bluuebunny/crossref_metadata_2025 huggingface dataset:
Phenotypic characterization of white cells in white cell‐reduced red cell concentrate using flow cytometry
Authors: Wenz B. , Burns E.R.
Published: March 2003

The residual white cell (WBC) content of donated units of red cell concentrate rendered WBC‐reduced by filtration through commercially available polyester filters was quantified and phenotypically analyzed. All studies were performed by flow cytometery. Quantification studies were performed with a DNA/RNA fluorophore, propidium iodide. WBC subset analyses were performed with fluorescence‐labeled monoclonal antibodies directed against various cluster differentiation (CD) loci. The results indicate that the filter removes in excess of 3 log10 total WBCs from the red cell components and depletes granulocytes to or beyond the specific assay's sensitivity of 3 log10. Total T and B cells, T4 and T8 lymphocytes, and monocytes are reduced by approximately 4 log10. These analyses provide plausible explanations for the clinical success of the filter and suggest other potential applications.
Platelet quality after 15‐day storage of platelet concentrates prepared from buffy coats and stored in a glucose‐free crystalloid medium
Authors: Bertolini F. , Rebulla P. , Porretti L. , Murphy S.
Published: March 2003

It has been reported previously that platelet concentrates (PCs) prepared from buffy coat pools diluted in a simple, glucose‐free medium (BC‐PCs) are effective in thrombocytopenic patients after 4 to 12 days of storage. Such preparations produce platelet increments similar to those of traditional PCs prepared from platelet‐rich plasma (PRP‐PCs) stored for 1 to 3 days. The purpose of this study was to obtain a series of in vitro measurements during storage to allow a more detailed characterization of BC‐PCs and a more detailed comparison of BC‐PCs with PRP‐PCs. At the beginning of storage, the level of (alpha granule membrane protein‐140 (GMP‐140), a marker of platelet activation, was significantly higher on PRP‐PC platelets, and BC‐PCs were superior in measurements reflecting platelet function, such as osmotic reversal, ATP release, and aggregation with collagen. Compared to PRP‐PCs, BC‐PCs were superior in the percentage of discs, total ATP, and glycoprotein lb expression by Day 7. This superiority became more striking on Day 11. Overall, 15‐day‐old BC‐PCs compared favorably to 7‐day old PRP‐PCs: BC‐PCs were superior in ATP release and aggregation with collagen, but they were not significantly different for all other measurements reflecting platelet quality and function. Thus, the quality of platelets in BC‐PCs was superior on Day 1, and this superiority progressed as storage continued. In addition, the metabolism of BC‐PCs was favorable. During 15 days of storage, pH and bicarbonate concentration did not change significantly In BC‐PCs; moreover, oxygen consumption was higher and glucose utilization and lactate accumulation were markedly slower than in PRP‐PCs. The results of additional experiments performed with pooled PRP‐PCs diluted In the glucose‐free medium and stored In the same fashion as BCPCs were Intermediate between those of experiments with BCPCs and PRP‐PCs, which suggests that the superior quality of BC‐PCs Is due In part to the buffy coat method of pre aration and In part to one or more components of the storage method. This approach to platelet procurement and storage has the disadvantage of a lower platelet yield. However, it can be concluded that it produces platelets of higher quality, contaminated with fewer white cells and less plasma and with better potential for storage than is possible with PRP‐PCs.
Severe immune‐mediated hemolytic anemia secondary to treatment with cefotetan
Authors: Gallagher N.I. , Schergen A.K. , Sokol‐Anderson M.L. , Sheahan E.J. , Chaplin H.
Published: March 2003

Severe acute hemolytic anemia developed in a woman following treatment with multiple antibiotics for possible postpartum uterine infection. On admission, the hemoglobin was 5 g per dL (50 g/L), the reticulocytes were 35 percent (0.350), the direct antiglobulin test was strongly positive for IgG and C3d (mixed fields), and the indirect antiglobulin test was negative. Serologic studies revealed antibody to cefotetan that reacted by both the immune complex and the drug adsorption mechanisms. Before the diagnosis of cefotetan‐related immune hemolysis was made, all medications had been discontinued, and the patient received 4 units of red cells and a short course of adrenocorticosteroids. Recovery was prompt and complete.
Transfusion‐associated graft‐versus‐host disease in a presumably immunocompetent patient after transfusion of stored packed red cells
Authors: SUZUKI K. , AKIYAMA H. , TAKAMOTO S. , MARUYAMA Y. , SAKAMAKI H. , AKAGI K. , MAEDA Y. , TAKENAKA M. , ONOZAWA Y.
Published: March 2003

A 54‐year‐old woman developed transfusion‐associated graft‐versus‐host disease (TA‐GVHD) after the transfusion of stored packed red cells obtained from unrelated donors. The patient was presumed to be immunocompetent. A diagnosis of TA‐GVHD was made by clinical features and postmortem pathologic findings. Sex chromatin analysis of the patient's lymphocytes demonstrated chimerism. HLA typing of the blood donors revealed one to be HLA‐homozygous for one of the patient's HLA haplotypes (A33‐B44‐Cblank). This case illustrates the risk in the general patient population of TA‐GVHD after routine blood transfusion therapy. Workers should be aware of this possibility and should continue searching for an efficient way to prevent it.
Inhibition of the classical activation pathway of complement‐mediated lysis by monoclonal antibodies to complement components C3c and C3d
Authors: Mushens R.E. , Bakacs T.
Published: March 2003

