bluuebunny/crossref_metadata_2025 huggingface dataset:
Preservation of red cell properties after virucidal phototreatment with dimethylmethylene blue
Authors: Wagner S.J. , Skripchenko A. , Robinette D. , Mallory D.A. , Cincotta L.
Published: March 2003

BACKGROUND: All published reports have described methods for virus photoinactivation which significantly alter red cell (RBC) properties during storage. In order to improve virucidal activity and reduce damage to RBCs, a series of phenothiazine derivatives were either synthesized or purified and screened for bacteriophage inactivation and red cell potassium efflux. One compound, 1,9‐dimethylmethylene blue (dimethyl‐methylene blue), had superior screening results and was chosen for further characterization.

STUDY DESIGN AND METHODS:White cell reduced RBC suspensions (30% hematocrit) were deliberately inoculated with extracellular virus or virus‐infected VERO cells, incubated with 4 microM dimethyl‐methylene blue and illuminated with cool‐white fluorescent light. Control and treated samples were titered for virus inactivation. In parallel studies, RBC suspensions were exposed to dimethylmethylene blue and light under identical conditions and assayed for in vitro RBC storage properties.

RESULTS:Phototreatment of RBC suspensions inactivated > 4.4 log10 of extracellular vesicular stomatitis virus (VSV), > 3.0 log10 of intracellular VSV, > 5.0 log10 of extracellular pseudorabies virus (PRV), > 4.8 log10 of intracellular PRV, > 4.7 log10 of extra‐cellular bovine virus diarrhea virus, 5.8 log10 of bacterio‐phage phi 6 and > 7 log10 of bacteriophage R17. Encephalo‐myocarditis virus, a nonenveloped picornavirus, was resistant to photoinactivation. Virucidal conditions resulted in no detectable IgG binding in 11 of 13 samples, unchanged RBC morphology, normal banding patterns of RBC membrane proteins on SDS PAGE, and unaltered characteristics of 12 of 13 RBC antigens during storage as measured by antibody titrations. In addition, minimal changes were observed in RBC osmotic fragility, lysis, potassium efflux, ATP and 2,3‐DPG levels, and the strength of one RBC antigen during storage of phototreated samples compared with controls. CONCLUSION: Dimethylmethylene blue photo‐treatment can inactivate several intracellular and extracellular model viruses under conditions which minimally alter RBC properties during 42 days storage at 1–6 degrees C.
The use of plasma as a replacement fluid in plasma exchange. Canadian Apheresis Group
Authors: Leblond P. F. , Rock G. , Herbert C. A. ,   Members of the Canadian Apheresis Group
Published: March 2003

BACKGROUND: The Canadian Apheresis Group has maintained a national registry of apheresis activities for the past 16 years. Since 1991, the use of plasma as a replacement fluid in plasma exchange has been recorded.

STUDY DESIGN AND METHODS: Six years of data from the registry on the use of plasma as a replacement fluid were analyzed.

RESULTS: Plasma was used in more than 25 percent of all plasma exchange procedures. Of 41,519 plasma exchange procedures reported, 11,970 used plasma alone or in combination with albumin. In 1991, 1026 (78%) of these procedures used plasma appropriately for either thrombotic thrombocytopenic purpura (TTP) or adult hemolytic uremic syndrome (HUS); between 1992 and 1996, these numbers were 1043 (81%), 1570 (86%), 1171 (87%), 2192 (92%), and 2741 (90%), respectively. In the remaining procedures, frozen or cryosupernatant plasma was administered to 326 patients for a total of 40 diseases other than TTP or HUS.

CONCLUSIONS: In those diseases for which plasma was administered as the sole replacement fluid, no disease appears to justify such treatment without the existence of an associated condition requiring specific replenishment of some plasma component. Further evaluation of the specific indications for the use of plasma as a replacement fluid in plasma exchange is required for diseases other than TTP or HUS.
Crossover use of donated blood for autologous transfusion: report of the Council on Scientific Affairs, American Medical Association
Authors: Blum L. N. , Allen J. R. , Genel M. , Howe J. P. ,   Council on Scientific Affairs American Medical Association
Published: March 2003

BACKGROUND: Controversy exists concerning whether the costs and potential risks outweigh the potential benefits of “crossover” use in the general blood supply of unutilized blood that was donated for autologous transfusion.

