 after 5 days of storage, with no pool having a pH below 6.2. In vitro platelet properties, such as morphology score, extent of shape change induced by ADP, total ATP, aggregation response to ADP and collagen, response to hypotonic stress, lactate dehydrogenase discharge, and beta‐thromboglobulin release, were similar for pools and control single PCs. In addition, comparable low levels of thymidine uptake were detected in the mononuclear leukocyte fraction of pooled and unpooled PCs that were stored for 5 days at 20 to 24 degrees C, which indicates that the mixing of lymphocytes in the pool did not stimulate in vitro immunologic reactions. Culturing of samples at the conclusion of storage did not show evidence of bacterial contamination. These studies demonstrate that pools of ABO‐identical PCs have satisfactory in vitro platelet properties after storage.
In vivo viability studies of two additive solutions in the postthaw preservation of red cells held for 3 weeks at 4°C
Authors: Moore G.L. , Hess J.R. , Ledford M.E.
Published: March 2003

An optimized additive solution was developed for the postthaw preservation of red cells that contained adenine, glucose, disodium phosphate, and citrate buffer. This solution, called AS‐17, was compared to AS‐3 solution in a clinical trial using 40 subjects (20 in each arm). Fresh‐frozen red cells were thawed and deglycerolized after 1 to 18 months and subjected to a second period of storage in either solution for up to 3 weeks at refrigerator temperatures. Both solutions yielded red cells with 24‐hour survivals in excess of 75 percent. Cells stored in AS‐3 for 21 days had a mean survival of 77 +/− 8 percent and cells stored in AS‐17 a mean survival of 79 +/− 11 percent. The AS‐17 solution resulted in improved maintenance of pH, p50, and 2,3 DPG compared to that with AS‐3, but both solutions appear adequate for 3 weeks of postthaw storage.
Quantitation of residual white cells in filtered blood components by polymerase chain reaction amplification of HLA DQ‐A DNA
Authors: Lee T.‐H. , Stromberg R. , Heitman J. , Tran K. , Busch M. P.
Published: March 2003

BACKGROUND: Over the past several years, blood filtration technology has improved dramatically, such that currently available experimental filters are capable of reducing white cells (WBCs) in blood components to less than 0.1 WBC per microL. These residual WBC concentrations are below the sensitivity of automated cell counters, as well as of large‐ volume (Nageotte) hemocytometers.

STUDY DESIGN AND METHODS: A quantitative polymerase chain reaction (PCR) amplification assay directed at HLA DQ‐A DNA sequences has been developed for the enumeration of WBCs in filtered blood. To ensure quantitative recovery of WBCs at very low residual cell concentrations, a direct red cell lysis and WBC concentration protocol using 0.5 mL of filtered blood was perfected. Amplified product is detected by oligomer hybridization using 32P‐labeled probes, with quantitation by image analysis of autoradiographic signals relative to a standardized dilution series processed in parallel.

RESULTS: Recovery of residual WBCs in filtrates was shown to be enhanced by the addition of xenogeneic WBCs or polystyrene beads, which served as “carrier” particles during red cell lysis and wash steps. A contribution of nuclear fragments in filtered blood to PCR signal in the range of 0.01 to 0.5 WBCs per microL was observed; a modified protocol was developed to minimize this effect. Parallel analysis of spiked dilution series and evaluations of 39 red cell components filtered through commercial filters indicated good correlation between PCR and standard Nageotte counts in the range of 0.1 to 10 WBCs per microL (r2 = 0.94); only PCR was able to detect residual WBCs in filtrates from prototype 6 log10 WBC‐reduction filters.

CONCLUSION: This assay should prove useful for the development and quality assurance of increasingly efficient WBC‐reduction filters.
Altered expression of adhesion molecules (L‐selectin and Mac‐1) on granulocytes during storage
Authors: Wikman A. , Lundahl J. , Fernvik E. , Shanwell A.
Published: March 2003

Background: The success of granulocyte transfusion therapy for neutropenic patients with sepsis is dependent on the number and quality of the granulocytes transfused. There is a progressive impairment in granulocyte function during storage.

Study Design and Methods: The effect of 1 to 2, 4, 24, and 48 hours' storage on receptor expression associated with granulocyte function has been analyzed.

Results: After 24 hours' storage, significant changes were found in the expression of receptors associated with adhesion to the endothelium: a decrease in L‐ selectin expression (p < 0.01) and an increase in Mac‐1 expression (p < 0.01). Receptors (CR1 and FcRIII), associated with adhesion to target, were either increased (CR1) or unaltered (FcRIII). The capacity to produce a reactive oxygen metabolite (hydrogen peroxide) remained essentially unchanged after 48 hours' storage. The ability of N‐formyl‐ methionyl‐leucyl‐phenyl alanine to mobilize Mac‐1 and CR1 was reduced after 48 hours' storage.

