bluuebunny/crossref_metadata_2025 huggingface dataset:
Inactivation of human immunodeficiency virus by gamma radiation and its effect on plasma and coagulation factors
Authors: Hiemstra H. , Tersmette M. , Vos A.H.V. , Over J. , van Berkel M. P. , de Bree H.
Published: March 2003

The inactivation of HIV by gamma‐radiation was studied in frozen and liquid plasma; a reduction of the virus titer of 5 to 6 logs was achieved at doses of 5 to 10 Mrad at ‐80 degrees C and 2.5 Mrad at 15 degrees C. The effect of irradiation on the biologic activity of a number of coagulation factors in plasma and in lyophilized concentrates of factor VIII (FVIII) and prothrombin complex was examined. A recovery of 85 percent of the biologic activity of therapeutic components present in frozen plasma and in lyophilized coagulation factor concentrates was reached at radiation doses as low as 1.5 and 0.5 Mrad, respectively. As derived from the first‐order radiation inactivation curves, the radiosensitive target size of HIV was estimated to be 1 to 3 MDa; the target size of FVIII was estimated to be 130 to 160 kDa. Gamma radiation must be disregarded as a method for the sterilization of plasma and plasma‐derived products, because of the low reduction of virus infectivity at radiation doses that still give acceptable recovery of biologic activity of plasma components.
In vivo and in vitro binding of C4 molecules on red cells: a correlation of numbers of molecules and agglutination
Authors: Giles C. M. , Davies K. A. , Walport M. J.
Published: March 2003

The fourth component of human complement (C4) is one that is essential to the antibody‐mediated classical activation pathway. C4d, present on all normal and most patient red cells (RBCs), may be detected by the human antisera anti‐Rodgers (Rg) and ‐Chido (Ch). A study has been made of the Rg/Ch antigens on normal and patient RBCs in an attempt to understand the mechanism by which C4 is bound to normal RBCs in the absence of RBC antibodies (Abs). Because RBCs from C1q‐deficient patients express Rg/Ch, it seems that C1q is not essential for C4 binding. Treatment of normal RBCs with proteolytic enzymes, including trypsin, eliminated positive reactions with anti‐Rg/Ch even though the C4d fragment is considered to be resistant to cleavage by trypsin. By correlating agglutination reactions with numbers of bound C4d and C3d molecules, it is evident that both C4d and C3d were affected by trypsin treatment and that anti‐Rg/Ch were not capable of agglutinating RBCs with less than 50 molecules of bound C4d. It is concluded that trypsin‐ sensitive and ‐insensitive RBC membrane structures may both act as acceptors for C4. RBCs with null phenotypes of the major blood group systems all expressed Rg/Ch antigens, so none of the structures that carry these antigens act preferentially as acceptors for C4.
Immunoadsorption for coagulation factor inhibitors
Authors: Uehlinger J. , Button G. R. , McCarthy J. , Forster A. , Watt R. , Aledort L. M.
Published: March 2003

Inhibitors to coagulation factors are among the most difficult problems in the management of coagulation disorders. Most presently available therapy does not assure hemostasis. An extracorporeal immunoadsorption system, which selectively binds IgG, was used to lower inhibitor levels in eight patients on 10 occasions. In this system, separated plasma is delivered to two staphylococcal protein A‐Sepharose columns, which are coupled to an elution monitor. Columns are eluted sequentially and regenerated to maximize IgG removal. Successful removal of the inhibitor was accomplished in all six hemophiliacs on seven occasions, as well as in a patient with acquired von Willebrand disease. All patients whose inhibitors were lowered to less than 10 Bethesda units achieved measurable factor levels when factor concentrate replacement was given. Immunoadsorption facilitates efficient removal of inhibitors, which allows factor replacement therapy.
Inactivation of viruses in platelet suspensions that retain their in vitro characteristics: comparison of psoralen‐ultraviolet A and merocyanine 540‐visible light methods
Authors: Dodd RY , Moroff G , Wagner S , Dabay MH , Dorfman E , George V , Ribeiro A , Shumaker J , Benade LE
Published: March 2003

The ability of two fundamentally different photochemical procedures to inactivate model viruses in platelet suspensions was compared. Merocyanine 540 (MC 540) with visible light was used as an example of an oxygen‐dependent chemical‐directed at the viral membrane, and aminomethyl trimethyl psoralen (AMT) with ultraviolet A light (UVA) was used as an example of a nucleic acid‐directed system. Antiviral conditions in petri dishes were identified and the effects of these procedures on platelet suspensions in plastic storage containers were studied. Concentrations of photochemicals in the 10 to 150 mumol range with 30 to 60 minutes of visible light (MC 540) or 1 to 2 minutes of UVA (AMT) readily inactivated 5 to 6 log10 of vesicular stomatitis virus (VSV) and other model viruses in platelet suspensions, provided the plasma concentration was reduced to about 15 percent by the use of a synthetic platelet storage medium. Extracellular pH, morphology scores, and aggregation response dropped markedly when platelets were treated with MC 540 and visible light. However, treatment with 136 mumol per L of AMT and 1 to 3 minutes of UVA could inactivate 5 log10 of VSV in platelet suspensions with retention of platelet characteristics for 4 days, particularly if oxygen levels were reduced during treatment. These studies demonstrate that AMT‐UVA treatment meets the initial requirements for virus inactivation in platelet suspensions.
Characteristics of elderly blood donors
Authors: Simon T.L. , Rhyney R.L. , Wayne S.J. , Garry P.J.
Published: March 2003