STUDY DESIGN AND METHODS: Published articles and reports were identified through systematic search of MEDLINE and review of references cited in previously identified articles, textbooks, and reports. Consultation was made with experts in blood donation and transfusion. Additional peer review was received from the American Medical Association (AMA) Council on Scientific Affairs

RESULTS: Concern over infectious disease transmission has led to increased interest in and support for autologous transfusion for individuals having planned surgeries. Different requirements exist for collection, labeling, and screening of blood to be used for autologous versus allogeneic transfusions; therefore, procedures for diverting autologous blood donations to the general blood supply involve considerable expense. Several cost‐effectiveness studies of autologous blood donation and transfusion conclude that currently this “crossover” appears to be an expensive procedure yielding little increased benefit from a societal perspective.

CONCLUSIONS: The recommendations in this report were adopted as AMA Policy at the AMA Annual Meeting in June 1997. The AMA does not encourage blood collection programs to “cross over” units donated for autologous use to the allogeneic blood supply. Practice guidelines are needed, and should be utilized to ensure parsimony in the use of autologous blood donations and transfusions.
Blood irradiation for intraoperative autotransfusion in cancer surgery: demonstration of efficient elimination of contaminating tumor cells
Authors: Hansen E. , Knuechel R. , Altmeppen J. , Taeger K.
Published: March 2003

BACKGROUND: Intraoperative blood salvage is contraindicated in cancer surgery because of contaminating tumor cells and the risk of systemic dissemination. On the basis of the radiosensitivity of cancer cells, irradiation of salvaged blood with 50 Gy is proposed as a way to allow return of salvaged blood.

STUDY DESIGN AND METHODS: Elimination of tumor cells by blood irradiation was studied in vitro with cells from 10 cell lines and from 14 tumor preparations after their addition to red cells in high numbers, or with blood shed during cancer surgery. Before and after gamma radiation, tumor cells were isolated by density gradient centrifugation and tested for their proliferative capacity in a cell colony assay. DNA metabolism was analyzed by incorporation of 5′ bromodesoxyuridine.

RESULTS: Survival curves of cells from various tumors confirmed D0 (the dose required to reduce the fraction of surviving cells to 37 percent of the original value) values in the range of 1.2 to 2.2 Gy. After irradiation of tumor cell‐contaminated blood with 50 Gy, no cell colony formation was observed, which indicates a reduction rate exceeding 10 log. Irradiated cancer cells showed viability, but no residual DNA metabolism.

CONCLUSION: The level of inactivation by a 50‐Gy dose far exceeds that needed to inactivate the number of proliferating tumor cells observed or expected in wound blood. These results provide the experimental basis for the clinical application of blood irradiation for intraoperative blood salvage in cancer surgery.
Cost of umbilical cord blood units released for transplantation
Authors: Sirchia G. , Rebulla P. , Tibaldi S. , Lecchi L.
Published: March 2003

BACKGROUND: A large number of institutions have started programs banking umbilical cord blood (UCB) for allogeneic unrelated‐donor and related‐donor transplantation. However, limited information is available on the financial issues surrounding these activities.

STUDY DESIGN AND METHODS: The aim of this study was to determine the fee per UCB unit released for transplantation that would allow cost recovery after 10 years. Three organizational models were considered suitable to provide units for five UCB transplants per 1 million population per year, a figure that would translate into an annual need for 280 units in Italy. Models A, B, and C included, respectively, seven networked banks, each with an inventory of 1,500 units; two networked banks, each with an inventory of 5,000 units; and one bank with an inventory of 10,000 units. It was estimated that it would take 3 years to develop the cryopreserved inventory and that approximately 3 percent of the inventory could be released and replaced each year during the 7‐year interval between the fourth and tenth years of activity. The data on the costs of labor, reagents and diagnostics, disposables, depreciation and maintenance, laboratory tests, and overhead, as well as the operational data used in the analysis were collected at the Milano Cord Blood Bank in 1996.

RESULTS: Fees of US $15,061, $12,666, and $11,602 per unit released during the fourth through the tenth years of activity allow full cost recovery (principle and interest) under Models A, B, and C, respectively.

