bluuebunny/crossref_metadata_2025 huggingface dataset:
Evidence that SE is distal to LU on chromosome 19q
Authors: Zelinski T. , Coghlan G. , Greenberg C. R. , McAlpine P. J. , Lewis M.
Published: March 2003

Analysis of a family informative for chromosome 19 loci establishes that the Lutheran blood group locus (LU) lies between the third component of human complement (C3) and the secretor loci (SE). Previously published lod scores for C3:LU are increased from 2.94 to 3.80. The linkage relationships (ẑ and Ô) between C3, LU, and SE are examined, and the proposed order and approximate genetic distances are determined to be: pter —C3 ‐ 10.6cM ‐ cen ‐ 7.4cM ‐ LU ‐ 9cM ‐ SE— qter.
Characteristics of a DNA probe (pa3′HVR) when used for paternity testing
Authors: Allen R. W. , Bliss B. , Pearson A.
Published: March 2003

DNA probes that detect polymorphic loci in the human genome are finding widespread application in many areas of genetic testing. Paternity testing represents one area for the application of probe technology; this report presents data obtained in a paternity testing program with a probe (pa3′HVR) derived from a locus (D16S85) approximately 8 kilobases (kb) downstream from the alpha globin gene complex on chromosome 16. The pa3′HVR probe used under stringent conditions of hybridization detects a highly polymorphic locus in chromosomal DNA digested with Pvu 2 restriction endonuclease. Alleles at the D16S85 locus were grouped into 58 size bins differing from one another by 100 base pairs in the black and white populations. The most common alleles detected in whites fell into the 2.3‐kb group with a collective frequency of 0.1849. In blacks, the most common allele group is 2.0 kb with a collective frequency of 0.1333. The probe was used for restriction fragment length polymorphism mapping in conjunction with standard paternity testing techniques in 100 paternity cases. Thirty direct exclusions were encountered in the 100 cases with standard testing methods, versus 27 exclusions with the pa3′HVR probe alone. Four exclusions detected with standard methods were not detected with the probe and one exclusion detected with the probe was missed by standard testing. The probability of excluding a falsely accused man by use of the pa3′HVR probe was approximately 90 percent. In cases where exclusions were not encountered, the data obtained with the pa3′HVR probe increased the paternity index calculated from standard testing by about 16‐fold. The authors conclude that the pa3′HVR probe used in conjunction with Pvu 2 restriction endonuclease can be extremely informative for paternity testing.
Evaluation of the polyethylene glycol‐indirect antiglobulin test for routine compatibility testing
Authors: Slater J. L. , Griswold D. J. , Wojtyniak L. S. , Reisling M. J.
Published: March 2003

All specimens received in the blood bank over a 5‐month period for crossmatch or group and screen requests were tested in parallel by a polyethylene glycol‐indirect antiglobulin test (PEG‐IAT) and a low‐ionic‐strength saline (LISS)‐IAT. The sera of 41 of 1471 patients had reactions, with 50 antibodies being detected. Ten antibodies reacted only on the PEG‐IAT and 14 only by the LISS‐IAT; the remaining 26 antibodies were detected by both methods. Of the antibodies that reacted only by the LISS‐IAT, one (anti‐Jka) was considered clinically significant, whereas five of the antibodies that reacted only by the PEG‐IAT (1 anti‐c, 2 ‐Fya, 1‐Jkb, and 1‐S) were considered significant. Two antibodies of questionable clinical significance were detected only by the PEG‐IAT. In 97 percent of the sera tested, no reaction was detected by either method. The PEG‐IAT is an acceptable technique for routine compatibility testing.
Utility of DRG and ICD‐9‐CM classification codes for the study of transfusion issues
Authors: Surgenor D. M. , Wallace E. L. , Churchill W. H. , Hao S. , Hale W. B. , Schnitzer J.
Published: March 2003

Red cell transfusions in all patients within specific medical or surgical diagnosis‐related groups (DRGs) and International Classification of Diseases (ICD‐9‐CM) classes were analyzed by a unique body of data that combined abstracted patient discharge records with the numbers of red cell units transfused. Informative measures of transfusion practice within an ICD‐9‐CM class were the proportion of patients transfused, the mean units transfused per patient, and the ratio of standard deviation to the mean of units transfused. Transfusion frequency plots (percentage of patients against units of red cells transfused per patient) revealed the existence of a modal transfusion frequency, as well as an asymmetric tail on the high frequency side. These and other features make it possible to characterize transfusion practice in specific ICD‐9‐CM classes. The mean units of red cells transfused for all patients in a DRG is a measure of blood resource utilization and should be useful in planning to meet future needs.
UMC, another Cromer‐related blood group antigen
Authors: Daniels G.L. , Okubo Y. , Yamaguchi H. , Seno T. , Ikuta M.
Published: March 2003

An antibody found in the serum of a Japanese blood donor detects a new high‐frequency red cell antigen named UMC. UMC is absent from Inab phenotype cells, weakly expressed on Dr(a‐) cells, and destroyed by α‐chymotrypsin but not trypsin treatment of red cells. UMC is, therefore, a Cromer‐related antigen. Immunoblotting with anti‐UMC showed that UMC antigen, in common with other Cromer‐related antigens, is carried on a red cell surface glycoprotein with an apparent molecular weight of 70,000, which presumably is the complement regulatory protein decayaccelerating factor.
In vivo red cell destruction by anti‐Lu6
Authors: Issitt P.D. , Valinsky J.E. , Marsh W. L. , DiNapoli J. , Gutgsell N. S.
Published: March 2003