The aging of our society will result in an increased demand for blood components, but it also has the potential to produce a large group of blood donors, the elderly. To study the effects of regular donation by older persons, a randomized, controlled trial is being conducted among 244 healthy, elderly volunteers. This report focuses on the efficacy of the recruiting efforts for that study and describes the resultant population in terms of their demographics, medical status, and donation safety. Of 325 potential subjects, 18 percent were disqualified and 7 percent refused entry into the study. After medical evaluation, only 2 persons were disqualified for conditions not detected by the usual blood services screening protocols. The resultant elderly donor population (n = 244) was well‐educated, middle‐income, and, for the most part, married. The group reported more past and present medical conditions, past surgical procedures, and current medications than would be expected in a younger donor group. Reactions to donation were infrequent and mild. With current screening and donation procedures, blood donation by the elderly appears to be safe and practical.
Presence of the red cell alloantibody anti‐E in an 11‐week‐old infant
Authors: DePalmam L. , Criss V.R. , Roseff S.D. , Luban N.L.C.
Published: March 2003

The development of red cell (RBC) alloantibodies in infants less than 4 months of age is believed to be rare. Though there are no well‐documented published accounts, the formation of alloanti‐E in a multiply transfused 11 ‐week‐old infant is reported here. The infant, blood group B, D +, developed necrotizing enterocolitis and renal failure requiring 31 transfusions of washed and unwashed RBCs (group B and group 0), as well as fresh‐frozen plasma and platelets. Six weeks after the first blood transfusion, alloanti‐E was detected. The anti‐E weakly agglutinated R2R2 screening RBCs at 37°C and sensitized these RBCs to react with anti‐IgG. The infant's RBCs were typed as E–. Passive transfer of alloanti‐E was ruled out by the negative antibody screening tests of each donor unit and the absence of any RBC alloantibodies in the mother's serum. Stored samples of the infant's sera were tested, and anti‐E was shown to be present approximately 11 days after exposure to a known E + RBC unit. The appearance of alloanti‐E in this time frame is consistent with a secondary immune response. Primary immunization most likely took place in the first 4 weeks of transfusion therapy.
Can the reading for serologic reactivity following 37 degrees C incubation be omitted?
Authors: JUDD W.J. , STEINER E.A. , OBERMAN H.A. , NANCE S.J.
Published: March 2003

The need to detect antibodies that agglutinate and/or hemolyze red cells (RBCs) directly at 37 degrees C, but do not react in subsequently performed indirect antiglobulin tests (IATs), is of concern relative to the streamlining and automation of antibody detection methods. To determine incidence and significance of such reactions, data from 87,480 tests, which used low‐ionic‐strength saline, 10‐minute incubation at 37 degrees C, and anti‐IgG, were analyzed for unexpected antibodies. There were 3590 positive tests, of which 475 showed reactions at 37 degrees C but not in subsequently performed IATs (37 + IAT‐). Of these, 196 reactions were due to autoantibodies or other factors usually considered insignificant with respect to the survival of transfused incompatible RBCs, 176 were due to alloantibodies of questionable clinical significance (M, Lea, P1, etc.), and 103 were associated with alloantibodies of potential clinical significance (63 E, 27 K, 5 Jka, 4 D, 3 cE, and 1 C). This latter reaction was seen in 72 patients, with two 37 + IAT‐antibodies occurring in each of 3 patients. Of the 75 potentially significant 37 + IAT‐antibodies, 57 were seen in patients recently exposed to homologous RBCs, 13 in patients with a history of transfusion and/or pregnancy, and 5 in patients with no known exposure to homologous RBCs. IAT reactivity was observed in subsequent samples with 27 of these antibodies. The predictive value of a 37 + IAT‐test was 21.7 percent for a potentially significant antibody. The incidence was 0.12 percent of all tests for unexpected antibodies. As 27 potentially significant 37 + IAT ‐ antibodies were found in subsequent samples to be IAT +, indicating a secondary (IgG) immune response, elimination of the 37°C reading for agglutination and hemolysis should not be undertaken lightly.
Separation of albumin from autologous plasma by heat
Authors: ZHOW Y. , ZHU M. , CHENG J. , YIAN B. , YIANG Y.
Published: March 2003

A simplified technique has been developed to separate albumin from the plasma of a patient undergoing treatment with plasma exchange. This albumin can be returned to the patient to maintain colloid osmotic pressure without risk of transmitting disease. Patient plasma is obtained by apheresis. It is heated at 70 degrees C for 60 minutes to denature and remove autoantibodies, immune complexes, and abnormal globulins. The resulting low‐concentration albumin solution (LCAS) retains its natural properties. Immunologic analysis by gel diffusion shows a reaction pattern of complete identity with that of human serum albumin. The albumin in LCAS is more than 90 percent of the remaining protein, as determined by scan of a cellulose acetate electrophoresis pattern. More than 97 percent of the albumin is in monomer form, as judged by polyacrylamide gel electrophoresis. No patient antibodies to neonantigens in the heat‐treated albumin can be identified by double radial immunodiffusion. More than 80 percent of the plasma albumin is recovered in LCAS. LCAS is safe and efficient for use in plasma exchange.
Age dependency of ABO histo‐blood group antibodies: reexamination of an old dogma
Authors: der Maur C. Auf , Hodel M. , Nydegger U.E. , Rieben R.
Published: March 2003