CONCLUSION: Although UCB procurement costs compare favorably with those of other hematopoietic cell sources, these results and the current fee of US $15,300 used in some institutions show that UCB is an expensive resource. Therefore, judicious planning of banking programs with high quality standards is necessary to prevent economic losses. The advantages of lower fees associated with the centralized banking approach of Model C should be balanced with the more flexible collection offered by Model A.
Toward standardization of CD34+ cell enumeration: an international study
Authors: Dzik W. , Sniecinski I. , Fischer J. ,   the Biomedical Excellence for Safer Transfusion Working Party
Published: March 2003

BACKGROUND: An international multicenter study, involving six sites in North America and six sites in Europe, was undertaken to assess the performance of standardized methods for the enumeration of CD34+ cells in peripheral blood over the dynamic range from 200 cells per μL to zero. Two commercially available techniques were studied, a flow cytometry method and a microvolume fluorimetry method.

STUDY DESIGN AND METHODS: Coded samples were centrally prepared and sent to test sites by overnight mail. Samples included internal replicates, linear dilutions, and specimens at the lower limit of detection. In addition, commercially available reagent positive control cells were sent to a subset of laboratories.

RESULTS: Over the sample range studied, the intersite precision among different laboratories was good with coefficients of variation ranging from 14 percent to 24 percent for microvolume fluorimetry and from 20 percent to 31 percent for flow cytometry. Intrasite precision ranged from 7 percent to 21 percent. Test linearity was excellent with sites demonstrating a mean r2 = 0.992 for microvolume fluorimetry and r2 = 0.984 for flow cytometry. The lower limit of detection was 5 CD34+ cells per μL for both commercial assays. Over the range of 5 to 50 CD34+ cells per μL, the microvolume fluorimetry assay reported slightly higher values than the flow cytometry assay. Preliminary analysis of reagent positive control cells showed very good precision and accuracy.

CONCLUSIONS: Standardization of CD34+ cells enumeration is improving and commercially available assays provide accurate and precise methods. More investigation of reagent positive control cells is warranted.
Expression of Rh30 and Rh‐related glycoproteins during erythroid differentiation in a two‐phase liquid culture system
Authors: Suyama K. , Li H. , Zhu A.
Published: March 2003

BACKGROUND: To gain insight into the formation of the Rh complex during erythroid differentiation, the ways in which Rh30 and Rh‐related glycoproteins, especially Rh50, were produced in a modified two‐phase liquid culture system were studied.

STUDY DESIGN AND METHODS: A mononuclear cell fraction from fresh peripheral blood was first cultured in a medium supplemented with conditioned medium collected from the culture of a bladder carcinoma cell line (5637) for 7 days. Nonadherent cells were then collected for culture in a secondary medium containing 2 U per mL of erythropoietin to initiate erythroid differentiation. The expression of Rh30 and Rh50 during secondary culture (16 days) was monitored by flow cytometry.

RESULTS: D+ cells appeared after Day 4 and increased to 70 percent by Day 8. On Day 12, 90 percent of the total cells became D+ and remained so until the end of the culture. A similar expression profile was obtained for Rh50. As determined from mean fluorescence intensities recorded in flow cytometry, the number of both D and Rh50 antigenic sites per cell increased as the differentiation progressed. Rh‐related glycoprotein, CD47, had expression patterns significantly different from those of Rh30 and Rh50. In addition, the cultured cells produced partially glycosylated protein (approx. 32 kDa) in Rh50.

CONCLUSION: Expressions of Rh30 and Rh50 occur simultaneously during erythroid differentiation, and both proteins are most actively synthesized at the last stage of the differentiation. In contrast, CD47 may be involved in expression of Rh30 in a different manner from Rh50. The two‐phase liquid culture system will be an excellent model for studying the interaction among the components of the Rh complex during protein synthesis and complex assembly on the cell membrane.
High rate of GB virus type C/HGV transmission from mother to infant: possible implications for the prevalence of infection in blood donors
Authors: Lefrère J.‐J. , Sender A. , Mercier B. , Mariotti M. , Pernot F. , Soulié J.‐C. , Malvoisin A. , Berry M. , Gabai A. , Lattes F. , Galiay J.‐C. , Pawlak C. , De Lachaux V. , Chauveau V. , Hreiche G. , Larsen M. , Férec C. , Parnet‐Mathieu F. , Roudot‐Thoraval F. , Brossard Y.
Published: March 2003

BACKGROUND: Because GB virus type C(GBV‐C)/HGV (GBV‐C/HGV) is blood‐borne and sexually transmitted, persons at risk of infection with such viruses have a high prevalence of GBV‐C/HGV markers. However, adults with no apparent risk factors, such as blood donors, frequently are positive for GBV‐C/HGV markers. Mother‐to‐infant transmission could explain this high prevalence, but it has been studied only through small series of GBV‐C/HGV‐infected mothers co‐infected with HCV or HIV.