An example is presented of an IgG1, anti‐Lu6, that reacted by indirect antiglobulin test and was capable of destroying antigen‐positive red cells in vivo. Two methods for the measurement of red cell survival, 51Cr labeling and flow cytometry, gave the same result: 20 percent of the test dose of Lu:6 red cells was destroyed in the first hour after injection and 80 percent in the first 24 hours. The clinical relevance of the antibody was correctly predicted by an in vitro monocyte monolayer assay. The finding that this example of anti‐Lu6 was clinically significant should not be taken to mean that all antibodies directed against high‐incidence Lutheran and Lutheran system‐related antigens will behave similarly. When such antibodies are encountered, in vivo and/or in vitro studies to assess their clinical significance are necessary before rare blood is used for transfusion.
Inhibition of an anti‐Pr1d cold agglutinin by citrate present in commercial red cell preservative solutions
Authors: Green E.D. , Curtis B.R. , Issitt P.D. , Gutgsell N.S. , Roelcke D. , Farrar R.P. , Chaplin H.
Published: March 2003

A patient with known cold autoimmune hemolyticanemia was admitted for surgery. Routine cold agglutinin evaluations, using commercial red cells (RBCs) in modified Alsever's preservative solution, revealed a cold agglutinin titer of 4 to 16. However, using RBCs washed four times with saline, a high‐titer (>2000 at 4° C) cold autoagglutinin was demonstrated. The cold agglutinin was shown to be an IgM kappa paraprotein with anti‐Pr1d specificity. The addition of Alsever's solution to washed RBCs inhibited the cold agglutinin. Each major component of Alsever's solution (neomycin, chloramphenicol, inosine, dextrose, and citrate) was tested individually; only citrate inhibited the patient's cold agglutinin. Various compounds structurally related to citrate were tested and found to cause various degrees of inhibition. The strongest inhibition correlated with the presence of either three carboxyl groups on molecules devoid of double‐bonded carbon atoms or two carboxyl groups in cis configuration. A panel of 54 cold agglutinins, including 7 with anti‐Pr specificity, was analyzed. None was significantly inhibited by Alsever's solution, although one with anti‐Pr2 specificity was weakly inhibited. In summary, these studies describe an anti‐Pr1d cold autoagglutinin that was inhibited by citrate in RBC preservative solutions. The failure to detect such a cold agglutinin can result from not washing RBCs free of citrate before testing.
The differentiation of delayed serologic and delayed hemolytic transfusion reactions: incidence, long‐term serologic findings, and clinical significance
Authors: Ness Paul M. , Shirey R. S. , Thoman S. K. , Buck S. A.
Published: March 2003

Delayed serologic transfusion reactions (DSTRs) and delayed hemolytic transfusion reactions (DHTRs) were studied in a large tertiary‐care hospital. A DSTR was defined by the posttransfusion finding of a positive direct antiglobulin test (DAT) and a newly developed alloantibody specificity. A DHTR was defined as a DSTR case that showed clinical and/or laboratory evidence of hemolysis. Thirty‐four cases of DSTR, 70 percent of which were due to anti‐E and/or ‐Jka, were documented prospectively over a 20‐month period. Retrospective review of the medical records found clinical evidence of hemolysis in only 6 (18%) of the 34. Thus, the incidence of DSTR was 1 (0.66%) of 151 recipients with posttransfusion samples available for testing, whereas the incidence of DHTR was only 1 (0.12%) of 854 patients tested. Fifteen of the 34 patients were followed for up to 174 days after reaction. Twelve of the 15 still demonstrated a positive DAT with anti‐ IgG only. Eluate studies indicated that the persistence of a positive DAT after DSTR or DHTR may involve several immunologic mechanisms, including the development of posttransfusion autoantibodies. This study indicates 1) that DSTRs are a frequent finding in multiply transfused patients, although most cases are benign and fail to meet rigid criteria for DHTR, and 2) that the persistence of a positive DAT after DSTR or DHTR is common.
Failure to detect evidence of human immunodeficiency virus type 1 (HIV‐ 1) infection by polymerase chain reaction assay in blood donors with isolated core antibodies (anti‐p24 or ‐p17) to HIV‐1
Authors: Mariotti M. , Rouger P. , Thauvin M. , Salmon C. , Lefrere J.J.
Published: March 2003

Individuals with isolated and persistent core antibodies (anti‐p24 or ‐ p17) to HIV‐1 are sometimes diagnosed through the systematic screening of blood donations. The significance of such serum reactivities remains unknown. Polymerase chain reaction (PCR) is a new technique allowing the direct detection of HIV‐1 DNA in blood samples. In this study, PCR was used to detect HIV‐1 DNA in 20 individuals with isolated and persistent core antibodies (14 anti‐p24 and 6 ‐p17), in seven sexual partners of these individuals, in 55 HIV‐1‐seropositive individuals (positive controls), and in 105 HIV‐1‐seronegative blood donors at low risk of HIV‐1 infection (negative controls). No HIV‐1 DNA was detected in individuals with isolated and persistent core antibodies, in their sexual partners, or in negative control individuals, although PCR was positive in 54 of 55 seropositive control individuals. These results strongly suggest that individuals with isolated and persistent core antibodies are not infected with HIV‐1.
The prevalence of antibody to HTLV‐I/II in United States plasma donors and in United States and French hemophiliacs
Authors: Canavaggio M. , Leckie G. , Allain J.P. , Steaffens J.W. , Laurian Y. , Brettler D. , Lee Helen
Published: March 2003

