bluuebunny/crossref_metadata_2025 huggingface dataset:
Red cell phenotyping using hexadimethrine bromide (Polybrene) in a microplate system
Authors: Anderson HJ , Patel S
Published: March 2003

To provide rapidly phenotyped units of blood, we adapted the hexadimethrine bromide (Polybrene) technique to microplate technology. Pilot samples from 282 donor units were phenotyped for antigens in the Rh, Kidd, Kell, Duffy, and Ss systems with a standard tube‐testing method and a Polybrene‐microplate (P‐MP) technique. Diluted antisera and a 1 percent red cell suspension were used to give P‐MP reactions that were accurate and easy to interpret. One microplate, containing 96 tests, was prepared and read within 15 minutes for P‐MP tests yielding direct agglutination (Rh, Jkb, Fya, Fyb), or within 19 minutes for P‐MP tests requiring an antiglobulin phase (Jka, K, S, s). No false‐positive results were found. No false‐negative reactions were found in typing for Rh, K, S, s, or Jka antigens. Phenotyping for Jkb and Duffy antigens gave a false‐negative rate of less than 0.015. Considerable savings in reagents were obtained through microplate miniaturization and through the enhancement of apparent antibody avidity in the Polybrene/low ionic medium which permitted dilution of reagent antisera. The P‐MP technique affords a rapid, accurate, simple, and inexpensive means of phenotyping large numbers of donor units.
Human erythrocytes shed exocytic vesicles in vivo
Authors: Dumaswala U. J. , Greenwalt T. J.
Published: March 2003

Studies were designed to demonstrate whether red cells (RBCs) lose membrane segments in vivo. Plasma separated from fresh blood collected in citrate‐phosphate‐dextrose‐adenine‐one was passed through 0.8 mu nitrocellulose filters to remove residual RBCs and formed elements. Exocytic vesicles were separated by centrifugation at 70,000 × g. Transmission electron microscopy showed vesicles 50 to 200 nm in diameter which contained hemoglobin. Sodium dodecyl sulfate‐ polyacrylamide gel electrophoresis of vesicle membranes showed that spectrin bands 1 and 2 and ankyrin band 2.1 were absent. These observations are the initial demonstration that RBCs shed exocytic vesicles in the normal circulation.
Unnecessary blood transfusions in elective colorectal cancer surgery
Authors: Tartter P. I. , Barron D. M.
Published: March 2003

In this 3‐year study, we analyzed the transfusion pattern of 288 patients undergoing potentially curative resection of colorectal cancer to determine the magnitude and causes of unnecessary blood transfusions. Transfusions were considered unnecessary if the preoperative hematocrit exceeded 36 percent or the discharge hematocrit exceeded 33 percent. Twenty‐five percent of the units of blood administered were unnecessary by these criteria. Excessive intraoperative transfusion (67 patients) and the practice of administering blood in pairs of units (33 patients) without reevaluating the hematocrit between successive units accounted for 90 percent of unnecessary transfusions. This study indicated that determination of the hematocrit immediately before administration of each unit of blood will reduce blood consumption of elective colon resections by 25 percent.
Limitations of the immediate spin crossmatch when used for detecting ABO incompatibility
Authors: Berry‐Dortch S. , Woodside C. H. , Boral L. I.
Published: March 2003

In the eleventh edition of the AABB's Standards for Blood Banks and Transfusion Services the requirement for an antiglobulin crossmatch was deleted if no clinically significant unexpected antibodies are detected in recipient serum testing and if there is no history of detection of such antibodies. Test methods that demonstrate ABO incompatibility remain a requirement; however, the means to accomplish this may prove controversial. The immediate spin crossmatch has been used for the purpose of detecting ABO incompatibility by many workers. Nonetheless, limitations of this technique became apparent in tests between A2B donor red cells and group B patient sera. The results of 204 of 531 immediate spin crossmatches, between these two blood groups, were found to be negative.
Effects of hydroxyethyl starch on fibrinogen, fibrin clot formation, and fibrinolysis
Authors: Strauss R. G. , Stump D. C. , Henriksen R. A. , Saunders R.
Published: March 2003

The effects of hydroxyethyl starch on the final stages of hemostasis were investigated in vivo and in vitro. When compared to control solutions of either 5 percent albumin or isotonic (0.9%) NaCl, 6 percent hydroxyethyl starch (HES) exerted several effects. Results of in vivo studies were as follows: following infusion of 1 liter of 6 percent HES into healthy subjects, fibrinogen and antithrombin‐III concentrations fell slightly due to plasma volume expansion and consequent dilution. Concentrations of fibrin monomer and fibrin‐ fibrinogen degradation products remained unchanged. Thrombin and reptilase clotting times were shortened to indicate rapid detection (and presumably accelerated formation) of fibrin clots. Urokinase‐ activated clot lysis times were shortened to suggest rapid fibrinolysis. Results of in vitro studies were similar. Shortened thrombin, reptilase, and urokinase‐activated clot lysis times were reproduced in vitro by mixing HES, but not albumin or NaCl, with normal plasma. Although these findings qualitatively are similar to those reported previously for dextran, the molecular mechanisms involved and the clinical importance, if any, of the hemostatic effects remain to be defined. Thus, it would be premature to conclude either that HES or dextran exert identical biological effects on hemostasis or that the two agents possess similar clinical properties. HES has an excellent safety record when it has been used during leukocytapheresis and for plasma volume expansion in recommended doses. Its effects when given in larger doses remain to be defined.
The effect of citrate, calcium, and magnesium ions on the potassium movement across the human platelet membrane
Authors: Weis‐Fogh U. S.
Published: March 2003

The intracellular K+ concentration in platelets is reduced during storage in citrated plasma. As shown previously, pH has a marked influence on this reduction. The effect of citrate and Ca2+ and Mg2+ on the K+ permeability of the membrane and on the active K+ transport across the membrane was investigated.

