bluuebunny/crossref_metadata_2025 huggingface dataset:
The storage of washed human erythrocytes in a protein‐free medium
Authors: Silbert J. A.
Published: December 2004

Protein‐free red blood cells were prepared by washing fresh citrate‐phosphate‐dextrose anticoagulated whole blood and resuspending the cells in a medium containing saline, adenine, phosphate, and glucose (APG). The protein concentration in the APG medium was reduced to 0.0024 percent of the original plasma protein. The 24‐hour red blood cell posttransfusion survival after 21 days of storage was 80 percent. Extracellular potassium concentrations were less than the CPD‐plasma control after 21 days of storage. Adenosine triphosphate (ATP) concentrations were lower after storage in APG, but intracellular ATP was not closely associated with posttransfusion survival. Hemolysis was slightly increased in the APG group. There appear to be no insurmountable barriers to the use of protein‐free media for the storage of washed red blood cells
Serologic and IgG subclass characterization of Cartwright (Yt) and Gerbich (Ge) antibodies
Authors: Vengelen‐Tyler V. , Morel P. A.
Published: December 2004

Examples of anti‐Yta and anti‐Ge were tested for reactivity with red cells treated with proteolytic enzymes and with antisera of known IgG specificity. Eight of 14 examples of anti‐Yta did not react as well with cells treated with either papain or ficin; treatment with trypsin did not reduce reactivity. Four examples of the anti‐Yta were IgG4; the others tested did not react with IgG subclass antisera. Of 13 examples of anti‐Ge, seven failed to react with red cells treated with trypsin, ficin, or papain. Nine examples were IgG1, one was IgG1 plus IgG3, and three did not react with IgG subclass antisera.
An anti‐neutrophil antibody associated with a propylthiouracil‐induced lupus‐like syndrome
Authors: Berkman E. M. , Orlin J. B. , Wolfsdorf J.
Published: December 2004

A 15‐year‐old woman with thyrotoxicosis controlled by propylthiouracil presented with chills, fever, splenomegaly, anemia, thrombocytopenia, leukopenia, hypergammaglobu‐linemia, immune complexes, a positive anti‐nuclear antibody test, and a cellular marrow with normal maturation. Anti‐neutrophil antibody was detected by cytotoxicity testing. The activity was restricted to the IgM fraction and was absorbed optimally at 4° C. The antibody activity was recovered in both heat and ether eluates made from granulocytes. Lymphocytes, platelets, and red blood cells failed to absorb reactivity. The antibody did not inhibit superoxide production or bacterial killing. Propylthiouracil was discontinued and all signs and symptoms resolved.
Complement activation by plasma separator membranes
Authors: McLeod B. C. , Viernes A. , Sassetti R. J.
Published: December 2004

Because of their structural similarity to dialysis membranes, six plasma separator membranes were evaluated for the ability to activate complement, as judged by immunoconversion of the third component of complement in crossed Immunoelectro‐phoresis. A polysulfone and two cellulose acetate membranes were relatively strong alternative pathway activators. Polypropylene, poly[vinylidine fluoride] and polyvinyl‐chloride derivative membranes were weak activators in some sera. Membrane activation was inhibited in citrate‐anticoagulated but not in heparin‐anticoagulated plasma. The results of such in vitro screening should be of value in selecting materials and anticoagulation regimens for membrane plasma separators.
Collection of platelets depleted of red and white cells with the “surge pump” adaptation of a blood cell separator
Authors: Hogge D. E. , Schiffer C. A.
Published: December 2004

The surge pump is a device which modifies platelet collection of a blood cell separator so that red and white blood cell contamination is minimized. Plasma collected from the donor is directed back into the centrifuge bowl at 200 ml per min, where it causes platelets to be floated off the red cell‐plasma interface and thus is collected as an almost pure platelet preparation. Fifty plateletapheresis procedures with the surge pump adaptation were compared to 50 procedures using the standard red cell method. Mean (±SD) white cell (>95% lymphocytes) contamination was 5.4 ±3.1 × 10s cells per collection with the surge pump and 63.5 ± 10 × 108 cells per collection with the standard red cell method (p < 0.0001). Mean collection hematocrit was 8.1 ± 2.6% with the standard method and <1% with the surge pump eliminating the need for crossmatch or centrifugation to remove red cells from ABO incompatible platelets. Surge pump collection produced a mean of 4.0 ± 1.6 times 10″ platelets compared to 5.0 ± 2.0 times 1011 platelets for the standard method (p<0.01). The mean time per run was 14.8 ± 2.4 min with the surge pump compared with 18.1 ± 3.3 min with the standard method (p< 0.001). Therefore, the platelet yield per minute of procedure time was comparable with both methods (surge pump, 37.3 ±11.7 × 10s platelets per min; standard method, 39.2 ± 14.3 X108 platelets per min.). Surge pump operation was learned easily by technologists and caused no donor complications. The surge pump is a simple and effective way of minimizing white and red blood cell contamination in platelet collections from the blood cell separator studied without compromising platelets yields.
Hemolytic disease of the newborn due to anti‐Lan
Authors: PAGE P. L.
Published: December 2004

Anti‐Lan caused mild hemolytic disease of the newborn in the second child of a previously transfused woman whose parents are first cousins. Based on the Lewis phenotype of the red cells of the proposita and her Lan‐negative sister, Len was shown to segregate independently of the Secretor laws
Anti‐M causing delayed hemolytic transfusion reaction
Authors: Alperin J. B. , Riglin H. , Branch D. R. , Gallagher M. T. , Petz L. D.
Published: December 2004