Antibody to HTLV‐I/II was detected in 19 (0.3%) of 6286 plasma donors from five regions of the United States (US). This seroprevalence rate is approximately 10 times that in whole blood donors. The regional distribution of infection was as follows: Southwest, 0.68 percent; Southeast, 0.45 percent; Midwest, 0.28 percent; Northwest, 0.1 percent; and Northeast, 0.0 percent. Rates of HTLV‐I/II infection in blacks (0.74%) and Hispanics (0.66%) were higher (both, p less than 0.001) than those in whites (0.08%). All 19 infected units were donated by subjects aged 30 or older, even though 52.9 percent of the donations came from persons less than 30 years old. Equal rates of HTLV‐I/II infection were found in men (0.31%) and women (0.29%). No HTLV‐I/II antibody was detected in 154 French and 25 US hemophiliacs who were transfused regularly with noninactivated plasma or its derivatives. This suggests that the transfusion of HTLV‐I/II‐seropositive plasma products does not transmit the viral infection.
Inhibition of human lymphocyte reactivity by plasma fibronectin in vitro
Authors: Schultz John C. , Shahidi N.T.
Published: March 2003

The effect of purified human plasma fibronectin (FN) on the reactivity of human lymphocyte‐rich mononuclear cells to mitogens and allogeneic cell interactions was studied. Concentrations of FN from 25 to 100 micrograms per 250 microL of culture consistently depressed phytohemagglutinin (PHA) responses. To exert an inhibitory effect, FN must be present within 20 hours after the addition of PHA to cells, and, therefore, it appears to interfere with early events in the transformation process. Increasing the concentration of PHA failed to reduce the inhibitory effect of FN, which suggests that the depressed response was not the result of FN‐PHA complex formation, which would reduce the amount of mitogen available for stimulation. This possibility was supported by the finding that FN also inhibited the mixed lymphocyte response (MLR), in a reaction that was not dependent on the activity of soluble antigen or mitogen. In contrast, the stimulation of lymphocytes to undergo transformation that is induced by the nonlectin mitogen, sodium periodate, was unaffected by FN. Periodate‐treated cells are, however, already stimulated to undergo transformation, prior to their exposure to FN. FN did not interfere with the activity of interleukin‐2, nor did it indirectly regulate lymphocyte responses by modifying the production and/or effect of humoral regulatory factors released from the adherent accessory cells (macrophages). These studies show that FN is a potent immunosuppressive agent in vitro.
Density gradient separation of peripheral blood stem cells: comparison of an automated cell processing device and manual methods
Authors: Hillyer C.D. , Tiegerman K.O. , Berkman Eugene M.
Published: March 2003

Peripheral blood stem cells were collected from normal donors by leukapheresis on a cell separator. The leukapheresis product contained 1.5 × 10(10) mononuclear cells (MNCs) and was divided into two aliquots that underwent either automated or manual density gradient separation with ficoll‐hypaque and subsequent washing. In the automated process, recovery of MNCs was 85 percent, reduction in platelet content was 64 percent, and the final hematocrit (Hct) was less than 1 percent. The manual separation resulted in 76‐percent MNC recovery, a 79‐percent reduction in platelet content, and a final Hct of less than 1 percent. The purified MNCs were then placed in methylcellulose culture at a concentration of 4 × 10(5) MNCs per mL. Quadruplicate 1‐mL aliquots were cultured, and colonies were counted and classified on Day 14. Comparison of automated and manual ficoll‐hypaque separations demonstrated no differences in the total, erythroid, or granulocyte‐ macrophage colony numbers. The cell processor used is fast, reliable, uncomplicated, and provides a sterile product containing progenitor cells that are not adversely affected by the automated ficoll‐hypaque separation.
Inhibition of in vitro adherence of antibody‐coated red cells to monocytes by the dialyzable leukocyte extract
Authors: Rebulla P. , Tedesco F. , Radillo O. , Battaglioli M. , Boeri M. , di San Secondo V. E. Rosso , Sirchia G.
Published: March 2003

The red cell‐monocyte assay (RMA), which has been used to evaluate the clinical significance of red cell (RBC) antibodies, was employed to test the effect of the dialyzable leukocyte extract (DLE) on in vitro adherence to monocytes of human RBCs coated with alloantibodies or autoantibodies. The total association index (TAI) of the RMA, expressing the number of RBCs adhering to or phagocytosed by 100 monocytes, indicated a potent inhibitory activity of DLE in the test system. TAI values of 100.4 +/− 20.1 (mean +/− SD) in the control sample, consisting of RBCs coated in vitro with anti‐D, dropped to 4.0 +/− 2.1 when DLE was present in the assay medium at a concentration of 0.5 U per mL. Similar results were obtained with RBCs coated with IgG antibodies in vivo. The inhibition was dose dependent and was associated with a thermolabile component of DLE. This study establishes that DLE can modulate monocyte function by inhibiting the recognition of IgG sensitized red cells.
Two cases of “hrB‐like” autoantibodies appearing as alloantibodies
Authors: Vengelen‐Tyler V. , Mogck N.
Published: March 2003

Two cases are described in which autoantibodies mimicked alloantibodies. The direct antiglobulin test (DAT) on the red cells (RBCs) from both patients was negative when routine manual hexadimethrine bromide (Polybrene) and enzyme‐linked antiglobulin techniques were used. The RBCs also did not react on direct bromelin and direct Polybrene tests. However, an “hrB‐like” antibody was eluted from the RBCs of both patients. The sera from these patients reacted with all e+ hrB+ RBCs but not with e+ hrB−, e−, or their own RBCs. The antibody in the serum of one patient was not adsorbed by R2R2 RBCs. Serologic tests initially suggested (by direct testing and adsorption studies) that the serum antibodies were alloantibodies rather than autoantibodies. RBCs taken from one patient, 8 months after her sample was first referred to our laboratory, reacted with a serum sample from her first admission. An RBC sample taken from the other patient, initially typed e+ and hrB− but 1 month later typed e+ and hrB+ by using the same anti‐hrB sera, was used to test the earlier samples.
Assessment of clinical significance of anti‐Ge in an untransfused man
Authors: Pearson H. A. , Richards V. L. , Wylie B. R. , Bruce D. , Watt J. M. , Wilkie D. , Kronenberg H.
Published: March 2003

