bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of a confidential method of excluding blood donors exposed to human immunodeficiency virus: studies on hepatitis and cytomegalovirus markers
Authors: MUSBACHER JACOB , CHIAVETTA JOANNE , NAIMAN RENEE , BUCHNER BARBARA , REEVES JACQUELINE , SCALIA VITO , HERST ROSLYN
Published: March 2003

A confidential self‐administered questionnaire was given to all blood donors prior to donation (n = 95,917). The questionnaire describes groups at increased risk of acquired immunodeficiency syndrome (AIDS) and requires the donor to designate his blood either for laboratory purposes or for transfusion. In a previous communication, we reported that donors in the former group had a much higher prevalence of antibody to human immunodeficiency virus (HIV) than age, sex and clinic matched controls or a group of “miscellaneous” donors who did not fill out the form properly. In this communication, we report results of tests for other viral markers performed on the three designation groups, namely laboratory‐designated, miscellaneous and controls. We found that the former two groups had a higher prevalence of antibody to hepatitis B surface antigen (anti‐HBs), hepatitis B core antigen (anti‐ HBc) and cytomegalovirus (anti‐CMV) than controls, but there were no differences in alanine aminotransferase (ALT) levels among the groups. In addition, the laboratory‐designated group had a higher prevalence of hepatitis B surface antigen (HBsAg) than the general donor population. These data indicate that a questionnaire designed to ascertain AIDS high‐risk donors is valuable in excluding donors who may be carriers of other viruses as well.
Prevalence of cytomegalovirus antibody in hemophiliacs and homosexuals infected with human immunodeficiency virus type 1
Authors: Jackson J.B. , Erice A. , Englund J.A. , Edson J.R. , Balfour H.H.
Published: March 2003

We determined the prevalence of antibody to cytomegalovirus (CMV) in the sera of non‐homosexual hemophilia patients and homosexual men infected with the human immunodeficiency virus type 1 (HIV‐1). CMV antibody testing by latex agglutination revealed 33 of 58 HIV‐1 infected hemophiliacs (57%) were antibody‐positive compared with 54 of 54 HIV‐1 infected asymptomatic non‐hemophiliac homosexuals (100%) (p less than .001). Nine of 15 hemophiliacs (60%) with symptomatic HIV‐1 infection were CMV antibody‐positive. We also tested 22 HIV‐1 antibody‐ negative hemophiliacs who had received non‐heat treated factor concentrates. 14 of these 22 (64%) were CMV antibody‐positive compared with 57% of HIV‐1 antibody‐positive hemophiliacs. We conclude 1) there is little correlation between transmission of HIV‐1 and CMV by factor concentrates, 2) the presence of CMV antibody does not appear to be associated with clinical stage of HIV‐1 infection in hemophiliacs, and 3) there may be a significant number of CMV antibody‐negative hemophiliacs with HIV‐1 infection at risk for primary infection and subsequent disease if CMV seronegative blood products are not provided for future transfusions.
Seroconversion to human immunodeficiency virus (HIV) in hemophiliacs
Authors: Andes W. A. , Daul C. B. , Deshazo R. D. , Palmer C. H.
Published: March 2003

The authors studied the natural history of human immunodeficiency virus (HIV) exposure in 187 hemophiliacs followed for an average of 45 months. Overall, 55 percent developed antibody specific for HIV and 21 percent developed persistent generalized lymphadenopathy. Most patients seroconverted sometime between early 1982 and the end of 1984. Four patients developed acquired immune deficiency syndrome (AIDS) and four seropositive patients developed idiopathic thrombocytopenia (ITP). One of the four patients who developed AIDS and three of the four with ITP had preexisting lymphadenopathy. None of the 10 patients with lymphadenopathy or the 20 asymptomatic patients was seropositive for human T‐lymphotropic virus, type I. Although seropositivity and lymphadenopathy have been found in many of the authors’ patients, few have developed clinical disease that can be related to HIV infection.
Autologous versus homologous donors
Authors: Kruskall Margot S. , Popovsky Mark A. , Pacini Donna G. , Donovan Lillian M. , Ransil Bernard J.
Published: March 2003

Autologous blood donations may provide a new source of blood when components not used by the donors are deemed suitable for homologous use. However, the risk of transfusion‐transmitted diseases form such donors has not been evaluated. We compared the prevalence of infectious markers and rate of abnormal responses to a confidential donor ballot in autologous donors from two blood collection programs, one blood center and one hospital‐based, to corresponding homologous blood programs. The incidence of abnormal test results in autologous donors for HIV antibodies (either Western blot confirmed or repeatedly reactive, unconfirmed), HBsAg, ALT, and anti‐HBc were not statistically different from homologous rates. The incidence of STS abnormalities in autologous donors was statistically significant, although all positive results were biologic false positives. The rate of abnormal responses to the confidential ballot was statistically significant only in autologous donors whose collections were already determined to be unsuitable for homologous use due to medical history problems. Although the data do not address infectious complications in transfusion recipients, this study offers no evidence that autologous blood components are less safe than their homologous equivalents.
Chemotaxin receptor cycling in fresh and stored granulocytes
Authors: Lane T.A. , Lamkin G.E.
Published: March 2003