A 52‐year‐old gravida 1, para 1 woman with M‐ red cells experienced a delayed hemolytic transfusion reaction and exhibited an anti‐M antibody following the infusion of four units of M+ red cells. Measurements of erythrocyte survival using 51Cr‐labeled donor M+ and M‐red cells and in vitro studies of monocyte‐macrophage phagocytosis of sensitized reagent red cells implicate anti‐M in the pathogenesis of hemolysis.
Auto‐anti‐A1 antibody in a patient with metastatic adenocarcinoma
Authors: Castella A. , LaBarge B. P. , Lauenstein K. J. , Davey F. R.
Published: December 2004

An autoantibody was found on the red cells of a patient who had never been transfused previously, but was ill with metastatic adenocarcinoma. The patient's blood group was At. In addition, the patient's serum and the eluate from the patient's red cells agglutinated A, but not A2, B, and O red cells. This auto‐A, antibody was reactive at a wide thermal range and was inactivated by dithiothreitol, suggesting the presence of an IgM immunoglobulin. Moreover, the antibody was not associated with a hemolytic anemia.
Rhesus alloimmunization following intensive plasma exchange
Authors: McBride J. A. , O'Hoski P. , Barnes C. C. , Spiak C. , Blajchman M. A.
Published: December 2004

A 17‐year‐old woman was admitted to the hospital for the treatment of rapidly progressive systemic lupus erythematosus. She failed to improve when treatment with cyclophosphamide and prednisone and, therefore, was treated with intensive plasma exchange. A total of 24 liters of plasma was exchanged during six separate procedures over an 8‐day period. The patient, who was blood group B Rh negative (Cde/cde), was found to have an IgGanti‐D antibody reacting at a titer of 16 by the indirect antiglobulin technique 6 weeks after the first plasma exchange procedure. The titer of this antibody subsequently rose to 512. This patient, who had neither been pregnant nor received any blood products other than the plasma used during the plasma exchange, was presumably immunized by Rh positive red cells or stroma present in the transfused plasma. It is estimated that the patient received approximately 1010 Rh positive cells, or approximately one ml of packed red cells‐a quantity sufficient to cause Rhesus alloimmunization.
Preparation and in vitro function of granulocyte concentrates for transfusion to neonates using the IBM 2991 blood processor
Authors: GOLDFINGER D. , MEDICI M.A. , HSI R. , McPHERSON J. , CONNELLY M.
Published: December 2004

ABSTRACT: Clinical studies have suggested that granulocyte transfusions may be of value in the treatment of septic neonatal patients who present with severe granulocytopenia. We have developed a protocol for the preparation of granulocyte concentrates from freshly collected units of whole blood, using an automated blood cell processor. The red cells were washed with saline. Then, the buffy coats were collected from the washed red cells and studied for their suitability as granulocyte concentrates for neonatal transfusion. The mean number of granulocytes per concentrate was 1.6 times 109 in a mean volume of 25 ml. Studies of granulocyte function, including viability, random mobility, chemotaxis, phagocytosis and nitro‐blue tetrazolium reduction, demonstrated that the granulocytes were functionally unimpaired following preparation of the concentrates. These studies suggest that concentrates of functional granulocytes, suitable for transfusion to neonatal patients, can be prepared from fresh units of whole blood, using a cell processor. This procedure is more cost‐effective than leukapheresis and allows for delivery of granulocytes for transfusion in a more timely fashion.
Isolation and cryopreservation of monocytes from plateletapheresis cellular residues
Authors: Hunt S. M. , Lionetti F. J. , Valeri C. R.
Published: December 2004

Human monocytes were isolated from the cellular residues remaining after plateletapheresis of donors using an automated blood cell processor. Mononuclear cells were obtained with density gradients and separated from lymphocytes by stepwise elutriation. The isolated cells were frozen using extracellular hydroxyethyl starch and intracellular dimethylsulfoxide.

In three procedures, approximately 1 × 10° monocytes were obtained. Ninety‐nine percent of isolated monocytes were viable in a fluorescein diacetate (FDA)‐ethidium bromide (EB) test. Myeloperoxidase‐positive cells were 95 percent and 90 percent in the two chambers. Ninety‐four percent of monocytes ingested five or more opsonized polycyclic hydrocarbon particles and 95 percent ingested one or more ethidium‐treated zymosan particles.

After storage in liquid nitrogen for up to 9 weeks, 99 percent of the cells were recovered after thawing. Of these, 95 percent were myeloperoxidase‐positive, 94 percent showed intact membranes in the FDA‐EB test, 95 percent ingested five or more opsonized polycyclic hydrocarbon particles, and 96 percent ingested one or more ethidium‐treated zymosan particles.
Standardization of IgG subclass antiserums for use with sensitized red cells
Authors: Postoway N. , Garratty G.
Published: December 2004

ABSTRACT: The only IgG subclass antiserums standardized for use with red cells are those sold by the Netherlands Red Cross. Their recommended method is a sedimentation antiglobulin test (AGT) not generally used in the USA. Some years ago we tested the antiserums for sensitivity and specificity using the routine centrifugatlon AGT and found the methods comparable. More recently, we have become concerned since the antiserums are being used by a variety of methods other than the recommended procedure. It is well‐known that it is difficult to prepare subclass antiserums without contaminating antibodies to other IgG markers. Thus, we thought it worthwhile to standardize the antiserums with particular emphasis on specificity using several different technics. We found that some methods that have been used to define the subclass of red cell‐bound IgG were inappropriate, and led to many false positive results.
A study of blood utilization in association with hysterectomy
Authors: Samra S.K. , Friedman B.A. , Beitler P.J.
Published: December 2004