A 19‐year‐old, untransfused Melanesian man from Papua New Guinea was admitted to the hospital for repair of an atrial septal defect. His serum contained an alloantibody that reacted strongly on the indirect antiglobulin test and was identified as anti‐Ge. Gerbich‐negative blood was transfused following urgent surgery. A 51Cr red cell survival study performed 2 weeks after surgery yielded zero survival of Gerbich‐ positive cells after 24 hours. A monocyte‐driven, antibody‐dependent, cell‐mediated cytotoxicity assay performed on both pretransfusion and posttransfusion serum samples and on concentrated serum showed less than 1 percent specific lysis of Gerbich‐positive cells. This did not correlate with the indication of clinical significance predicted by the 51Cr study. Red cell adherence and phagocytosis, not evident in a monocyte monolayer assay using native serum, were demonstrable in 16 percent of monocytes by the use of concentrated serum.
White cell depletion of red cell and pooled random‐donor platelet concentrates by filtration and residual lymphocyte subset analysis
Authors: Freedman J. , Blanchette V. , Hornstein A. , Farkas S. , Milner R. , Adams M. , Lim F. C. , Garvey M. B. , Hannach B.
Published: March 2003

This study comparing the relative white cell (WBC)‐depleting efficiency of single and double filtration used two filters and new, sensitive, and reliable methods for performing WBC counts on WBC‐depleted blood products. A single filtration of red cell (RBC) concentrates with a cotton‐wool filter reduced WBC content by 98.64 percent, but the range of residual WBCs was wide, and many filtered units still contained more than the theoretical immunizing dose of 5 to 10 × 106 WBCs. A second filtration, however, always produced RBC units that had < 5 × 106 WBCs. Although the degree of WBC depletion observed after a single filtration of a 6‐unit pool of random‐donor platelet concentrates was greater with a polyester filter than with the cotton‐ wool filter (98.92 vs. 98.14% reduction, respectively, when mean prefiltration WBC count was less than 600 × 106, in both cases, 25 percent of filtered products still contained greater than 5 × 106 WBCs; a second filtration (with the cotton‐wool filter), however, produced units that were always below the immunizing dose. All double‐ filtered platelet concentrates had less than 106, and one‐half had less than 104 residual WBCs. Platelet loss was similar with both filters (+/− 16% loss with one filtration). The effectiveness of the filters in producing products that were WBC‐depleted below the immunizing dose was dependent on the prefiltration WBC content (but not on the age of the units), and it may be worthwhile to employ methods to ensure that total prefiltration WBC count of the product is < 400 × 106. Although the polyester filter was more efficient at WBC depletion overall, flow cytometric analysis of mononuclear cell subpopulations in components before and after filtration showed that it resulted in a greater number of residual dendritic cells, and that filtration with the cotton‐wool filter resulted in the same or a lesser amount of dendritic cells being transfused to the patient.
Sia‐b1 and I antigens recognized by Mycoplasma pneumoniae‐induced human cold agglutinins
Authors: Roelcke D. , Kreft H. , Northoff H. , Gallasch E.
Published: March 2003

Postinfection cold agglutinins (CAs) with anti‐Sia‐b1 (i.e., anti‐Sialo‐ branched) specificity frequently occurring together with anti‐I CAs recognize antigenic determinants that are present on Oh red cells and are partially destroyed by endo‐β‐galactosidase on the red cell surface. They differ markedly from the monoclonal anti‐Sia‐b1 FI CA, which recognizes an epitope not expressed on Oh cells, and resistant to the enzyme. On the other side, Sia‐b1 determinants reacting with postinfection CAs share the characteristics of I determinants, with the exception that Sia‐b1 determinants require sialyl groups as an immunodominant component. Five sera containing coexisting anti‐Sia‐b1 and anti‐I CAs were tested against Oh and enzyme‐treated group O red cells. The results are consistent with a postinfection autoimmune response against a sialo‐type 2 structure common for Sia‐b1 and I determinants, which could serve as a receptor for Mycoplasma pneumoniae.
Virucidal levels of ozone induce hemolysis and hemoglobin degradation
Authors: Wagner S.J. , Wagner K.F. , Friedman L.I. , Benade L.F.
Published: March 2003

The animal virus, vesicular stomatitis virus (VSV), and the bacterial virus, phi 6, were inactivated by greater than 4 log10 in response to incubation with 13 to 14 mL of 1.4 mmol per L (65 micrograms/mL) to 1.6 mmol per L (75 micrograms/mL) of overlaid ozone in virus‐spiked, dilute, red cell suspensions. Virus inactivation was greatly inhibited when ozone was overlaid in the presence of high‐hematocrit red cells or, to a lesser degree, high levels of plasma. At hematocrits at which 5 to 6 log10 of VSV were inactivated, ozone caused 30‐percent hemolysis, as measured by the loss of total cellular hemoglobin. Unexpectedly, this level of hemolysis could not be observed in supernatants because of the ozone‐induced destruction (bleaching) of extracellular hemoglobin. These results suggest that ozone may have little biological specificity for damaging viruses over red cells.
Immune‐mediated hemolysis associated with the administration of a radiographic contrast medium
Authors: Nordhagne R. , Vik H. , Wolthuis K. , Bohn H.P. , Urdahl P. , Michaelsen T.E
Published: March 2003

A female patient developed serious hemolysis in association with the injection of a radiographic contrast medium (RCM). Serologic investigation of her serum suggested complement‐mediated hemolysis, induced by an RCM‐dependent IgM antibody in her serum. The antibody was of high titer and low avidity. The antibody showed cross‐reactions with related radiographic contrast media and reacted only with group I adult RBCs.
Paired comparison of platelet concentrates prepared from platelet‐rich plasma and buffy coats using a new technique with111In and51Cr
Authors: Keegan T. , Heaton A. , Holme S. , Owens M. , Nelson E. , Carmen R.
Published: March 2003