Platelets were incubated in dialyzed plasma at 37 degrees C, pH 5.5 to 7.9, with and without added Ca2+ and Mg2+ and citrate. After 60‐ to 120‐minute incubation, a steady state was reached, and ouabain was added to inhibit the active K+ transport. In the pH interval 5.5 to 6.2 citrate, Ca2+ and Mg2+ had no influence on either the active K+ transport or the K+ permeability. Between pH 7.1 and 7.9, Ca2+ and Mg2+ decreased the permeability, whereas citrate increased it. At pH 7.9, citrate had a depressing effect on the active transport. These results indicate that a low citrate concentration and a pH below 7 are important for maintaining a high intracellular K+ of platelets during storage in plasma.
“New” autoantibody specificity in autoimmune hemolytic anemia defined with red cells treated with 2‐aminoethylisothiouronium bromide and a dithiothreitol‐papain solution
Authors: Marsh W. L. , DiNapoli J. , ØYen R. , Pacanowski L. , Kessler L. A.
Published: March 2003

Specificity of IgG autoantibodies that react with random red cell samples but not with cells treated with 2‐aminoethylisothiouronium bromide (AET)‐previously was related to the Kell blood group system. We now describe an IgG autoantibody, from a patient who died with autoimmune hemolytic anemia, that reacted with random red cell samples, Ko cells, and with cells treated with a dithiothreitol‐papain solution, but did not react with AET‐treated cells or with Rhnull cells. Serological reactivity of the autoantibody is unlike that of IgG autoantibodies that do not react only with Rhnull cells. It appears that the reactive antigen is not part of the Kell or Rh systems, but is probably modified by the membrane anomaly present in Rhnull red cells.
Autoimmune hemolytic anemia with concurrence of warm and cold red cell autoantibodies and a warm hemolysin
Authors: Freedman J. , Lim F. C. , Musclow E. , Fernandes B. , Rother I.
Published: March 2003

This report describes the laboratory findings and clinical course of a patient with thrombophlebitis, venous gangrene, and autoimmune hemolytic anemia. Three concomitant red cell autoantibody activities were detected: a low‐titer, high‐thermal‐amplitude, IgM anti‐I cold agglutinin; an IgG warm ‘incomplete’ panagglutinating autoantibody; and an IgM warm hemolysin.
Anti‐Tcb, an antibody that defines a red cell antigen antithetical to Tca
Authors: Lacey P. A. , Block U. T. , Laird‐Fryer B. J. , Moulds J. J. , Bryant L. R. , Giandelone J. A. , Linnemeyer D. R.
Published: March 2003

A patient's serum, used as a source of anti‐Goa typing serum, was found to contain a second antibody that detected a ‘new” red cell antigen present in approximately 5 percent of blacks, but not yet found in Caucasians. Among blacks, the ‘new’ antigen shows an antithetical relationship to the high frequency antigen Tca, as evidenced by tests on eight unrelated Tc(a‐) black women and the families of two of these propositi. The antigen, designated Tcb, has not been found on the red cells of Tc(a‐) persons whose racial origins differ from the propositi of this report, which suggests that more than one genetic mechanism can produce the Tc(a‐) phenotype. The presence of both anti‐Goa and anti‐ Tcb in the same serum appears to be a coincidental and unrelated finding.
Estimation of transfused red cell survival using an enzyme‐linked antiglobulin test
Authors: Kickler T. S. , Smith B. , Bell W. , Drew H. , Baldwin M. , Ness P. M.
Published: March 2003

An enzyme‐linked antiglobulin test (ELAT) method was developed to estimate survival of transfused red cells. This procedure is based on a principle analogous to that of the Ashby technique were antigenically distinct red cells are transfused and their survival studied. We compared the ELAT survival to the 51Chromium method (51Cr) in four patients. Three patients with hypoproliferative anemias showed T 1/2 by ELAT of 17.5, 18, and 17 days versus 18.5, 20, and 19 days by the 51Cr method. A fourth patient with traumatic cardiac hemolysis had two studies performed. In this case, the ELAT showed a T 1/2 of 10 and 8.1 days while 51Cr T 1/2 values were 11 and 10.5 days. The ELAT method for measuring red cell survival yielded data which agreed closely with the results of the 51Cr method. Although 51Cr is the accepted method for red cell survival, the ELAT method can be used to estimate transfused red cell survival.
Cold reacting antibodies recognizing antigens dependent on N− acetylneuraminic acid
Authors: Staub C. A.
Published: March 2003

Serum from a 75‐year‐old man with lymphoproliferative disease was found to contain a persistent cold agglutinin meeting most of the characteristics of examples of anti‐Gd reported previously. It differed, however, in that it contained IgG and IgM components and its reactivity was not inhibited by sialyllactose. A search of random serums containing cold agglutinins was undertaken to determine the incidence of antibodies to antigens dependent on N‐acetylneuraminic acid. Two further examples were found in 47 serums containing cold agglutinins. One of these also demonstrated a Gd or “Gd‐like” specificity. The other appeared to be an anti‐Pr.
Platelet factor 4 release from the platelets stored in platelet concentrates
Authors: Shimizu T. , Ishikawa Y. , Morishima Y. , Fukuda T. , Kato K.
Published: March 2003