We performed a retrospective study of 167 women undergoing hysterectomy in a large tertiary‐care teaching hospital in order to assess intraoperative blood requirements and to study the decision‐making process that occurs when blood is transfused intraoperatively. Recommended preoperative blood orders for hysterectomy patients based on data from the study are provided. Although “type and screen” is commonly considered to be an adequate preoperative order for patients undergoing vaginal hysterectomy, our data indicate that 2 units may be a more appropriate order when the procedure is combined with pelvic floor repair. The preoperative hematocrit, the estimated intraoperative blood loss, and the duration of the hysterectomy were integral to the decision‐making process. If women undergoing hysterectomy were transfused intraoperatively only when their hematocrit actually fell to 30 percent, and not in anticipation of such an event, unnecessary transfusions would be reduced.
Use of recombinant human antithrombin in patients with congenital antithrombin deficiency undergoing surgical procedures
Authors: Konkle Barbara A. , Bauer Kenneth A. , Weinstein Robert , Greist Anne , Holmes Houston E. , Bonfiglio Judith
Published: December 2004

BACKGROUND : Hereditary antithrombin (AT) deficiency is associated with a significant risk of venous thromboembolism. Patients with this disorder frequently require long‐term anticoagulation. Discontinuation of anticoagulation for childbirth or surgery may carry a substantial thrombotic risk. For this reason, replacement with AT concentrate has been used when anticoagulation is interrupted. A new recombinant human AT concentrate, produced using transgenic technology, has recently been developed.

STUDY DESIGN AND METHODS : Human recombinant AT (rhAT) was provided by GTC Biotherapeutics, Inc. on a compassionate‐use basis for five patients with hereditary AT deficiency who underwent six surgical procedures. Patients were treated perioperatively. Dosing was determined individually by the investigators with a goal of maintaining an AT activity of 80 to 150 percent.

RESULTS : There was no clinical evidence of thrombosis or bleeding. Four of the five patients had postoperative duplex ultrasound studies of the lower extremities, which showed no evidence of acute thrombosis. Four patients were tested for antirecombinant rhAT antibodies postoperatively and were negative.

CONCLUSION : These case reports indicate that rhAT can provide effective support for AT‐deficient patients who undergo surgery. Further study of this product is needed to define optimal dosing and further assess clinical response.
GUTI: a new antigen in the Cromer blood group system
Authors: Storry Jill R. , Sausais Laima , Hue‐Roye Kim , Mudiwa Flora , Ferrer Zennie , Blajchman Morris A. , Lublin Douglas M. , Ma Bei‐Wen , Miquel Juan F. , Nervi Flavio , Pereira Jaime , Reid Marion E.
Published: December 2004

BACKGROUND : The Cromer blood group system consists of seven high‐incidence and three low‐incidence antigens carried on decay‐accelerating factor (DAF). This report describes the identification and characterization of a new Cromer high‐incidence antigen, named GUTI.

STUDY DESIGN AND METHODS
: RT‐PCR and sequence analysis were performed on cDNA prepared from a Chilean donor whose serum contained the alloantibody (anti‐GUTI). Based on the observed point mutation, a PCR‐RFLP assay using
Mae II was developed. To map the epitope, DAF‐deletion mutants were tested by immunoblotting with anti‐GUTI.

RESULTS
: Sequence analysis revealed a substitution of 719G>A in
DAF
in the proband. The proband's parents and two daughters were heterozygotes for 719G>A, one sister whose RBCs typed GUTI– was homozygous for 719A, and one sister had the wild‐type
DAF (719G). Seven additional heterozygote samples were identified among 214 Chileans. No heterozygotes were found in 197 New York donors. Analysis using DAF‐deletion mutants showed the antigenic determinant to be within short consensus repeat (SCR) 4.

CONCLUSION : This study describes a novel high‐ incidence antigen (GUTI) in the Cromer blood group system characterized by the amino acid arginine at position 206 in SCR4 of DAF. The GUTI‐negative proband has a substitution mutation that predicts for histidine at this position.
Expression of human neutrophil antigen‐2a (NB1) is increased in pregnancy
Authors: Caruccio Lorraine , Bettinotti Maria , Matsuo Kazuhiko , Sharon Victoria , Stroncek David
Published: December 2004

BACKGROUND : The expression of human neutrophil antigen‐2a (HNA‐2a) or NB1 is highly variable. This study investigated variations in neutrophil expression of HNA‐2a by comparing the expression of epitopes recognized by three HNA‐2a‐specific CD177 antibodies among healthy adults and pregnant women.

STUDY DESIGN AND METHODS : Flow cytometry was used to compare the neutrophil reactions of three CD177 antibodies (MEM‐166, TAG4, and 7D8) among 52 healthy adults. Because HNA‐2a expression is greater in women than men, neutrophils were studied from 21 women early in pregnancy and 23 women late in pregnancy.

RESULTS : All three CD177 antibodies reacted with two populations of neutrophils: a brightly staining population and a dimly staining population. Among the 52 healthy adults, there was no difference in the mean size of the brightly reactive population of neutrophils recognized by each antibody (46 ± 23% for MEM‐166, 49 ± 23% for TAG4, and 49 ± 23% for 7D8), and all three antibodies were specific for HNA‐2a. For all three antibodies the size of the antigen bright neutrophil population was greater on neutrophils collected both early and late in pregnancy than in healthy adults. There was no difference in the size of the antigen bright population among neutrophils tested early and late in pregnancy.