Eight epitope‐mapped monoclonal antibodies (MoAbs) to complement component C3d and five to complement component C3c were investigated to determine whether they could inhibit the classical activation pathway of complement‐mediated lysis (CML) by using blood group AB red cells sensitized by A or B MoAbs. Three IgM C3d MoAbs and one IgG1 C3c MoAb were able to inhibit CML in a dose‐dependent manner. In the presence of excess complement, no inhibition was observed. The greatest inhibition was observed with two high‐affinity IgM antibodies that were specific for epitope 1 on the C3d component. Some inhibition was observed with a high‐affinity IgM antibody specific for epitope 3 of the C3d component and alwo with a lower‐affinity IgG antibody specific for epitope 1 of the C3c component. The results indicate that some complement MoAbs have the capacity to distinguish between conformationally and/or functionally different forms of red cell‐bound C3.
Storage of platelets in additive solutions: a new method for storage using sodium chloride solution
Authors: Gulliksson H. , Sallander S. , Pedajas I. , Christenson M. , Wiechel B.
Published: March 2003

The in vitro effect of 6‐day storage of platelets prepared from 6 pooled buffy coat (BC) units and stored in a platelet storage medium containing approximately 40 percent CPD‐plasma and 60 percent platelet additive solution (PAS) was evaluated. PAS is composed of sodium and potassium chloride, citrate, phosphate, and mannitol. The total count of platelets per pooled unit included in the in vitro studies (n = 25) was 376 ± 59 × 109 (mean ± SD). The present study included three steps. 1. Evaluation of platelet storage in one (n=7) and two (n=6) 1000‐mL polyolefin containers using PAS. During storage in one container, significantly lower values were found for pH, pO2, glucose, ATP, and the ratio of ATP to AMP+ADP+ATP. The values for mean platelet volume, pCO2, lactate, and extracellular adenylate kinase activity were significantly higher. These results indicate that storage in only one polyolefin container is not appropriate for maintaining satisfactory platelet quality. During storage in two polyolefin containers, a remarkably decreased lactate production (0.07 ± 0.02 mmol/day/1011 platelets) was noted. 2. PAS was substituted for saline during 6‐day storage in two 1000‐mL polyolefin containers (n = 12). The composition of the platelet preparations was the same in all other respects. Similar in vitro results were noted with PAS and saline, which indicated that PAS has no specific effect on the storage of platelets different from that of saline. 3. A paired patient transfusion study (n=8) gave corrected count increments at 1 hour following transfusion of 20.4 ± 6.6 (BCs in saline) and 19.0 ± 7.9 (standard platelet concentrates prepared from platelet‐rich plasma [PCs]) and at 12 to 24 hours of 10.9 ± 4.0 (BCs) and 7.2 ± 5.4 (PCs). No significant differences were found. The results of this study indicate that saline in combination with specific storage containers can be used for the 5‐day storage of platelets prepared from pooled BCs.
Enzyme‐linked immunosorbent assay for the detection of substances that carry blood group A specificity
Authors: Montano R.F. , Romano E.L. , Moase E. , Cooper D.K. , Ye Y. , Taylor P. , Soyano A. , Tam Y.
Published: March 2003

An enzyme‐linked immunosorbent assay (ELISA) was developed to estimate the amount of material carrying blood group A activity in biologic samples. A soluble synthetic form of the A antigenic determinant (A trisaccharide, ATS) conjugated to peroxidase competes with the blood group A substance present in a biologic sample for anti‐A attached to a solid phase by a second antibody coating the plastic micro‐wells. A reference curve is constructed by using known quantities of ATS to compete with a fixed amount of ATS‐peroxidase conjugate. The A substance activity in a sample is obtained by extrapolating the degree of inhibition of the binding of the ATS‐peroxidase conjugate to an equivalent amount of ATS in the reference curve. The assay is reproducible, specific, and sensitive. It has been used in pharmacologic studies to estimate the concentration of ATS in the blood and urine of rats, rabbits, and baboons and in a study with human samples, testing the potential clinical use of ATS to neutralize anti‐A when therapeutically indicated. It is also useful for the detection of ABO natural products in secretions, thus allowing the accurate classification of secretor and nonsecretor individuals.
Fibrin glue from single‐donation autologous plasmapheresis
Authors: Casali B. , Rodeghiero F. , Tosetto A. , Palmieri B. , Immovilli R. , Ghedini C. , Rivasi P.
Published: March 2003

A method is described for preparing fibrin glue from fibrinogen obtained by double cryoprecipitation of plasma collected by plasmapheresis. Plasma (average volume, 528.8 +/− 86 mL), collected from 12 plasmapheresis donors was cryoprecipitated twice. When the cryoprecipitated pellet was resuspended in 0.5 to 2 mL of saline (total volume of saline‐resuspended fibrinogen: 5 mL), the average yield of fibrinogen was 78.4 +/− 18.3 mg per mL. This is comparable to commercial preparations, is suitable for clinical use, and offers greater safety at a reduced cost.
Prestorage removal of Yersinia enterocolitica from red cells with white cell‐reduction filters
Authors: Kim D.M. , Brecher M.E. , Bland L.A. , Estes T.J. , McAllister S.K. , Aguero S.M. , Carmen R.A. , Nelson E.J.
Published: March 2003