Many consider that granular components of platelets should remain intact during storage of platelet concentrates (PCs) in order to participate in hemostasis when infused. The present investigations were done to determine factors affecting the release of an alpha‐granular component, platelet factor 4 (PF‐4), from platelets stored as PCs. Sedimentation of the pelleted platelet button and stronger flat‐bed agitation induced a significant PF‐4 release. After 1 day of storage at 22 degrees C, the percent PF‐4 release correlated roughly with a pH fall in PCs (r = 0.520, y = 0.014 + 7.204). Rates of pH fall were roughly proportional to the platelet count. In addition, there was a positive relationship between PF‐4 release and the platelet count per unit volume (r = 0.615, y = 0.045 + 1.327). The results indicated that one of the critical factors which determines the amount of PF‐4 release during storage is concentrated pH. Lowering the platelet count per unit volume in PCs is better for the maintenance of granular components in platelets during storage because of the intact platelet metabolism.
Analysis of saline‐washed red cells for transfusion to neonatal patients
Authors: Sanner H. E. , Wooten M. J.
Published: March 2003

A protocol was developed under which washed red cells were used to provide small‐volume transfusions for neonatal patients (under 4 months of age). Two pediatric units, each containing approximately 135 ml of red cells, were prepared from a freshly collected unit of group O Rh‐ negative whole blood. Units not more than 6 days old were washed with 1l of 0.9 percent NaCl in a blood cell processor. Aliquots of the washed red cells were drawn into labeled syringes for transfusion to neonatal patients. Data collected from 40 units of washed red cells prepared according to this protocol showed 83 percent red cell recovery, and satisfactory reduction in white cells, plasma proteins, extracellular potassium, red cell metabolic waste products, and anticoagulant. Over 5 years, a total of 2085 different neonatal patients received 7875 separate transfusions prepared from 1860 units of washed red cells. The clinical response was excellent, and no suspected reactions were reported.
The effect of In(Lu) on some high‐frequency antigens
Authors: Daniels G. L. , Shaw M. A. , Lomas C. G. , Leak M. R. , Tippett P.
Published: March 2003

Antibodies to various high‐frequency antigens were used to test red cells of members of 12 families of Lu(a‐b‐) propositi carrying the In(Lu) gene. Kna, Kna‐like, McCa, and Sia usually are expressed more weakly on cells of the Lu(a‐b‐) members than on those of their relatives who were not Lu(a‐b‐). In most families, In(Lu) also suppresses Yka and Csa expression. Some antiserums are more efficient than others of the same specificity in demonstrating the weakened expression. The expression of Ch and Rg antigens on red cells is not affected by In(Lu).
Isolation of a Kell‐reactive protein from red cell membranes
Authors: Wallas C. , Simon R. , Sharpe M. A. , Byler C.
Published: March 2003

A red cell membrane protein which exhibits Kell blood group antigen activity has been identified with a purified anti‐Kell bound to a Protein‐A agarose column and eluting with lithium diiodosalicylate (LIS). Although anti‐Kell as well as the Kell‐reactive membrane protein were eluted from the column, the eluate was capable of reducing the titer of added anti‐Kell from 64 to 4. In addition, the eluate was shown to possess Kell reactivity by binding I125 Protein A after incubation with anti‐Kell. Electrophoresis (SDS gel polyacrylamide 5–20% gradient) showed a band at approximately 90,000 daltons when solubilized membranes from Kell‐positive red cells were used but not when membranes from dithiothreitol‐ and papain‐treated Kell‐positive red cells or Kell‐negative red cells were used. A band isolated with unreduced conditions was capable of neutralizing anti‐Kell.
A serious source of error in antiglobulin testing
Authors: Bruce M. , Watt A. H. , Hare W. , Blue A. , Mitchell R.
Published: March 2003

The investigation of a failure of proficiency showed that certain saline solutions are inappropriate for use in blood group serology tests. In particular, it was found that solutions of unexpectedly low pH and/or those autoclaved and stored in plastic containers could severely compromise the sensitivity of the antiglobulin test when used as wash solutions. The observed loss of sensitivity ranged from a reduction in titration score to a complete failure in the detection of clinically significant blood group antibodies. It is suggested that careful consideration should be given to the source, pH, and storage container of saline solutions intended for use in serological tests and that improved standardization and sensitivity could be achieved by using phosphate‐buffered saline pH 7.0 to 7.2 for all such purposes. It is recommended that unbuffered saline solutions of pH less than 6.0 should not be used for serological testing.
The incidence of development of irregular red cell antibodies in patients with sickle cell anemia
Authors: Sarnaik S. , Schornack J. , Lusher J. M.
Published: March 2003

Two hundred and forty‐five patients with SS hemoglobinopathy were screened for alloimmunization; they had all been transfused previously. Their median age was 10 years. Nineteen patients (7.75%) were found to be sensitized to various red cell antigens. The median number of transfusions received by sensitized patients was 23 compared to three in the unsensitized group, indicating an increased risk with more transfusion exposures. Thirteen sensitized patients were re‐exposed to transfusion and four (30%) developed new alloantibodies. Once immunized, the risk of developing more alloantibodies on re‐exposure did not increase with increasing number of exposures. The most frequently observed alloantibody was anti‐K, accounting for 38 percent of all instances. Anti‐Lea and ‐Leb were next in frequency (24%); Rh antibodies were seen in 14 percent of cases. Children with sickle cell anemia who were multiply transfused had a low frequency of alloimmunization. Responders had an increased rate of further sensitization which did not increase with number of subsequent exposures. Phenotypically matched blood is probably not warranted in most patients receiving transfusions for sickle cell anemia.
Du confirmation
Authors: Schmidt P. J. , Klein R. E. , Sherwood W. C.
Published: March 2003

The need for repeat antiglobulin testing to detect the Du phenotype in donor blood already labeled as Rh‐negative was challenged. The confirmatory testing by hospital transfusion services rejected only 47 units in 2.6 million tested. The confirmatory antiglobulin test for Du currently required of hospital transfusion services could be omitted.
Bacterial proliferation in platelet concentrates
Authors: HEAL J.M. , SINGAL S. , SARDISCO E. , MAYER T.
Published: March 2003