STUDY DESIGN AND METHODS: To determine the rate of mother‐to‐infant transmission of GBV‐C/HGV RNA in women who are HCV‐ or HIV‐negative, a prospective study was performed in a cohort of 288 mothers screened for viral RNA and in the infants born to GBV‐C/HGV‐infected mothers.

RESULTS: Thirteen mothers (4.5%) were found positive for GBV‐C/HGV RNA. Of the infants in whom at least one blood sample was collected between the third and the ninth months of life, 89 percent were positive for viral RNA. The majority of these newborns were negative for GBV‐C/HGV RNA at birth and positive after the third month. The viral RNA titers of infants born to GBV‐C/HGV‐infected mothers appeared as elevated as those of their mothers. All the GBV‐C/HGV‐infected infants remained positive for viral RNA during the entire study period. No clinical events possibly linked to a primary GBV‐C/HGV infection were reported in infants. Serum ALT level and blood count remained within normal values throughout the follow‐up of all GBV‐C/HGV‐infected infants.

CONCLUSION: The frequency of mother‐to‐infant GBV‐C/HGV transmission is elevated and could explain the high prevalence of GBV‐C/HGV markers (viral RNA and E2 antibody) in adults at low risk for blood‐borne or sexually transmitted viruses, such as blood donors.
HCV RNA in blood donors with isolated reactivities by third‐generation RIBA
Authors: Lemaire J.M. , Courouce A.M. , Defer C. , Bouchardeau F. , Coste J. , Agulles O. , Cantaloube J.F. , Barlet V. , Barin F. ,   for the Viral Hepatitis Study Group of the French Society of Blood Transfusion
Published: March 2003

BACKGROUND: The objective of this collaborative study was to learn the proportion of HCV RNA‐positive samples obtained from a population of donors with isolated anti‐HCV reactivities by third‐generation RIBA (RIBA‐3) (indeterminate results).

STUDY DESIGN AND METHODS: During a 2‐year period, 11 blood transfusion centers kept all samples with indeterminate RIBA‐3 results to test them by PCR, using both local and commercial techniques.

RESULTS: Of the 758 RIBA‐3 indeterminate samples, 10 (1.3%) were positive for HCV RNA: 3.3 percent (6/180) and 1.3 percent (4/317) of samples with anti‐core or anti‐NS3 reactivity, respectively, and none of the 52 and 209 samples with anti‐NS4 or anti‐NS5 reactivity, respectively. HCV RNA‐positive donors with anti‐core reactivity were infected with different subtypes (1 with HCV subtype 1b, 1 with 2, 1 with 2a/2c, 2 with 3a, and 1 with 5a), and a follow‐up indicated a chronic‐carrier state in two of the six donors. Acute hepatitis was diagnosed in three of the four donors with anti‐NS3 reactivity alone. Two of these three were IV drug users and were infected with subtype 1a.

CONCLUSION: HCV RNA‐positive donors with indeterminate results in RIBA‐3 are extremely rare, but they do exist. They were observed only when either anti‐core or anti‐NS3 was present. With such a RIBA‐3 profile, PCR testing remains necessary to reveal an eventual acute or chronic HCV infection.
Clinical illness due to parvovirus B19 infection after infusion of solvent/detergent‐treated pooled plasma
Authors: Koenigbauer Ulrike F. , Eastlund Ted , Day John W.
Published: March 2003

BACKGROUND: Lipid‐enveloped viruses such as HIV, HBV, and HCV can be inactivated by treatment with solvents and detergents. HAV and human parvovirus B19 lack lipid envelopes and are not inactivated. Solvent/detergent‐treated pooled plasma (S/D plasma) contains neutralizing antibodies, but it is not known whether the parvovirus B19 antibody content is sufficient to prevent transmission of the disease. A patient is described who developed a clinical illness due to parvovirus B19 infection after the infusion of S/D plasma.