Two techniques for the preparation of platelet concentrate (PC), the standard platelet‐rich plasma (PRP) and buffy coat (BC) methods, were compared in nine paired studies with regard to platelet harvest, white cell (WBC) contamination, and PC quality after 5 days of 22°C storage. Platelet harvest using the BC method averaged approximately 56 percent of the whole blood level (6.2 × 1010/concentrate), which was less than the 76 percent achieved with the PRP‐PC method (8.7 × 1010/concentrate). An additional 5 units collected into an experimental siphon bag for BC‐PC processing showed improved platelet harvest (6.7 × 1010/concentrate, or approx. 70% of whole blood). WBCs remaining in the BC‐PC averaged 0.19 × 108per unit compared to 3.6 × 108per unit for PRP‐PC. Buffy coat processing produced red cell (RBC) units with 50 percent of the WBC contamination of conventionally prepared units (9.8 ± 6.2 × 108/unit vs. 18.9 ± 7.1 × 108/unit). The siphon bag further reduced WBC levels in the AS‐3 RBC units (6.4 ± 3.7 × 108/unit). In vitro studies performed on Days 1 and 5 after collection showed no significant differences in platelet metabolic and biologic function or cell integrity. β‐thromboglobulin and surface glycoprotein levels, indicators of platelet activation and membrane alteration, respectively, did not differ significantly in the PRP‐PC and BC‐PC; nor was lactate production higher in PRP‐PC, despite the substantially higher WBC counts. Autologous in vivo platelet viability determinations were performed by using concurrent transfusion of111In‐labeled freshly drawn platelets and51Cr‐labeled stored platelets. Paired f test analysis of BC‐PC versus PRP‐PC indicated no significant differences in platelet recovery and survival after 5 days of 22°C storage in polyolefin containers. Therefore, these studies confirm the equivalence of PC quality, comparable platelet harvest with the siphon bag, and decreased WBC contamination with the BC method.
A latex particle assay for platelet‐associated IgG
Authors: Ramos R.R. , Curtis B.R. , Chaplin H.
Published: March 2003

A modified, sensitive, solid‐phase assay for platelet‐associated IgG is reported. Direct comparisons were made between a 125I monoclonal radioimmunoassay (RIA) and the polyclonal antibody latex particle assay. In 209 simultaneous comparisons with the RIA, the sensitivity of the latex test was 91 percent, specificity was 100 percent, and overall efficiency was 97 percent. Commencing with platelet‐rich plasma, the direct latex particle test for platelet‐bound IgG requires 45 minutes; 90 minutes are required to crossmatch one patient with 12 donors. The advantages of the latex assay are absence of radioactivity, stability of reagents, economy, speed, specificity, and sensitivity.
Growth of Yersinia species in artificially contaminated blood bags
Authors: Franzin L. , Gioannini P.
Published: March 2003

Transfusion blood bags were inoculated with less than 0.1 colony‐ forming units (CFU) of Yersinia enterocolitica (serotypes O:3, O:5, and O:9) and Yersinia pseudotuberculosis I per mL and stored at 4 ° C for 41 days. During storage, samples were collected periodically and colony counts of serial dilutions were performed by plating on tryptic soy agar. After a lag‐phase, the growth of Yersinia species from less than 0.1 CFU per mL to 10(9) CFU per mL was observed in 3 of 10 bags. The data showed that prolonged storage of contaminated blood bags allows Y. enterocolitica and Y. pseudotuberculosis to multiply to high levels.
Risk factors associated with hepatitis B or C markers or elevated alanine aminotransferase level among blood donors on a tropical island: the Guadeloupe experience
Authors: FEST T. , VIEL J.F. , AGIS F. , COFFE C. , DUPOND J.L. , HERVE P.
Published: March 2003

Donated blood is currently screened for hepatitis B surface antigen (HBsAg), antibody to hepatitis B core antigen (anti‐HBc), antibody to hepatitis C virus (anti‐HCV), and alanine aminotransferase (ALT) levels to prevent posttransfusion hepatitis. A prospective study of 2368 blood donors was carried out in Guadeloupe (French West Indies) with a view to determining the risk factors associated with serologic abnormalities. Blood donors included in the study had to complete a questionnaire. Statistical analysis was performed on the data thus obtained: 571 donations (24%) were positive for at least one of the four analyzed markers. The results were that 3.2 percent were positive for HBsAg, 22 percent for anti‐HBc, and 0.8 percent for anti‐HCV, and 1.4 percent had ALT > or = 45 IU per L. A good correlation was found between anti‐HCV and elevated ALT. Transfusion history and two socioeconomic categories (working class, military personnel) were found to be risk factors. Other risk factors were lifelong residence in Guadeloupe (with risk increasing with the number of years), birthplace and current residence in the southern part of the island, and the existence of gastrointestinal discomfort unrelated to viral hepatitis (odds ratio = 2.98). The results of this study illustrate the difficulty of implementing a preventive policy against posttransfusion hepatitis in a tropical area. The unique epidemiologic situation of Guadeloupe as regards hepatitis B virus has led to more restrictive criteria for the acceptance of blood donors.
HLA antibody enhancement by double addition of serum: use in platelet donor selection
Authors: BRYANT P.C. , VAYNTRUB T.A. , SCHRANDT H.A. , KALIL J.E. , GRUMET F.C.
Published: March 2003