Current textbooks for transfusion medicine state that anti‐A and/or anti‐B (anti‐A/B) agglutination titers–and thus the respective antibody concentrations–reach their maximum in individuals 5 to 10 years old and then gradually decline with the increasing age of the individual. This statement is largely based on a study by Thomsen and Kettel that dates to 1929. In the present article, ABO antibodies in sera of 175 healthy persons aged 61 to 97 years, as well as sera of 170 newborn infants and children aged 0 to 17 years, were analyzed. Microhemagglutination tests were performed with all sera and complemented by ABO enzyme‐linked immunosorbent assays to measure the immunoglobulin class (IgM, IgG, and IgA) of the anti‐A/B. As in a previous study using sera of persons aged 20 to 67 years, individual differences exceeded age‐related changes for all variables. Median values of IgG and IgA anti‐A/B were elevated in elderly persons of blood group O, whereas no significant changes were observed in other variables. In particular, the decrease in agglutination titers with the increasing age of the individuals was far less pronounced than previously described; even in sera of persons aged 90 to 97 years, median agglutination titers of 128 were found. Results in the sera of children confirm previously reported data that agglutination titers and IgM anti‐A/B reached adult levels at the age of 5 to 10 years.
Lack of clinical significance of “enzyme‐only” red cell alloantibodies
Authors: Issitt P.D , Combs M.R. , Bredehoeft S.J. , Campbell M.L. , Heimer M. , Joyner L. , Lorentsen L. , Remley C. , Bullock S. , Bumgarner J. , Zakeriniasar Mitra , Kirkland Amelia W. , Melroy Helen M. , Millikin Donna M.
Published: March 2003

In a retrospective study on samples from 10,000 recently transfused patients, 35 samples were found to contain an antibody that reacted with ficin‐treated red cells but was not demonstrable by low‐ionic‐ strength saline solution and indirect antiglobulin test (LISS‐IAT). In those 35 patients, the specificity of the antibody was such that each patient would have been transfused with antigen‐negative blood had the antibody reacted in LISS‐IAT. Tests on red cells from the units already transfused showed that 19 patients had among them received, by chance, 32 antigen‐positive and 74 antigen‐negative units. The remaining 16 patients had among them received 57 units that were, again by chance, all antigen negative. One patient given antigen‐positive blood suffered a delayed transfusion reaction; in two others the antibodies became LISS‐IAT active after transfusion. However, similar changes to the LISS‐ IAT‐active state were seen with two antibodies of patients given only antigen‐negative blood. Also found in the 10,000 patients were 28 clinically insignificant antibodies, 77 sera in which the antibody was too weak to identify, and 216 autoantibodies that reacted only with ficin‐treated red cells. These data support a belief, generally held in the United States but not necessarily elsewhere, that the use of protease‐treated red cells for routine pretransfusion tests creates far more work than the accrued benefits justify.
The National Marrow Donor Program
Authors: Stroncek D. , Bartsch G. , Perkins H.A. , Randall B.L. , Hansen J.A. , McCullough J.
Published: March 2003

As the number of successful marrow transplants has increased, the lack of HLA‐identical sibling donors for 60 to 70 percent of transplant candidates has become a serious problem. Pilot studies established that marrow transplantation between phenotypically HLA‐identical, unrelated individuals can be accomplished successfully. Therefore, the National Marrow Donor Program was established to develop a large file of volunteer marrow donors and to serve as a center for the coordination of the donor search and donor‐recipient matching processes. By November 1991, 63 months after the program was established, 457,205 potential marrow donors typed for HLA‐A and ‐B antigens had agreed to be listed in the marrow donor registry. A donor search had been initiated for 8481 patients. At least one potential donor matched for at least three of the four HLA‐A and ‐B antigens was located for 99.8 percent of patients. Among the 3156 searches that were completed, 940 (29.8%) resulted in a transplant. The median time in which to locate a matched donor, complete all predonation evaluations, and obtain donor consent was 208 days. The most common diagnosis in patients who underwent transplantation was chronic myelogenous leukemia (42.0%). When this analysis was completed in November 1991, the National Marrow Donor Program was operating a national network of 99 donor centers and 53 transplant centers. The donor file was increasing rapidly, and a follow‐ up system was in place to determine the effects of donation on the donors and the outcome in the patients who underwent transplantation. This national network of donor and transplant centers exists and is now facilitating unrelated‐donor marrow transplants. The National Marrow Donor Program made it possible to locate donors for many patients in need of a transplant and helped to determine the role of unrelated‐ donor marrow transplants in the treatment of many diseases.
Exposure of newborn infants to di‐(2‐ethylhexyl)‐phthalate and 2‐ ethylhexanoic acid following exchange transfusion with polyvinylchloride catheters
Authors: Plonait S.L. , Nau H. , Maier R.F. , Wittfoht W. , Obladen M.
Published: March 2003

Infants in the neonatal intensive care unit are regularly exposed to the plasticizer di‐(2‐ethylhexyl)‐phthalate (DEHP) following exchange transfusion or extracorporeal membrane oxygenation. Whether such exposure leads to increased morbidity is not known, although elevated levels of DEHP have been associated with necrotizing enterocolitis and cholestasis. The hypothesis that infants undergoing exchange transfusion are exposed to toxic levels of DEHP and the presumed metabolite 2‐ethylhexanoic acid (EHXA) was tested by measuring serum levels of DEHP in 16 newborn infants (gas‐liquid‐chromatography) and urine concentrations of EHXA in 6 of these infants (gas chromatography‐ mass spectrometry). DEHP levels were undetectable (< 1 microgram/mL) before exchange but ranged from 6.1 to 21.6 micrograms per mL of serum (average, 12.5 +/− 6.2 micrograms/mL) after a single exchange transfusion. DEHP uptake did not result in cholestasis. EHXA peak levels were 127 to 416 ng per mL of urine, with a median of 174 ng per mL. Concentrations of EHXA were lower than anticipated, which indicates that EHXA is not a major metabolite in the neonatal infant.
Column agglutination technology: the antiglobulin test
Authors: Reis K.J. , Chachowski R. , Cupido A. , Davies D. , Jakway J. , Setcavage T.M.
Published: March 2003