The authors reported previously that stored granulocytes (PMN) had decreased receptor affinity (Kd) for and chemotaxis (CTX) to the chemotactic peptide F‐Met‐Leu‐Phe (FMLP), but the evidence did not favor a significant role for altered FMLP receptor affinity in causing diminished CTX of stored PMN. Since recruitment and/or recycling of FMLP receptors is required for normal CTX, the hypothesis that stored PMN might have abnormal FMLP receptor cycling was tested. The effect of storage on the proportion of high‐ and low‐affinity FMLP receptors was also investigated. Units of PMN were tested within 4 hours of collection and after 24 and 48 hours of storage at 22° C, unagitated, in 150‐ml transfer packs. In comparison to fresh PMN, there was no alteration in the Kd of the high‐affinity FMLP receptor of PMN stored for 24 to 48 hours; however, the Kd of the low‐affinity receptor increased (fresh PMN = 36 ± 5 nM; 24 hours = 107 ± 19; 48 hours = 121 ± 22; p < 0.01 for both 24 and 48 h versus fresh PMN). Likewise, while the number of high‐affinity receptors increased (fresh PMN = 25,000 ± 6,000 receptors/PMN; 24 hours = 95,000 ± 21,000; 48 hours = 161,000 ± 40,000; p < 0.01 for both 24 and 48 hours versus fresh PMN). No abnormality was found in the ability of stored PMN to down‐regulate FMLP receptors in the presence of ligand or to reexpress FMLP receptors after a 15‐minute incubation in the absence of ligand. In contrast, the initial rate of FMLP receptor reappearance after down‐regulation was significantly impaired in stored PMN (fresh PMN = 1320 ± 330 receptors/PMN; 24 hours = 180 ± 80; 48 hours = 260 ± 100; p < 0.05 for both 24 and 48 hours versus fresh PMN). These studies thus confirm the authors' earlier findings of decreased FMLP receptor affinity in stored PMN and suggest that this change is due to an alteration in the Kd of the low‐affinity receptor. The overall significance of this and the defect in reexpression of FMLP receptors in stored PMN is unclear but may contribute to defective CTX of stored PMN.
Assessment of HIV‐1 screening test sensitivities using serially diluted positive sera can give misleading results
Authors: Thorn R. M. , Braman V. , Stella M. , Yi A.
Published: March 2003

ABSTRACT: Three FDA licensed HIV‐1 viral lysate and two nonlicensed recombinant antigen assays were used to evaluate six serially diluted plasma samples and 2 highly characterized seroconversion series. The sensitivity as measured by serial dilution did not necessarily correlate with the sensitivity as measured by seroconversion performance with the lysate and the recombinant assays. It is concluded that national licensing agencies should arrange to share seroconversion panels to evaluate accurately the sensitivity of new HIV‐1 screening tests.
Successful allogeneic cryopreserved marrow transplantation
Authors: LASKY L.C. , VAN BUREN N. , WEISDORF D.J. , FILIPOVICH A. , MCGLAVE P. , KERSEY J.H. , MCCULLOUGH J. , RAMSAY N. K. C. , BLAZAR B. R.
Published: March 2003

ABSTRACT: Cryopreservation has been used extensively in autologous marrow transplantation (BMT), but there has been limited use in allogeneic BMT. We describe here 6 cases of successful engraftment following allogeneic BMT with cryopreserved marrow. Patients suffered from Wiscott‐Aldrich syndrome, osteopetrosis, aplastic anemia, and acute lymphocytic, acute non‐lymphocytic, and chronic myelogenous leukemia, and ranged in age from 5 mos to 35 yrs. Marrow was collected using standard techniques. In one case T‐cells were removed to prevent graft‐vs‐host disease. Marrow was frozen for a variety of reasons. Buffy coat cells were frozen at controlled rate in 10% DMSO, and stored in liquid nitrogen for 6 to 49 d. Engraftment (WBC > 1000/uL × 3 d) occurred from 13 to 37 d post BMT. In 4 of 4 cases in which data are available, donor origin of engraftment was documented, 1 with cytogenetics, 2 with red cell typing, and 4 with restriction fragment length polymorphisms. 3 patients are alive and well 21, 21, and 42 months post BMT. These results suggest frozen marrow can be successfully used for allogeneic BMT
Extended survival of neocytes produced by a new system
Authors: Simon T. L. , Sohmer P. , Nelson E. J.
Published: March 2003

Red cells (RBCs) prepared by a new system using centrifugation to produce neocyte enrichment were studied in two laboratories. The system used a blood bag with a geometric configuration such that younger, less dense cells could be separated from older, denser cells. Phthalate ester density gradient curves determined that neocyte enrichment was 81.3 percent in one laboratory and 82 percent in the other. RBC viability was studied by 51Cr autologous transfusion in normal volunteer donors. A randomized, paired design was used in which each donor was transfused once each with neocytes and with RBCs of all ages. The mean control half‐life was 34.8 ± 5.4 days in one laboratory and 34.0 ± 3.6 days in the other. The mean half‐life of the neocyte‐enriched RBCs was 45.2 ± 8.2 days in one laboratory and 45.1 ± 4.4 days in the other. This represented a more than 30 percent increase in half‐life for the neocyte‐enriched RBCs, a significant difference. This new system, a two‐bag set that costs $15, allows the simple, efficient separation of neocyte‐enriched RBCs that would have a longer half‐life and could reduce the transfusion requirement in patients receiving chronic transfusion therapy.
HIV 2 infection among blood donors and other subjects in France
Authors: COUROUCE A.M. ,   THE “RETROVIRUS” STUDY GROUP OF THE FRENCH SOCIETY OF BLOOD TRANSFUSION: F. BARIN, J. BAUDELOT, S. CHAMARET, M. GUEGUEN, C. JANOT, J. M. LEMAIRE, M. MANIEZ, F. MESNIER, L. MOUILLOT, L. NOEL, M. L. NORTH, C. ROUZIOUX AND W. SMILOVICI
Published: March 2003

Abstract: During a 3 year period, from August 1985 to August 1988, 18 HIV 2‐infected blood donors were detected in France as a result of systematic HIV 1 screening. These sera were characterized as HIV 2 by specific Western blot and synthetic peptides. Within the same period, 40 other HIV 2 infected subjects were identified by our study group, independently of blood donations. Thirty of these 58 subjects living in France originate from West Africa (8 blood donors), 3 (1 blood donor) are Portuguese men who had lived in West Africa and 25 (9 blood donors) are of French origin; among these, 16 have had a known close contact with West Africa.