ABSTRACT: Growth curves of bacteria in platelet concentrates were studied to determine whether increasing the shelf‐life of platelets from 3 to 7 days might have contributed to the increased number of deaths caused by contaminated platelets that have been reported to the FDA since 1980. Platelets inoculated with 103 organisms or more showed logarithmic bacterial growth throughout the 7 days of storage or until the platelets became visibly abnormal. With an inoculum of 102 organisms or less, proliferation patterns were variable: 20 percent (%) showed uninhibited logarithmic growth and 50 percent (%) remained sterile throughout the 7 days. In another 30 percent (%), bacterial growth was temporarily suppressed for 5 to 6 days before bacteria actively proliferated. These data support the hypothesis that bacterial contamination of platelets not clinically significant at 3 days of storage might become so during the current 7 day shelf life.
Serologic characteristics of a further example of anti‐Xga and the frequency of Xga in North London blood donors
Authors: Devenish A. , Burslem M. F. , Morris R. , Contreras M.
Published: March 2003

An example of an IgG anti‐Xga of subclasses IgG1 and IgG2 that fixed complement was detected in a 92‐year‐old man. The serum was used to test 4000 routine British blood donors. The incidence of Xga was 0.677 in men and 0.895 in women.
Anti‐Rh33, the second separable example, also made by a person who made anti‐D and has C+ red cells
Authors: Issitt P. D. , Wren M. R. , McDowell M. A. , Strohm P. L. , Roberts T. M.
Published: March 2003

A complex serologic investigation resulted in identification of the second example of separable anti‐Rh33. A patient who had been transfused frequently and who had red cells that lacked a portion or portions of the D mosaic of antigens made anti‐D against that portion of D missing from her red cells, anti‐c, anti‐V, anti‐Rh33, anti‐K, and an autoantibody that mimicked the reactions of anti‐D. Differential adsorption experiments showed that the anti‐Rh33 was separable from the other antibodies present. The serologic findings in this case and in the study in which anti‐Rh33 was first found show some sort of relationship between D and Rh33. First, one of the genes, RoHar, that makes Rh33 makes a form of D that is difficult to detect. Second, the only other gene thus far known to make Rh33, DIV (C)—, encodes for a form of D from which portions of the D mosaic are missing. Third, both examples of separable anti‐Rh33 were made by women who made anti‐D and whose red cells were C+.
Frequency of PLA1 in blacks
Authors: Ramsey G. , Salamon D. J.
Published: March 2003

Racial frequencies of platelet‐specific antigens in racial groups other than whites have not been studied. The prevalence of PLA1 was determined in white and black blood donors. Two hundred forty‐two of 243 blacks were PLA1‐positive (99.6%), compared with 242 of 250 whites (96.8%) (p = 0.021). This black population has the highest frequency of PLA1 yet reported.
Cryopreservation of hematopoietic progenitor cells collected by hemapheresis
Authors: Heal J.M. , Brightman A.
Published: March 2003

Colony‐forming units‐granulocyte/monocyte (CFU‐GM) harvested from normal donors as a byproduct of plateletapheresis can be cryopreserved successfully with 10 percent dimethyl sulfoxide with the use of standard protocols that have been developed for freezing bone marrow. Short‐term storage of CFU‐GM in polypropylene vials in the liquid phase of liquid nitrogen yielded a recovery rate of 88 ± 5 percent (mean ± SEM). Significant loss of committed progenitor cells was not detected until 1 year after freezing (65 ± 5%). A comparison of CFU‐GM recovery with the polyolefin bag technique (107 ± 13%) was not statistically different from that obtained with polypropylene vials (78 ± 7%). Although the freezing bags are more expensive and prone to fracture than the vials, they are easier to handle, store, and thaw in the laboratory, and reinfusion to the patient is more convenient. Cryopreservation of CFU‐GM harvested from peripheral blood appears feasible in either of two freezing systems.
B antigen acquired by normal A1 red cells exposed to a patient's serum
Authors: Stayboldt C. , Rearden A. , Lane T.A.
Published: March 2003

The authors report a patient with the acquired B antigen whose serum contained a factor capable of inducing the acquired B phenotype on normal A1 red cells in vitro. The patient's red cells were not, however, agglutinated by a monoclonal anti‐B. These findings and the abolition of the acquired B antigen by treatment of the patient's red cells with acetic anhydride support the deacetylation theory of the genesis of the acquired B antigen.
Evidence that the Lub blood group antigen is located on red cell membrane glycoproteins of 85 and 78 kd
Authors: Parsons S.F. , Mallinson G. , Judson P.A. , Anstee D.J. , Tanner M.J. , Daniels G.L.
Published: March 2003

A murine monoclonal antibody of specificity anti‐Lub was produced. Immunoblotting of the electrophoretically separated components of membranes from Lu(b+) red cells with the monoclonal antibody identified two glycoproteins of relative molecular mass 85 and 78 kd, respectively. The expression of Lub antigenic activity on these glycoprotein components was shown to be dependent on the presence of one or more N‐glycosidically linked oligosaccharides and on the presence of disulphide bonding.
Low ionic antiglobulin tests
Authors: Ahn J. H. , Kochwa S. , Rosenfield R. E.
Published: March 2003