Conclusion: Since the regulation of adhesion molecules is important for the recruitment of granulocytes into an inflammatory site, the observed in vitro changes in L‐selectin and Mac‐ 1 expression during storage may be of importance for the quality of granulocyte concentrates.
Iron and erythropoietin measurement in autologous blood donors with anemia: implications for management
Authors: Tasaki T. , Ohto H. , Noguchi M. , Motoki R. , Kikuchi S. , Sato A. , Hoshino S.
Published: March 2003

Background: The importance of autologous blood donation for elective surgery is recognized, and the method is being used at many hospitals. Not all patients are able to deposit a sufficient amount of blood before surgery because they cannot recover rapidly enough from phlebotomy‐induced anemia. The ability to donate sufficient blood for autologous use was studied in patients who are particularly susceptible to phlebotomy‐induced anemia.

Study Design and Methods: Of 840 patients who donated blood for autologous use in elective surgery from November 1987 through May 1993, 20 with rheumatoid arthritis, 24 with iron deficiency anemia, and 37 aged 65 years and above with normocytic anemia were compared with 24 nonanemic elderly patients who donated a total of 1000 mL of blood for autologous use. Patients received iron sulfate orally and donated blood once a week until operation.

Results: The amount of blood collected before surgery per control patient was more than that in others. Consequently, there was a tendency to allogeneic blood transfusion in patients with rheumatoid arthritis or elderly patients. The ferritin levels in controls and in patients with iron deficiency anemia during the donation period were almost within the normal range in spite of iron supplementation, which implied a good utilization of iron sulfate for erythropoiesis. On the other hand, the rise in ferritin levels in the elderly and in patients with rheumatoid arthritis suggested inappropriate iron availability for erythropoiesis and resulted in an increase in iron storage. Since an adequate endogenous erythropoietin response to phlebotomy‐induced anemia was not observed in these patients, impaired erythropoietin production was considered one of the reasons for anemia.

Conclusion: Patients with iron deficiency anemia are able to continue donating blood for autologous use so long as they have sufficient iron supplementation. However, the elderly or those with rheumatoid arthritis occasionally fail to donate a sufficient volume of blood before surgery as a result of phlebotomy‐induced anemia, which is caused in turn by impaired erythropoietin production.
Confirmation of hepatitis C infection: a comparison of five immunoblot assays
Authors: Zaaijer H.L. , Vrielink H. , van Exel‐Oehlers PJ , Cuypers H.T. , Lelie P.N.
Published: March 2003

BACKGROUND: Recently, new immunoblot assays for the detection of antibodies to hepatitis C virus (HCV) became available.

STUDY DESIGN AND METHODS: The performance of five confirmatory anti‐HCV immunoblot assays was studied with samples with known HCV antibody and HCV RNA status. The assays were a third‐generation strip recombinant immunoblot assay (RIBA‐3, Chiron Corp., Emeryville, CA), a second‐generation HCV blot (DB‐2 blot, Diagnostic Biotechnology, Singapore), the Wellcozyme HCV Western blot (Murex blot, Murex Diagnostics, Dartford, UK), an immunodot HCV assay (Matrix, Abbott Laboratories, Chicago, IL), and the third‐generation HCV line immunoassay (Liatek‐III, Organon Teknika, Boxtel, The Netherlands).

RESULTS: Sensitivity on samples from 48 HCV RNA‐positive, second‐generation RIBA (RIBA‐2)‐positive persons and specificity on samples from 31 low‐risk donors was 96 percent or better for all assays. The sensitivity on 31 HCV RNA‐positive, RIBA‐2‐ indeterminate samples was as follows: Liatek‐III, 94 percent; RIBA‐3, 90 percent; Murex blot, 61 percent; Matrix, 55 percent; and DB‐2 blot, 39 percent. In testing 39 HCV RNA‐negative, RIBA‐2‐indeterminate donor samples, the percentage found to be negative was Liatek‐III, 77 percent; RIBA‐3, 67 percent; Murex blot, 49 percent; DB‐2 blot, 33 percent; and Matrix, 15 percent. The order of sensitivity on four HCV seroconversion series was (from high to low): RIBA‐3, Liatek‐III, DB‐2 blot, Murex blot, and Matrix; the differences were small.