CONCLUSION : HNA‐2a antigen expression increases in pregnancy. Some of the variations in neutrophil expression of HNA‐2a among individuals is likely due to differences in gene regulation or differences in post‐translational protein modifications rather than gene polymorphisms.
Use of sentinel sites for daily monitoring of the US blood supply
Authors: Nightingale Stephen , Wanamaker Virginia , Silverman Barbara , McCurdy Paul , McMurtry Lawrence , Quarles Philip , Sandler S. Gerald , Triulzi Darrell , Whitsett Carolyn , Hillyer Christopher , McCarthy Leo , Goldfinger Dennis , Satcher David
Published: December 2004

BACKGROUND : This report describes the first year of a government‐sponsored program that uses daily reports from 29 sentinel sites to monitor the capacity of the US blood supply to meet demand.

STUDY DESIGN AND METHODS : From August 15, 2001, to August 14, 2002, 29 sentinel sites provided daily reports of the number of units of RBCs in inventory, transfused, exported, and outdated by ABO and Rh, and platelets by random or apheresis donor. Days supply of each component category was calculated as the number of units in inventory reported on a day divided by the sum of units transfused, exported, and outdated on that day. Sites also provided daily responses to questions about threatened or actual shortages.

RESULTS : The median of the days supply of RBCs at the 26 hospital transfusion services was 7.2 days. However, median days supply varied substantially by site and by day of the week. A+, O+, and O– units accounted for 30, 35, and 12 percent of total inventory and were maintained at a median supply of 7.4, 6.4, and 9.5 days, respectively. Reports of threatened RBC shortages peaked in early January 2002 and again in early July 2002. The July 2002 peak was about twice the January 2002 peak. Inventories at community‐based centers were similar to those at hospital transfusion services. Hospitals maintained only a 1‐day supply of platelets. Eight percent of random and 4 percent of apheresis platelets were outdated. There were 20 reports that surgery had to be postponed or canceled because platelets were unavailable.

CONCLUSIONS : Inventories of RBCs maintained at the participating sites were sufficient, with only one brief exception, to meet local demand during the first year of this monitoring program. The weekly rate of threatened shortage reports was more sensitive than days inventory as a predictor of actual shortages of RBCs. Unlike RBCs, platelet days supply, reports of threatened or actual platelet shortages, and platelet outdate rates did not vary seasonally.
A simple cryopreservation method for dendritic cells and cells used in their derivation and functional assessment
Authors: Celluzzi Christina M. , Welbon Craig
Published: December 2004

BACKGROUND: Cryopreservation and storage permitting multiple treatments with single donations is of practical importance to cellular therapies. HES and DMSO, used successfully in simple clinical procedures for freezing marrow and peripheral blood progenitor cells at –80°C, was tested on antigen‐presenting dendritic cells (DCs) and cells used in their derivation.

STUDY DESIGN AND METHODS: DCs cultured in serum‐free media from adherent or CD14+ apheresis MNCs (n = 36) in the presence of GM‐CSF + IL4 ± TNFα were frozen and stored at –80°C in 6‐percent HES, 5‐percent DMSO, and 4‐percent HSA. Apheresis MNCs, CD14+ monocytes, and lymphocytes were similarly frozen and later thawed for culture. Cells were assayed for viability, DC phenotype, mixed lymphocyte reaction, and antigen presentation before and 3, 6, 9, 12 or more months after freezing.

RESULTS: DCs retained viability (82 ± 2.3%) for at least 24 months. Mature and immature phenotype and function were preserved. Thawed MNCs and CD14+ cells differentiated to DCs and lymphocytes maintained high functional viability (92 ± 3%) comparable to prefreeze levels.

CONCLUSION: A simple –80°C freezing and storage method that combines extracellular (HES) and intracellular (DMSO) agents is practical and preserves functional viability of DCs, MNCs, CD14+ monocytes, and lymphocytes.
Analysis of PBPC cell yields during large‐volume leukapheresis of subjects with a poor mobilization response to filgrastim
Authors: Moncada Victoria , Bolan Charles , Yau Yu Ying , Leitman Susan F.
Published: December 2004

BACKGROUND: The circulating CD34 count is a reliable predictor of peripheral blood progenitor cell (PBPC) yields in subjects with a vigorous mobilization response to G‐CSF, however, the value of this parameter in poor mobilizers is uncharacterized.

STUDY DESIGN AND METHODS: Consecutive PBPC procedures (n = 81) with preapheresis CD34 counts less than 20 per μL (poor mobilizers) were compared with an equal number of good mobilizers (preapheresis CD34 counts ≥20/μL). G‐CSF was administered at standard doses (10 μg/kg/day).

RESULTS: CD34 yields correlated strongly with preapheresis CD34 counts in both good and poor mobilizers and were higher in allogeneic than autologous donors. For a standard 75‐kg recipient, a CD34 cell dose of greater than 2 × 106 per kg was never achieved in less than two 15‐L procedures if the preapheresis CD34 count was less than 8 per μL. Preapheresis WBC and MNC counts were lower in poor than in good mobilizers (28.4 vs. 43.0 and 3.25 vs. 5.01 × 103/μL, respectively, p < 0.0001). Total WBC counts correlated more strongly with preapheresis CD34 counts, total CD34 yields, and CD34 yields per L processed in good mobilizers (R = 0.50, R = 0.44, and R = 0.42, respectively) than in poor mobilizers (R = 0.22, R = 0.02, and R = 0.01, respectively), whereas total MNC counts correlated more strongly with these parameters in poor (R = 0.38, R = 0.23, and R = 0.27, respectively) than in good mobilizers (R = 0.04, R = 0.13, and R = 0.16, respectively).