Seven serologic procedures were studied to determine their respective value in compatibility and screening tests. All seven were significantly improved by the use of 4 volumes of serum, rather than 1, with 1 volume of red cell suspension, and a low‐ionic antiglobin test (LIAGT) was distinctly superior to the other six procedures evaluated. In this test, during the incubation of serum and cells at 37 degrees for 20 minutes, ionic concentration was reduced 62 percent. However, after removal of all supernatant, the red cells were washed three times with an isotonic solution that provided 80 percent reduction in ionic concentration, and the washed cells were tested for their agglutinability with low‐ionic (80% ionic reduction) anti‐IgG antiglobin reagent. This modified LIAGT was usually more, and apparently never less, sensitive than a test described earlier and is expected to be associated with much less nonspecificity. The extreme sensitivity of LIAGT for many long‐term frozen stored alloantiserums is a retained property of the modified test and has been associated with IgG aggregation during storage.
Comparison of a commercial hexadimethrine bromide method and low‐ionic‐ strength solution for antibody detection with special reference to anti‐ K
Authors: Letendre P. L. , Williams M. A. , Ferguson D. J.
Published: March 2003

The sensitivities of manual low‐ionic hexadimethrine bromide (Polybrene, LIP) and low‐ionic Polybrene indirect antiglobulin tests (LIPAT) were compared with those of a manual low‐ionic‐strength indirect antiglobulin test (LISS) by using a commercial Polybrene kit. One hundred antibodies were coded, titrated, and tested in parallel. LIP did not detect 36 antibodies: 31 anti‐K, two anti‐E, two anti‐Fya, and one anti‐Jka. LIPAT did not detect seven anti‐K, two anti‐E, and two anti‐Jka. The combination of LIP and LIPAT did not detect two anti‐ E that were reactive only in a two‐stage enzyme test and seven anti‐K that had titers of 2 or lower by LISS. LISS detected all antibodies except for the two enzyme‐reactive anti‐E. There were no significant differences in the titers of 63 percent of the antibodies studied. For 54 percent of the antibodies in the Kell system, LISS produced significantly higher titers; for 25 percent of antibodies in the Rh system, LIP did so. The poor sensitivity of the Polybrene kit for anti‐ K makes it unsuitable as a primary method for antibody screening.
Exercise and circulating hematopoietic progenitor cells (CFU‐GM) in humans
Authors: Brightman A. , Heal J. M.
Published: March 2003

In order to determine whether the rise in blood circulating committed hematopoeitic progenitor cells (CFU‐GM) was sustained long enough after exercise to be a practical adjunct to increasing stem cell yields by hemapheresis, the authors evaluated two exercise protocols in 15 normal donors. Immediately after a brief period of intense exercise, there was a significant increase in the absolute number of blood CFU‐GM from 164 +/− 27 to 240 +/− 46 per ml. However, all values returned to baseline within 15 minutes. Prolonging the exercise did not enhance either the circulating CFU‐GM concentration or the duration of the rise. If blood hematopoietic stem cells respond to exercise similarly, such a transient increase would not appear to be of use in stem cell harvesting.
Oxidative product formation in irradiated neutrophils
Authors: Wolber R. A. , Duque R. E. , Robinson J. Paul , Oberman H. A.
Published: March 2003

The effect of irradiation on neutrophil oxidative function was evaluated using a flow cytometric assay of intracellular hydrogen peroxide (H2O2) production. This assay quantitates the H2O2‐dependent conversion of the nonfluorescent compound, 2′‐7′‐dichlorofluorescein (DCFH), into fluorescent 2′‐7′‐dichlorofluorescein (DCF) on a single‐ cell basis. Intracellular H2O2 production in response to stimulation with phorbol myristate acetate was not affected by neutrophil irradiation at doses up to 2500 rad. In addition, irradiation of intracellular DCFH and aqueous 2′‐7′‐dichlorofluorescein diacetate (DCFH‐DA) resulted in DCF production, which suggested that oxidative molecules produced by aqueous radiolysis were detected by this assay. This study indicates that radiation doses of 1500 to 2500 rad, which are sufficient to prevent induction of graft‐versus‐host disease by transfused blood components, are not deleterious to neutrophil oxidative metabolism.
Chromatographic removal of hepatitis B virus from a factor IX concentrate
Authors: Einarsson M. , Iwarson S. , Smallwood L. , Snoy P. , Gerety R. J.
Published: March 2003

Non‐A, non‐B hepatitis virus can be removed from a factor IX concentrate by a hydrophobic chromatographic step added to the ordinary fractionation process. The efficacy of this procedure for removal of hepatitis B virus (HBV) was evaluated in chimpanzees. A well‐defined hepatitis B virus (HBV) inoculum was added to a factor IX preparation and this preparation was subjected to chromatography with octanohydrazide‐Sepharose 4B at a high salt concentration and then injected intravenously into two chimpanzees. A control chimpanzee was inoculated with the part of the factor IX/HBV preparation that had not been chromatographed. The two chimpanzees that received the treated material remained free of any serologic or biochemical evidence of hepatitis B infection during a 12‐month follow‐up, whereas the control chimpanzee had hepatitis B. After a later HBV challenge, the two healthy animals also had hepatitis B. The hydrophobic binding procedure seems to be useful for the adsorption of viral agents in blood components.
A heat‐treated factor VIII concentrate prepared by controlled‐pore glass adsorption chromatography
Authors: Benny A. G. , Scott R. H. , Woodfield D. G.
Published: March 2003

Four years' experience with a method for preparing a high‐purity, low‐ fibrinogen, heat‐treated factor VIII concentrate is reported. The process, batch adsorption of a cryoprecipitate extract with controlled‐ pore glass granules, removes 77 percent of the cryoprecipitate fibrinogen, resulting in a final concentrate‐specific activity of 0.74 IU factor VIII per mg of protein at a yield of 194 IU factor VIII per kg of starting plasma. Heat treatment of the lyophilized concentrate for 72 hours at 60 degrees C results in less than 10 percent loss of factor VIII activity. This process does not require expensive fractionation equipment, is suitable for small‐to medium‐scale batch concentrate production and could be adopted by moderately well‐equipped regional blood processing laboratories for the decentralized production of a high‐quality, heat‐treated factor VIII concentrate.
Temporary deferral and donor return
Authors: Piliavin J. A.
Published: March 2003