CASE REPORT: A 36‐year‐old woman with myasthenia gravis underwent five plasma exchange procedures from January 15 to January 25, 1999, using albumin, except for 5 units of SD plasma given because of a low fibrinogen level. Four of the 5 units were implicated in a recall after high levels of parvovirus B19 DNA were found in several lots. Two weeks after the infusion, the patient developed fatigue, a rash, and severe polyarthralgias. Parvovirus B19 IgG and IgM antibody titers were consistent with an apos infection.

CONCLUSION: Clinically apparent parvovirus B19 infection can follow the use of S/D plasma that contains high levels of parvovirus B19 DNA.
A cell‐kinetic model of CD34+ cell mobilization and harvest: development of a predictive algorithm for CD34+ cell yield in PBPC collections
Authors: Humpe Andreas , Riggert Joachim , Meineke Ingolf , Kurz Martin , Eil Anita , Storkebaum Beate , Binder Claudia , Munzel Ulrich , Funke Ingrid , Höcker Paul , Wiesneth Markus , Köhler Michael
Published: March 2003

BACKGROUND: Mobilization and homing of PBPCs are still poorly understood. Thus, a sufficient algorithm for the prediction of PBPC yield in apheresis procedures does not yet exist.

STUDY DESIGN AND METHODS: The decline of CD34+ cells in the peripheral blood during apheresis and their simultaneous increase in the collection bag were determined in a prospective study of 18 consecutive apheresis procedures. A cell‐kinetic, four‐compartment model describing these changes was developed. Retrospective data from 136 apheresis procedures served to further improve this model. A predictive algorithm for the yield was developed that considered the sex, weight, and height of the patient, the number of CD34+ cells in peripheral blood before apheresis, the inlet flow, and the duration of the apheresis. The accuracy of this algorithm was evaluated by comparison of the predicted and the observed yields of CD34+ cells in 105 prospective autologous and 148 retrospective allogeneic apheresis procedures.

RESULTS: The correlation between predicted and observed yields was good for the autologous and allogeneic groups with a correlation coefficient (r) of 0.8979 and 0.8311 (p<0.0001), respectively. The regression is described by the equations log (measured value [m]) = 1.0118 + 0.8595 × log (predicted value [p]) for the autologous and log (m) = 2.226 + 0.7559 × log (p) for the allogeneic group. The respective equations for the zero‐point regression are log (m) = 1.014 × log (p) and log (m) = 1.026 × log (p). The probability that the measured value was 90 percent or more of the predicted value was 83.8 percent for the autologous and 90.5 percent for the allogeneic apheresis procedures.

CONCLUSION: The predictive accuracy of the algorithm and the slope of the zero‐point regression curve were higher for allogeneic than autologous PBPC collections. The predictive algorithm may be a useful tool in PBPC harvest, enabling the adaptation of the size of the apheresis to the needs of each patient.
Efficacy of a new collection procedure for preventing bacterial contamination of whole‐blood donations
Authors: Bruneau Cecile , Perez Paul , Chassaigne Maurice , Allouch Pierre , Audurier Andre , Gulian Christine , Janus Guy , Boulard Gilbert , De Micco Philippe , Salmi L.R. , Noel Luc
Published: March 2003

BACKGROUND : Transfusion‐related bacterial contamination is a serious problem. The introduction of bacteria into donations at the collection stage seems frequent, despite well‐conducted phlebotomy site preparation. Additional preventive measures are required.

STUDY DESIGN AND METHODS : The aim of this study was to assess the potential efficacy of excluding the first 15 mL of blood to reduce the bacterial contamination of donations. A special device allowed the aseptic collection of two samples at the beginning of donation: S1 (first 15 mL) and S2 (next 15 mL). Bacteriologic cultures of S1 and S2 were performed by using an automated system. The procedure′s efficacy was measured by the proportion of positive donations in S1 that were then negative in S2.

RESULTS : S1 and/or S2 were positive in 76 (2.2%) of 3385 donations. In about three‐fourths of the culture‐positive donations, contamination was detected in the first 15‐mL sample only. Gram‐positive cocci accounted for 81 percent of species, gram‐positive bacilli for 14 percent, and gram‐negative bacilli for 5 percent. The new procedure would have prevented the introduction of bacteria in 55 donations, reducing to 0.6 percent the risk of contamination from the first 15 mL collected.