CONCLUSION: CD34 cell yields correlate strongly with preapheresis CD34 counts. Based on this analysis, a CD34 count greater than or equal to 8 per μL is the threshold for performing PBPC collections in our institution.
Frequency of HBV DNA detection in US blood donors testing positive for the presence of anti‐HBc: implications for transfusion transmission and donor screening
Authors: Kleinman Steven H. , Kuhns Mary C. , Todd Deborah S. , Glynn Simone A. , McNamara Anne , DiMarco Anthony , For The Retrovirus Epidemiology Donor Study Michael P. Busch
Published: December 2004

BACKGROUND: An estimate of the rate of HBV DNA‐positive, anti‐HBc‐positive units is important for evaluating the need for anti‐HBc donor screening, especially in the context of HBV NAT.

STUDY DESIGN AND METHODS: HBsAg EIA‐nonreactive, anti‐HBc‐reactive (Corzyme, Abbott Laboratories) specimens were retrieved from a repository and were retested for anti‐HBc (with PRISM HBcore, Abbott Laboratories, currently under FDA review) and anti‐HBs (with PRISM Ausab, Abbott Laboratories, research assay). HBV DNA testing using a PCR assay with a greater than 95 percent detection rate of less than 50 copies per mL was performed on a subset of specimens that were PRISM HBcore‐reactive and were anti‐HBs‐ negative or reactive at less than 100 IU per L.

RESULTS: A total of 395 of 1231 specimens eligible by our serologic criteria were tested by PCR. Four anti‐HBs‐negative specimens were PCR‐positive with estimated HBV DNA copy numbers of 10 per 30 copies per mL in two specimens and 50 to 100 copies per mL in two others. The HBV DNA detection rate in anti‐HBs‐negative specimens was 3.7 percent, and the projected rate among all Corzyme‐reactive specimens was 0.24 percent, leading to an estimated yield of 1 HBV DNA‐positive, anti‐HBc‐positive unit in 49,000 units that were otherwise eligible for transfusion (95% CI, 1 in 16,600‐1 in 152,600).

CONCLUSIONS: Anti‐HBc screening detects HBsAg EIA‐negative, HBV‐infected donors at a rate comparable to the estimated residual risk for HBV window‐period infections. The low viral load in the HBV DNA‐positive samples suggests that minipool NAT will not detect most potentially infectious units from anti‐HBc‐positive donors.
The cost‐effectiveness of NAT for HIV, HCV, and HBV in whole‐blood donations
Authors: Jackson B.R. , Busch M.P. , Stramer S.L. , AuBuchon J.P.
Published: December 2004

BACKGROUND: The risk of viral infection associated with blood transfusion is lower than ever before because of aggressive screening and testing practices. NAT technology has lowered that risk even further but at an additional cost to the health‐care system.

STUDY DESIGN AND METHODS: Marginal cost‐effectiveness of NAT for HIV, HCV, and HBV in whole‐blood donations was calculated with a previously published Markov decision model. This model was updated with disease incidence data from all 2001 American Red Cross whole‐blood donations as well as window‐period data from the Retrovirus Epidemiology Donor Study (REDS).

RESULTS: Whole‐blood donation NAT for HIV and HCV is expected to cost between $155 million (minipool NAT) and $428 million (single‐donation NAT) per year in the US and avert 4 to 7 HIV infections and 56 to 59 HCV infections. Adding HBV NAT would be expected to avert 9 to 37 HBV infections at an additional cost of between $39 million and $130 million per year. Overall, NAT would cost between $4.7 million and $11.2 million per quality‐adjusted life‐year saved. Discontinuing HIV p24 antigen and HBc testing would offset this somewhat.

CONCLUSIONS: The cost‐effectiveness of whole‐blood NAT is poor. The testing cost would need to decrease significantly to bring the cost‐effectiveness in line with most other accepted medical practices.
Cord blood processing with an automated and functionally closed system
Authors: Zingsem Jürgen , Strasser Erwin , Weisbach Volker , Zimmermann Robert , Ringwald Jürgen , Goecke Tamme , Beckmann Matthias Wilhelm , Eckstein Reinhold
Published: December 2004

BACKGROUND: Umbilical cord blood processing with standard centrifugation techniques is performed in open systems and results in varying cell and volume recoveries.

STUDY DESIGN AND METHODS: Forty umbilical cord blood donations were randomly assigned to processing either with a microprocessor‐controlled cell separator equipped with closed disposables or with a manual separation procedure in blood bags. The collection efficiency of nucleated cells, MNCs, RBCs, and CD34+ cells and the processing time were analyzed.

RESULTS: Using the cell processor, mean collection efficiencies were 78.6 ± 24.9 percent for nucleated cells, 77.4 ± 27.8 percent for MNCs, 55.5 ± 14.6 percent for RBCs, and 83.6 ± 32.5 percent for CD34+ cells, while they were 73.1 ± 13.2 percent for nucleated cells, 78.1 ± 14.9 percent for MNCs, 26.0 ± 12.2 percent for RBCs, and 77.0 ± 17.6 percent for CD34+ cells when using the standard centrifugation technique. The processing time was about 20 minutes for automated processing and 60 to 80 minutes for the standard centrifugation technique.