The identification of HLA class I alloantibodies is important for organ transplantation and platelet transfusion in alloimmunized patients. Because microcytotoxicity testing against frozen trays of lymphocyte panels is rapid and efficient for determining specificities of unknown antibodies, a simple method was devised to increase test sensitivity to weak antibodies. Standard anti‐human globulin (AHG)‐facilitated microcytotoxicity was modified by the insertion of a double addition‐of‐ serum (DAS) step, and reagent and patient's sera were evaluated by both methods. DAS modification increased antibody titers and, more significantly, made the identification of weak specificities easier because of the twofold to threefold increase in reactivity rates (29‐ 42% for AHG vs. 75–82% for DAS) of panel cells that were expected to be positive, while low (approx. 1%) “extra” reaction rates were maintained for cells that were expected to be negative. DAS was relatively unaffected by variations in serum volumes or target cell preparation, and its use did not significantly increase test time or costs. In a program of platelet donor selection driven by donor antibody rather than donor‐recipient antigen matching, DAS greatly facilitated platelet transfusion support for alloimmunized patients.
A serologic relationship among the NFLD, BOW, and Wu red cell antigens
Authors: KAITA H. , LUBENKO A. , MOULDS M. , LEWIS M.
Published: March 2003

A survey of sera containing antibodies to multiple low‐incidence antigens revealed a variety of patterns of reactions with NFLD+, BOW+, and Wu+ red cell samples. Although NFLD, BOW, and Wu are distinct antigenic determinants (International Society of Blood Transfusion numbers 700.37, 700.46, and 700.13, respectively), their ability to absorb and elute “crossreactive” antibodies indicates a serologic and possibly a genetic relationship among the three.
Expression of blood group antigens on red cell microvesicles
Authors: ORESKOVIC R.T. , DUMASWALA U.J. , GREENWALT T.J.
Published: March 2003

The purpose of this study was to determine whether epitopes of the A, B, D, Fya, M, N, S, s, and K blood group antigens are present on microvesicle membranes shed by red cells during storage. Vesicles were isolated from outdated units of blood having and lacking the specified antigens. Diluted antisera were absorbed with fixed quantities of vesicles from red cells with the test antigen and red cells lacking that antigen (controls). The adsorbed and unadsorbed antisera were titrated and scored by using panel cells from persons known to be heterozygous for all the non‐AB antigens. The mean titration scores following adsorption with the vesicles from A, B, D, M+N‐, M‐N+, S+s‐, S‐s+, and Fy(a+b‐) units were appreciably lower than the control scores (0, 0, 3, 2, 2, 0, 4, and 4 vs. 19, 23, 34, 13, 12, 16, 18, and 29, respectively), which indicated the presence of these epitopes on the membrane of shed vesicles. The results following adsorption with K:1,2 vesicles were equivocal.
A therapeutic, highly purified factor XI concentrate from human plasma
Authors: BURNOUF‐RADOSEVICH M. , BURNOUF T.
Published: March 2003

A highly purified factor XI (FXI) concentrate was prepared from human plasma by a process comprising a filter adsorption step and chromatography on a cation exchange resin. The freeze‐dried FXI, which solubilized quickly, had high specific activity (130–150 U/mg protein), high potency (approx. 100 U/mL), and excellent stability for at least 24 hours at room temperature in the liquid state. The overall recovery was about 220 U of FXI per liter of plasma. Minor protein contaminants (C1‐inhibitor, fibronectin, IgG, and alpha‐2‐macroglobulin) were found to be between 0.13 and 0.46 mg per 1000 U of FXI. Fibrinogen and relevant coagulation factors (factors II, V, VII, IX, X, XII, XIII, and VIII/von Willebrand factor) were undetectable, as evidenced by immunologic and immunoelectrophoretic data. Components of the kinin system were present in trace amounts or were undetectable. No evidence of activated factors such as factors Xa and IXa was found. Proteolytic activity, as assessed by S‐2288 chromogenic substrate, was negligible and thrombin was undetectable. A solvent‐detergent treatment was included prior to chromatographic purification to enhance viral safety against lipid‐enveloped viruses. In vitro and in vivo animal studies demonstrated the absence of thrombogenic, hypotensive, or toxic effects. No thrombogenic activity was found in the Wessler model in rabbits at doses of 900 to 1100 U of FXI per kg of body weight. This FXI preparation could be beneficial in substitution therapy of congenital or acquired FXI deficiency, especially as a way to avoid the use of fresh‐frozen plasma.
Transfusion therapy: improved patient care and resource utilization
Authors: Rosen N.R. , Bates L.H. , Herod G.
Published: March 2003

Improving the quality of medical care while reducing costs is one of the major challenges facing the health care system in the United States. At a 1020‐bed, tertiary‐care, teaching hospital, the Transfusion Committee modified transfusion practice by establishing new transfusion guidelines based upon national standards rather than local practices and by implementing educational and monitoring systems. Over a 3‐year period, the number of transfusions decreased, the types of transfused components changed, and the waste due to unused components decreased. From the baseline of Fiscal Year (FY) 1989 (89), the number of exposures to components from allogeneic blood donors for the patient population decreased by 11,015 in FY 90, 14,067 in FY 91, and 16,990 in FY 92, thereby decreasing the risk of transfusion‐transmitted disease, transfusion reaction, and alloimmunization. As compared to costs in FY 89, the altered transfusion practices resulted in cost savings of $376,269 in FY 90, $566,375 in FY 91, and $684,704 in FY 92. Over the 3‐ year period, exposures to components from allogeneic blood donors for the patient population were reduced by 42,072, and the total cost savings was $1,627,348. The methodology and results should be reproducible at other hospitals.
Rh phenotypes of Chinese blood donors in Hong Kong, with special reference to weak D antigens
Authors: Mak K.H. , Yan K.F. , Cheng S.S. , Yuen M.Y.
Published: March 2003

