bluuebunny/crossref_metadata_2025 huggingface dataset:
A quick and efficient way to pool platelets
Authors: Gorgone B. C. , Coliano F. R. , Tuttle S. A. , Ervin T. J.
Published: December 2004

An easy and efficient method for pooling multiple platelet concentrates is presented. By adaption of a laboratory vacuum system, the process of pooling platelet units was significantly shortened without platelet loss. In our laboratory time study, the use of this pooling system resulted in time savings of 27.5 percent and 35.9 percent during pooling of four‐unit and six‐unit platelet packs, respectively. Platelet loss by the vacuum method was not distinguishable from that of the hand method (p = 0.4 by two‐sample t test)
The effect of irradiation on platelet arachidonic acid metabolism
Authors: Stuart M. J.
Published: December 2004

Irradiation of platelets in vitro to 2000 rads did not alter arachidonic acid metabolism through the cyclooxygenase pathway. Production of platelet thromboxane B2 and 12‐hydroxy‐5,8–10‐heptadecatrienoic acid remained similar in control and paired irradiated platelets. In paired samples the platelet lipoxygenase metabolite 12‐hydroxy‐5,8,10,14‐eicosatetraenoic acid was significantly increased postirradiation to 42 ± 2.8 percent when compared to a control value of 34.1 ± 3.6 percent (p < 0.02). Although this latter product has been demonstrated to be chemotactic for neutrophils, its role in platelet function has not been specifically determined.
Tca: a high‐frequency blood group antigen
Authors: Laird‐Fryer B. , Dukes C. V. , Lawson J. , Moulds J. J. , Walker E. M. , Glassman A. B.
Published: December 2004

An IgG antibody defining a high‐frequency red cell antigen was detected in the sera of two unrelated, nontransfused black women GT (gravida 1, para 0) and DLC (gravida 4, para 2). Inheritance of the antigen was established by the family study of DLC. The clinical significance of the antibody is not yet known. The antibody has been designated anti‐Tca, and the corresponding antigen, Tca. No Tc(a‐) individuals were found among 5000 Caucasians, 5000 Japanese, or 450 blacks; however, anti‐Tca was identified in the sera of two other unrelated black women (LZ and WM). A relationship between Tca and Cra has been proposed by other serologists, but further evidence is needed to confirm the association.
Anti‐Wj: an autoantibody that defines a high‐incidence antigen modified by the In(Lu) gene
Authors: Marsh W. L. , Brown P. J. , DiNapoli J. , Beck M. L. , Wood M. , Wojcicki R. , de la Camara C.
Published: December 2004

An IgG autoantibody, named anti‐Wj, reacts with all random and most selected red cell samples. It does not agglutinate cells of Lu(a‐b‐) type caused by the In(Lu) dominant inhibitor gene, but cells of recessive Lu(a‐b‐) type are reactive. These data establish that synthesis of the Wj antigen is suppressed by the In(Lu) gene, but it is not a part of the Lutheran blood group system. The Wj antigen is not well developed on fetal red cells. This example of auto anti‐Wj is a non‐complement‐binding IgGI protein that did not appear to cause accelerated in vivo destruction of the patient's red cells.
A slide test for detecting neutrophil‐associated complement components
Authors: Harmon D. C. , Weitzman S. A. , Stossel T. P.
Published: December 2004

Complement components deposited on neutrophils were detected by a new test that uses staphylococcal protein A and antisera against complement. Serum from three of 51 patients thought to have immune neutropenia deposited complement but not IgG on their own and normal neutrophils. Some cases of immune neutropenia may be caused primarily by complement‐mediated neutrophil destruction.
A technetium‐99m red cell survival technique for in vivo compatibility testing
Authors: Holt J. T. , Spitalnik S. L. , McMican A. E. , Wilson G. , Blumberg N.
Published: December 2004

A technique for labeling red cells with Technetium‐99m was developed using in vitro reduction by stannous pyrophosphate. The use of Technetium‐99m and Chromium‐51 enabled simultaneous determination of 60‐minute autologous and heterologous red cell survival using small aliquots. In vivo pretransfusion red cell compatibility in a patient with a panagglutinin was identical to that with 51Cr‐labeled cells. Simultaneous testing of two heterologous donor units or testing multiple units in the course of a few days was also feasible. In nine studies (one patient and six healthy controls) no significant elution of the 99mTc was observed over 1 hour.
Use of the manual Polybrene test in the routine hospital laboratory
Authors: Fisher G. A.
Published: December 2004

The manual Polybrene® test described by Lalezari and Jiang was tested in parallel with saline, papain, and antiglobulin tests routinely used in this laboratory for antibody detection and identification and compatibility testing. Among 5381 patient sera tested, 264 antibodies were detected in 244 patients. In all, 6353 units of blood were crossmatched. The manual Polybrene test was generally more sensitive than the other three tests and was much faster. In order to detect anti‐K, a supplementary antiglobulin phase was needed. The manual Polybrene test was less useful in detecting IgM antibodies.
Serum potassium changes following packed red cell transfusions in newborn infants
Authors: Batton D. G. , Maisels M. J. , Shulman G.
Published: December 2004

We studied the effect of packed red cell transfusions on serum potassium levels in sick newborn infants. In 11 transfusions the infants' serum potassium levels fell from a mean of 5.1 ± 1.2 mEq per I (SD) to 4.9 ± 1.2 mEq per I within 1 hour of the transfusion. Plasma potassium in stored packed red cells rose markedly within 48 hours of drawing. In spite of this, the transfusion of 10 ml per kg of packed red cells did not affect the serum potassium concentration in the newborn infant.
Anti‐Cra: family study and survival of chromium‐labeled incompatible red cells in a Spanish‐American patient
Authors: Smith K. J. , Coonce L. S. , South S. F. , Troup G. M.
Published: December 2004