Deferral from donation is usually for conditions that make the donor only temporarily unable to give. Based on questionnaires given to 1247 donors with subsequent follow‐up of their donation records for 6 months, permanent psychological effects of deferral are suggested. Only one of 36 deferred first‐time donors (2.8%) compared with 27.3 percent of those who were not deferred returned for a second donation during the next 6 months. Among most first‐time donors, temporary deferral may be interpreted psychologically as providing a permanent excuse for not donating.
Th activation in congenital hypoplastic anemia
Authors: Herman J. H. , Shirey R. S. , Smith B. , Kickler T. S. , Ness P. M.
Published: March 2003

The authors identified persistent Th activation in five of seven children (71.4%) diagnosed as having Fanconi's anemia or Diamond‐ Blackfan syndrome. Th reactivity was no longer present in one patient after bone marrow transplantation. Tests on family members and other patients with bone marrow dysfunctions of childhood showed no Th activation. Less than 1 percent of healthy children or blood donors had Th activation. Patients with a variety of hemolytic and hypoplastic conditions also had a low incidence of Th activation. However, 13.5 percent of cord blood specimens demonstrated Th reactivity. This study indicates that Th activation may be a red cell developmental marker present in congenital hypoplastic anemias and also expressed on newborn red cells.
Predicting the clinical significance of red cell alloantibodies using a monocyte monolayer assay
Authors: Nance S.J. , Arndt P. , Garratty G.
Published: March 2003

Few data have been published that correlate in vitro monocyte monolayer assays (MMA) and red cell (RBC) survival in patients with alloantibodies of unknown significance. Over the past 6 years we gathered clinical correlations in 12 patients with the following antibodies: anti‐Lan (three patients), ‐Ge (three patients), ‐Yta (five patients), and ‐Ytb (one patient). RBC survival was estimated using 51Cr studies in seven patients and follow‐up of transfusion of incompatible blood in the other five. Six patients with no evidence of RBC destruction had negative MMA findings (anti‐Lan [one patient], ‐Ge [two patients], and ‐Yta [three patients]). Five patients with evidence of in vivo RBC destruction had significant MMA results. The two clinically significant anti‐Lans required fresh serum to give a meaningful MMA result. One patient (anti‐Ytb) had an MMA result of borderline significance‐normal 51Cr RBC survival at 1 hour‐but a reduced T50Cr. The MMA we used appeared to predict the clinical outcome of transfusion in every patient with antibodies to high‐frequency antigens whom we tested.
The Yt blood groups in Israeli Jews, Arabs, and Druse
Authors: Levene C. , Bar‐Shany S. , Manny N. , Moulds J.J. , Cohen T.
Published: March 2003

The Yta and Ytb allelic frequencies were determined by examining the red cells of 1683 blood samples from Israeli Jews, Arabs, and Druse with anti‐Yta and ‐Ytb. The Ytb allelic frequencies ranged between 0.1005 and 0.1522 in the Jewish communities and were 0.1294 and 0.1429 in the Arab and Druse communities, respectively. These are the highest Ytb allelic frequencies observed so far in any population tested, so the Yt blood group system can be used as a genetic marker in these populations. No factors were recognized that may have influenced the selection for the Ytb allele.
Selective elution of HLA antigens and β2‐microglobulin from human platelets by chloroquine diphosphate
Authors: Kao KJ.
Published: March 2003

To determine whether chloroquine can specifically elute HLA antigens and β2‐microglobulin (β2‐M) from the platelet surface, quantitative immunofluorescence flow cytometry and monoclonal antibodies were used to show that HLA antigens and β2‐M were proportionally eluted from the platelet surface without affecting the membrane glycoproteins IIb and IIIa. Second, an autoradiogram of electrophoresed I125‐labeled platelets showed that only β2‐M but not other I125‐labeled membrane proteins could be eluted. Although HLA antigens were poorly labeled by I125 and could not be detected on the autoradiogram, the eluted HLA antigens could be detected by anti‐HLA monoclonal antibody and immunoblotting techniques. No loss of plasma membrane integrity was observed by transmission electron microscopy after chloroquine treatment of platelets. The results indicate that chloroquine selectively elutes HLA antigens and their noncovalently associated β2‐M without affecting other integral platelet membrane proteins.
Hybrid sialoglycoprotein content of St(a+) red cells
Authors: Rearden A.
Published: March 2003

St(a+) red cells contain a ficin‐resistant hybrid sialoglycoprotein (SGP) consisting of the amino terminus of Ss sialoglycoprotein (Ss SGP) and the carboxyl terminus of MN sialoglycoprotein (MN SGP). Ficin‐ modified St(a+) but not St(a−) red cells were agglutinated by monoclonal antibodies NN5 (anti‐N) and 31 (detecting a sialic acid‐ dependent determinant on the SGPs). SDS‐polyacrylamide gel electrophoresis showed three periodic acid‐Schiff‐staining protein bands in ficin‐modified St(a+) but not St(a−) red cell‐membranes; these bands bound monoclonal antibodies NN5 and 31 on Western blots. Using Scatchard analysis of 125I‐NN5 IgG binding to M+N− red cells, estimates were obtained of 25,000 Ss SGP molecules per red cell per s allele, 33,000 Ss SGP molecules per red cell per S allele, and 75,000 hybrid SGP molecules per red cell per hybrid allele. These results are consistent with the expression of the hybrid SGP gene at a level intermediate between the MN SGP gene and the Ss SGP gene. Monoclonal antibodies NN5 and 31 may be useful in screening ficin‐treated red cells for hybrid SGP variants.
Bulk cryopreservation of lymphocytes in glycerol
Authors: Areman E.M. , Simonis T.B. , Carter C.S. , Read E.J. , Klein H.G.
Published: March 2003