A new system for typing and screening blood, based on the sieving effect of glass bead microparticles, has been developed. The test is performed in a microcolumn in which the red cell agglutinates are trapped in the glass bead matrix during centrifugation, and unagglutinated cells form a pellet at the bottom of the column. Anti‐human globulin reagents were incorporated in the diluent and the new test system, column agglutination technology, was compared to conventional tube tests and low‐ionic‐strength method. Sera and plasmas (228 samples) were screened for red cell antibodies with two anti‐human globulin reagents: one containing only anti‐IgG and the other containing both anti‐IgG and anti‐C3b, ‐C3d. After initial testing, there was 94‐percent agreement between column agglutination technology and tube tests, and after repeat testing, there was 97‐percent agreement. The column agglutination technology anti‐human globulin test eliminates the need to wash red cells, which decreases the overall test time. The test is easy to perform, and the results are more objective than those with tube and microplate methods.
Acute changes in systemic blood pressure and urine output of conscious rats following exchange transfusion with diaspirin‐crosslinked hemoglobin solution
Authors: Keipert P.E. , Gonzales A. , Gomez C.L. , MacDonald V.W. , Hess J.R. , Winslow R.M.
Published: March 2003

This report describes acute changes in systemic blood pressure and urine output observed after a 50‐percent isovolemic exchange transfusion (ET) with diaspirin‐crosslinked hemoglobin (alpha alpha Hb). Stroma‐free Hb was crosslinked between the alpha chains by using a 14C‐labeled diaspirin, bis(3,5‐dibromosalicyl)fumarate. Forty conscious, chronically cannulated rats underwent ET with 14C‐labeled alpha alpha Hb solution (8.0 g/dL [80 g/L]). This resulted in systemic hypertension for 3 to 4 hours after ET (mean arterial pressure rose from 120 to 145 torr at 1 to 2 hours after ET) and mild bradycardia for 2 to 3 hours (heart rate decreased from 420 to 335 beats/min [bpm] before stabilizing at 360 +/− 10 bpm). This was accompanied by significant diuresis immediately after ET (5‐ 6‐fold increase in urine output, which normalized after 12 hours), and mild hemoglobinuria. The total amount of Hb recovered in the urine was < 5 percent of the injected dose. Reversed‐phase high‐performance liquid chromatography and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis confirmed the presence of crosslinked alpha alpha Hb molecules in the urine. Renal excretion of radioactivity was significantly greater, with 20 percent of total radioactivity being eliminated within 24 hours. The plasma half‐life for alpha alpha Hb was 5 hours (administered dose, 2.4 g Hb/kg body weight). Thus, infusion of alpha alpha Hb caused a transient systemic hypertension, and intramolecular crosslinking alone was not enough to exclude completely the filtration of alpha alpha Hb by the kidneys.
Results of 1‐year screening of donors in The Netherlands for human T− lymphotropic virus (HTLV) type I: significance of Western blot patterns for confirmation of HTLV infection
Authors: ZAAIJER H.L. , CUYPERS H.T.M. , DE WIT C. DUDOK , LELIE P.N.
Published: March 2003

BACKGROUND: Between January 1993 and January 1994, Dutch blood banks screened approximately 674,000 volunteer donors for the presence of antibodies to human T‐lymphotropic virus type I (HTLV‐I). STUDY DESIGN AND METHODS: Confirmatory testing was performed on samples from 870 different anti‐HTLV‐I‐reactive donors (0.13% of the total tested). RESULTS: According to the authors' Western blot (WB) interpretation criteria, 15 (0.002%) donors tested HTLV‐I‐positive in the WB; 201 tested negative, and 654 (75% of donors reacting on enzyme‐linked immunosorbent assay) tested indeterminate. Fresh samples from 234 of 870 anti‐HTLV‐reactive donors were tested for HTLV‐I and type II (HTLV‐ II) DNA by polymerase chain reaction: all 15 WB‐positive donors tested positive for HTLV‐I DNA; 206 WB‐indeterminate and 13 WB‐negative donors tested negative for HTLV‐I and ‐II DNA. Application of the manufacturer's (World Health Organization‐based) guidelines for WB interpretation would have resulted in the misclassification of 48 (23%) of 206 polymerase chain reaction‐negative donors as HTLV‐I/II‐positive. Risk factors were present in 14 of 15 HTLV‐I‐infected donors: 8 had a partner from an HTLV‐I‐endemic area, 4 were from HTLV‐I‐endemic countries, and 2 had received blood transfusions. CONCLUSION: HTLV‐I and ‐II infection is rare among Dutch blood donors. HTLV screening will prevent few cases of HTLV‐related disease, but it will prevent a further spread of the virus by transfusion. In a low‐risk population, conservative guidelines for WB interpretation unnecessarily generate an excess of false‐positive results.
Waldenström's macroglobulinemia with an IgM paraprotein that is both a cold agglutinin and a cryoglobulin and has a suppressive effect on progenitor cell growth
Authors: ZAGO‐NOVAREIT M. , KHURI F. , MILLER K.B. , BERKMAN E.M.
Published: March 2003