Prestorage removal of phagocytic white cells (WBCs) may increase the survivability of contaminating bacteria in units of stored red cells. Fourteen units of whole blood were inoculated with 65 colony‐forming units per mL of Yersinia enterocolitica (serotype O:3) and processed into AS‐3‐preserved red cells. Five red cell units were filtered with a prototype third‐generation filter and five red cell units with a second generation filter. WBC reduction was performed on the day of collection. Four red cell units were not filtered. Three noninoculated whole blood units served as negative controls; two were filtered (one with each type of WBC‐reduction filter) and one remained unfiltered. All red cell units were then stored at 4 degrees C for 42 days. One of the five filtered red cell units (20%) in each filter group supported growth of Y. enterocolitica. In contrast, 4 (100%) of 4 unfiltered inoculated red cell units had growth (p = 0.04). Overall, 2 (20%) of 10 units of WBC‐reduced red cells supported the growth of Y. enterocolitica, as compared to 100 percent of unfiltered red cell units after inoculation (p = 0.015). Bacterial contamination was not detected in any of the three noninoculated units. It can be concluded that prestorage WBC filtration significantly reduces the potential for growth of Y. enterocolitica in red cells stored at 4 degrees C for 42 days.
Removal of Yersinia enterocolitica from AS‐1 red cells
Authors: Buchholz D.H. , AuBuchon J.P. , Snyder E.L. , Kandler R. , Edberg S. , Piscitelli V. , Pickard C. , Napychank P.
Published: March 2003

The growth of Yersinia enterocolitica in AS‐1 red cells was investigated so as to study the organism's proliferation kinetics and to evaluate the effect of prestorage white cell (WBC) reduction on bacterial multiplication. Twenty‐four 2‐unit pools of ABO‐compatible whole blood were prepared and inoculated with Y. enterocolitica to final concentrations ranging from 0.3 to 132 organisms per mL. After inoculation, pools were split equally, AS‐1 red cells were prepared, and 1 unit of each pair (test unit) was WBC‐reduced with a WBC‐ reduction filter. Quantitative bacterial cultures of both WBC‐reduced and control units were performed at several points throughout preparation and storage. Less than 10 percent of the inoculated organisms was recovered from blood samples taken after a 7‐hour room‐ temperature holding period. By the end of 42 days of storage, Y. enterocolitica was recovered from unfiltered red cells in 2 of the 6 units inoculated at the lowest levels (0.3 and 0.7 organism/mL), from 8 of the 12 units inoculated at the intermediate levels (2.8, 5.2, 30.7, and 43 organisms/mL) and from 6 of the 6 units inoculated at the highest levels (98.8 and 132 organisms/mL). Positive cultures were seen as early as Day 7. In contrast, filtered units inoculated at all levels ≥ to 98.8 organisms per mL (21/21 units) were sterile at the end of the 42‐day storage period, while 2 units (2/3) inoculated at 132 organisms per mL showed growth despite filtration. Thus, passage of deliberately contaminated AS‐1 red cells tarough this filter appeared to reduce significantly the likelihood of growth of Y. enterocolitica during subsequent storage, in spite of the removal of large numbers of WBCs. Because the concentration of bacteria in asymptomatic carrier donors is thought to be very low, prestorage filtration of blood shortly following col‐ lection may represent a means by which the risk of subsequent proliferation of Y.enterocolitm may be reduced or avoided as well as a means of providing a WBC‐ reduced blood component.
Experimental animal model of refractoriness to donor platelets: the effect of plasma removal and the extent of white cell reduction on allogeneic alloimmunization
Authors: Bordin J.O. , Bardossy L. , Blajchman M.A.
Published: March 2003

Platelet transfusion effectiveness may be limited in multiply transfused patients by the development of the refractory state. White cell (WBC)‐reduction filters with variable efficiency (1–3 log10 reduction) are available and have been shown to be effective in reducing the incidence of platelet alloimmunization. However, the threshold number of WBCs below which alloimmunization would no longer occur is yet to be determined. A previously established animal model was used to examine the relative efficiency of second‐ and third‐ generation filters in reducing the frequency of refractoriness to allogeneic platelets. In this model, California Black rabbits are used as blood donors and New Zealand White rabbits as transfusion recipients. Eight weekly transfusions of either second‐generation or third‐generation WBC‐reduced blood resulted in no difference between the two groups in mean platelet survival and rate of refractoriness to allogeneic platelets. To evaluate the possible incremental benefit of removing supernatant plasma to prevent platelet refractoriness, experiments were performed in which groups of animals were given transfusion(s) with red cell suspensions that had been WBC‐reduced or both plasma‐depleted and WBC‐reduced. A significantly lower rate of allogeneic platelet refractoriness was seen in the rabbits that received WBC‐reduced and plasma‐depleted red cells than in those that received red cells that had been WBC‐reduced only. These data provide evidence that the combined use of plasma depletion and WBC reduction can decrease still further the frequency of refractoriness produced by allogeneic blood transfusions.
Chagas' disease diagnosis: evaluation of several tests in blood bank screening
Authors: Carvalho M.R. , Krieger M.A. , Almeida E. , Oelemann W. , Shikanai‐Yassuda M.A. , Ferreira A.W. , Pereira J.B. , Saez‐Alquezar A. , Dorlhiac‐Llacer P.E. , Chamone D.F. , Goldenberg S.
Published: March 2003

Blood transfusion is one of the principal routes of transmission of Chagas' disease, a major endemic disease in Latin America. Methods for blood screening are not accurate and may yield false results that lead to high social and economic costs. This study compares two methods of diagnosing Chagas' disease (indirect immunofluorescence and hemagglutination) and several enzyme‐linked immunosorbent assays (ELISAs) with regard to specificity and sensitivity, by using human sera with known serologic and parasitologic characteristics, as well as samples with discrepant results on conventional serologic tests. An ELISA using recombinant antigens showed no cross‐reactivity with sera that were positive for other diseases. All evaluated ELISAs performed well, and their use may lead to a reduction of more than 50 percent in the number of discordant sera. Further improvements are needed in view of the complexity of the serologic diagnosis of Chagas' disease.
Comparison of plateletpheresis with a standard and an improved collection device
Authors: Maresh S. , Randels M.J. , Strauss R.G. , Winegarden D. , Ludwig G.A.
Published: March 2003