Among Hong Kong Chinese blood donors, 99.71 percent were found to be D+. Of these, 55.02 percent were of the phenotype CCDee. The Du phenotype was found to be present in 0.016 percent. Among the 0.27 percent who were apparently D‐, 0.079 percent were of the Del phenotype, while the remaining 0.19 percent were “true D‐,” as defined by a nonreactive eluate obtained by an adsorption and elution procedure using anti‐D. The ccdee phenotype constitutes 56.77 percent of the “apparent D‐” and 80.24 percent of the true D‐. Data show that anti‐D rarely occurs in Hong Kong Chinese, and it is postulated that this could be due to the presence of a very weak form of the D antigen among many of those who appear to be D‐.
Lack of detection of human immunodeficiency virus type 1 DNA by polymerase chain reaction in the plasma and lymphocytes of seronegative exposed hemophiliacs
Authors: Henrard DR , Laurian Y. , Mehaffey W.F. , Allain J.P.
Published: March 2003

Direct detection of human immunodeficiency virus type 1 (HIV‐1) DNA in serum or plasma samples has been reported in seronegative as well as seropositive individuals. An alkaline lysis procedure was adapted for polymerase chain reaction (PCR) analysis of plasma specimens. Eighty‐ five seronegative hemophiliacs, 52 of whom had been exposed to HIV‐ contaminated blood components, and 19 seronegative at‐risk individuals were studied. Each sample was extracted and amplified with SK38/39 gag primers at least three times. Seventy‐six samples (72%) were consistently negative for HIV‐1 DNA, 24 (22%) were positive only once, and 4 (3%) were positive twice. Genomic DNA from peripheral mononuclear cells was prepared from 12 of 76 negative samples, 18 of 24 samples that were positive once, and 2 of 4 samples that were positive twice and analyzed with both gag and long terminal repeat primers. None (0/32) of these cellular DNAs were positive for HIV‐1, which suggests that these seronegative exposed hemophiliacs were not latently infected with HIV‐1. In contrast, all (10/10) control cells from seropositive patients were positive with both primer pairs. The detection of HIV‐1 DNA in serum or plasma may be prone to a high level of false‐positive PCR signals and should be interpreted with caution.
Predicting hemolytic disease of the newborn: a comparison of the monocyte monolayer assay and the chemiluminescence test
Authors: Lucas G.F. , Hadley A.G. , Nance S.J. , Garratty G.
Published: March 2003

The ability of the monocyte monolayer assay (MMA) and the chemiluminescence test (CLT) to predict the clinical significance of alloantibodies associated with hemolytic disease of the newborn (HDN) was assessed by the use of 22 well‐characterized antisera—predominantly anti‐D—from alloimmunized pregnant women. Seventeen sera were obtained before delivery from women whose infants were antigen positive for the antibody specificities identified in the maternal serum. With testing of these 17 sera by MMA, 10 results were in agreement with the presence or absence of HDN, but there were 5 false‐positive and 2 false‐negative results. With the CLT, 16 results were in agreement with the presence or absence of HDN, and there was 1 false‐negative result. Five sera were obtained from women whose infants were antigen negative for the antibody specificities identified in the maternal serum. The CLT and the MMA were both subject to false‐positive results with these sera. These results suggest that the CLT may be more valuable than the MMA as a noninvasive test for predicting the clinical significance of alloantibodies in HDN.
Differential effect of cryopreservation on natural killer cell and lymphokine‐activated killer cell activities
Authors: Martí F. , Miralles A. , Peiró M. , Amill B. , de Dalmases C. , Piñol G. , Rueda F. , García J.
Published: March 2003

The use of lymphokine‐activated killer (LAK) cell therapy in delayed treatment requires the use of cryopreserved effector cells. The purpose of this study was to determine the optimal cryopreservation protocol for the maintenance of cytotoxic activity in mononuclear cells (MNCs). MNCs were cryopreserved with dimethyl sulfoxide or 1,2‐propanediol before and after 3 days of culture with recombinant interleukin 2. The effects of cryopreservation on cell recovery, LAK cell and natural killer (NK) cell cytotoxic activities, and surface antigen markers were studied. Recovery of nonactivated MNCs was higher with 1,2‐propanediol than with dimethyl sulfoxide (p < 0.05). Cytotoxic activities, measured with a 51Cr release assay, significantly decreased after thawing, on both activated cells (76.3%; range, 35.8–92.2%) and fresh cells (54.6%; range, 17.5–75.4%). A 6‐day kinetic test was used to compare the cytotoxic activity of cryopreserved and fresh cells. The results showed different patterns for NK cells (cryopreserved cells had lower levels of activity than fresh cells) and LAK cells (cryopreserved cells had higher levels of activity than fresh cells). Phenotype changes of effector cells in culture, with and without cryopreservation, were monitored by flow cytometry using monoclonal antibodies. These results were compared with changes in the cytotoxicity of cells with and without cryopreservation. After thawing, there was a decrease in MNCs expressing CD14 and CD56. Recovery of the CD56 marker correlates with increased cytotoxic activity. Despite some loss of NK cell activity, it is concluded that MNCs may be successfully cryopreserved before their use in immunotherapeutic treatment.
Factors affecting Yersinia enterocolitica (serotype O:8) viability in deliberately inoculated blood
Authors: Wagner S.J. , Robinette D. , Dodd R.
Published: March 2003