BACKGROUND: A patient with Waldenstrom's macroglobulinemia was admitted to the hospital with fever, leg pain, and dyspnea. The patient had gas gangrene of the left leg that required above‐the‐knee amputation. Plasmapheresis was instituted to treat hyperviscosity. STUDY DESIGN AND METHODS: The patient's serum contained an IgM‐kappa paraprotein, a cryoglobulin, and a cold agglutinin. The serum was studied. RESULTS: The patient's red cells typed as A1, Rh‐positive. The direct antiglobulin test was negative. The serum contained a cold agglutinin with anti‐Pr cold agglutinin specificity (titer 4096). Maximal thermal range was 30 degrees C. Following dithiothreitol treatment, the cold agglutinin activity disappeared. The serum IgM concentration in the tested sample was 62.3 g per L. The cold agglutinin titer in the supernatant after removal of the cryoglobulin was 256, and the IgM level was 0.31 g per L. Redissolving the cryoglobulin in a equivalent volume of saline resulted in a cold agglutinin titer of 4096 and an IgM level of 68.4 g per L. These results indicate that the cryoglobulin and the cold agglutinin are the same paraprotein. Serum protein electrophoresis using agarose gel and immunofixation of the serum revealed an IgM‐kappa monoclonal band. Progenitor cell assays were performed by adding the patient's serum at final concentrations of 0, 1, 5 and 10 percent (vol/vol) to patient's and normal donor's peripheral blood mononuclear cells. Inhibition of burst‐forming units‐ erythroid and colony‐forming units‐granulocyte/macrophage by the patient's serum was demonstrated. Appropriate controls and the use of the serum of another patient with Waldenstrom's macroglobulinemia did not suppress progenitor cell growth. The patient's serum inhibited colony formation in a dose‐response fashion. CONCLUSION: Reports of cryoprecipitable cold agglutinins are rare. This case is unusual because the IgM‐kappa paraprotein was also a cold agglutinin with anti‐ Pr specificity and erythroid and granulocyte‐macrophage progenitor cell‐ suppressive properties.
Low retention of white cell fragments by polyester fiber white cell‐ reduction platelet filters
Authors: Ramos Ricardo R. , Curtis Brian R. , Duffy Brian F. , Chaplin H
Published: March 2003

BACKGROUND: There is intense interest in the potential of current white cell (WBC)‐reduction filters to prevent the alloimmunization of patients by the residual donor WBCs in filtered blood components transfused to them. Little attention has been paid to the capacity of current synthetic fiber filters to remove WBC membrane fragments bearing detectable leukocyte antigens.

STUDY DESIGN AND METHODS: Fluorescein isothiocyanate‐conjugated monoclonal antibody to a universal WBC membrane antigen (CD45) and high‐speed centrifugation coupled with ficoll‐hypaque differential sedimentation were used to quantitate low‐density WBC fragments in single‐donor platelet components before and after filtration to determine if a polyester fiber filter retained WBC fragments.

RESULTS: Prefiltration measurements in 25 single‐donor platelet components indicated that WBC fragments increased with length of storage up to 5 days at room temperature (p < 0.0001). When fragments in eight components were measured before and after filtration, absolute values for differences were insignificant (p = 0.15). CONCLUSION: WBC fragments were poorly retained by these polyester fiber WBC‐reduction filters. The antigenicity of WBC fragments could contribute to the WBC alloimmunization of some recipients of WBC‐reduced blood components.
Platelet concentrates stored for 5 days in a reduced volume of plasma maintain hemostatic function and viability
Authors: Ali A.M. , Warkentin T.E. , Bardossy L. , Goldsmith C.H. , Blajchman MA
Published: March 2003

BACKGROUND: Platelet concentrates prepared from whole blood are generally suspended in a standard volume of 50 to 60 mL of plasma and can be stored thus at 20 to 24 degrees C for up to 5 days. In vitro studies suggested that this plasma volume could be reduced to 30 to 35 mL without impairing platelet function.

STUDY DESIGN AND METHODS: This study evaluated whether platelets stored for 5 days in a reduced volume (30‐35 mL) of plasma maintained their in vivo viability, hemostatic function, and recovery in recipients. Paired autologous platelet survival studies were done in 20 adult volunteers to assess platelet viability. A rabbit ear bleeding‐time model was used to compare the hemostatic effectiveness of human platelet concentrates stored for 5 days in the standard or reduced volume of plasma. Platelet recovery was compared in thrombocytopenic hospital patients.

RESULTS: Paired platelet survival studies indicated no significant difference between the values in platelet concentrates stored for 5 days in the reduced volume of plasma and the values in those stored in the standard volume. In the animal model, there was no significant difference in the bleeding times achieved by either set of platelet concentrates. The platelet count increments in thrombocytopenic patients were measured. The platelet count increments in patients who received reduced‐volume platelet concentrates were as good as the increments achieved in patients given standard‐volume concentrates.

CONCLUSION: The in vivo viability, recovery, and hemostatic function of platelets collected in polyvinylchloride plastic containers and stored in 30 to 35 mL of plasma for 5 days are maintained as well as those of platelets stored in 50 to 60 mL of plasma.
Protective effect of blood transfusions on postoperative recurrence of Crohn's disease in parous women
Authors: Silvis R. , Steup W.H. , Brand A. , Zwinderman K.A. , Lamers C.B. , Griffioen G. , Gooszen H.G.
Published: March 2003

Background: Perioperative blood transfusion (BT) appeared to have adverse effects on survival after surgery for malignant tumors while pretransplantation BT suppressed allograft rejection. Interest grew in the effect of BT on postoperative recurrence of Crohn's disease.

Study Design and methods: To determine the effect of perioperative BT on the recurrence of Crohn's disease after primary surgery, the medical histories of 148 patients with Crohn's disease, 62 males and 86 females (49 nonparous and 37 parous), were reviewed. Eighty‐seven patients received perioperative BT.