When HIV 2‐infected subjects were asymptomatic, cross reactivity between antibodies to HIV 2 and HIV 1 proteins was generally observed with the two‐step ELISA assays using total HIV 1 proteins; it was poor with the competitive assays and variable with the assays using recombinant proteins. When the subjects had signs of immunodeficiency, cross reactivity decreased.

The data confirm that HIV 2 is not widespread in France and that most of HIV 2‐infected but asymptomatic subjects are recognized by several HIV 1 ELISA assays. Accordingly, systematic screening for HIV 2 with an additional test cannot be recommended at the present time, but a combined HIV 1 and HIV 2 test will be useful when available.
A double buffy coat method for red cell removal from ABO‐incompatible marrow
Authors: Rosenfeld C. S. , Tedrow H. , Boegel F. , Gremba C. , Shadduck R. K.
Published: March 2003

Hemolytic reactions caused by transfusion of ABO‐incompatible marrow can be ameliorated by either reduction of isohemagglutinins in the recipient or depletion of incompatible red cells from the harvested marrow. This article describes a rapid and reliable method for removal of incompatible marrow red cells on a blood cell processor using a double buffy coat technique. In five allogeneic bone marrow transplants, the maximum value of transfused incompatible red cells was 8.8 ml. There was no evidence of a hemolytic transfusion reaction in any patient. The median cell recovery for nucleated marrow cells and progenitor cells was 77 and 104 percent, respectively. Engraftment occurred at a median of 13 days (range, 11–21 days) after transplantation. The double buffy coat method of red cell depletion is an acceptable method for processing ABO‐incompatible marrow.
Long‐term use of antecubital veins for plasma exchange
Authors: Noseworthy J. H. , Shumak K. H. , Vandervoort M. K. ,   Canadian Cooperative Multiple Sclerosis Study Group
Published: March 2003

True or sham plasma exchange was done weekly for 20 weeks in patients in two of the randomization groups in a prospective, blind clinical trial of experimental treatments for multiple sclerosis. Because patients could be randomized to receive sham plasma exchange and placebo medications, it was decided when the trial was designed that the use of fistulae, arteriovenous shunts, venous cutdowns, or other aggressive forms of venous access would not be permitted for any patient. Accordingly, patients judged to have inadequate superficial antecubital veins were ineligible for the trial. To date, only 13 (4.4%) of 294 patients considered for entry into the trial have been rejected on these grounds. In only 4 of the 93 patients undergoing exchange was it necessary to discontinue plasma exchange because of inadequate venous access. In 79.3 percent of the 1207 exchanges done in these patients, there were no problems of any kind with venous access. In 5.4 percent of these 1207 exchanges, it was necessary to terminate the procedure prematurely because of difficulties with patients' veins. Thus, the great majority of patients free of serious systemic illness (other than chronic progressive multiple sclerosis) can undergo weekly plasma exchange for up to 20 weeks using superficial antecubital veins without the need to resort to more invasive methods of venous access.
5‐Day storage of platelets collected on a blood cell separator
Authors: Rock G. , Senack E. , Tittley P.
Published: March 2003

One of the earliest devices available for plateletpheresis is the Haemonetics system; this machine has been updated recently to permit the use of software in a closed system and thus storage of the collected platelets beyond 24 hours. The authors examined the in vitro and in vivo function of platelets collected on the Haemonetics AutoSurge machine and stored for 5 days in two separate 1000‐mL CLX bags. The average count per bag was 1.8 ± 0.4 × 1011 in approximately 200 mL of plasma. Immediately following collection, the platelet response to ADP and epinephrine represented 78 and 35 percent, respectively, of the preapheresis values. Aggregation to single stimuli subsequently decreased to 29 and 0 percent, respectively by Day 5. This response is equal to or better than the response reported with the Fenwal CS‐3000, the only other plateletpheresis device routinely used for long‐term storage. The pH of the preparations was well maintained throughout storage, and there was little alteration in hypotonic shock response or serotonin uptake. Serotonin release decreased consistently. The morphology scores indicated good maintenance of shape immediately following collection; this subsequently decreased after 5 days of storage. Bacterial cultures were negative in all instances. The 51Cr in vivo survival and recovery was good with 65.5 ± 7.1 percent recovery and an average survival of 7.3 ± 1.3 days (multiple hit; n = 5). The data indicate that storage of the Haemonetics plateletpheresis product is feasible and that the product is as good as others currently available. TRANSFUSION 1989;29:626–628.
Evaluation of platelets prepared by apheresis and stored for 5 days. In vitro and in vivo studies
Authors: Shanwell A. , Gulliksson H. , Berg B. K. , Jansson B. A. , Svensson L. A.
Published: March 2003