A 22‐year‐old Spanish‐American woman with juvenile rheumatoid arthritis developed anti‐Cra after transfusion during colectomy. No Cra negative family members were found among 13 relatives, including four siblings and both parents. Chromium‐labeled red cell survival studies showed a T½ of 14 days with Cra positive cells. Two units of Cra positive blood were transfused uneventfully for bleeding after ileorectal anastomosis.
In vitro evaluation of a pediatric microaggregate blood filter
Authors: LONGHURST D.M. , GOOCH W.M. , CASTILLO R.A.
Published: December 2004

ABSTRACT: Use of a pediatric stainless steel microaggregate screen filter to filter whole blood, gravity‐sedimented packed cells, and centrifuged packed cells (cPC) from 20 healthy adults resulted in excessive levels of hemolysis (to 1477.9 mg/dl plasma hemoglobin with 5‐day‐old cPC) and unacceptable frequency of occlusion (10 to 40 percent).
Fatal hemolytic disease of a newborn due to anti‐D in an Rh‐positive Du variant mother*
Authors: Lacey P. A. , Caskey C. R. , Werner D. J. , Moulds J. J.
Published: December 2004

This report presents an account of fatal hemolytic disease of the newborn (HDN) due to anti‐D in a mother whose red cells showed the phenotypic characteristics of Rh positive Du variant. The proposita's third pregnancy was uneventful until the eighth month, when she presented with an unusually large abdomen. A sonogram showed fetal hydrops, and amniocentesis yielded a Δ OD450 reading in Zone 3. Anti‐D with a titer of 4096 was identified in the mother's serum. Following delivery by cesarean section, the premature infant did not respond to resuscitation. The autopsy findings were consistent with Rh erythroblastosis fetalis. The proposita has been classified as a Category VI Du variant with anti‐D in her serum.
Storage of platelets collected by apheresis
Authors: Rock G. A. , Blanchette V. S. , Wong S. C.
Published: December 2004

Increasingly, platelets obtained by apheresis must be shipped some distance prior to infusion. We have studied the effect of storage and the size of the storage container on in vitro platelet function over a 48‐hour period. Five donors each donated twice at 2‐week intervals with one donation of 200 ml taken into a 300‐ml bag and the other into a 2000‐ml bag. The platelets collected into the 2000‐ml bag maintained pH well over 48 hours. The Po2 stayed at a high level, and the Pco2 decreased by 48 hours. The lactate accumulation was 38 mg per dl, and the aggregation and hypotonic shock response were maintained well. In comparison, platelets stored in the 300‐ml bag had a rapid decrease in pH accompanied by a marked rise in Pco2. Lactate accumulation was also high at 124 mg per dl at 48 hours. Aggregation, 14C‐serotonin uptake, hypotonic shock response, and platelet recovery were reduced. Exposure to a high concentration of the plasticizer, di‐2‐ethylhexyl phthalate, equivalent to that which occurs in the 2000‐ml bag did not affect platelet function. Storage of the apheresis platelets in a double volume of plasma in the 2000‐ml bag likewise had little effect. The data indicate that, as measured by in vitro response, apheresis platelets can be maintained for up to 48 hours if stored in a 2000‐ml bag.
The surge technique: a method to increase purity of platelet concentrates obtained by centrifugal apheresis
Authors: Schoendorfer D. W. , Hansen L. E. , Kenney D. M.
Published: December 2004

A surge technique has been developed to increase the purity of high‐yield platelet concentrations prepared on a blood processor with the Latham bowl. The surge technique combines elutriation with centrifugal separation, utilizing plasma recirculated from the plasma/air bag back into the centrifuge bowl to elute platelets from the red cell mass. Platelet concentrates prepared by surge collection with six separation cycles (n = 22), contained an average of 3.9±1.4X1011 platelets, with0.15±0.11 X109 leukocytes, and red cells below the level of detection. The surge technique reduces collection time by 4 minutes per cycle and eliminates the need for a secondary centrifugation; thus, 96 minutes donor processing time would permit eight separation cycles. Platelets collected by the surge technique exhibited unaltered morphology and capacity to take up radioactively labeled serotonin in vitro compared to pre‐apheresis controls. Results from in vitro functional studies also indicate that the capacity of platelets collected with the surge technique to respond to various concentrations of adenosine diphosphate, collagen, and thrombin by aggregation and secretion of both serotonin (dense bodies) and β‐thromboglobulin (α‐granules) was not significantly different (p ≤ 0.05) from that of pre‐apheresis controls.
Iron balance in superdonors
Authors: Monsen E. R. , Critchlow C. W. , Finch C. A. , Donohue D. M.
Published: December 2004

Volunteers who are able to donate blood frequently without becoming anemic provide a unique opportunity to observe iron balance under conditions of controlled blood loss. The 88 men and 88 women studied had given a mean of 17 donations (range 10–24) over 4 years. Twenty‐seven percent of men and 45 percent of women took iron‐containing supplements. In these “superdonors,” estimated iron loss averaged 3.8 mg/day in men, 3.4 mg/day in menstruating women and 3.0 mg/day in nonmenstruating women. Dietary iron consumption estimated from the dietary history averaged 17 mg/day for men and 13 mg/day for women; in general, the superdonors consumed a superior diet. Sixty‐three percent of the unsupplemented individuals had ferritin values less than 20 μg/l. Assuming that stores were constant, these individuals would have to absorb 25 percent of their dietary iron to meet total iron loss. Supplemented men and women taking an average of 24 and 34 mg of additional iron per day, respectively, showed relatively little difference in iron balance from unsupplemented individuals; 58 percent had serum ferritin values of less than 20 μg/l, and 28 percent were equal to or less than 12 μg/l. The low level of absorption (6–9%) in supplemented individuals suggests that the iron availability of iron‐containing preparations ingested may have been poor.
Is a room‐temperature crossmatch necessary for the detection of ABO errors?
Authors: Trudeau L. R. , Judd W. J. , Butch S. H. , Oberman H. A.
Published: December 2004