CONCLUSION : Although the final effect on blood component bacterial contamination rates cannot be derived from the study, excluding the first 15 mL of blood may reduce the rate of bacterial contamination in donations.
The potential impact of incentives on future blood donation behavior
Authors: Sanchez Ana M. , Ameti Dannie I. , Schreiber George B. , Thomson Ruth A. , Lo Annie , Bethel James , Williams Alan E. ,   for the Retrovirus Epidemiology Donor Study
Published: March 2003

BACKGROUND: It is important to assess the potential efficacy and safety of offering donation incentives as part of recruitment and retention programs.

STUDY DESIGN AND METHODS : In 1995, 7489 allogeneic donors responded to an anonymous mail survey that inquired about demographics, donation history, infectious disease risks, and the potential appeal of incentives.

RESULTS: The projected net effect of offering blood credits and medical testing would be to motivate, respectively, 58 percent and 46 percent of donors to return, whereas offering an item of limited value would motivate 20 percent to do so. First‐time and younger donors reported more frequently than repeat or older donors that incentives would appeal to them. Donors attracted by cash were 60 percent more likely to have a risk for transfusion‐transmissible infections (p = 0.03). Although not statistically significant, the odds of being an at‐risk donor were higher among individuals attracted by tickets to events (OR 1.5) and extra time off work (OR 1.2).

CONCLUSION: These findings suggest that offering blood credits and (though to a lesser extent) items of limited value could be safe and effective strategies for retaining donors. Although medical tests were found to have broad appeal, studies are needed to identify tests in which donors would be most interested.
Effect of gamma radiation on the in vitro aggregability of WBC‐reduced apheresis platelets
Authors: Zimmermann Robert , Schmidt Stefan , Zingsem Jürgen , Glaser Anke , Weisbach Volker , Ruf Andreas , Eckstein Reinhold
Published: March 2003

BACKGROUND: The effect of gamma radiation on single‐donor apheresis platelet concentrates (SDPs) has been elucidated only incompletely. The only existing report on the function of SDPs stored in the irradiated state found a deterioration in the in vitro aggregability at the end of shelf life in SDPs divided before irradiation with 1500 cGy.

STUDY DESIGN AND METHODS: The in vitro properties of platelets were examined in four series of irradiated and control platelets, each obtained from the same 15 donors. Irradiation with 3000 cGy was performed on Days 0, 3, and 5. Cellular content, aggregability by ADP alone or ADP and epinephrine, spontaneous and induced CD62 expression, β‐thromboglobulin release, glucose consumption, lactate production, and pH were measured immediately after preparation and on Days 3 and 5 after donation.

RESULTS: Comparable in vitro properties were measured in irradiated and control platelets, whether irradiation was performed on Day 3 or Day 5. However, in platelets irradiated on Day 0, we found a significantly better in vitro aggregability by 20 μM ADP immediately after irradiation and by 10 μM ADP and 2 μM epinephrine at the end of shelf life than was found in the other groups (Day 5 results: Day 0 irradiation: 75 ± 32%; Day 3 irradiation: 45 ± 45%; Day 5 irradiation: 47 ± 41%; control: 40 ± 24%; p<0.05).

CONCLUSION: Gamma radiation had no adverse effect on platelet quality in extremely WBC‐reduced SDPs. On the contrary, a slight, but significantly better in vitro aggregability was found in SDPs irradiated before storage than in platelets irradiated later during storage and in unirradiated platelets. This increased in vitro aggregability persisted until the end of shelf life.
Mathematical modeling and computer simulation of erythrocytapheresis for SCD
Authors: Nifong Thomas P. , Bongiovanni Michael B. , Gerhard Glenn S.
Published: March 2003

BACKGROUND: Erythrocytapheresis is used to prevent acute chest syndrome and stroke in patients with sickle cell disease (SCD). However, such regimens are associated with significant risks, such as iron overload and potential exposure to transfusion‐transmitted infectious diseases. Computer modeling of erythrocytapheresis procedures may help optimize treatments and minimize risks.