CONCLUSION: Detection of HCV antibodies was not refined by the addition of new HCV antigens (NS5, E2/NS1), but by improved classical antigens (core, NS3, NS4). Replacement of the commonly used RIBA‐2 will resolve the status of a high proportion of RIBA‐2‐indeterminate samples.
Lack of transmission of neutrophil and platelet antibodies by intravenous immunoglobulin in bone marrow transplant patients
Authors: KLUMPP T.R. , HERMAN J.H. , MANGAN K.F. , SCHNELL M.K. , GOLDBERG S.L. , MACWNALD J.S.
Published: March 2003

BACKGROUND: Several studies have demonstrated that the administration of intravenous immunoglobulin (IVIG) may be followed by the transient appearance of positive red cell antibody screens, positive direct antiglobulin tests, and, occasionally, frank hemolysis. However, little information is available regarding the possibility that IVIG could transmit neutrophil and/or platelet antibodies.

STUDY DESIGN AND METHODS: Serum samples were obtained both immediately before and immediately after the administration of 12 separate lots of commercially available IVIG to bone marrow transplant patients. RESULTS: None of the patients were shown by standard granulocyte immunofluorescence testing to have acquired neutrophil antibodies. Four of the 12 postinfusion sera were positive for platelet antibodies in standard platelet suspension immunofluorescence testing, but in all four instances the corresponding preinfusion serum was positive as well. CONCLUSION: The risk of acquiring neutrophil and/or platelet antibodies after the administration of commercially available IVIG appears to be low.
Quencher‐enhanced specificity of psoralen‐photosensitized virus inactivation in platelet concentrates
Authors: MARGOLIS‐NUNNO H. , ROBINSON R. , BEN‐HUR E. , HOROWITZ B.
Published: March 2003

BACKGROUND: Treatment with psoralens and UVA (PUVA) has been shown to be efficacious in eliminating the risk of virus transmission by platelet concentrates (PCs). It has previously been demonstrated that, during the inactivation of cell‐free vesicular stomatitis virus (VSV) by aminomethyltrimethylpsoralen (AMT) and UVA in PCs, platelet function could be protected either by oxygen removal before irradiation or by inclusion of a type I free radical quencher, such as mannitol.

STUDY DESIGN AND METHODS: Under previous PUVA treatment conditions for PCs (25 micrograms/mL AMT; 30 min UVA at 7 mW/cm2; 2 mM [2 mmol/L] mannitol), more than 6 log10 of added cell‐free VSV was completely inactivated. In the current study, various PUVA conditions are evaluated for efficacy in inactivating other viral forms that could be present in PCs. Maintenance of platelet integrity (i.e., platelet number, solution pH, and aggregation response during initial storage after treatment) and kill of cell‐associated VSV are examined.

RESULTS: While cell‐free viruses were inactivated efficiently under previous PUVA conditions, cell‐associated VSV and the non‐lipid‐enveloped bacteriophage M13 were not. Effective inactivation of these viruses was achieved by raising the concentration of AMT to 50 micrograms per mL and extending the period of irradiation to 90 minutes (39 J/cm2). However, for maintenance of platelet integrity under these conditions, the prior removal of oxygen or the inclusion of compounds known to quench both type I and type II photoreactants (e.g., flavonoids such as rutin) was required.

CONCLUSION: These findings suggest that the viral safety of PCs may be enhanced through treatment with AMT and UVA in the presence of flavonoids, and that flavonoid use may prove beneficial in other systems where oxygen‐mediated damage occurs
The collection and evaluation of peripheral blood progenitor cells sufficient for repetitive cycles of high‐dose chemotherapy support
Authors: Benjamin R.J. , Linsley L. , Axelrod J.D. , Churchill W.H. , Sieff C. , Shulman L.N. , Elias A. , Ayash L. , Malachowski M.E. , Uhl L. , Gaynes Lisa , McCauley Mary , Thompson Lynn , Mazanet Rosemary , Antman Karen , Schnipper Lionel E. , Tepler Isadore , Antin J.H. , Wheeler Catherine
Published: March 2003

BACKGROUND: The development of an optimized peripheral blood progenitor cell (PBPC) harvest protocol to provide support for repetitive chemotherapy cycles is described.

STUDY DESIGN AND METHODS: PBPCs mobilized by cyclophosphamide plus granulocyte‐colony‐stimulating factor (G‐CSF) were studied in 163 leukapheresis harvests from 26 lymphoma patients. Harvested cells were transfused with two chemotherapy cycles and with an autologous bone marrow transplant. Progenitor cell content was examined in the context of hematopoietic engraftment.

RESULTS: Mobilization allowed the harvest of large numbers of PBPCs. Peak harvests tended to occur after the recovering white cell count exceeded 10 × 10(9) per L. CD34+ lymphomononuclear cell (MNC) and colony‐forming units‐granulocyte‐macrophage (CFU‐GM) counts correlated poorly, but both measures peaked within 24 hours of each other in 21 of 26 patients, which demonstrated PBPC mobilization. Engraftment of platelets (> 50×10(9)/L) and granulocytes (> 500×10(6)/L) was achieved in a median of 20.5 and 16 days, respectively. A minimum number of progenitors necessary to ensure engraftment could be derived.