The detection of anti‐A and anti‐B isohemagglutinins by low‐ionic‐strength saline tests at 37°C and by the indirect antiglobulin technique, without an “immediate'spin” or room‐temperature phase, has been studied. Using such a procedure, all but one of 2746 patient blood samples reacted in accordance with ABO type when tested against A2 and B red cells. However, the discrepant sample also was nonreactive when tested by “immediate‐spin” technique against saline‐suspended A2 red cells. Our findings indicate that compatibility tests performed at 37°C in low‐ionic‐strength saline are as sensitive as “immediate‐spin” tests with saline‐suspended red cells for the detection of ABO errors. Performing serologic tests for unexpected alloantibodies and donor‐recipient compatibility without an “immediate‐spin” or room‐temperature phase abbreviates pretransfusion testing and reduces the detection of clinically insignificant alloantibodies solely reactive at room temperature.
Identification and quantitation of the protein content of blood group antibody eluates prepared by the rapid acid‐stromal method
Authors: Rearden A. , Chiu P. , Elmajian D.
Published: December 2004

Anti‐D sensitized red cells were converted to stroma, either by digitonin or hypotonic lysis, and eluates prepared by the modified rapid acid‐stromal method. SDS‐PAGE and both rocket‐ and crossed‐immunoelectrophoresis showed that, of the total protein content of the eluates, 11 to 25 percent was IgG, 18 to 45 percent was globin, and 34 to 71 percent was red cell membrane protein. Eluates were free of albumin, transferrin, and other non‐IgG serum proteins; they did not contain proteolytic activity, and eluates were noninhibitory to the growth of committed erythroid progenitors. The rapid acid‐stromal method produced high‐titered blood group antibody eluates free of contaminating immune or nonimmune activity which were suitable for assay of blood group antigen expression by hemopoietic stem cells and committed progenitors.
Prepooled cryoprecipltate for treatment of hemophilia A*
Authors: MENITOVE J.E. , SPERO J.A. , RICHAROS W.A. , LEWIS J.H. , MONTGOMERY R.R. , GILL J.C. , HASIBA U. , WHEELER D.V. , SHERWIN J.A. , RUSSELL‐VASKOV S. , MARLAR R.A. , MAXWELL N.G. , ASTER R.H.
Published: December 2004

ABSTRACT: Lyophllized factor VIII concentrates, because of their convenience, are the most commonly used products for providing antihemophilic factor to patients with hemophilia A. Heightened concerns about associated complications indicate that cryoprecipitate usage will increase in the immediate future. We present a protocol for making cryoprecipitate more usable by “prepooling” individual units of cryoprecipitate in an “open” system. Our experience indicates this approach is safe, effective and acceptable to patients, including those on home therapy programs.
Comparison of antibody elution techniques by enzyme‐linked antiglobulin test
Authors: Gibble J. W. , Salamon J. L. , Ness P. M.
Published: December 2004

The enzyme‐linked antiglobulin test was used to determine the percentage of antibody removed from sensitized red cells by five elution methods: Rubin ether, xylene, digitonin‐acid, glycine, and heat. Antibodies examined in the study included anti‐D, ‐c, ‐E, and ‐K. With two examples of anti‐D, more antibody was eluted by the Rubin ether method (45.5% average) than the xylene (38%) or the digitonin acid method (35%) (p < 0.05); the glycine (8%) and heat method (15%) were less efficient. With one example of anti‐E and one example of anti‐c, more antibody was eluted by the ether method. The percentage of anti‐K recovery, however, was greater with the digitonin acid method (45%) than with the ether (30%). We found the enzyme‐linked antiglobulin test method useful in the quantitative evaluation of elution procedures. TRANSFUSION1983;23:300–304.
Enzyme‐linked immunosorbent microtiter assay for red cell serology
Authors: Reppun T. , McIlravy D. , Sherman L. A.
Published: December 2004

Enzyme‐linked immunoabsorbent assays (EIA) permit quantitation and automation of reactions. Previous reports have also shown improved sensitivity for red cell antibodies. We have developed a microtiter technique for EIA, thus decreasing the amounts of reagents required. EDTA was used rather than NaOH to stop the enzyme reaction allowing more precise timing of reactions and improved quantitation of large batches of tests. As in prior reports, the sensitivity was slightly greater than the antiglobulin test, but occasional red cell antibodies failed to show specificity by EIA. The use of microtiter plates and EDTA decreased the steps and reagents required and has the potential for automation of at least the optical reading of such assays in the blood bank.
A computer system designed for a hospital transfusion service
Authors: Reich L. M. , Mitchell M. J. B. , Jambois W. H. , Hoffer J. L. , Reps D. N. , Mayer K.
Published: December 2004

A computer system that enhances the efficiency of a hospital transfusion service is described. It reduces the possibility of clerical errors; shortens the time elapsed between the ordering and the delivery of blood products; decreases the outdating of these units; and provides easy access to medical, demographic, and statistical information.
An individual with McLeod syndrome and the Kell blood group antigen K(K1)
Authors: Marsh W. L. , Schnipper E. F. , Johnson C. L. , Mueller K. A. , Schwartz S. A.
Published: December 2004