Results: Overall, perioperative BT showed no effect on recurrence. Patients with Crohn's disease limited to the ileum had a better prognosis with regard to recurrence than did patients with Crohn's disease located in the colon or located in both ileum and colon, but the difference was not significant. Perioperative transfusion seemed to protect against recurrent disease after colon resection, which might be explained by the fact that colon resections, which often necessitate perioperative BT, generally result in a shorter bowel segment at risk for recurrent disease. Overall, parous women showed a worse prognosis than nonparous females and men (p = 0.022). Transfusions had a beneficial effect in parous women (p = 0.068) and, after correction for type of operation, this beneficial effect was significant (p = 0.026). After perioperative BT, parous women had a similar prognosis with respect to recurrent Crohn's disease as nonparous females and men.

Conclusion: Perioperative BT has a beneficial effect on the postoperative recurrence of Crohn's disease in parous women.
Seroprevalence of human T‐lymphotropic virus, hepatitis C virus, and human immunodeficiency virus in Asian American potential bone marrow donors
Authors: Sun M. , Chao K.H. , Modahl L.E. , Leoung G. , Khayam‐Bashi H. , Ng V.L. , Murphy E.L.
Published: March 2003

Background: Asian Americans are generally underrepresented both as volunteer blood and bone marrow donors.

Study Design and Methods: To investigate the risk of transfusion transmission of viruses that is associated with increasing participation by Asian American donors, antibodies to human T‐lymphotropic virus (HTLV), hepatitis C, and human immunodeficiency virus in Asian American volunteers recruited as potential bone marrow donors were measured. A total of 1354 Asian Americans were enrolled in the study, of whom 54 percent were Chinese, 26 percent Japanese, 9 percent Filipino, 4 percent Korean, 3 percent Indian, and 5 percent of other Asian or mixed Asian and other ethnicity. The majority of the study population was aged 20 through 49 and of high socioeconomic status, as indicated by education and income. Viral antibodies were measured with both screening enzyme‐linked immunosorbent assays and supplemental testing, and polymerase chain reaction was used to resolve discrepant HTLV results.

Results: Confirmed seroprevalence rates for HTLV were 0.15 percent with one manufacturer's Western blot and 0.3 percent with the other; however, no sample was positive for HTLV types I or II in polymerase chain reaction. Confirmed seroprevalence to hepatitis C virus was 0.5 percent. No subject was seropositive for human immunodeficiency virus.

Conclusion: On the basis of the moderate size and high education level of this study population, it is concluded that Asian American volunteer bone marrow donors do not pose a greater risk for transmission of HTLV type I or II, human immunodeficiency virus, or hepatitis C virus than does the average American blood donor.
Detection and characterization of hepatitis C virus RNA in immune globulins
Authors: Yu M.Y. , Mason B.L. , Tankersley D.L.
Published: March 2003

BACKGROUND: Hepatitis C virus (HCV) RNA was measured in immune globulins and its chemical and physical properties were characterized.

STUDY DESIGN AND METHODS: The study examined 69 immune globulin lots from 7 manufacturers, including 44 intravenous and 25 intramuscular immune globulin preparations. In addition, 8 experimental intravenous immune globulin preparations were investigated. Detection and quantitation of HCV RNA were achieved by reverse transcription and nested polymerase chain reaction at limiting dilution. A multi‐antigen anti‐HCV enzyme immunoassay was also used to test these immune globulins.

RESULTS: The highest level of HCV RNA was found in an experimental immune globulin lot derived from a plasma pool made up of 186 anti‐c100‐3‐reactive units. HCV RNA was detected only in 1 of 7 manufacturers' experimental intravenous immune globulin preparations derived from a pool made up of 2887 anti‐c100‐3‐negative units. It was also detected in commercial intravenous immune globulin lots prepared by the same manufacturer from source plasma, but not from recovered plasma. More than half of the commercial intramuscular immune globulin lots, including specific immune globulin products, were HCV RNA positive. All immune globulin products examined were reactive for anti‐ HCV. Certain similarities were found for HCV RNA present in an immune globulin product and plasma. Ethanol at 20 or 25 percent had no effect upon the buoyant density of HCV RNA.

CONCLUSION: Many immune globulin preparations contained HCV RNA, with levels depending upon both the type of starting plasma and the manufacturing process. Exposure to ethanol did not appear to affect the physical characteristics of HCV RNA.
At(a−) phenotype: description of a family and reduced survival of At(a+) red cells in a proposita with anti‐Ata
Authors: Sweeney Joseph D. , Holme Stein , McCall Lee Ann , Huett Donna , Storry Jill , Reid Marion
Published: March 2003

BACKGROUND: There is a paucity of data on the August (At) blood group antigen and clinical significance of anti‐Ata. STUDY DESIGN AND METHODS: A proposita with the At(a‐) phenotype was identified by the finding of anti‐Ata in the cord blood eluate of her fifth live infant. Family members were studied, and a small aliquot of 51Cr‐labeled At(a+) red cells was transfused to determine survival. RESULTS: There was no evidence of hemolytic disease of the newborn, as determined by the normal hemoglobin and bilirubin and normal clinical conditions. Six of seven siblings were tested, and two At(a‐) female siblings were identified. In contrast to the proposita, neither sister had detectable anti‐Ata in her serum, although each has had only one pregnancy. A monocyte monolayer assay performed on serum from the proposita gave a result of 20‐percent hemolysis (normal, <3%), which is consistent with a clinically significant antibody. Transfusion of a small volume of allogeneic red cells that were phenotypically matched with the proposita, except for Ata, resulted in a 1‐hour survival of 95 percent, but a 24‐hour survival of only 18 percent, of the transfused cells. The survival pattern was exponential, which is characteristic of a non‐ complement‐binding IgG antibody. CONCLUSION: Despite the absence of hemolytic disease of the newborn, this example of anti‐Ata would appear to be a clinically significant antibody for the purposes of transfusion practice. Therefore, approaches to the management of clinical situations in which transfusion is required or likely should focus on the availability of autologous cells or frozen allogeneic At(a‐) red cells.
Quantitation of CD62, soluble CD62, and lysosome‐associated membrane proteins 1 and 2 for evaluation of the quality of stored platelet concentrates
Authors: Divers S. G. , Kannan K. , Stewart R. M. , Betzing K. W. , Dempsey D. , Fukuda M. , Chervenak R. , Holcombe R. F.
Published: March 2003