CONCLUSION: Using the new cell processor, the collection efficiencies for nucleated cells, MNCs, and CD34+ cells are similar to those obtained by established centrifugation techniques while the RBC reduction is less effective. The main advantages of the new systems are the closed system, the more standardized processing procedure, and a significantly shorter processing time.
Retrospective targeted HCV lookback using centralized contracted notification service
Authors: Saxena Sunita , Shulman Ira A. , Pachciarz Judith , Downey Nancy , Figueroa Priscilla I.
Published: December 2004

BACKGROUND: Mandated HCV 2.0 lookback significantly challenged the human and financial resources available to the six Los Angeles County Department of Health Services (DHS) hospital blood banks. To comply with FDA requirements, DHS developed a centralized process that utilized the services of a contract vendor for performing HCV lookback.

STUDY DESIGN AND METHOD: A DHS public health nurse acted as HCV lookback coordinator and as liaison with the vendor. Cases were electronically forwarded to the vendor, who then reviewed the patient's chart to obtain information necessary for tracking the patient through commercial databases. The vendor was responsible for notifying the recipient for pretest counseling and for providing documentation of all efforts.

RESULTS: In total, 411 recipients were identified, of which, 168 cases were completed by the hospitals, 243 were forwarded to the vendor, 50 percent were deceased, 21 percent were contacted, and 28 percent could not be contacted, and 1 percent could not be located on any databases. The vendor contacted 35 of 66 cases that the hospitals had unsuccessfully attempted to contact. Of the cases forwarded to the vendor, 82 percent were completed on time. Delays were attributable to the vendor in less than 2 percent of cases. The cost per case forwarded to the vendor was $322.

CONCLUSION: Utilizing a professional service to conduct HCV recipient notifications is an effective strategy for meeting FDA deadlines and for freeing transfusion service staff to perform critical functions. It remains to be seen whether such strategies offer a cost savings.
Collection of MNCs and progenitor cells by two separators for PBPC transplantation: a randomized crossover trial
Authors: Ikeda Kazuhiko , Ohto Hitoshi , Nemoto Kenji , Yamamoto Go , Kato Katsuhiko , Ogata Takashi , Shichishima Tsutomu , Maruyama Yukio
Published: December 2004

BACKGROUND: Efficient collection of progenitor cells is essential for PBPC transplantation. Two apheresis machines (Amicus, Baxter Healthcare; and Spectra, Gambro BCT, software version 4.7) were compared prospectively by a crossover trial.

STUDY DESIGN AND METHODS: Apheresis collections were performed for two consecutive days on patients for autologous and donors for allogeneic PBPC transplantation. The patients and donors, receiving a G‐CSF, were randomized into two groups. In Group I, PBPCs were collected by the Amicus on the 1st day and the Spectra on the 2nd day, and the reverse order was used with Group II. A total of 60 apheresis procedures of 30 (16 in Group I and 14 in Group II) among 40 patients and donors enrolled were performed and evaluated.

RESULTS: The nucleated cell counts, MNC counts, CD34+ PBPC counts, and amounts of CFU‐GM collected per procedure were similar with the Amicus and the Spectra. On the other hand, the decrease of peripheral blood platelet counts of patients and donors was more prominent from using Spectra than Amicus (p < 0.0001). Components collected by the Amicus had fewer platelets than those collected by the Spectra (p < 0.0001). The efficiencies of collecting nucleated cells, MNCs, and CD34+ PBPCs were not different between the machines (p > 0.05). However, the efficiency of collecting platelets was significantly higher with Spectra than with Amicus (p < 0.0001). The Amicus took longer than the Spectra to process the same volume (p < 0.05).

CONCLUSION: Amicus is superior to Spectra in avoiding apheresis‐induced thrombocytopenia caused by platelets contaminating the collected samples. Therefore, the Amicus is useful for patients with thrombocytopenia or with a less‐than‐normal platelet count.
Barcode technology: its role in increasing the safety of blood transfusion
Authors: Turner C.L. , Casbard A.C. , Murphy M.F.
Published: December 2004

BACKGROUND:  Incorrect blood component transfusion is the most frequent serious incident associated with transfusion. Errors responsible for these incidents frequently involve patient misidentification.

STUDY DESIGN AND METHODS:  This study evaluated a barcode patient identification system involving hand‐held computers for blood sample collection for compatibility testing and the administration of blood. Audit of practice was carried out before and after its introduction.

RESULTS:  The baseline audit revealed poor practice, particularly in patient identification. Significant improvements were found in the procedure for the administration of blood following the introduction of barcode patient identification, including an improvement from 11.8 to 100 percent in the correct verbal identification of patients (p ≤ 0.001). Similar significant improvements were found in matching verbally stated identification details with details on patient identification wristbands, in correct patient identification before the collection of blood samples, and in the proportion of correctly labeled samples. Staff found the barcode identification system easy to operate and preferred it to standard procedures.

CONCLUSIONS:  A barcode patient identification system was found to simplify the clinical transfusion process and improve practice. These results provide support for further work on the development of such systems for transfusion and for other hospital procedures requiring patient identification.
Reversibility of severe metabolic stress in stored platelets after in vitro plasma rescue or in vivo transfusion: restoration of secretory function and maintenance of platelet survival
Authors: Rinder Henry M. , Snyder Edward L. , Tracey Jayne B. , Dincecco Dorothy , Wang Chao , Baril Laurence , Rinder Christine S. , Smith Brian R.
Published: December 2004

BACKGROUND:  Determining reversible aspects of the platelet storage lesion may result in improved function and survival of transfused platelets.