McLeod syndrome is an X‐linked condition in which individuals of McLeod blood group phenotype have weak Kell antigens, acanthocytic red cells, and a muscular disorder. We now report a family in which two brothers have McLeod syndrome. One is K:‐1, while the other is the first known K:1 person with McLeod syndrome. The K1 gene in the latter is expressed weakly and was inherited from the father, in whom it is expressed normally. The brothers have the same clinical and laboratory manifestations of McLeod syndrome but have different Kell genes. Therefore, the Kell gene is unlikely to have any positive input into development of McLeod syndrome; its role is one of passive involvement in which its expression is modified.
Changes in protein‐bound, complex‐bound, and ionized calcium related to parathyroid hormone levels in healthy donors during plateletapheresis
Authors: Toffaletti J.
Published: December 2004

In order to relate changes in protein‐bound and complex‐bound calcium to levels of citrate and secretion of parathyroid hormone during mild hypocalcemia, total, ultrafiltrable, and ionized calcium were measured in 13 apparently healthy donors during plateletapheresis. Carboxyl‐terminal parathyroid hormone was measured in seven of the donors, and both amino‐terminal and intact parathyroid hormone assays were done on one donor. Mean concentrations of total and ionized calcium decreased during most of the procedure, as did total protein, albumin, and phosphorus. Complexed calcium increased, which was attributed to the large increases in citrate levels. Protein‐bound calcium decreased more than other forms of calcium, even allowing for the decrease expected from hemodilution. The carboxyl‐terminal parathyroid hormone response varied among different donors, but on the average, decreased slightly before gradually rising during the citrate‐induced hypocalcemia. These findings suggest that calcium citrate does not affect the secretion of parathyroid hormone, and that at least a portion of protein‐bound calcium is readily exchangeable and possibly influenced by the hormone.
Characterization of biochemical changes occurring during storage of red cells
Authors: Moroff G. , Dende D.
Published: December 2004

Citrate‐phosphate‐dextrose‐adenine (CPDA‐1), containing 0.25 mM adenine (final concentration) and 25 percent more glucose than citrate‐phosphate‐dextrose (CPD), has extended the allowable storage time for red cells to 35 days. Studies were conducted to understand better the characteristics of stored CPDA‐1 red cells in relation to the properties of stored CPD red cells. Units with hematocrits near 80 percent showed the following: First, adenosine triphosphate (ATP) and total adenine nucleotide levels of red cells stored with CPDA‐1 remained essentially constant during the first 3 weeks of storage after which the levels decreased; with red cells stored with CPD, ATP, and adenine nucleotide, levels were decreased even after 1 week of storage. Second, the pattern of the fall in 2,3‐diphosphoglycerate was similar in red cells stored with CPD and CPDA‐1. Third, changes in plasma and red cell levels of sodium and potassium, and in plasma ammonia levels, were comparable in CPD and CPDA‐1 units; changes in cation levels were most pronounced during the initial 2 weeks of storage. Fourth, hemolysis was much greater in units stored in CPDA‐1 for 35 days than in units stored in CPD for 21 days. Fifth, residual glucose concentrations were adequate in units drawn in CPDA‐1 and stored for 35 days. We conclude that the changes in the biochemical characteristics of units of red cells stored with CPD and CPDA‐1 are similar in most instances with the notable exception of the better maintenance of adenosine triphosphate levels in red cells stored with CPDA‐1.
Range forb lectins
Authors: Hardman J. T. , Beck M. L. , Owensby C. E.
Published: December 2004

In this study we tested the seeds of uncultivated plants for lectin activity. Extracts prepared from the seeds of 167 forb species were tested against human red cell samples. Extracts of 54 species agglutinated unmodified or enzyme‐modified red cells. In nine species, the hemagglutinating lectin activity could be neutralized by N‐acetyl‐D‐galactosamine, mannose, or glucose.
Parathyroid response during therapeutic plasma exchange
Authors: BONGIOVANNI M.B. , STRAUSS J.E. , ZISELMAN E.M. , WURZEL H.A.
Published: December 2004

ABSTRACT: Significant hypocalcemia is infrequently encountered during therapeutic plasma exchange. An increase in parathyroid hormone secretion during the procedure may be responsible for maintaining the calcium levels.
Testing for the D zygosity with three different methods revealed altered Rhesus boxes and a new weak D type
Authors: Perco Paul , Shao Chao‐Peng , Mayr Wolfgang Richard , Panzer Simon , Legler Tobias Jörg
Published: December 2004

BACKGROUND
: The discrimination of D+/D+ from D+/D– partners of D– mothers with anti‐D is important to estimate the risk for HDN. This may be achieved if the presence or absence of the hybrid
Rhesus box in the father can be demonstrated.

STUDY DESIGN AND METHODS
: A new PCR‐SSP method specific for the hybrid
Rhesus box comprising an internal amplification control was compared with two published PCR‐based methods (PCR‐SSP and PCR‐RFLP) in 83 D+, 13 D–, and 37 weak D samples.

RESULTS
: The deletion of
RHD
was detectable in all D– and weak D samples. By all three methods, concordant results were obtained in 82 of 83 D+ samples, with one sample showing discrepant results. The control band in the PCR‐RFLP method, specific for the downstream
Rhesus box,
was missing in two weak D samples, namely a weak D type 4.0 and a novel weak D type dubbed weak D type 29. Further investigations revealed an altered downstream
Rhesus box
in the weak D type 29 sample. In the weak D type 4.0 sample, no amplicon was achieved with any primer specific for the upstream and downstream
Rhesus box.

CONCLUSION
: A PCR‐SSP method with internal control was established for the detection of the hybrid
Rhesus box.
Polymorphisms in the downstream
Rhesus box
may interfere with the detection of
RHD.
The development of a quantitative ELISA for antibodies against human platelet antigen type 1a
Authors: Bessos Hagop , Perez Silvia , Armstrong‐Fisher Sylvia , Urbaniak Stan , Turner Marc
Published: December 2004

BACKGROUND : Severe neonatal alloimmune thrombocytopenia is often due to antibodies against human platelet antigen type 1a (HPA‐1a). The aim of this study was to develop a quantitative ELISA for the measurement of antibodies against HPA‐1a.