To evaluate the effect of storage on apheresis platelets collected with a closed‐system blood cell separator, an in vitro investigation was performed, with measurements of pH, lactate, ATP, the ratio of ATP to the total adenine nucleotide content, and adenylate kinase. Unmodified apheresis platelets and apheresis platelets with plasma added were compared with conventional platelets stored in PL‐1240 or PL‐732 plastic containers. During 6 days of storage, there were similar changes in all variables with one exception: the extracellular activity of adenylate kinase was lower in apheresis platelets with plasma than in the other three groups (p < 0.01). In vivo studies were carried out with 111Indium‐labeled autologous platelets in eight volunteers. Apheresis platelets with 100 mL of plasma added were stored in two 1000‐mL containers (PL‐732) at 22° C during agitation. Platelets from one of the containers were labeled with 111Indium and transfused into the volunteer within 24 hours. Platelets from the other container were labeled after 5 days of storage and transfused into the same donor. There were no significant differences between apheresis platelets stored for 1 day and those stored for 5 days: the mean percentage of recovery was 58.4 and 57.6 percent, t½ was 69 and 67 hours, and the survival time was 5.5 and 5.6 days, respectively.
The maintenance of platelet properties upon limited discontinuation of agitation during storage
Authors: Moroff G. , George V.M.
Published: March 2003

Platelet concentrates are routinely stored with continuous agitation but may need to be maintained without agitation for substantial time periods. Studies were conducted in vitro to assess the retention of platelet properties after the discontinuation of agitation. Platelets were maintained without agitation in an insulated cardboard container. In one study, platelet concentrates were kept at 20 to 24°C for the entire 24‐hour period. In another study, they were kept at 37°C for 6 hours with subsequent storage at ambient room temperature for the remainder of the 24‐hour holding period. Under the simulated shipping conditions, discontinuation of agitation for 24 hours between Days 2 and 3 of a 7‐day storage period minimally influenced the maintenance of a series of in vitro platelet properties. The maintenance of platelet concentrates at 20 to 24°C, sitting undisturbed on a bench top for 9 hours, after storage for 6 days with continuous agitation, also had no damaging influence.
Life‐threatening, antiglobulin test‐negative, acute autoimmune hemolytic anemia due to a non‐complement‐activating IgG1 kappa cold antibody with Pra specificity
Authors: Curtis B.R. , Lamon J. , Roelcke D. , Chaplin Hugh
Published: March 2003

A 21‐year‐old man with fulminant cold autoantibody hemolytic anemia (CAHA) was hospitalized with hemoglobinemia, hemoglobinuria, hemoglobin concentration of 3.3 gm per dL, a negative direct antiglobulin test (DAT) with polyspecific and anti‐C3d reagents, a negative Donath‐ Landsteiner test, and a cold agglutinin titer of 80. He failed to respond to corticosteroids, multiple plasma exchanges, and cyclophosphamide; he required 54 transfusions in 10 days to maintain a hemoglobin concentration of 6.0 to 10.0 g per dL. He improved dramatically after a splenectomy was performed. The wide‐thermal‐ amplitude cold agglutinin proved to be an IgG1 kappa antibody with Pra specificity. The patient's serum exhibited normal complement activation. When the DAT was carried out at 0 to 4 degrees C, the result was strongly positive for IgG; the indirect antiglobulin test at 0 to 4 degrees C was positive with the patient's serum diluted 1 in 640. Within 6 months, he was in complete remission and receiving no therapy. As compared with eight patients with CAHA that was exclusively IgG‐mediated, this patient is remarkable for his requirement for many transfusions and for DATs that were consistently negative for C3d.
Relationship of transfusion and infectious complications after gastric carcinoma operations
Authors: Pinto V. , Baldonedo R. , Nicolas C. , Barez A. , Perez A. , Aza J.
Published: March 2003

To determine the effect of transfusion on the incidence of postoperative infection, a retrospective cohort study of 196 patients who underwent surgery for gastric carcinoma in the period from 1985 through 1989 was carried out. Seventy‐one patients (36.2%) developed postoperative septic complications; they had received an average of 4.2 blood units, as compared with 2.7 units received by patients not affected (p < 0.0053). The hypothesis of dose‐response relationship is supported by the Mantel‐Haenszel test, as applied to the overall results (p < 0.01) and the results grouped by duration of operation (p < 0.02). Furthermore, logistic regression analysis shows transfusion to be an independent risk factor in the incidence of infection (p < 0.01), as are antibiotic prophylaxis (p < 0.015), urinary tract catheterization (p < 0.002), and the duration of surgery (p < 0.027). This significance is attained after adjustment for age, gender, period of evolution of symptoms; preoperative infection(s), number of white cells, hemoglobin level and total proteins on diagnosis, location of tumor, tumor, nodes, and metastasis staging, operative technique, drainage of the area of operation, enteral nutrition, and the histologic studies and macroscopic appearance of the tumor. This study is further evidence that transfusion may cause an increased incidence of postoperative infection.
Selective in vivo removal of rheumatoid factor by an extracorporeal treatment device in rheumatoid arthritis patients
Authors: Lazarus H.M. , Cohen S. B. , Clegg D. O. , Menitove J. E. , Sorin S. B. , Hinkle S. , Markenson J. A. , Saal S. , Goodnough L. T. , Fleischmann R. M.
Published: March 2003