STUDY DESIGN AND METHODS:  Using a model of high‐dose (apheresis‐derived) platelet concentrates (PC), functional changes imposed by transient adverse metabolic conditions (pH < 6.0 for 1‐2 hr) that could be reversed by autologous plasma rescue followed by standard platelet storage were investigated. Whole‐blood‐derived PCs were transfused into a small number of normal volunteers to determine platelet recovery and survival.

RESULTS:  Without rescue, high‐dose PCs developed severe in vitro functional derangements at the time of the pH nadir including 1) loss of resting morphology; 2) complete abrogation of osmotic recovery and platelet aggregation and glycoprotein IIb/IIIa up‐regulation to agonist; and 3) decreased α‐granule release. By contrast, spontaneous and agonist‐induced binding of annexin V were unaffected by adverse metabolic conditions. Plasma rescue to an optimal pH improved morphology scores, stabilized osmotic recovery, and completely restored platelet secretory responses, as measured by aggregation, glycoprotein IIb/IIIa up‐regulation, and α‐granule release. In a limited number of studies, plasma rescue was accompanied by preserved in vivo platelet recovery and survival after autologous transfusion after 5 days of storage.

CONCLUSION:  Transient derangement of platelet metabolism, which does not increase membrane phosphatidylserine exposure, causes in vitro functional abnormalities that are fully reversed or stabilized by metabolic rescue. Preliminary data suggest that such rescued platelets may have normal posttransfusion recovery and survival.
The RHCE allele ceRT: D epitope 6 expression does not require D‐specific amino acids
Authors: Wagner Franz F. , Ladewig Birgit , Flegel Willy A.
Published: December 2004

BACKGROUND:  False‐positive D typing in patients may lead to anti‐D immunization caused by D+ transfusions or by omission of anti‐D prophylaxis. Known causes of such errors are RhCE variants carrying RhD‐specific amino acids and cold agglutinin activity of some frequently used monoclonal anti‐D.

STUDY DESIGN AND METHODS:  The molecular basis of eight samples referred because of “false‐positive” reactions with some commercial monoclonal anti‐D was investigated by PCR and nucleotide sequencing from genomic DNA. PCR with sequence‐specific priming was developed to specifically detect the underlying aberrant RHCE allele. The D epitope profile of the allele was determined by serology.

RESULTS:  The aberrant reactivity of the samples was caused by the RHCE allele RHCE(R154T) that occurred in a cde haplotype. The phenotype dubbed ceRT expressed the important D epitope 6, which is the target epitope of most monoclonal anti‐D used in routine typing.

DISCUSSION:  The characterization of ceRT demonstrated a previously unknown mechanism of antigen D expression that does not require any D‐specific amino acid. At least for some D epitopes, D‐like structures may be mimicked by RhCE proteins carrying amino acid substitutions not representative for RhD.
Protein composition and activation markers in plasma collected by three apheresis procedures
Authors: Burnouf Thierry , Kappelsberger Claudia , Frank Kerstin , Burkhardt Thomas
Published: December 2004

BACKGROUND: Scientific and technical advances made in transfusion medicine sustain the need for more comprehensive understanding of the impact of collection procedures on the quality of plasma for fractionation and for transfusion. This prospective work evaluated protein composition and markers of activation in plasma donations collected with three different automatic collection procedures (performed on Haemonetics machines), including a new procedure using a high‐separation core‐molded bowl.

STUDY DESIGN AND METHODS: A total of 90 collection procedures have been performed from a population of 37 donors, under comprehensively standardized conditions. Plasma aliquots were taken from the plasma units within 30 minutes of the end of the collection procedures and immediately frozen at −70°C. Content in an extended range of proteins and of markers of activation of the coagulation and fibrinolytic systems has been measured using standard in vitro testing methods.

RESULTS: Plasma donations had normal mean total protein, IgG, IgM, and fibrinogen content. The mean levels in coagulation FV, FVII, FVIII, and FXI and in antithrombin were above the standard international requirements. There was no sign of activation of the hemostasis system, as assessed by activated FVII, thrombin antithrombin complex, Prothrombin fragment 1+2, and D‐dimers. Activated complement component C3 and C5 were low.

CONCLUSION: Data indicates the good and consistent protein composition of plasma obtained by those automatic apheresis procedures. In particular, the new high‐separation core procedure yields a high‐quality plasma meeting requirements for transfusion and fractionation.
Effect of work duration on physiological and rating scale of perceived exertion responses during self‐paced interval training
Authors: Seiler Stephen , Sjursen Jarl Espen
Published: December 2004

This study compared running velocity, physiological responses, and perceived exertion during self‐paced interval training bouts differing only in work bout duration. Twelve well‐trained runners (nine males, three females, 28±5 years, VO2 max 65±6 mL min−1 kg−1) performed preliminary testing followed by four “high‐intensity” interval sessions (Latin squares, 1 session week−1 over 4 weeks) consisting of 24×1, 12×2, 6×4, or 4×6‐min running bouts with a 1:1 work‐to‐rest interval (total session duration 48 min). The average running velocity decreased (93%, 88%, 86%, 84% vVO2 max, P<0.01) with increasing work duration. Peak VO2 averaged about 92±4% of VO2 max for 2‐, 4‐, and 6‐min intervals compared with only 82±5% for 1‐min bouts (P<0.001). Six of 12 athletes achieved their highest average VO2 and heart rate during 4‐min intervals. The average RPEpeak (rating scale of perceived exertion) was ∼17±1 for all four interval sessions. RPE increased by 2–4 U during an interval training session. The mean lactate concentration was similar across sessions (4.3±1.1–4.6±1.5 mmol L−1). Under self‐paced conditions, well‐trained runners perform “high‐intensity” intervals at an RPE of ∼17, independent of interval duration. The optimal interval duration for eliciting a high physiological load is 3–5 min under these training conditions. Increases in RPE during an interval bout are not associated with increasing blood lactate concentration.
Out with a Bang! Tetrahymena as a Model System to Study Secretory Granule Biogenesis
Authors: Turkewitz Aaron P.
Published: December 2004