STUDY DESIGN AND METHODS : HPA‐1a glycoprotein (GP) IIb‐IIIa was immobilized and mixed with recalcified anti‐HPA‐1a‐positive plasma overnight at 4°C. The beads were washed, the antibodies against HPA‐1a were eluted, and the eluate pH level was promptly adjusted. The purified antibodies were dialyzed and used for the development of an ELISA incorporating HPA‐1a‐coated plates.

RESULTS
: Serial doubling dilutions of the purified antibodies resulted in consistent sigmoid standard curves with a sensitivity of 0.5 μg per mL. To determine the reproducibility of the ELISA, antibodies against HPA‐1a in five plasma samples (Samples A‐E) were measured at serial doubling dilutions in four separate assays. Three of the samples (Samples A‐C) contained antibodies against HPA‐1a. The mean amounts in μg per mL (
± SD, percentage of CV) obtained in the four assays were as follows: Sample A, 133 (9.4, 7.1%); Sample B, 16.5 (1.7, 10%); and Sample C, 8 (0.8, 10%). The amounts in the two antibody‐negative controls (Samples D and E) were consistently less than 0.2 μg per mL.

CONCLUSION : Using immobilized HPA‐1a1a, antibodies against HPA‐1a has been purified, and a quick and simple quantitative assay has been developed.
The impact of intensive serial plasmapheresis and iron supplementation on iron metabolism and Hb concentration in menstruating women: a prospective randomized placebo‐controlled double‐blind study
Authors: Bier‐Ulrich Anne‐Marie , Haubelt Hannelore , Anders Christof , Nagel Dietmar , Schneider Steffen , Siegler Karl E. , Seiler Dieter , Hellstern Peter
Published: December 2004

BACKGROUND : Iron (Fe) depletion is common among regular whole‐blood donors, but can be prevented through regular oral Fe supplementation. Little is known, however, about the Fe metabolism of donors undergoing intensive plasmapheresis. These donors lose considerable amounts of blood drawn for laboratory analyses and remaining in the disposable plastic sets.

STUDY DESIGN AND METHODS : Menstruating women were enrolled in a prospective placebo‐ controlled double‐blind study. One hundred women were randomly allocated to receive either 100 mg of elemental Fe per day or placebo over 24 weeks and asked to donate plasma at 1‐week intervals. Hb was determined before each plasmapheresis. Ferritin, transferrin, and Fe concentration and reticulocyte count were measured every 4 weeks.

RESULTS : Thirty donors in the placebo group and 29 receiving Fe completed the study. The total mean blood loss was 526 mL in the placebo group and 546 mL in the Fe arm (p = 0.271). The number of donations with Hb values lower than 12.5 g per dL requiring prolongation of the time interval until the next plasmapheresis was significantly greater in the placebo arm. In the placebo group (n = 30), ferritin levels began to decline significantly 4 weeks after entry. When the study was completed, Hb concentration and reticulocyte count also were found to be significantly lower in the placebo group than in the Fe study arm (p = 0.028 and p = 0.036, respectively). Hb, ferritin, and transferrin levels and reticulocyte counts did not change significantly in the Fe group during the observation period.

CONCLUSION : Menstruating women undergoing regular plasmapheresis at short intervals are prone to develop Fe depletion. This can be prevented by regular Fe intake. Laboratory analyses in product plasma instead of serum gained from whole‐blood samples could be an alternative to reduce blood loss.
Factors that influence platelet recovery after transfusion: resolving donor quality from ABO compatibility
Authors: Jiménez Teresa M. , Patel Sachin B. , Pineda Alvaro A. , Tefferi Ayalew , Owen Whyte G.
Published: December 2004

BACKGROUND : A system was established to examine the extent to which the apheresis donor determines platelet recovery after transfusion, to measure the impact of ABO identity, and to predict outcome by evaluating the donor.

STUDY DESIGN AND METHODS : The percentage of platelet recovery was measured after prophylactic transfusion of apheresis units divided from single donors to paired recipients with uncomplicated thrombocytopenia secondary to leukemia chemotherapy. Platelet microaggregation induced by citrate was measured at the time of apheresis.

RESULTS : Platelet recoveries in paired recipients correlated strongly when both transfusions were ABO‐ identical. When one recipient was ABO‐identical and the other was ABO‐nonidentical, nonidentical transfusions yielded one‐third the recovery of ABO‐identical transfusions. In ABO‐identical transfusions, platelet recovery in donors having microaggregates in the before‐apheresis ACD sample was one‐third that in donors without microaggregates. This difference was observed at 1 and 24 hours. Expression of P‐selectin in the apheresis units at the time of transfusion correlated well with ACD microaggregates in the before‐apheresis sample.

CONCLUSION : When transfusions of platelets are ABO‐identical, donor quality dominates recovery in circulation. Donor quality is predicted by a rapid and simple assay of citrate‐induced microaggregation performed at the time of apheresis. When donor quality is factored out, ABO identity prevails.
Frequency and duration of plasma CMV viremia in seroconverting blood donors and recipients
Authors: Drew W. Lawrence , Tegtmeier Gary , Alter Harvey J. , Laycock Megan E. , Miner Richard C. , Busch Michael P.
Published: December 2004

BACKGROUND : Both CMV‐seronegative blood and unscreened, filtered blood carry a low but definite risk of transmitting CMV infection. To explain this residual risk, evidence of cell‐free viremia was sought in seroconverting and seroprevalent blood donors and seroconverting transfusion recipients by means of a plasma‐based assay for CMV DNA.