A prospective phase II trial was conducted to assess the feasibility, tolerance, and efficacy of a device designed for selective removal of rheumatoid factor from the plasma of rheumatoid arthritis patients. The device contained terpolymer hydrogel‐coated plates with chemically attached, aggregated human immunoglobulin G, and it operated as an immunoaffinity column. Sixty‐one patients aged 25 to 73 underwent weekly plasmapheresis treatments (the primary therapy phase). During the trial, patients continued current rheumatoid arthritis medications without dose adjustments. All patients received two to six treatments (primary therapy). Responding patients were eligible to continue apheresis treatment every 2 to 6 weeks (maintenance therapy). No serious, untoward side effects were noted in the course of this study; of 640 treatments, only 2 (in different patients) were aborted, one because of complaints of dizziness and angioedema and the other because of chest tightness and shortness of breath. Except for a significant (p < 0.05) decrease in serum iron, no significant changes in complete blood count, serum electrolytes, renal and hepatic function tests, or serum C3 and C4 were noted. Although the trial was not designed to determine clinical efficacy, patients noted less morning stiffness, longer time to onset of fatigue, and improved global pain assessment (p < 0.004); significant objective improvements were noted in joint pain, tenderness, swelling, and the number of affected joints (p < 0.001). One‐half of the treated patients had at least a 50 percent improvement in objective measures of antirheumatic activity. Although serum rheumatoid factor could be recovered in vitro in large quantities, no correlation was noted between changes in serum rheumatoid factor concentration and clinical response. This therapeutic modality was safe, well tolerated, and associated with clinical improvement in some patients. It merits further evaluation in a prospective phase III clinical trial.
Reliable confirmation and quantitation of human immunodeficiency virus type 1 antibody using a recombinant‐antigen immunoblot assay
Authors: Busch M. P. , Amad Z. El , McHugh T. M. , Chien D. , Polito A. J.
Published: March 2003

The recombinant DNA‐derived, human immunodeficiency virus (HIV) antigen‐ based immunoblot assay (RIBA‐HIV216) is a new supplemental (confirmatory) test developed to detect antibodies to HIV‐1. The assay employs four recombinant viral antigens, corresponding to HIV‐1 p24, p31, p41 and gp120 proteins, in an immunoblot format. With this assay, HIV‐1 antigen reactivity was detected in all 683 infected patient serum or plasma specimens evaluated; 665 (97.6%) of these sera met the criteria for a positive interpretation, and 18 (2.6%) were classified as indeterminate. All 683 samples reacted with the recombinant gp41‐ equivalent protein. The first sequential enzyme immunoassay (EIA)‐ reactive samples collected from 33 seroconverting homosexual men reacted on RIBA‐HIV216. Eleven (1.1%) of 999 EIA‐negative blood donor sera reacted weakly with a single recombinant antigen (p24 or p31), whereas 13 to 48 percent had indeterminate reactions on viral lysate Western blots. One (1.5%) of 66 EIA‐positive, Western blot‐negative blood donor samples and 19 (29%) of 66 EIA‐positive, Western blot‐ indeterminate blood donor samples scored indeterminate on RIBA‐HIV216. Nonspecific reactivity was seen with only 1 (0.8%) of 114 patient sera containing possible interfering antibodies, whereas 33 percent of these samples had indeterminate reactions on Western blot and 35 percent had equivocal reactions on immunofluorescence assay (IFA). We conclude that the RIBA‐HIV216 is approximately as sensitive as and significantly more specific than virus‐derived Western blot and IFA. The RIBA‐HIV216 also allows for semiquantitation of specific antibodies that may be of value in clinical staging and therapeutic monitoring.
A rare‐event analysis model for quantifying white cells in white cell‐ depleted blood
Authors: Wenz B. , Burns E. R. , Lee V. , Miller W. K.
Published: March 2003

An analysis model to detect and quantify white cells (WBCs) in red cell concentrates (RBCC) drawn from units of blood that are highly depleted of WBCs is described. WBC detection is performed by fluorescence analysis of 50 microL of RBCC labeled with propidium iodide, a DNA/RNA fluorophore. Quantification is performed by regression analysis of standard dilutions of RBCC in substantially WBC‐free red cells. This RBCC diluent is obtained by filtration of blood through a new medium. The method proves to be precise (CV = 7%), efficient (± 30 min/aliquot), and linear (r = 0.99) to 6 log10 WBC depletion of the native product. The current technique is preferable to those suggested previously, such as ficoll concentration, which requires the sacrifice of the unit of blood for counting purposes, and to earlier fluorescence analysis techniques that do not employ WBC‐free red cell diluents. The latter do not monitor extremely low concentrations of WBCs because they lack adequate signal‐to‐noise discrimination. The sensitivity of the described method allows for monitoring of WBC depletion procedures with greater efficiency than is currently available commercially.
Effect of delayed centrifugation or reading on the detection of ABO incompatibility by the immediate‐spin crossmatch
Authors: Shulman I. A. , Calderon C.
Published: March 2003

The immediate‐spin (IS) crossmatch is a method that detects ABO incompatibility. However, under certain circumstances, this test may show unwanted negative or weak results, even though the red cells (RBCs) and serum being tested are ABO incompatible with each other. The present study investigated the potential effect of delayed centrifugation or reading on the IS crossmatch test performance. When the centrifugation step of the IS crossmatch between group O sera and group A1 RBCs was delayed for 2 minutes, 5 of 200 crossmatches showed no agglutination with only trace (n = 2) or moderate (n = 3) hemolysis, and one crossmatch showed only weak, macroscopic agglutination, but moderate hemolysis. All six sera contained A antibodies that were lytic in vitro and, therefore, probably were capable of causing in vivo hemolysis of transfused A1 RBCs. Delaying the reading of the IS crossmatch test for 2 minutes had no apparent effect on test performance. These data demonstrate the importance of technologists’ recognition of hemolysis as a positive result on IS crossmatches, especially if the performance of the centrifugation step of the test is delayed. Furthermore, the unwanted negative agglutination results were abolished by suspending the group A1 RBCs in saline containing EDTA. The authors’ laboratory has modified its IS crossmatch procedure so that donor RBCs are routinely suspended in saline containing EDTA before testing. This procedural change should increase the safety of the IS crossmatch.
Tissue transplant‐transmitted infections
Authors: Kakaiya R. , Miller W. V. , Gudino M. D.
Published: March 2003