CONCLUSION: Cyclophosphamide and G‐CSF allowed the harvest of sufficient PBPCs to support multiple rounds of chemotherapy. Harvest should commence when the recovery white cell count exceeds 10×10(9) per L. PBPC harvest CD34+MNC counts are as useful as CFU‐GM results in the assessment of PBPC content, and they may allow harvest protocols to be tailored to individual patients.
Elimination of potential mutagenicity in platelet concentrates that are virally inactivated with psoralens and ultraviolet A light
Authors: Margolis‐Nunno H. , Robinson R. , Ben‐Hur E. , Chin S. , Orme T. , Horowitz B.
Published: March 2003

BACKGROUND: For virus sterilization of platelet concentrates (PCs), treatment with aminomethyltrimethyl psoralen (AMT) and long‐wavelength ultraviolet A light (UVA) has shown efficacy. It has been found that treatment with 50 micrograms per mL of AMT and 38 J per cm2 of UVA in the presence of 0.35‐mM rutin efficiently kills viruses while maintaining platelet integrity. There is, however, concern about the mutagenic potential of psoralens and UVA (PUVA)‐treated PCs.

STUDY DESIGN AND METHODS: Adsorption of PUVA‐treated PCs with a hydrophobic resin containing C18 as the ligand was used for AMT removal, which was quantitated by the use of radioactive AMT. PUVA‐treated PCs, with and without C18 treatment, were examined for solution pH and platelet aggregation response to agonists. In addition, residual AMT activity was determined by AMT's virucidal activity or incorporation into cellular DNA upon a second UVA irradiation and by its mutagenic potential in the Ames test.

RESULTS: After PUVA treatment of PCs, residual AMT retained virucidal and adduct‐forming ability upon re‐ exposure to UVA, but activities were less than those observed originally. As has been found previously, AMT had mutagenic potential following incubation in the dark with rat liver S9 microsomal enzymes. The PUVA treatment reduced this potential by 90 percent. C18 adsorption following PUVA treatment had no negative effect on platelet integrity and eliminated 50 percent of the added radioactive AMT. In addition, all detectable virucidal, nucleic acid‐modifying, and mutagenic activities of AMT‐treated PCs were removed by C18.

CONCLUSION: These results suggest that hydrophobic resin adsorption of PUVA‐treated PCs will conveniently remove functional psoralens and eliminates their mutagenic potential.
Long‐term follow‐up of and infectivity in blood donors with hepatitis C antibodies and persistently normal alanine aminotransferase levels
Authors: Rossini A. , Gazzola G.B. , Ravaggi A. , Agostinelli E. , Biasi L. , Albertini A. , Radaeli E. , Cariani E.
Published: March 2003

BACKGROUND: Little is known about the prevalence of serum hepatitis C virus (HCV) RNA in blood donors with HCV antibodies and persistently normal alanine aminotransferase (ALT) levels. STUDY DESIGN AND METHODS: Thirty‐nine anti‐HCV‐positive donors with normal ALT on four determinations at 3‐month intervals were further tested monthly for 6 months, and they had normal ALT values. The presence of HCV RNA was determined in these 39 donors. RESULTS: Serum HCV RNA was detected in 16 of 39 donors, 14 of 14 who reacted on second‐generation recombinant immunoblot assay (RIBA‐2) and 2 of 15 who were indeterminate. None of the 10 RIBA‐2‐nonreactive donors had evidence of viremia. The 15 RIBA‐2‐ indeterminate samples were tested with third‐generation RIBA (RIBA‐3); the results showed reactivity in 5 (including the 2 HCV RNA positive), an indeterminate pattern in 7, and nonreactivity in 3 (all RNA negative). Among HCV RNA‐positive subjects, mean age (p < 0.05), mean ALT (p < 0.001), signal‐to‐cutoff (S:CO) ratio on second‐generation enzyme‐linked immunosorbent assay (p < 0.001), and gamma globulin levels (p < 0.05) were higher than those among HCV RNA‐negative subjects. During 6 additional months of ALT monitoring, completed by 36 of 39 donors, increased values were detected in 6 (5 HCV RNA positive). In 4 of those 6, however, ALT levels were less than 1.5‐fold the upper normal limit. HCV RNA results were unchanged at the end of 1‐year follow‐up. CONCLUSION: Forty‐one percent of anti‐HCV‐positive donors with persistently normal ALT had active HCV infection. Long‐term ALT monitoring allowed the detection of significantly increased enzyme values in only 2 of 16 viremic donors. Reactivity on RIBA‐2 or ‐3, greater age, mean ALT levels in the upper range of normal, higher S:CO ratio on second‐generation enzyme‐linked immunosorbent assay, and higher gamma globulin levels were predictive of viremia.