STUDY DESIGN AND METHODS : A CMV DNA PCR assay (COBAS Amplicor CMV Monitor, Roche) was used to detect CMV DNA in 384 paired plasma samples from 192 donors who seroconverted to anti‐CMV, 488 anti‐CMV EIA‐positive samples from 60 seroprevalent donors, and 113 serial samples from 11 seroconverting recipients with posttransfusion CMV hepatitis.

RESULTS : Three of 384 samples from 192 seroconverting donors had low levels of plasma CMV DNA (400‐1600 copies/mL); one donor was positive before seroconversion, and the other two, after seroconversion. None of the 488 serial samples from 60 anti‐CMV‐ positive donors contained CMV DNA in plasma. Three of 11 recipients demonstrated transient plasma viremia that temporally coincided with seroconversion.

CONCLUSIONS : Plasma CMV DNA was detected in a small percentage of seroconverting blood donors and a larger percentage of recipients but was undetectable in seroprevalent donors. Plasma viremia in seroconverting donors may partially explain the low residual risk of CMV transmission by both CMV‐seronegative and WBC‐reduced seropositive blood.
CD34+CD38– is a good predictive marker of cloning ability and expansion potential of CD34+ cord blood cells
Authors: Encabo Araceli , Mateu Emilia , Carbonell‐Uberos Francisco , Miñana María Dolores
Published: December 2004

BACKGROUND : Ex vivo expansion of HPCs is an attractive approach to overcoming the current limitations of human cord blood transplantation. It is important not only to define the optimal culture conditions but also to know the number of progenitor cells that can be obtained. CD34+ cells have a great variability in their cloning capacity and in their ability to expand HPCs. This study was carried out to assess whether this variability could be due to intrinsic or extrinsic factors.

STUDY DESIGN AND METHODS : CD34+ cells were analyzed for the expression of CD38, CD133, and CD117 and cultured in serum‐free culture medium with four cytokine combinations: SCF plus thrombopoietin plus flt3 ligand (STF), STF plus IL‐3, STF plus IL‐6, and STF plus IL‐6 plus IL‐3. After a 1‐week culture, the numbers of CD34+ cells and CFUs were determined.

RESULTS : The variability observed both in the cloning ability of CD34+ isolated cells and in their expansion capacity was inversely related to the frequency of the more immature CD34+CD38– cells. When more mature CD34+CD38+ cells were present within CD34+‐isolated cells, a higher cloning ability, measured as CFUs, and a higher expansion capacity were observed.

CONCLUSION : Enumeration of CD34+CD38– cells is correlated with the number of committed progenitors and the capacity of generating CD34+ cells, an important parameter if expansion protocols must be used in clinical transplantation.
Rapid and reliable genotyping of human platelet antigen (HPA)‐1, ‐2, ‐3, ‐4, and ‐5 a/b and Gov a/b by melting curve analysis
Authors: Randen Ingrid , Sørensen Kirsten , Killie Mette K. , Kjeldsen‐Kragh Jens
Published: December 2004

BACKGROUND: The probability for occurrence of neonatal alloimmune thrombocytopenic purpura (NAITP) depends largely on the frequency of each individual phenotype in various populations. In caucasians, antibodies to human platelet antigen (HPA)‐1a are the major cause of neonatal alloimmune thrombocytopenic purpura, whereas in the Japanese population, antibodies to HPA‐4b is most frequently involved in NAITP. Conventional PCR techniques for platelet antigen genotyping rely on sequence‐specific primers (SSPs) and detection by gel electrophoresis, a method which is laborious and time consuming. New PCR technology, measuring the match of a hybridization probe with its target and thereby allowing simultaneous detection of both alleles, provides an efficient tool for genotyping of the HPA systems.

STUDY DESIGN AND METHODS: A total of 105 healthy blood donors were genotyped for HPA‐1, ‐2, ‐3, ‐4, and ‐5 a/b and Gov a/b with new primers and probes designed for mutation detection by melting curve analysis (using LightCycler technology). Donor DNA was independently genotyped by an allele‐specific assay, using SSPs, in a reference laboratory.

RESULTS: There was full concordance between the two genotyping methods, and genotype frequencies were comparable with previous studies in caucasians.

CONCLUSION: We present rapid and reliable detection systems for HPA‐1, ‐2, ‐3, ‐4, and ‐5 a/b and Gov a/b based on mutation detection of both alleles simultaneously by melting curve analysis. As the Gov system has been reported to have similar frequency of involvement in alloimmune thrombocytopenia as HPA‐5, the opportunity for genotyping should aid the diagnosis of such patients.
Fluctuation of HCV viral load before seroconversion in a healthy volunteer blood donor
Authors: Fang Chyang T. , Tobler Leslie H. , Haesche Carmen , Busch Michael P. , Phelps Bruce , Leparc German
Published: December 2004

BACKGROUND: Newly implemented NAT has been shown to be able to effectively identify HCV‐positive blood donated during the preseroconversion period.

STUDY DESIGN AND METHODS: EDTA‐plasma pools of 24 donations were tested using an HIV‐1/HCV multiplex NAT under an FDA‐approved IND application. Samples in a positive pool were retested individually. Positive samples were further tested by two discriminatory assays to determine specific viral reactivity. Upon obtaining informed consent, seronegative donors with positive NAT results were enrolled into a follow‐up study for risk factor analysis and laboratory testing.

RESULTS: A donation by a 29 year old female was identified as HCV NAT‐positive with negative serology and an elevated ALT. Her two previous donations, 5 and 12 months earlier, were both seronegative and with normal ALT. Her husband tested positive for HCV RNA. The donor remained seronegative for at least 36 days. The index donation had a viral RNA concentration of >500,000 copies per mL while the first seropositive sample was NAT‐negative. Laboratory data on serial follow‐up samples showed 100‐ to 1,000‐fold fluctuations in viral load during a period of 48 days prior to seroconversion.