Tissue bankers, as well as those transplanting tissues, have been sensitized to the possibility of transmission of fatal infection via tissue transplants, particularly following recent reports of a few cases of AIDS or HIV infection from bone, semen, and skin grafts. It is beyond the scope of this review to describe the steps taken by tissue banks to enhance the safety of tissue transplants. Of note is the fact that a number of new donor screening tests, such as those for antibody to HIV, HBcAg, hepatitis C virus, and human T cell lymphotropic virus type I, have recently been implemented. In addition, rapid advances in the medical history screening of tissue donors and tissue procurement, processing, and preservation continue. Viral inactivation studies are also being undertaken. All these measures are being introduced to increase the safety of tissue transplants.
Polymerase chain reaction and allele‐specific oligonucleotides in paternity testing of the deceased
Authors: Howard P. L. , Collins C. C. , Heintz N. H.
Published: March 2003

The assignment of paternity when the alleged father has died is now possible by use of a variety of allele‐specific oligonucleotides after amplification of genomic DNA by the polymerase chain reaction (PCR). Issues relating to the inheritance of estates may be decided on fact rather than allegation. PCR‐based genotyping of DQ alpha haplotypes from paraffin‐embedded tissue of the deceased was used to prove non‐ paternity in the case reported here. Because the child was female, it was also possible to confirm the exclusion by using a second polymorphic site located in the factor VIII gene on the × chromosome.
Antibodies to hepatitis C virus in autologous blood donors
Authors: Conover P.T. , Fang C.T. , Lam E. , Hirschler N.V. , Jackson J.B. , Yomtovian R.A.
Published: March 2003

Hepatitis C virus (HCV) is the major cause of posttransfusion hepatitis. Two anti‐HCV enzyme immunoassay (EIA) kits and one recombinant immunoblot assay (RIBA) were used to test serum samples of 1476 donations from 692 autologous blood donors to assess the prevalence of anti‐HCV and its relationship to transfusion history. Of all autologous blood donations, 23 (1.6%) reacted when tested with one EIA kit and 29 (2.0%) reacted when tested by the other EIA kit. Of the autologous donors, 12 (1.78%) reacted by the first EIA kit and 14 (2.02%) by the second. Discrepancies in the EIA results from different donations by the same donor were seen in seven donors. The RIBA was positive or indeterminate in 33 percent of the EIA‐reactive donations and in 41 percent of EIA‐reactive donors. All RIBA‐positive and ‐ indeterminate samples reacted with both EIA kits. There was no significant difference in the EIA‐reactive rates of autologous and first‐time homologous blood donors. Previously transfused autologous blood donors had a higher anti‐HCV EIA‐reactive rate than nontransfused autologous donors, but the difference was not significant. In regard to hepatitis C, the use of autologous blood for homologous transfusion appears to be as safe as the use of blood from first‐time homologous donors. Universal testing of previously transfused patients for hepatitis C appears premature at this time. Discrepant anti‐HCV EIA results from different donations from the same individual have implications regarding donor deferral.
Prevalence of antibody to Trypanosoma cruzi among blood donors in Los Angeles, California
Authors: Kerndt P.R. , Waskin H.A. , Kirchhoff L.V. , Steurer F. , Waterman S.H. , Nelson J.M. , Gellert G.A. , Shulman I.A.
Published: March 2003

Transfusion‐associated Chagas' disease is a serious public health problem in Central and South America. With the recent influx of immigrants from Chagas' disease‐endemic areas, concern about the risk of disease from blood transfusion has increased in the United States. To assess the prevalence of Trypanosoma cruzi infection in one area, 1024 consecutive blood donations from 988 voluntary blood donors at a medical center in Los Angeles County were screened serologically. The median age of donors screened was 32.5 years; 53.4 percent were male, and 38.4 percent were born in Chagas' disease‐endemic countries. All donor sera were tested by complement fixation (CF) and indirect immunofluorescence (IIF) tests. A radioimmunoprecipitation assay (RIPA) was also done on all sera from CF‐ or IIF‐reactive donors and an equal number of sera from nonreactive donors. A second serum specimen was obtained, and interviews were completed for 18 (67%) of 27 donors with an initial CF titer greater than or equal to 8 or an IIF titer greater than or equal to 64. The overall seroreactivity (by CF and IIF) was 1.1 percent (11/988). One donor (0.1%) had antibody specific to the 72‐ and 90‐kDa antigens of T. cruzi on RIPA. Seven recipients of blood components from the seroreactive donors were located and were seronegative at 3 to 6 months. Seroreactive donors were 3.6 times more likely to have been born or to have resided in Mexico or Central America, 8.7 times more likely to have donated blood in the past, and 11.8 times more likely to have a history of malaria prophylaxis or treatment.
Comparative evaluation of supplemental hepatitis C virus antibody test systems
Authors: Evans C.S. , Tobler L. , Polito A. , Stewart J. , Chien D. , Wilber J. , Quan S. , Delaney S. , Kuo G. , Busch M.P.
Published: March 2003