The authors describe a method for freezing large amounts of peripheral blood lymphocytes (PBL) in a 20 percent glycerol solution. Between 0.6 and 4.3 × 10(9) cells in autologous plasma were frozen in polyethylene freezing bags in a final volume of 50 ml. The recovery after thawing averaged 89 +/− 14 percent with a mean viability by trypan blue dye exclusion of 80 +/− 7 percent (n = 11). In aliquots of fresh and frozen‐ thawed PBL from the same subjects radiolabeled with 111In, the radiolabeling efficiency for both fresh and thawed cells was 49 +/− 15 percent (p = 0.98, n = 5). The mitogen mean stimulation indices for glycerol‐frozen cells (471 with phytohemagglutinin‐M, 176 with pokeweed mitogen, and 380 with concanavalin A) were superior to those of cells frozen by a standard technique with dimethylsulfoxide (DMSO) (141, 47, and 123; p less than 0.05) and comparable to those of fresh PBL (403, 75, and 147). In a mixed lymphocyte culture, glycerol‐frozen PBL showed significantly greater responsiveness to a pool of stimulator cells than did PBL frozen in DMSO (p = 0.03). Thawed cells are viable and functional as demonstrated by their response to mitogens and their ability to stimulate and respond in mixed lymphocyte culture.
Effect of citrate anticoagulants on factor VIII levels in plasma
Authors: Rock G. , Tittley P. , Fuller V.
Published: March 2003

The citrate anticoagulants used during blood collection have been developed for their benefits to red cells. The concentrations in which they are used are strictly regulated in the United States: citrate‐ phosphate‐dextrose‐adenine (CPDA) is used in a 1:8 ratio for the collection of whole blood, whereas 4 percent sodium citrate (NaCit) is used in a 1:10 ratio for manual plasmapheresis. Acid‐citrate‐dextrose formula A (ACD‐A) or formula B (ACD‐B) and NaCit are commonly used in a 1:12 or 1:15 ratio during automated plasmapheresis. These anticoagulants have different initial and final pH values and citrate concentrations and different effects on the recovery of factor VIII (FVIII) in the plasma. NaCit has a higher initial pH (6.64) than ACD‐A (4.98), ACD‐B (5.60), or CPDA (5.12). The effects of these different anticoagulants on plasma constituents obtained from six healthy subjects were studied. In standard citrate concentrations, the FVIII level was significantly lower (p < 0.05) in the NaCit used for manual plasmapheresis than in either of the ACD solutions used in automated plasmapheresis (104 U/dl vs. 153 and 160 U/dl). When various ratios of NaCit to blood were used, the pH increased from 7.62 at a 1:10 dilution to 7.65 at a 1:50 dilution. As expected, a progressive decrease in anticoagulant level was associated with an increase in ionized calcium and also in the level of FVIII, with the latter values rising from 104 U per dl at 1:10 to 137 at 1:20 and 148 U per dl at 1:30. Clot formation was detected only at a ratio of 1:35. The results indicate that maximal FVIII recovery occurred at anticoagulant concentrations far below those used in current practice in North America. Reduction of anticoagulant‐to‐blood ratios to half of the currently accepted values for ACD‐A, CPDA, and NaCit would increase the amount of FVIII available for fractionation.
Reduction in the toxicity of a component of an artificial blood substitute by supercritical fluid fractionation
Authors: Lane T.A. , Krukonis V.
Published: March 2003

Artificial blood substitutes (ABS) containing the surfactant Pluronic F68 (F68) are reported to be cytotoxic to cell lines. Because of F68's reported impairment of granulocyte function, it was hypothesized that F68 was also responsible for the cytotoxicity of ABS, possibly as a result of a separable fraction of the F68, separation of which could be achieved using supercritical fluid fractionation (SFF). SFF employs gases, such as carbon dioxide, under high pressure to dissolve the parent compound, a process that is followed by step‐by‐step precipitation and recovery of the dissolved material. The toxicity of F68 to human and animal cells was investigated by culturing Hela and B16 cells in the presence or absence of F68. Cells were grown for 4 days, harvested, and counted. Hela and B16 growth were markedly inhibited by F68. Four lots of F68 at 20 mg per ml inhibited Hela growth by 54 ± 10 percent (p < 0.05). In four dose‐response experiments, three lots of F68 caused 50 percent inhibition of cell growth at 25 ± 17 mg per ml. Four lots of F68 were also processed by SFF. Hela cells were grown in the presence of control media, parent (unextracted) F68, the early fractions of SFF extraction (XT), and the residual (partially purified) (RES F68). All parent F68, XT, and RES F68 were at 20 mg per ml. In 11 studies, the XT were 77 ± 25 percent more toxic than the parent F68 (p < 0.02), and the RES F68 were 30 ± 11 percent less toxic than parent F68 (p < 0.02). SFF of F68 also significantly diminished or eliminated F68‐induced inhibition of granulocyte chemotaxis. It is concluded that the cytotoxicity of ABS, as previously reported, is due to F68; that F68 exhibits marked, dose‐dependent toxicity to growth of human and animal cells; that SFF separated a highly toxic fraction of parent F68; and that removal of this fraction significantly diminished the cytotoxicity and chemotaxis‐inhibiting properties of F68. Thus, it may be possible to generate a nontoxic F68 polymer using SFF or a similar technology.
Risk of HIV infection in recipients of untested blood from donors now anti‐HIV‐positive
Authors: Grindon A. J. , Critchley S. E. , Ward J. W.
Published: March 2003