CONCLUSIONS: This case suggests that, at least in some newly infected individuals, the HCV viral load can fluctuate dramatically prior to seroconversion, and that NAT, even on individual samples, will not totally prevent HCV transmission.
WBC reduction in cryopreserved RBC units
Authors: Arnaud Françoise G. , Meryman Harold T.
Published: December 2004

BACKGROUND: WBC reduction of blood components by filtration is widely practiced to decrease the incidence of alloimmunization. Freezing RBCs reduces the WBC load but is insufficient to achieve the currently recommended US limit of 5 × 106 cells per unit.

STUDY DESIGN AND METHODS: Blood units were WBC reduced by filtration or by buffy‐coat (BC) removal and then frozen in the presence of a high‐glycerol concentration. The count of residual WBCs was determined by flow cytometry after deglycerolization.

RESULTS: Without WBC reduction, the total number of WBCs present after freezing and thawing was 11.5 ± 9.2 × 106 WBCs per unit (n = 18). Particulate residues from monocytes and neutrophils that were detected in the remaining cell populations were positive for CD66b, CD3, CD14, and CD41. Removal of 40 mL of BC at the time of blood collection lowered the number of WBCs after freezing and deglycerolization to 1.9 ± 1.20 × 106 per unit (n = 11). Similar results were obtained when only 20 mL of BC was removed using a modified blood‐bag design. Unfiltered RBC units that were stored for 15 days at 4°C after BC removal contained fewer than 5 × 106 WBCs after deglycerolization. Units WBC reduced by filtration before freezing had no detectable WBCs after thawing and washing (n = 14) and did not contain particulate residues. Filtration after deglycerolization was effective in reducing the WBC count below 106, although some debris was still present.

CONCLUSION: RBC freezing alone will not reduce residual counts to recommended levels. However, initial removal of BC can provide an economical alternative to WBC filtration for cryopreserved units. Units that were not WBC reduced before freezing can be filtered after deglycerolization when needed.
Effects of X‐ray radiation on the rheologic properties of platelets and lymphocytes
Authors: Thomas Suny , Bolch Wesley , Kao Kuo Jang , Bova Frank , Tran‐Son‐Tay Roger
Published: December 2004

BACKGROUND: Blood is often irradiated before transfusion for severely immunocompromised patients to prevent a potentially fatal complication of transfusion‐ associated GVHD.

STUDY DESIGN AND METHODS: This study evaluates the effects of X‐ray radiation on platelet and lymphocyte rheology because the ability of these blood cells to deform is vital to their flow throughout the microvascular system. Micropipette aspiration experiments were conducted on platelets and lymphocytes exposed to X‐ray radiation doses of 0 (control), 25, and 50 Gy.

RESULTS: A significant increase in the Young modulus of elasticity was observed between control platelets and irradiated platelets at 25 Gy (p = 0.02) and 50 Gy (p = 0.03). Percent cell activation increased significantly in 25 Gy‐irradiated platelets (p = 0.008). In addition, lymphocytes irradiated at 25 Gy have a higher viscosity than controls (p < 0.02). A significantly larger number of activated cells is found in the 50 Gy‐irradiated lymphocyte population (p < 0.04).

CONCLUSION: The changes in the deformability and activation of irradiated platelets and lymphocytes may reduce local blood flow and lead to intermittent blockage, which may cause a change of blood flow in microvasculatures.
Production of recombinant murine‐human chimeric IgM and IgG anti‐Jsb for use in the clinical laboratory
Authors: Huang T.J. , Reid M.E. , Halverson G.R. , Yazdanbakhsh K.
Published: December 2004

BACKGROUND: Directly agglutinating MoAbs are more useful than IgG MoAbs of murine origin for typing RBCs from donors and patients. The molecular manipulation and conversion of a murine IgG MoAb into mouse‐ human chimeric IgM and IgG antibodies are described.

STUDY DESIGN AND METHODS: cDNA encoding the variable heavy‐ and light‐chain genes of a murine hybridoma anti‐Jsb cell line (MIMA‐8) were cloned into human IgM or IgG expression vectors, which were then separately stably transfected into SP2/0‐Ag14 B‐cells. The secreted antibodies were screened by ELISA and analyzed by flow cytometry and hemagglutination.

RESULTS: Forty percent (16 of 40) of the stable clones secreted IgM and 66 percent (12 of 18) of the stable clones secreted IgG. The chimeric IgM from the highest expressing clone reacted 4+ in LISS at room temperature. The chimeric IgG from one clone reacted 4+ by the IAT, resembling the specificity of the original murine antibody. Both manipulated MoAbs reacted specifically with RBCs as assessed by flow cytometry.

CONCLUSION: Human‐mouse chimeric IgM and IgG from a murine IgG MoAb anti‐Jsb has been successfully engineered for use in the clinical laboratory. This approach can potentially be used to manipulate other murine MoAbs to blood group antigens into more clinically useful human isotypes.
Longitudinal thrombopoietin plasma concentrations in fetuses with alloimmune thrombocytopenia treated with intrauterine PLT transfusions
Authors: Cremer Malte , Dame Christof , Schaeffer Hans J. , Giers Guenther , Bartmann Peter , Bald Rainer
Published: December 2004

BACKGROUND:  The purpose of this study was to describe longitudinal thrombopoietin (TPO) plasma concentrations in fetuses with fetomaternal alloimmune thrombocytopenia (FMAIT).