The release of polypeptides in response to extracellular cues is a notable feature of endocrine, exocrine and neuronal cells, and is based on regulated exocytosis via dense‐core secretory granules. There is interest in this mode of secretion because of its importance in human physiology and also because regulated exocytosis reflects a complex pathway of membrane traffic that includes compartment‐specific reversible macromolecular assembly, coat‐independent vesicle budding, maturation/remodeling of both lumenal and membrane constituents, and stimulus‐dependent membrane fusion. Secretory granules are absent in most unicellular model organisms but are highly developed in the Ciliates, which therefore offer attractive systems to study these phenomena. In Tetrahymena thermophila, biochemical and genetic approaches have begun yielding insights into issues ranging from control of granule core assembly, based on reverse genetic analysis of granule cargo, to questions about factors involved in granule biogenesis, based on random mutational approaches.
Characterization of purified intraembryonic hematopoietic stem cells as a tool to define their site of origin
Authors: Bertrand Julien Y. , Giroux Sébastien , Golub Rachel , Klaine Michèle , Jalil Abdelali , Boucontet Laurent , Godin Isabelle , Cumano Ana
Published: December 2004

Little is known about hematopoietic stem cell (HSC) development from mesoderm. To gain more information on the intraembryonic HSC site of origin, we purified multipotent hematopoietic progenitors from the aorta–gonads–mesonephros (AGM) of mice. This population, expressing c-Kit, AA4.1, CD31, and CD41, but not Flk1, and mainly negative for CD45, proved capable of long-term reconstitution in sublethally irradiated Rag2γc
-/-
recipients. We assigned the expression of GATA-2, GATA-3, and lmo2 to AGM-HSC, whereas erythromyeloid progenitors express only GATA-2. This unique combination of surface markers and transcription factors could be allocated in the AGM to the intraaortic clusters and the subaortic patches underlying aortic endothelial cells. Taken together, those data indicate that embryonic HSCs (
i
) differ from their fetal liver and adult counterpart by the low expression of CD45, (
ii
) do not colocalize with aortic endothelial cells as previously thought, and (
iii
) are localized, at 10.5 days postcoïtum, in the splanchnic mesoderm underlying aortic endothelial cells, within GATA-3
+
CD31
+
cell clusters.
Power gain exhibited by motile mechanosensory neurons in
Drosophila
ears
Authors: Göpfert M. C. , Humphris A. D. L. , Albert J. T. , Robert D. , Hendrich O.
Published: December 2004

In insects and vertebrates alike, hearing is assisted by the motility of mechanosensory cells. Much like pushing a swing augments its swing, this cellular motility is thought to actively augment vibrations inside the ear, thus amplifying the ear's mechanical input. Power gain is the hallmark of such active amplification, yet whether and how much energy motile mechanosensory cells contribute within intact auditory systems has remained uncertain. Here, we assess the mechanical energy provided by motile mechanosensory neurons in the antennal hearing organs of
Drosophila melanogaster
by analyzing the fluctuations of the sound receiver to which these neurons connect. By using dead WT flies and live mutants (

tilB
2

,

btv
5P1

, and

nompA
2

) with defective neurons as a background, we show that the intact, motile neurons do exhibit power gain. In WT flies, the neurons lift the receiver's mean total energy by 19 zJ, which corresponds to 4.6 times the energy of the receiver's Brownian motion. Larger energy contributions (200 zJ) associate with self-sustained oscillations, suggesting that the neurons adjust their energy expenditure to optimize the receiver's sensitivity to sound. We conclude that motile mechanosensory cells provide active amplification; in
Drosophila
, mechanical energy contributed by these cells boosts the vibrations that enter the ear.
HTLV-I Tax directly binds the Cdc20-associated anaphase-promoting complex and activates it ahead of schedule
Authors: Liu Baoying , Hong Sohee , Tang Zhanyun , Yu Hongtao , Giam Chou-Zen
Published: December 2004

Expression of the human T lymphotropic virus type I (HTLV-I) transactivator/oncoprotein, Tax, leads to faulty mitosis as reflected by chromosome aneuploidy, cytokinesis failure, and formation of micro- and multinucleated cells. Here we show that HTLV-I-transformed T cells progress through S/G
2
/M phases of the cell cycle with a delay. This delay is correlated with a decrease in the levels of cyclin A, cyclin B1, and securin. In
tax
-expressing cells, the Cdc20-associated anaphase promoting complex (APC
Cdc20
), an E3 ubiquitin ligase that controls metaphase to anaphase transition, becomes active before cellular entry into mitosis as evidenced by premature cyclin B1 polyubiquitination and degradation during S/G
2
. Consistent with the notion that Tax activates APC
Cdc20
directly, Tax is found to coimmunoprecipitate with Cdc20 and Cdc27/APC3. The APC
Cdc20
activity prematurely activated by Tax remains sensitive to spindle checkpoint inhibition. Unscheduled activation of APC
Cdc20
by Tax provides an explanation for the mitotic abnormalities in HTLV-I-infected cells and is likely to play an important role in the development of adult T cell leukemia.