A blood cell separator with a specialized separation chamber ([TNX‐6]CS‐ 3000 Plus) was developed for the collection of platelet concentrates with higher platelet yields and lower white cell contamination than obtained with the standard blood cell separator (CS‐3000). To compare these devices, normal donors were scheduled for paired plateletpheresis procedures spaced 4 weeks apart, with one procedure using the CS‐3000 Plus and the other using the CS‐3000. Overall, the platelet yield per unit (mean ± SEM) was 4.3 ± 0.1 × 10(11) with the CS‐3000 Plus versus 3.7 ± 0.1 × 10(11) with the CS‐3000 (p < 0.001), and the white cell contamination per unit (mean ± SEM) with the former was 2.4 ± 0.7 × 10(6) versus 84.1 ± 21.1 × 10(6) with the latter (p < 0.001). The sequence of procedures (i.e., the order in which the devices were paired) was selected randomly, and similar results were found regardless of sequence. When donors with predonation platelet counts of > or = 200 × 10(9) per L (n = 21) were studied separately, 76 percent of the collections by the CS‐3000 Plus contained > or = 4 × 10(11) platelets versus 34 percent of those by the CS‐3000 (p < 0.01), and 93 percent of the collections by the former contained < 5 × 10(6) white cells (69% contained < 1 × 10(6)) versus 0 percent of those by the latter (p < 0.01). Thus, platelet collections with the TNX‐6 chamber consistently demonstrated high platelet yields and strikingly low white cell contamination–qualities that justify converting standard devices to devices with a TNX‐6 chamber.
Simultaneous confirmation and differentiation of human T‐lymphotropic virus types I and II infection by modified western blot containing recombinant envelope glycoproteins
Authors: Brodine S.K. , Kaime E.M. , Roberts C. , Turnicky R.P. , Lal R.B.
Published: March 2003

A modified Western blot (WB) that includes both shared (r21e) and unique recombinant envelope proteins from human T‐lymphotropic virus (HTLV) type I (rgp46I) and type II (rgp46II) was compared to conventional HTLV serologic tests in 379 United States blood donors and individuals residing in diverse geographic regions, and the specimens were categorized as positive (n = 158), indeterminate (n = 158), or negative (n = 63) for HTLV infection. Of the 158 HTLV‐I/II‐positive specimens (66 requiring radioimmunoprecipitation assay [RIPA] for confirmation), 156 reacted concordantly with r21e, gag, and either rgp46I or rgp46II, thus eliminating the need for RIPA in all but two specimens and yielding a test sensitivity of 98.7 percent. Of the 158 indeterminate and 63 negative specimens, none reacted with r21e and rgp46I or rgp46II, yielding a test specificity of 100 percent. Furthermore, analysis of an additional 184 consecutive specimens from a retrovirology reference laboratory demonstrated that the modified WB correctly identified 27 of 28 HTLV‐I specimens and all 13 HTLV‐II specimens, with a test sensitivity of 97.6 percent. None of specimens that were indeterminate or nonreactive in conventional WB and/or RIPA and none of the screening enzyme immunoassay‐negative specimens reacted with r21e and either rgp46I or rgp46II, for a test specificity of 100 percent. Thus, the modified WB appears to be highly sensitive and specific for simultaneous detection and discrimination of HTLV‐I from HTLV‐II and has the advantage of being a one‐step assay that is easily performed in all types of laboratory settings and allows rapid, reliable, and standardized testing for HTLV‐I/II infection.
Human immunodeficiency virus type 1 nucleic acids detected before p24 antigenemia in a blood donor
Authors: de Saussure P. , Yerly S. , Tullen E. , Perrin L. H.
Published: March 2003

A blood donation from a 46‐year‐old homosexual man was discarded because of elevated alanine aminotransferase levels. Thirteen days later, the patient presented with symptomatic primary human immunodeficiency virus type 1 (HIV‐1) infection. Virologic investigations were performed retrospectively on blood samples (including the donated blood) obtained before the symptoms. The HIV‐1 genome was present, either integrated in mononuclear cell DNA or circulating in plasma, 39 days before the appearance of p24 antigen and 65 days before the appearance of HIV‐1 or HIV type 2 antibody. It is concluded that p24 antigenemia is present during only a fraction of the seronegative “window” period. This case illustrates the risk of infection associated with blood transfusion in spite of HIV‐1 antibody testing and stresses the need to improve nontechnical exclusion procedures as well as non‐antibody‐based diagnostic tests.
Roles of acetate and phosphate in the successful storage of platelet concentrates prepared with an acetate‐containing additive solution
Authors: Shimizu T. , Murphy S.
Published: March 2003