Implementation of routine blood donor screening using anti‐hepatitis C virus (HCV) enzyme immunoassay (EIA) has resulted in an urgent need for well‐characterized supplemental assays to confirm the presence of HCV antibodies. A comparative study of four commercially available supplemental assays is reported here: first‐ and second‐generation versions of a strip recombinant immunoblot assay (RIBA‐1 and RIBA‐2), an HCV neutralization EIA, and HCV neutralization plus synthetic peptide EIA. Three hundred sixty‐seven blood donor specimens that were repeatedly reactive on HCV EIA were studied. Most specimens (93%) were also evaluated by radioimmunoassay (RIA) with a six‐antigen panel, and 60 selected specimens were tested for HCV RNA by the polymerase chain reaction (PCR). RIBA‐1 and RIBA‐2 gave concordant results with 86 percent of specimens, while an additional 13 percent were correctly classified by RIBA‐2 but not RIBA‐1. Neutralization EIA alone correctly identified 94 percent of the study group, while the remaining 6 percent required the peptide EIA or the combined neutralization‐peptide assay system for correct classification. The RIBA‐2 and neutralization‐peptide assay systems yielded identical results for 86 percent of specimens, and these results were supported by RIA and selected PCR testing. Only 2 specimens (0.5%) were frankly discrepant, while 51 specimens were indeterminate on either (47) or both (4) assays. When either the RIBA‐2 or neutralization‐peptide assay yielded an indeterminate interpretation, the other system correctly classified the specimen (based on concordance with RIA and PCR data) in a high proportion (92%) of cases. The overall high degree of concordance between RIBA‐2 and neutralization‐peptide assays, as well as the correlating RIA and PCR results, supports the validity and utility of these supplemental anti‐HCV test systems.
Quantitation of red cell‐bound IgG by an enzyme‐linked antiglobulin test in human immunodeficiency virus‐infected persons
Authors: Bordin J.O. , Kerbuary J. , Souza‐Pinto J.C. , Conti E. , Accetturi C.A. , Kishiwada M.A.M. , Novo N.F. , Castelo A.
Published: March 2003

Anemia, thrombocytopenia, and neutropenia have been observed in patients with acquired immune deficiency syndrome (AIDS) and AIDS‐related complex. To investigate whether red cells (RBCs) of patients with human immunodeficiency virus infection were coated with IgG and/or complement (C3), blood samples of 239 patients were tested. The prevalence of a positive direct antiglobulin test on RBCs was 16.7 percent. By use of an enzyme‐linked antiglobulin test (ELAT) to measure more accurately the number of IgG molecules per RBC in a group of 67 patients, 30 of the 67 individuals were observed to have increased numbers (mean, 155) compared to normal controls and to patients with hypergammaglobulinemia due to multiple myeloma or chronic liver disease. Hemoglobin level was correlated with the number of IgG molecules per RBC (p = 0.008), but no correlation could be demonstrated between those numbers and serum immunoglobulin (p = 0.10) or circulating immune complexes (p = 0.38). Our results with ELAT suggest that some AIDS patients may have specific binding of IgG on the surface of their RBCs, rather than nonspecific uptake; further clinical correlations are necessary to confirm these findings.
Five‐day storage of platelets in a non‐diethylhexyl phthalate‐plasticized container
Authors: SNYDER E.L. , ASTER R.H. , HEATON A. , GRODE G. , NAPYCHANK P. , KAGEN L. , JEFFERIES L.C. , HEDBERG S. , BUCHHOLZ D.H.
Published: March 2003

A non‐diethylhexyl phthalate (DEHP)‐plasticized blood bag for 5‐day storage of random‐donor platelet concentrates has been developed. The plastic bag is composed of polyvinylchloride plastic with a butyryl trihexyl citrate plasticizer. The suitability of this plastic for the storage of platelet concentrates for use in clinical transfusion practice was evaluated. In vitro storage studies showed no significant differences at Day 5 for a series of in vitro assays (test plastic vs. control plastic) including pH (7.31 vs. 7.44), lactate dehydrogenase discharge (21.8 vs. 17.1%), pO2 (103 vs. 120 torr), osmotic recovery (52 vs. 57%), and morphology score (527 vs. 516). For paired radiolabeled recovery and survival data from autologous blood donors, results showed equivalence between the test plastic and two control plastics. A small but significant difference between test and control plastics in regard to survival was found by using a linear computer model, but not with a gamma function (multiple‐hit) model. For paired transfusions to thrombocytopenic patients, the corrected count increments at 1 to 4 hours (test vs. control) were 13,534 versus 15,494 (p > 0.05, NS). Similar results were seen for corrected count increments determined at 12 to 24 hours. It can be concluded that platelets stored in the test plastic are acceptable for use in clinical practice.
In vitro changes in platelet function and metabolism following increasing doses of ultraviolet‐B irradiation
Authors: Johnson R.B. , Napychank P.A. , Murphy S. , Snyder E.L.
Published: March 2003