Interpretation of in vitro experiments using Yersinia enterocolitica in blood components requires information on factors affecting the organism's survival. Several factors were found to influence the survival of Y. enterocolitica (serotype O:8) in blood components. A 20‐ minute room‐temperature incubation with plasma‐containing components resulted in approximately 2 log10 inactivation. Inactivation could be prevented by preincubation treatment of the plasma at 55°C for 1 hour, which suggests the involvement of heat‐labile plasma factors. No antibacterial activity was observed in washed red cells during the 20‐minute room‐temperature incubation. However, Y. enterocolitica colony‐forming units declined by up to 2 log10 in washed red cells during the first days of 4 degrees C storage. Use of a white cell‐ reduction filter on freshly inoculated samples removed approximately 1 log10 of the organism regardless of whether bacteria were suspended in saline or washed red cells. Thus, bacterial levels may be affected by plasma, cellular components, and white cell‐reduction filters. However, caution should be exercised in interpreting in vitro spiking studies designed to investigate the potential benefits of white cell reduction to eliminate the growth of Y. enterocolitica because of potential differences between naturally infected and experimentally inoculated blood.
Frequency of platelet‐specific antigens among Indonesians
Authors: Santoso S. , Santoso S. , Kiefel V. , Masri R. , Mueller‐Eckhardt C.
Published: March 2003

This study reports the frequencies of the alloantigens of four major platelet‐specific alloantigen systems among Indonesians. One hundred and sixty‐eight unrelated Indonesian blood donors were phenotyped for the alloantigens of the Zw (PIA, HPA‐1), Bak (HPA‐3), Yuk (Pen, HPA‐4), and Br (HPA‐5) systems by use of a glycoprotein‐specific immunoassay. All were positive for the alloantigens Zwa, Yukb, and Brb. Three (1.79%) and 1 (0.59%) of the 168 donors were positive in testing for Zwb and Yuk(a) antigens, respectively. Fifteen (9.26%) of 162 Indonesians had Br(a) antigens. Of the 166 donors tested, 121 (72.89%) were Bak(a) positive and 134 (80.72%) were Bakb positive. In addition, the phenotype frequency of Nak(a) was determined by using monoclonal antibody OKM5 in a platelet enzyme‐linked immunosorbent assay. Its frequency in the present cohort was 95.83 percent (161/168). This study confirms the differences in platelet antigen distributions in Asians and whites. Both glycoprotein IV deficiency and the Yuk polymorphism are also found among Indonesians.
A prospective study of symptomatic bacteremia following platelet transfusion and of its management
Authors: Chiu E. K. W. , Yuen K. Y. , Lie A. K. W. , Liang R. , Lau Y. L. , Lee A. C. W. , Kwong Y. L. , Wong S. , Ng M. H. , Chan T. K.
Published: March 2003

BACKGROUND: The danger of bacteremia due to contaminated platelets is not well known. There are also no established guidelines for the management of febrile reactions after platelet transfusion.

STUDY DESIGN AND METHODS: To determine the risk of symptomatic bacteremia after platelet transfusion, 3584 platelet transfusions given to 161 patients after bone marrow transplantation were prospectively studied. Platelet bags were routinely refrigerated for 24 hours after transfusion. Septic work‐up was initiated for a temperature rise of more than 2 degrees C above the pretransfusion value within 24 hours of platelet transfusion or a temperature rise of more than 1 degree C that was associated with chills and rigor. Diagnosis of bacteremia after platelet transfusion was made only when the pairs of isolates from the blood and the platelet bags were identical with respect to their biochemical profile, antibiotic sensitivity, serotyping, or ribotyping.

RESULTS: Thirty‐seven febrile reactions, as defined above, occurred. Bacteremia subsequent to platelet transfusion was diagnosed in 10 cases. There was a 27‐percent chance (95% CI, 15–43%) that these febrile reactions represented bacteremia that resulted from platelet transfusion. For a subgroup of 19 patients with a temperature rise of more than 2 degrees C, the risk of bacteremia was 42 percent (95% CI, 23–64%). Septic shock occurred in 4 of the 10 bacteremic patients. A rapid diagnosis was possible because the involved bacteria were demonstrated by direct Gram stain of the samples taken from the platelet bags of all 10 patients.

CONCLUSION: Significant febrile reactions after platelet transfusion are highly likely to be indicative of bacteremia. Routine retention of platelet bags for subsequent microbiologic study was useful in the investigation of these febrile reactions. Empiric antibiotic therapy is indicated.
Monocyte chemoattractant protein production in red cell incompatibility
Authors: Davenport RD , Burdick M , Strieter RM , Kunkel SL
Published: March 2003

BACKGROUND: Mononuclear phagocytes play a central role in hemolytic transfusion reactions by erythrophagocytosis and production of inflammatory mediators. Factors that affect the number or function of monocyters would be expected to alter the clinical course of hemolytic transfusion reactions, and thus the production of monocyte chemoattractant protein‐1 (MCP‐1), a recently described chemotactic and activating cytokine specific for monocytes, was investigated in two distinct settings of red cell (RBC) incompatibility.

STUDY DESIGN AND METHODS: Fresh heparinized whole blood was incubated with ABO‐ compatible or ‐incompatible RBCs. Isolated peripheral blood mononuclear cells were incubated with anti‐D‐coated or uncoated RBCs. MCP‐1 was measured in the plasma or culture medium by enzyme‐linked immunosorbent assay. MCP‐1 gene expression was detected by Northern blot analysis of buffy coat or mononuclear cell total RNA. RESULTS: Significant levels of MCP‐1 protein in plasma or medium were detected 24 hours after the addition of incompatible RBCs, but not in the first 6 hours. Nonimmune hemolysis of added RBCs did not stimulate MCP‐1 production. The inactivation of complement by heat treatment of plasma prior to the addition of RBCs to whole blood did not prevent MCP‐1 production. Nor did neutralizing antibodies to tumor necrosis factor prevent MCP‐1 production in ABO incompatibility. MCP‐1 production was associated with increased steady‐state levels of white cell MCP‐1 mRNA, which occurred more rapidly in ABO than Rh incompatibility. CONCLUSION: The monocyte‐ specific chemotactic cytokine MCP‐1 is produced by peripheral blood leukocytes in response to RBC incompatibility. MCP‐1 may act in a positive feedback loop to recruit and activate monocytes during hemolytic transfusion reactions, thus contributing to the maintenance of these reactions.