The development of a synthetic medium for platelet storage is an important goal in transfusion medicine. Its use would make large volumes of plasma available for fractionation and might improve the quality of platelets after storage. Several investigators have described successful storage in media containing acetate. The previous work of the authors showed that platelet concentrates (PCs) can be stored successfully for 5 days at 22 degrees C by using an additive solution (Seto sol) to replace 80 to 95 percent of the plasma usually employed as a suspending medium. Seto sol contains 23 mM (23 mmol/L) sodium acetate and 25 mM (25 mmol/L) sodium phosphate. The roles of acetate and phosphate in achieving successful platelet storage were studied in the work reported here. The concentration of acetate decreased linearly for 7 days at 0.61 +/− 0.11 mumol per day per 10(9) platelets in parallel with the disappearance of 1–14C or 2–14C acetate. There was no disappearance of tritiated acetate from PCs or of 1–14C acetate from platelet‐free mixtures of plasma and Seto sol, which suggests that the disappearance of 14C acetate from PCs reflected oxidation to CO2, which could leave PCs through the walls of the plastic container. Since O2 consumption was 1.47 mumol per day per 10(9) platelets, and the oxidation of a molecule of acetate requires 2 molecules of oxygen, acetate oxidation accounted for approximately 85 percent of oxygen consumption by platelets. The pH of PCs stored in Seto sol was nearly constant for 7 days, whereas, without acetate, it fell to 6.4 +/− 0.1 on Day 5.It is known that the oxidation of one molecule of acetate produces one molecule of CO, and one molecule of bicarbonate. Indeed, in PCs in Set0 sol, there was an elevation of bicarbonate concentration of 1.4 mEq per L by Day 3, which suggests bicarbonate production. Thus, acetate acts as both a substrate for oxidative meta‐ bolism and a source of buffer. However, in 5 of 11 PCs stored in Set0 sol without phosphate, the pH dropped to 6.2 within 3 days. PCs that showed a drop in pH were those in which the content of plasma carryover during PC preparation was less than 12.5 percent of the suspending medium. It appears that a buffer system in addition to that provided by acetate is necessary to the maintenance of platelet integrity during storage when plasma carryover is low.
Storage of single‐donor platelet concentrates: metabolic and functional changes
Authors: Bock M. , Glaser A. , Pfosser A. , Schleuning M. , Heim M.U. , Mempel W.
Published: March 2003

During the last decade, the trend toward intensifying chemotherapeutic regimens in patients with hematologic malignancies rapidly increased the demand for single‐donor platelet concentrates (PCs). The logistics of such supply, however, necessitated the storage of these blood components prior to transfusion. Today, most blood centers use di(2‐ ethylhexyl)phthalate‐free blood bags, which are assumed to allow a storage period of up to 5 days. This report describes biochemical and functional changes of stored single‐donor PCs, which may influence the expected quality of PCs. The acid‐base status is characterized by an initial respiratory alkalosis compensated by a metabolic acidosis. Changes in extracellular electrolyte, lactate dehydrogenase, glucose, lactate, elastase, and complement levels, as well as in the release of alpha granule content and the initial activation of plasma coagulation, are demonstrated. These changes result in a functional impairment of stored PCs as reflected by thromboxane and serotonin release reaction and by aggregation and in vitro bleeding time studies. In contrast, in vivo recovery and survival rates have been reported to be unaffected. Whether the good recovery and survival rates are caused by a rejuvenescence of stored PCs in vivo or are due to injured circulating platelets has not yet been proven.
Significance of antibody to hepatitis B core antigen in blood donors as determined by their serologic response to hepatitis B vaccine
Authors: Aoki S.K. , Finegold D. , Kuramoto I.K. , Douville C. , Richards C. , Randell R. , Fernando L. , Holland P.V. , Zeldis J.B.
Published: March 2003

Because large numbers of volunteer blood donors may be disqualified for “false‐positive” results on tests for antibody to hepatitis B core antigen (anti‐HBc), a more specific definition of anti‐HBc enzyme immunoassay (EIA)‐reactive was evaluated, including only those donor samples that were “strongly” reactive (sample‐to‐cutoff absorbance ratio, < 0.45). Results using this definition and other anti‐HBc test methods were compared to the serologic response (antibody to hepatitis B surface antigen [anti‐HBsAg]) to hepatitis B vaccination. Fifty‐eight volunteer blood donors who had previously been deferred as donors, because of reactive anti‐HBc tests (all other blood screening tests were negative, including those for HBsAg and anti‐HBsAg) on two occasions, were vaccinated for hepatitis B. It was assumed that an anamnestic response to vaccine indicated past infection with hepatitis B, while a primary response to vaccine indicated lack of past infection. One (2%) of 43 donors with a historically “weak” anti‐HBc (reactive absorbance ratio, > or = 0.45) had an anamnestic response to vaccine, compared to 8 (53%) of 15 with historically “strong” anti‐HBc (reactive absorbance ratio, < 0.45) (p < 0.005). Anti‐HBc testing using the microparticle EIA method also correlated well with hepatitis B vaccination results. The use of a narrower definition of “reactive” for anti‐HBc EIA testing yielded much more specific, but slightly less sensitive, results.
Effective use of a strategy using assigned red cell units to limit donor exposure for neonatal patients
Authors: Cook S. , Gunter J. , Wissel M.
Published: March 2003

Critically ill neonatal infants receive frequent small‐volume red cell (RBC) transfusions for replacement of blood drawn for laboratory analysis or for treatment of symptomatic anemia secondary to underlying medical conditions and/or a relative bleeding diathesis. Retrospective review of transfusion practice in a hospital revealed that the donor exposure‐to‐transfusion ratio was 1:1.3. In an effort to limit donor exposure and decrease the risk of transfusion‐transmitted disease, a sterile connection device was used for multiple small‐aliquot preparations. Three neonatal infants were given part of the same RBC unit, and the assigned RBC unit was used only until it reached 14 days of age. These criteria resulted in 49‐percent reduction in donor exposure for neonatal infants weighting less than 1500 g and 27‐percent reduction for those weighing more than 1500 g. The donor exposure‐to‐ transfusion ratio decreased to 1:2.5. RBC waste from syringe aliquot preparation and residual volume at Day 15 were a mean of 32 percent of the total unit volume. Of 722 transfusion occurrences, there were no reported adverse effects due to elevated potassium subsequent to transfusion of 14‐day‐old RBCs. This limited‐donor‐exposure strategy effectively meets the needs of a transfusion service and will reduce the donor exposures of neonatal infants in similar institutions.
Large‐volume hemocytometer chamber for accurate counting of white cells (WBCs) in WBC‐reduced platelets: validation and application for quality control of WBC‐reduced platelets prepared by apheresis and filtration
Authors: Lutz P. , Dzik W.H.
Published: March 2003