Ultraviolet‐B (UV‐B) irradiation of platelet concentrates (PCs) may prevent the development of posttransfusion HLA alloimmunization. This study evaluated the effect of increasing doses of UV‐B radiation on stored PCs. Pooled PCs were irradiated at UV‐B doses of 600, 2400 or 10,000 mJ per cm2 and stored up to 96 hours under standard blood bank conditions. Compared to nonirradiated room‐temperature and 37 degrees C controls, the irradiated units showed no significant changes in platelet count, white cell count, discharge of lactate dehydrogenase, release of beta‐thromboglobulin, metabolism of ATP, ADP, ammonia, glutamine, glutamate, hypoxanthine, pCO2, or pO2 at any time of storage following any of the three UV‐B doses. However, after a dose of 10,000 mJ per cm2, there were significant decreases in in vitro assays of platelet function‐specifically, osmotic recovery and morphology score. Some metabolic systems were also affected by the 10,000 mJ per cm2 radiation dose, as shown by a decline in pH and bicarbonate and an increase in glucose consumption and lactate production (p < 0.05). The changes in these latter assays appeared only after 96 hours of postirradiation storage. Such changes were not seen in either the room‐ temperature or 37 degrees C control groups. Thus, heat generated during irradiation, per se, did not appear responsible for the observed in vitro changes in platelet function and metabolism. On the basis of the assays analyzed, it is concluded that UV‐B irradiation of PCs at doses up to 10,000 mJ per cm2 does not induce significant metabolic or functional derangements following short‐term storage (24–48 hours).(ABSTRACT TRUNCATED AT 250 WORDS)It is likely, how‐ever, that hi dose UV‐B irradiation followed by long‐term (96‐hour) stora e will adversely adct some platelet metabolic and functional properties.
Detection of cytomegalovirus in blood donors by the polymerase chain reaction
Authors: Smith K.L. , Kulski J.K. , Cobain T. , Dunstan R.A.
Published: March 2003

A polymerase chain reaction (PCR) assay was developed and optimized to detect cytomegalovirus (CMV) DNA in the blood of 86 normal donors who had originally tested seropositive for CMV. Evidence of previous or current infection with CMV was determined by rescreening of the blood for CMV antibodies and by detecting the presence of infectious virus in the white cells by cell culture. DNA was extracted from the blood of donors by a manual or an automated method and amplified by PCR using primers from the major immediate early gene of CMV DNA. The amplified product was detected by visualization of a fluorescent 435‐base pair DNA band in an electrophoretic agarose gel after ethidium bromide staining and confirmed by slot‐blot DNA hybridization using an oligonucleotide probe with complementarity for the major immediate early gene. Seven (8%) of the 86 donors were positive for CMV DNA in both fluorescence and hybridization studies. These donors were also antibody positive. While 74 (86%) of the 86 donors were positive for the presence of CMV antibodies in enzyme‐linked immunosorbent assay, none was positive for virus in cell culture. PCR has the potential to be an effective and reliable procedure for the detection of CMV DNA in donor blood, but further study is required for this technique to be used for diagnostic or routine screening purposes.
Hepatitis C virus RNA in serum of blood donors with or without elevated transaminase levels
Authors: Romeo R. , Thiers V. , Driss F. , Berthelot P. , Nalpas B. , Brechot C.
Published: March 2003

The pattern of hepatitis C virus (HCV) viremia in blood donors who are positive for antibody to HCV (anti‐HCV) according to the level of transaminase activity is unclear. A polymerase chain reaction‐based HCV RNA detection method was used to study two clearly defined groups of anti‐HCV‐positive blood donors with repeatedly normal (n = 27) and elevated (n = 17) alanine aminotransferase (ALT) levels. HCV RNA was detected in only 4 of 27 blood donors with normal ALT values and 15 of 17 with elevated ALT values. These results indicate that anti‐HCV‐positive blood donors with normal ALT levels constitute a heterogeneous group, as HCV viremia is detectable in only a small proportion of cases. Polymerase chain reaction should be useful in the surveillance of anti‐HCV‐positive blood donors with normal ALT levels, by identifying those who might benefit from further investigation and treatment.
Analysis of the expression of neutrophil‐specific antigen NB1: characterization of neutrophils that react with but are not agglutinated by anti‐NB1
Authors: Stroncek D.F. , Plachta L.B. , Herr G.P. , Dalmasso A.P.
Published: March 2003

The neutrophil‐specific NB antigen system has been serologically characterized with human alloantisera. Two alleles, NB1 and NB2, have been described. NB1 is expressed on a subpopulation of peripheral blood neutrophils in 97 percent of healthy donors. Human alloantibodies have been used to identify the 58‐ to 64‐kDa glycoprotein (GP) on which NB1 is located. NB1 can usually be detected by both a granulocyte immunofluorescence (GIF) assay and a granulocyte agglutination (GA) assay, but neutrophils from some donors have been found to react with anti‐NB1 in GIF but not in GA assays. To determine if the latter neutrophils express NB1 and the corresponding 58‐ to 64‐kDa GP, these neutrophils were probed with rabbit and human sera specific for NB1. First, the proportion of neutrophils that express NB1 was quantitated. Neutrophils from donors that typed as NB1‐positive in both GA and GIF assays were analyzed by flow cytometry with antisera to NB1. Human and rabbit anti‐NB1 reacted with 71 ± 17 percent and 70 ± 17 percent of neutrophils, respectively. There was no difference in the expression of NB1 in NB1‐homozygous and NB1‐heterozygous individuals. In contrast, significantly fewer neutrophils from four donors that typed as NB1‐positive in GIF assay but not GA assay reacted with human (27 ± 12%; p < 0.001) and rabbit (26 ± 12%; p < 0.001) anti‐NB1. When neutrophils from these same four donors were probed with rabbit and human anti‐NB1 by immunoblotting and immunoprecipitation, the 58‐ to 64‐kDa GP was identified. No difference in the electrophoretic mobility of the 58‐ to 64‐kDa GP could be detected in donors with low or high percentages of NB1‐positive neutrophils. These studies show that some neutrophils that are not agglutinated by NB1 antibody do express NBI and the 58‐ to 64‐kDa GP.