STUDY DESIGN AND METHODS:  Group 1 was the control group, 8 fetuses with normal hematopoiesis. Group 2 consisted of 4 nonthrombocytopenic fetuses with fetomaternal human PLT antigen incompatibility. Group 3 consisted of 14 fetuses with prenatal‐diagnosed severe FMAIT owing to human PLT antigen‐1a incompatibility. Fetal PLT counts, MoAb‐specific immobilization of PLT antigen score, and TPO plasma concentrations were measured in a total number of 94 serial samples taken by cordocentesis before intrauterine PLT transfusion.

RESULTS:  Normal fetal TPO plasma concentrations ranged between 15 and 119 pg per mL (Group 1 median, 67 pg/mL). In fetuses with risk of FMAIT but normal PLT counts, TPO concentrations were normal (Group 2 median, 72 pg/mL; range, <15‐158 pg/mL). In FMAIT with thrombocytopenia, the median TPO concentration was significantly higher than in Groups 1 and 2 (Group 3 median, 172 pg/mL; range, 15‐623 pg/mL; p < 0.001). In the longitudinal analysis, TPO concentrations remained constant (n = 8), peaked only transiently (n = 3), or increased at the end of gestation (n = 3). Elevated TPO concentrations (592 and 623 pg/mL) were detected in one patient, who already had intracranial hemorrhage in utero.

CONCLUSION:  TPO concentrations are normal or slightly elevated in FMAIT. Further clinical information can be provided by the longitudinal analysis of TPO concentrations in severe FMAIT.
A unique 502C>T mutation in exon 7 of ABO gene associated with the Bel phenotype in Taiwan
Authors: Lin P.‐H. , Li L. , Lin‐Tsai S.‐J. , Lin K.‐T. , Chen J.‐M. , Chu D.‐C.
Published: December 2004

BACKGROUND:  The ABO system includes many variant subgroups. Some of them are difficult to identify serologically, leading to mistyping of blood groups. For example, Bel is often typed as O blood group.

STUDY DESIGN AND METHODS:  DNA sequencing and a molecular approach were explored to accurately determine the genotypes of Bel subgroups. Seven Bel blood donors and 106 individuals with other blood groups were analyzed serologically and molecularly.

RESULTS:  The serologic results of these seven Bel blood donors showed that their RBCs do not react with anti‐B or anti‐A,B, and their B antigen was detected by adsorption and elution methods. Sequencing results for exons 6 and 7 of ABO genes showed a new Bel allele with a C>T substitution at nucleotide position 502 in exon 7 of the ABO gene in all seven cases but not in other blood groups. Consequently, an amplification‐created restriction site protocol was designed to detect the 502C>T genotype in Bel subgroup cases.

CONCLUSION:  A novel 502C>T mutation was found in the Bel subgroup in Taiwan and successfully developed a rapid and accurate molecular protocol to detect this mutation. To our knowledge, the new Bel allele that was found is unique in Taiwanese residents.
A simple controlled‐rate freezing method without a rate‐controlled programmed freezer provides optimal conditions for both large‐scale and small‐scale cryopreservation of umbilical cord blood cells
Authors: Itoh Tsuneo , Minegishi Masayoshi , Fushimi Junko , Takahashi Hiroyuki , Kudo Yoshinori , Suzuki Akira , Narita Ayuko , Sato Yuko , Akagi Kozo , Wada Yuichi , Saito Akira , Kikuchi Miwako , Okamura Kunihiro , Kaku Mitsuo , Tsuchiya Shigeru
Published: December 2004

BACKGROUND:  Umbilical cord blood (CB) is being used as a source of alternative HPCs for transplantation with increasing frequency. The goal of CB banks for unrelated transplantation is to provide good quality‐controlled CB units that can be transplanted for HPCs into the largest possible number of patients.

STUDY DESIGN AND METHODS:  Large CB samples in freezing bags wrapped with insulators and small samples in cryotubes placed into double styrene‐foam boxes were cryopreserved at −85°C without a rate‐controlled freezing machine, followed by storage in the liquid phase of nitrogen. After thawing these cells, the viability and recovery of cells, as well as the recovery rate of HPCs such as CD34+ cells, CFU‐GM, and total CFU were evaluated.

RESULTS:  Measurement of the freezing rate in CB bags and cryotubes demonstrated that this simple method for cryopreservation of CB cells provided optimal conditions for both large‐scale and small‐scale cryopreservation. Recovery of CB progenitor cells after cryopreservation was also shown to be potentially acceptable when evaluated with CD34+ cells, CFU‐GM, and total CFU. These results were comparable to the method using a rate‐controlled programmed freezer.

CONCLUSIONS:  A simple method for cryopreservation of CB cells without a rate‐controlled programmed freezer could provide a sufficient‐enough potential for the transplantability of HPCs after thawing.
Acquired deficit of forebrain glucocorticoid receptor produces depression-like changes in adrenal axis regulation and behavior
Authors: Boyle Maureen P. , Brewer Judson A. , Funatsu Michiyo , Wozniak David F. , Tsien Joe Z. , Izumi Yukitoshi , Muglia Louis J.
Published: December 2004

Dysregulation of the hypothalamic–pituitary–adrenal (HPA) axis is a hallmark of major depressive disorder. A number of studies have shown that this dysregulation is correlated with impaired forebrain glucocorticoid receptor (GR) function. To determine whether a primary, acquired deficit in forebrain GR signaling is an etiologic factor in the pathogenesis of depression, we generated a line of mice with time-dependent, forebrain-specific disruption of GR (FBGRKO). These mice develop a number of both physiological and behavioral abnormalities that mimic major depressive disorder in humans, including hyperactivity of the HPA axis, impaired negative feedback regulation of the HPA axis and, increased depression-like behavior. Importantly, a number of these abnormalities are normalized by chronic treatment with the tricyclic antidepressant, imipramine. Our findings suggest that imipramine's proposed activities on forebrain GR function are not essential for its antidepressant effects, and that alteration in GR expression may play a causative role in disease onset of major depressive disorder.