A detailed description is provided of a method using a large‐volume (50 microL) hemocytometer to count very low numbers of white cells (WBCs) in platelet components. A method employing a Nageotte hemocytometer uses crystal violet stain and a standard microscope with a reading time of 5 to 10 minutes. The method is validated by using serial dilutions of known concentrations of WBCs in platelets and by correlation with a flow cytometric technique. The interobserver coefficient of variation was 11.9 percent for WBC concentrations > 2 per microL. Use of the method for the evaluation of 203 WBC‐reduced platelet components prepared by apheresis or by filtration revealed that over 94 percent of components had WBC content < 5 × 10(6). This method could easily be applied in the routine quality control of WBC‐reduced platelet components and in clinical studies employing these components.
A rapid molecular diagnosis of posttransfusion graft‐versus‐host disease by polymerase chain reaction
Authors: Hayakawa S. , Chishima F. , Sakata H. , Fujii K. , Ohtani K. , Kurashina K. , Hayakawa J. , Suzuki K. , Nakabayas H. , Esumi M. , Nemoto N. , Sakurai I. , Satoh K.
Published: March 2003

A woman with recurrent Paget's disease of the vulva developed acute graft‐versus‐host disease (GVHD) 12 days after radical surgery and massive blood transfusion. Molecular diagnosis of lymphocyte chimerism in the peripheral blood was made by polymerase chain reaction (PCR) directed against a Y chromosome‐specific sex‐determining region Y (SRY) gene. PCR with skin biopsy after onset of GVHD also revealed infiltration of SRY‐positive donor lymphocytes. The diagnosis was confirmed by HLA‐DNA typing with PCR‐sequence‐specific oligonucleotide that revealed the presence of complex HLA‐DR chimerism in the peripheral lymphocytes collected after onset of GVHD. The use of SRY‐ directed PCR is a rapid technique for the early diagnosis of acute posttransfusion GVHD in female patients.
Use of a licensed electrolyte solution as an alternative to tissue culture medium for bone marrow collection
Authors: Areman E.M. , Dickerson S.A. , Kotula P.L. , Spitzer T.R. , Sacher R.A.
Published: March 2003

Bone marrow for transplantation is traditionally collected into tissue culture medium with heparin. A licensed electrolyte solution (Plasma‐ Lyte A [PLA]) was used as a substitute for tissue culture medium in the harvesting of 28 bone marrows, 17 autologous and 11 allogeneic, which were subsequently transplanted. Data that were analyzed from the 25 evaluable patients consisted of the numbers of cells and colony‐forming units in the transplanted marrow as well as the time to neutrophil and platelet engraftment. These results were compared with those in the 30 (26 evaluable) preceding transplanted marrows that were collected into a tissue culture medium (RPMI‐1640 [RPMI]). The autologous marrow transplant patients in both the PLA and RPMI groups reached a neutrophil count of > or = 0.5 × 109 per L a mean of 19 days following transplantation. The patients who underwent transplantation with allogeneic bone marrow collected in RPMI achieved > or = 0.5 × 109 per L of neutrophils an average of 20 days following transplantation, while those who received marrow collected in PLA achieved engraftment of neutrophils to that level in a mean of 21 days. Because in vitro and in vivo results with RPMI and PLA are similar in this study, further work using a licensed solution for clinical bone marrow transplantation is indicated.
Transmission of human T‐lymphotrophic virus type I infection to a neonatal infant by transfusion of washed and irradiated red cells
Authors: DePalma L. , Luban N.L.
Published: March 2003

Human T‐lymphotropic virus type I and/or II (HTLV‐I/II) may be transmitted by the transfusion of blood and blood components. Several factors are critical to the efficiency of transmission. These include the number of contaminating white cells, the component volume, and the age of the component. After look‐back notification, there was an investigation of the HTLV‐I/II serostatus of three patients who had received blood from a donor now found to be positive for HTLV‐I antibodies by enzyme immunoassay and Western blot. The donor red cell unit was group O negative and cytomegalovirus antibody negative; it was washed and irradiated at 2800 cGy and aliquoted into six small‐volume transfusions for four neonatal infants. Three of the four patients were available for testing 3.5 years after their exposure. The fourth neonatal infant died on Day 11 of life. The three tested infants received 14, 14, and 44 mL of component, respectively. HTLV‐I seroconversion was documented by enzyme immunoassay and Western blot (p19, p24) and occurred only in the patient receiving 44 mL. On the basis of quality control data, it is estimated that the affected infant received 8 × 10(7) white cells in the 44‐mL aliquot. Washing and irradiation will not necessarily eliminate HTLV‐I transmission.
Platelet crossmatches of single‐donor platelet concentrates using a latex agglutination assay
Authors: Ogden D.M. , Asfour A. , Koller C. , Lichtiger B.
Published: March 2003