BACKGROUND: Platelets become activated during storage, which results in secretion of granules, vesiculation of microparticles, secretion of protein, and a number of other biochemical and morphologic processes that decrease the utility of platelet concentrates stored for transfusion. STUDY DESIGN AND METHODS: To evaluate the quality of stored platelet concentrates, the cell surface expression of specific activation‐dependent antigens (CD62 and lysosome‐associated membrane proteins 1 and 2 [LAMP‐1, LAMP‐2]) on platelets stored in a hospital blood bank over a 7‐day period was examined. Relative microparticle counts and the expression of CD62 by microparticles, as well as platelet concentrate supernatant levels of soluble CD62, were determined. RESULTS: The percentage of platelets expressing CD62 increased significantly from Day 1 to Day 5 (p < 0.05) of storage; the mean fluorescence values for CD62 did not. In contrast, the mean fluorescence values of LAMP‐1 and LAMP‐2 rose significantly (p < 0.01 and p < 0.05, respectively) between Days 1 and 5. Significant declines in CD62, LAMP‐1, and LAMP‐2 percent expression and mean fluorescence were seen on Day 6 of storage (p < 0.001). Microparticle numbers increased significantly during storage and correlated with levels of CD62 protein (free and membrane‐bound) (r = 0.95 vs. Day 2, p < 0.05; r = 0.88 vs. Day 5, p < 0.05). CONCLUSION: Flow cytometric evaluations of the expression of cell surface CD62, LAMP‐1, and LAMP‐2 are complementary tests that, especially when used in conjunction with the quantitation of CD62 protein, provided a simple and effective means of evaluating the quality of platelet concentrates stored for transfusion.
Lea and Leb tissue glycosphingolipids
Authors: Hauser R.
Published: March 2003

BACKGROUND: The Lea and Leb antigens are synthesized by cells making exocrine secretions, but it is not well‐defined where plasma and red cell Le(a) and Le(b) antigens are made.

STUDY DESIGN AND METHODS: This study uses chromatographic and immunochemical methods to extract Lea and Leb glycolipids from human tissues.

RESULTS: Lea and Leb antigens may be synthesized by both epithelial and mesodermal tissues. Expression of Lewis transferase activity was demonstrated in various tissues, confirming the epistatic interaction by Le and H genes to form the Leb antigen.

CONCLUSION: The Lea and Leb substances found in plasma might originate from the epithelial cells of the digestive tract.
Posttransfusion purpura‐like syndrome associated with CD36 (Naka) isoimmunization
Authors: Bierling P. , Godeau B. , Fromont P. , Bettaieb A. , Debili N. , El‐Kassar N. , Rouby J.J. , Vainchenker W. , Duedari N.
Published: March 2003

BACKGROUND: CD36 deficiency, which could lead to CD36 isoimmunization, has been reported in the Japanese population. CD36 isoantibody has been involved in platelet transfusion refractoriness. CASE REPORT: A 50‐year‐ old woman originally from Corsica developed severe acute thrombocytopenia after massive transfusion. She was found to be CD36 deficient, and platelet immunoassays revealed a CD36 (Naka) platelet isoantibody. Although the involvement of another mechanism could not be entirely ruled out, the thrombocytopenia was attributed to posttransfusion purpura‐like syndrome. The antibody was also involved in platelet transfusion refractoriness. CD36 deficiency was present in two members of the patient's family as well. Flow cytometry studies demonstrated the absence of CD36 expression on the surface of blood monocytes and cultured erythroblasts and megakaryocytes from one of the two CD36‐deficient family members studied, but, in the absence of previous immunization, these CD36‐deficient patients were not isoimmunized. In contrast, CD36 deficiency was not found in a population of 808 healthy blood donors in the Paris, France, area.

CONCLUSION: CD36 isoantibody might be involved in some cases of posttransfusion purpura and platelet transfusion refractoriness. These findings also confirm the extremely low frequency of CD36 deficiency among whites.
Large‐volume leukapheresis in pediatric patients: processing more blood diminishes the apparent magnitude of intra‐apheresis recruitment
Authors: Gorlin J.B. , Vamvakas E.C. , Cooke E. , Galacki D. , Geha R. , Humphreys D. , Kent P.
Published: March 2003

Background: Recruitment of progenitors during a large‐volume collection, as defined by increasing relative and absolute numbers of progenitors (colony‐forming units‐granulocyte‐macrophage [CFU‐GM] of CD34+ cells), has been reported previously.

Study Design and Methods: To ascertain whether intra‐apheresis recruitment occurs in pediatric patients who have undergone mobilization with chemotherapy and granulocyte‐colony‐stimulating factor (G‐CSF), each hour's portion of a 4‐hour leukapheresis was collected into separate bags, and assessed by complete blood count, CFU‐GM, and CD34+ cell assays. Seven pediatric patients (median age, 7; range, 2–19) were studied in connection with 2 to 4 collections each, for a total of 21 collections (with hourly samples). The collections lasted for 4 hours, at an inlet rate of 1 to 3 mL per kg per minute, for daily processing totals of 5 to 12 blood volumes. (One blood volume [mL] is estimated by the patient's weight in kg × 70 mL/kg.) Smaller (younger) patients had inlet rates exceeding 2 mL per kg per minute, and larger (older) patients had rates of 1 to 1.5 mL per kg per minute. CFU‐GM and CD34+ cell counts obtained each hour of the collection and divided by the first hour's value were compared by nonparametric repeated‐measures ANOVA.