Recipients of untested blood from donors who at a subsequent donation were positive for HIV antibody by enzyme immunoassay (EIA) were evaluated, whether the result on Western blot (WB) assay was negative (EIA+/WB‐) or positive (EIA+/WB+). For 109 EIA+/WB‐ donors, 78 recipients were tested for HIV antibody, and 3 (4%) were positive. Two of the three anti‐HIV‐positive recipients had clotting disorders, and the other had been massively transfused; in each of these three cases, subsequent test data exonerated the EIA+/WB− donor. For 101 current EIA+/WB+ donors, 35 recipients were tested for HIV antibody, and 13 (37%) were positive. For donors subsequently found to be EIA+/WB+, the rate of isolation of HIV was the same whether the recipients were anti‐ HIV‐positive or anti‐HIV‐negative (each, 5/6). While recipients of blood from donors subsequently found to be EIA+/WB+ were at substantial risk for HIV infection, regardless of the donor's subsequent HIV culture result, risk of HIV infection was not demonstrated for recipients of blood from donors later found to be EIA+/WB‐.
A family showing inheritance of the Inab phenotype
Authors: Lin R. C. , Herman J. , Henry L. , Daniels G. L.
Published: March 2003

An 86‐year‐old white American woman was found to have a very rare red cell phenotype, the Inab phenotype. Her cells were Cr(a−), Tc(a−b−c−), Dr(a−), Es(a−), WES(a−b−), and IFC‐. Unlike the two other Inab phenotype patients, she has no history of protein‐losing enteropathy or any similar intestinal disorder. One of her four siblings also has the Inab phenotype, providing the first evidence that this phenotype may be inherited. Serum inhibitions showed that anit‐Cra, ‐TCa, ‐Dra, ‐WESb, and ‐IFC are partially inhibited by sera from three siblings with the common red cell phenotype but not by serum from the Inab phenotype sibling. The serum of the proposita contained an antibody to a high‐ frequency antigen that showed characteristics of being a Cromer‐related antibody.
Antibody to c antigen consequent to renal transplantation
Authors: Hjelle B. , Donegan E. , Cruz J. , Stites D.
Published: March 2003

A case of apparent red cell (RBC) autoimmunity due to anti‐c is described in the recipient of a cadaveric renal transplant. Although the patient suffered little hemolysis as a result, a strong direct anti‐ c agglutinin was detectable on the patient's RBCs 8 days after transplantation and in his serum only 4 days after transplantation. We believe that the antibody was produced by passively transferred B lymphocytes in the donor kidney, and that the recipient may have been protected from more serious hemolysis by immunosuppressive agents administered at the time of transplantation. The donor received multiple transfusions before transplantation, but his serum lacked anti‐ c in a screen performed shortly before his death. This case raises the possibility that donor alloantibodies to other Rh antigens on recipient RBCs can arise after transplantation, if the donor has been transfused.
Platelet transfusion refractoriness associated with two rare platelet‐ specific alloantibodies (anti‐Baka and anti‐PlA2) and multiple HLA antibodies
Authors: Langenscheidt F. , Kiefel V. , Santoso S. , Mueller‐Eckhardt C.
Published: March 2003

A 65‐year‐old patient with pancytopenia resulting from osteomyelosclerosis became refractory to platelet transfusions during long‐time transfusion support. He developed two rare, platelet‐specific antibodies (anti‐PlA2 and ‐Baka) disguised by strong, multispecific HLA antibodies. The specificity of the platelet‐specific antibodies was detected by a newly designed enzyme‐linked immunosorbent assay using glycoprotein‐specific monoclonal antibodies for immobilization of platelet antigens.
Monitoring of platelet morphology during storage of platelet concentrates
Authors: Fijnheer R. , Pietersz R. N. I. , Korte D. de , Roos D.
Published: March 2003

During storage, human platelet concentrates progressively lose the capacity to survive and function in vivo after transfusion. A shape transformation from disc to sphere is the most reliable in vitro determinant for the loss of the in vivo survival of platelets. To find an objective measurement for platelet morphology, we studied the effect of anticoagulant, temperature, and storage on the apparent median platelet volume (MPV) as determined by a particle counter and on changes in platelet shape as measured by light transmission at different stirring rates in an aggregometer, by swirling patterns, and by light microscopy. Changes in MPV, light transmission, and morphology score by light microscopy were observed within 1 minute after collection of blood in CPDA. As compared to blood immediately fixed on withdrawal, in CPDA blood, the MPV increased from 4.1 to 5.7 fl, and light transmission difference decreased from 22 to 7 percent. A partial restoration of these determinants was found when the whole blood was incubated for 30 minutes at 37°C, before preparation of platelet‐rich plasma. In the first 5 days of platelet storage, the MPV increased from 4.6 to 5.0 fl; thereafter, it started to decrease. An increase in fragmented platelets after 5 days was observed on light microscopy. The light transmission difference showed a slow disc‐to‐sphere transformation during storage. This transformation accelerated from Day 5 to Day 7; after 11 days, only spheres were detected. After 7 days the swirting pattern scores were still in accordance with the presence of discs, whereas the other structure‐associated determinants showed already spheric and even fragmented platelets. In the first 5 days, short incubation at 37°C largely restored the disc shape of the platelets, but this effect diminished after prolonged storage. Light transmission at different stirring rates is the most reliable and simplest assay for measuring platelet shape changes during storage. However, because of its noninvasive nature, the measurement by swirling patterns is useful in blood bank practice during the first 5 days of storage.