A latex agglutination assay was evaluated for the purpose of identifying compatible platelet donors for alloimmunized recipients. Assay reagents were prepared by adsorbing detergent‐solubilized, donor‐specific platelets to polystyrene latex beads. Semiquantitative results for up to 30 donors can be completed in less than 1 hour. These reagents retained their immunoreactivity for at least 3.5 months. A retrospective study has established the assay's upper limit of compatibility. The prospective study evaluated transfusions to a group of multiply transfused patients. Part I evaluated 143 crossmatched, single‐donor platelet transfusions given to 50 patients. In 96 percent of the cases, a positive crossmatch was associated with an unsuccessful transfusion outcome; in 84 percent of the transfusions, a negative crossmatch predicted a satisfactory platelet increment. The overall predictability, sensitivity, and specificity were 87, 62, and 99 percent, respectively. Part II evaluated 105 transfusions given to the 43 patients (of 50) in whom no incidence of fever, sepsis, or bleeding could be documented. A positive crossmatch was 96‐percent efficient in predicting an unsuccessful transfusion, whereas a negative crossmatch was associated with an adequate platelet increment following 89 percent of the transfusions. The overall predictability was 91 percent, the sensitivity was 72 percent, and the specificity was 99 percent. Within‐run and between‐run variations were 6.3 and 6.2 percent, respectively. These results demonstrate that detergent‐solubilized platelet antigens, immobilized on latex particles, can be used in a cost‐effective crossmatching procedure.
Effect of storage time on clinical efficacy of single‐donor platelet units
Authors: Leach M.F. , AuBuchon J.P.
Published: March 2003

Decrements in platelet function and recovery are known to accumulate during the 5‐day storage period, and the clinical response to platelets transfused after several days' storage has been suggested by some researchers to be less than that seen with platelets stored for shorter periods. The clinical response to single‐donor platelet (SDP) units (as measured by corrected count increments [CCIs] and intertransfusion intervals) was investigated in autologous bone marrow transplant patients. Twenty‐seven consecutive autologous bone marrow transplant patients with a variety of hematologic and solid organ malignancies were evaluated for posttransfusion CCIs after 419 SDP transfusions of units stored for 1 to 5 days. Patients were not excluded from the study because of clinical condition, such as fever or sepsis. The mean 15‐minute posttransfusion CCI for SDP units stored for only 1 day (11,006 ± 5,157) was no different than that for units stored for 5 days (10,225 ± 4,481; p > 0.05); 24‐hour posttransfusion CCIs were also not different if the SDP unit had been stored for 1 day or 5 days (6229 ± 4489 vs. 4786 ± 2759; p > 0.05) or for any intermediate period. Nor were intertransfusion intervals affected by storage time. While platelets may exhibit a progressive lesion during the 5‐day storage period, these changes do not result in a decreased clinical response.
Changes in intention to donate blood under a hypothetical condition of reduced confidentiality
Authors: Banks H.D. , Williams A.E. , Nass C.C. , Gimble J.
Published: March 2003

Lawsuits related to transfusion‐associated human immunodeficiency virus infections have increasingly resulted in requests for the release of confidential information about volunteer blood donors. Concern that loss of confidentiality might change blood donor behavior led to a survey of donors at collection sites within an American Red Cross Blood Services Region. Of the 361 respondents, 50.3 percent (181) indicated reduced intent to provide accurate medical and personal history information under conditions of reduced confidentiality. Ten percent (34) indicated that they were not sure whether they would or would not donate blood in the future under this condition. The results indicate that the possibility of release of donors' medical and personal information may have a negative effect on the safety and adequacy of the nation's volunteer blood supply.
Strategies to recruit plateletpheresis donors from a registry of HLA‐typed, unrelated, bone marrow donors
Authors: Bolgiano D.C. , Smith S. , Slichter S.J.
Published: March 2003

Rising demand for single‐donor platelet components—from random donors, to maintain platelet inventories, or from HLA‐compatible donors, to support alloimmune platelet‐refractory patients—necessitated increasing the size of a community plateletpheresis donor registry. This study compares two strategies for recruiting whole‐blood donors into a plateletpheresis program. The whole‐blood donors who were asked to participate in this study had recently joined an unrelated bone marrow donor registry and had been HLA‐typed as part of that process. An in‐person recruitment strategy, which was time‐intensive for the apheresis donor coordinator, served as the standard. A by‐mail strategy involved the mailing of recruitment materials to marrow‐donor registry participants. Marrow‐donor registry participants were approached about apheresis participation after they had indicated an interest in the plateletpheresis program by returning a tear‐off section of an informational brochure that was sent to them along with their marrow‐donor registry materials. A total of 852 marrow‐donor registry participants were randomly assigned to one of two recruitment strategies, and the recruitment rates were the same (46%) for both methods. In addition, levels of apheresis participation and attrition rates of donors recruited by either strategy were comparable. Thus, the simple strategy of mailing information about a plateletpheresis program is a very cost‐effective method of recruiting donors.
Microwave dissociation of antigen‐antibody complexes: a new elution technique to permit phenotyping of antibody‐coated red cells
Authors: McCullough J.S. , Torloni A.S. , Brecher M.E. , Tribble L.J. , Hill M.G.
Published: March 2003