Results: Second‐, third‐ and fourth‐hour CD34+ progenitor cell counts were arithmetically higher than first‐hour counts, but the trend did not reach significance (p = 0.1561). Second‐hour counts were higher than first‐hour counts in the overall analysis (mean ± standard error [SE], 1.00 and 1.39 ± 0.1, respectively; p = 0.0525) and in children older than 5 years (1.00 vs. 1.70 ± 0.30, respectively; p = 0.0259), but not in children younger than 5 years (p = 0.8125). CFU‐GM counts did not differ among the 4 hours of collection (p = 0.1717) or between the first and second hour (p = 0.9587).

Conclusion: In larger (older) patients, from whom fewer blood volumes were collected, there is a trend toward intra‐apheresis recruitment, although less than reported previously. In the smaller (younger) patients, from whom more blood volumes were collected, no trend was observed. Lack of (or submaximal) prior mobilization in previously reported studies may have facilitated intracollection recruitment. Alternatively, the larger number of blood volumes collected from the smaller (younger) patients may have masked intra‐apheresis recruitment. The study documents the feasibility of large‐volume, 4‐hour leukapheresis in pediatric patients.
Comparison of random‐donor platelet concentrates prepared from whole blood units and platelets prepared from single‐donor apheresis collections
Authors: Sloand E. M. , Yu M. , Klein H. G.
Published: March 2003

BACKGROUND: The use of fresh platelets results in better posttransfusion recovery and survival than does the use of platelets that have been stored before transfusion. Activation of platelets during preparation and storage may be one of the factors responsible for a number of storage‐related changes in platelet membrane proteins. Blood centers commonly prepare platelet concentrates from both multiple units of whole blood and single‐donor plateletpheresis collections. STUDY DESIGN AND METHODS: Seventeen plateletpheresis concentrates, anticoagulated with ACD, were compared to platelets prepared from whole blood from the same donor that was anticoagulated with CPDA‐1 (random‐ donor platelets). After preparation, plateletpheresis and random‐donor platelets were stored in plastic storage bags at 22 degrees C for 5 days. Platelet surface glycoproteins were examined by flow cytometry after platelets were fixed in dilute plasma with 1‐percent formaldehyde and stained with fluorescein isothiocyanate‐labeled monoclonal antibodies CD42b (anti‐glycoprotein [GP]lb), CD41a (anti‐GPllb/llla), and CD62 (anti‐P‐selectin). RESULTS: The binding of anti‐CD42b was greater in plateletpheresis concentrates than in random‐donor platelets on Days 3 and 5 (p < 0.01) of storage; binding of anti‐CD62 was greater in the random‐donor concentrates (p < 0.01) on Days 3 and 5. Plateletpheresis concentrate aggregation responses were greater on Day 5 (p < 0.01). To determine if the type of anticoagulant and the method of mixture with blood contributed to these changes, 10 samples were split into aliquots and prepared in two separate ways: One group of samples was prepared by allowing anticoagulant (ACD) and blood to flow into the tube at a rate of 3 microL per second, and the other group of samples was prepared by allowing blood to flow into tubes containing a measured amount of CPDA‐1. The first samples bound more anti‐CD42b than the second samples (p < 0.01). The second group of samples contained significantly more microvesicles that bound anti‐CD41a than did the first group (p < 0.01). Samples prepared by the first method but anticoagulated with CPDA‐1 contained more microvesicles but had the same amount of anti‐CD42b binding as did similarly prepared samples anticoagulated with ACD (p < 0.05). CONCLUSION: Platelet concentrates prepared from single units of whole blood and anticoagulated with CPDA‐ 1 bind less anti‐CD42b and more anti‐CD62 than do platelets obtained by apheresis. These differences may be attributed to platelet sedimentation and the transient exposure of some of the platelets in the blood that is first collected during whole‐blood donation to high concentrations of anticoagulant.
Evaluation of methods for preparing and thawing cryopreserved CD34+ and CD34− cell lines for use as reagents in flow cytometry of hematopoietic progenitor cells
Authors: Bowman C. A. , Yu M. , Cottler‐Fox M.
Published: March 2003

BACKGROUND: Flow cytometry is used to quantitate CD34+ hematopoietic progenitor cells for transplantation. The present study evaluates methods for preparing and thawing cryopreserved CD34+ and CD34‐ cell lines for use as flow cytometry reagents. STUDY DESIGN AND METHODS: The human myeloid leukemic cell lines KG 1 a (CD34+) and K562 (CD34‐) were grown in culture under standard conditions and then prepared on ficoll gradients of different densities to determine which gave the component that was most reproducible. After ficoll preparation, the cells were frozen in standard cryopreservation media and four methods of thawing were examined. Determination of the method that gave the cell component that was most reproducible was based on viability, percentage of cell recovery, and maintenance of CD34 antigenicity status. RESULTS: Ficoll gradient preparation improved the ease of flow cytometry analysis when original viability was low, and it produced a more uniform cell population. However, it resulted in significant cell loss for both cell lines. While the cell recovery for K562 cells was not significantly different with any of the densities of ficoll, recovery was significantly better for KG 1 a cells with ficoll at a specific gravity of 1.077. Of the thawing methods examined, all three that involved a rapid thaw at 37 degrees C were statistically equivalent to each other and were better than thawing at 4 degrees C. CONCLUSION: With a standardized method of preparing cell lines as reagents for quality control purposes, data comparison among cell processing laboratories may more readily be initiated. Such cell lines could also be useful as a teaching tool for flow cytometry and in proficiency testing.