To evaluate the effectiveness of microwave irradiation in dissociating IgG from red cells (RBCs), the use of chloroquine diphosphate (CDP) was compared to that of microwaves. Fifteen paired samples of RBCs from 15 patients with positive direct antiglobulin tests (DATs) were treated with both CDP and microwave radiation. Total microwave exposure times ranged from 20 to 100 seconds. Posttreatment DATs were performed, and the reaction grades of the posttreatment DATs were compared. RBC phenotyping was also performed on repeatedly microwaved RBCs to demonstrate possible effects on RBC antigen expression. Microwaves successfully reduced the reaction grade of the DAT in 14 of 15 samples; CDP reduced the reaction grade in 12 of 15 samples. In samples with a DAT of 2+ or greater (n = 13), the microwave method yielded a greater reduction in DAT strength in six cases (results in the other 7 cases were identical with both methods) (p = 0.01). Five of eight cases with a DAT of 3+ showed a greater reduction in the DAT with microwave treatment than with CDP treatment; results in the remaining three cases were identical (p = 0.03). RBC antigenicity remained unchanged after exposure to microwave radiation (A, B, C, c, D, E, e, Fya, Fyb, Jka, Jkb, K, k, S, and s). Microwave treatment required less than 10 minutes per sample, while CDP treatment required 30 to 120 minutes per sample (mean, 88 min). The microwave technique of antigen‐antibody dissociation from RBCs provides a rapid and accurate method of facilitating the phenotyping of RBCs coated with warm autoantibodies and is superior to other methods, which destroy RBC antigens. The microwave procedure is preferable to CDP treatment in patients with a DAT of 2+ or greater or when rapid results are desired.
The value of variable number of tandem‐repeat polymorphisms in cases of disputed paternity not resolved by conventional markers: two case reports
Authors: Schlaphoff T.E. , Reavis S.C. , Rousseau J. , Creemers P.C. , du Toit E.D.
Published: March 2003

Disputed paternity cases are routinely tested in the authors' laboratory for red cell antigen, plasma protein, red cell enzyme, and HLA polymorphisms. This report concerns two cases in which the above test results made exclusion of paternity doubtful. In one case, exclusion of paternity was based on one discrepancy in the Duffy blood group system only, a unique situation in the investigators' experience of more than 2500 cases; the investigators were, therefore, reluctant to use this as the only evidence of exclusion. In the other case, it was necessary to postulate the presence of a rare haplotype, MSu, in the MNS blood group system to explain paternity. It was therefore decided to investigate allelic variable number of tandem‐repeat (VNTR) DNA polymorphisms in these disputed paternity trios. VNTR DNA typing convincingly excluded these accused men from paternity.
Effect of storage and ultraviolet B irradiation on CD14‐bearing antigen‐ presenting cells (monocytes) in platelet concentrates
Authors: FIEBIG E. , LANE T.A.
Published: March 2003

BACKGROUND: Ultraviolet B (UVB) irradiation of platelet concentrate (PCs) reduces platelet alloimmunization, but the mechanism of the effect is unclear. Evidence suggests that UVB may downregulate the expression of surface adhesion molecules on passenger antigen‐ presenting cells in PCs.

STUDY DESIGN AND METHODS: The effect of blood bank storage, platelet preparation from whole blood, and UVB irradiation on the quantitative expression of intercellular adhesion molecule‐1 (ICAM‐1, or CD54), HLA‐DR, CD45, and CD11c on CD14‐positive antigen‐presenting cells (monocytes) was studied by using two‐color flow cytometry.

RESULTS: Blood bank storage for 4 days resulted in upregulation of ICAM‐1 and HLA‐DR and downregulation of CD14 but left the expression of CD11c and CD45 unchanged. Preparation of PCs from fresh whole blood was associated with a rapid increase in CD11c without upregulation of ICAM‐1 and HLA‐DR. UVB irradiation before storage inhibited the upregulation of ICAM‐1 and HLA‐DR, resulted in accelerated downregulation of CD14, and was associated with increased loss of monocytes. Agitation of the PC bag during irradiation was of critical importance, since omission of agitation resulted in largely uninhibited upregulation of ICAM‐1 but was still associated with significantly higher cell loss than that seen in unirradiated controls. CONCLUSION: UVB exposure nonspecifically affects monocytes in PCs, resulting in downregulation of surface molecules that are important for antigen presentation, as well as in significant cell loss
Quality control of red cell filtration at the patient's bedside
Authors: Sirchia G , Rebulla P , Parravicini A , Marangoni F , Cortelezzi A , Stefania A
Published: March 2003

BACKGROUND: Concern has been raised about the quality of white cell (WBC) reduction in blood components when it is performed by filtration at the patient's bedside. Thus, the quality of red cell (RBC) filtration performed at the bedside through two new flatbed WBC‐ reduction filters was evaluated.

STUDY DESIGN AND METHODS: In the laboratory, 25 and 10 RBC units suspended in additive solution were stored for 1 to 2 and 14 to 21 days, respectively, and filtered through each filter. At the end of filtration, an automated complete blood count and a manual WBC count (Nageotte chamber) were determined in two samples collected from 1) a segment clamped at 5 and 25 cm below the filter along the line connecting prefiltration and postfiltration bags and 2) the postfiltration bag. In addition, 10 of the 11 nurses of the hematology outpatient clinic administered to hematologic patients 25 RBC units stored for 1 to 2 days through each type of filter. At the end of transfusion, a segment was collected from the transfusion set and a WBC count was performed as described above. No filter priming or rinsing with saline was done.

RESULTS: WBC counts obtained after laboratory filtration in the segments were similar to those obtained from the postfiltration bags and from the segments collected at the bedside in all cases, with the exception of 14‐ to 21‐day‐old RBCs filtered in the laboratory through one of the filters, which produced slightly higher WBC counts in the segments than were seen in the postfiltration bags. The difference was not significant. In no case was the count in the postfiltration bag higher than that in the segment. Nurse training was easy, and bedside filtration was associated with no untoward effects.

CONCLUSION: The RBC filtration at the patient's bedside can be equal in quality to that performed in the laboratory, at least in such clinical settings as hematology and oncology departments in which blood transfusion is common practice, and if simple training is provided to the nursing staff. Under the conditions of this study, it seems that quality control of RBC bedside filtration is feasible and simple.