STUDY DESIGN AND METHODS: Mathematical models based upon material balance equations and patient‐specific statistical analyses were developed to estimate HbS levels immediately after erythrocytapheresis and immediately before the next treatment. The equations were incorporated into a software application that was used to model the effects of various treatment values on four patients treated with 90 erythrocytapheresis procedures.

RESULTS: Immediate postprocedure HbS values were accurately estimated with correlations between measured and calculated values ranging from R2 = 0.83 to 0.96. Estimates of HbS just before the next treatment correlated well in three patients (R2 = 0.71 to 0.83) but poorly in one (R2 = 0.28 to 0.46). Varying the treatment values by computer simulation led to a wide variation in the number of RBC units and the net RBC volume transfused.

CONCLUSION: Computer modeling of erythrocytapher‐esis can be used to optimize chronic treatment regimens for SCD patients and potentially to minimize the risks of overtransfusion.
Effect of rejuvenation and frozenstorage on 42‐day‐old AS‐1 RBCs
Authors: Szymanski Irma O. , Teno Richard A. , Lockwood William B. , Hudgens Russell , Johnson George S.
Published: March 2003

BACKGROUND: The FDA has approved a 42‐day storage period for RBCs stored in ADSOL (AS‐1). This study was undertaken to provide data for the FDA about the feasibility of salvaging AS‐1 RBCs at the end of their storage period by rejuvenation and freezing.

STUDY DESIGN AND METHOD: The investigation, consisting of a study (n = 10) and control (n = 6) arm, was carried out in two centers. In both centers, eight healthy volunteers donated a unit (450 mL) of whole blood. The RBC concentrates were stored at 4°C in AS‐1 for 42 days. The study units were rejuvenated, whereas the control units were not. All units were stored frozen at —80°C, then deglycerolized and kept for an additional 24 hours at 4°C.

RESULTS: After the 42‐day storage period, ATP had declined to 62 percent of the original value, 2,3 DPG was zero, and MCV was significantly larger than that of fresh RBCs. Following rejuvenation and deglycerolization, the mean ATP level was 141 percent, the mean 2,3 DPG level was 109 percent, and the MCV was normal. The freeze‐thaw‐wash recovery of the rejuvenated and nonrejuvenated RBCs was similar, 88.4 and 84.0 percent, respectively. There was no difference in hypoxanthine, inosine, and uric acid levels in the rejuvenated and nonrejuvenated units, which indicated that the chemicals in the rejuvenation solution and their by‐products had been removed during processing. In both centers, the mean 24‐hour survival of rejuvenated, deglycerolized RBCs exceeded 75 percent, whereas that of nonrejuvenated RBCs did not. The long‐term survival rates of viable study and control RBCs were similar.

CONCLUSION: Forty‐two‐day‐old AS‐1 RBCs that have been rejuvenated and then frozen have more than 75 percent viability and normal oxygen delivery function. Rejuvenation of RBCs does not introduce additional safety hazards to blood transfusion.
Outcomes of a program to evaluate mother and baby 6 months after umbilical cord blood donation
Authors: Lecchi Lucilla , Rebulla Paolo , Ratti Ilaria , Magri Mara , Garcea Filomena , Marangoni Francesca , Bertelé Tommaso , Giordano Rosaria , Sirchia Girolamo
Published: March 2003

BACKGROUND: A routine program of evaluating mothers and infants 6 months after umbilical cord blood donation was started at the Milano Cord Blood Bank (MCBB) in 1996. This study evaluated the main outcomes of this program.

STUDY DESIGN AND METHODS: All mothers donating cord blood at this bank from February 1996 through May 1999 were invited to visit the bank or the collection suite 6 months after delivery to report on the health condition of their babies and to provide a fresh blood sample for repeat basal serologic tests (HBsAg, anti‐HCV, anti‐HIV‐1/2, and syphilis). A bank volunteer contacted the mothers by telephone to schedule their visits just before the expiration of the 6‐month period. Before collection of the new sample, a trained operator interviewed the mothers to review the mother's medical history information collected at donation and to obtain the baby's postnatal medical history.

RESULTS: Of the 2450 mothers enrolled in the study, 2315 (94.5%) attended the bank in agreement with the program, 4 promised to attend, 95 could not be traced, 26 declined the invitation, and 10 were unable to attend. Of the 135 mothers who could not be traced, 29 (21.4%) belonged to non‐European ethnic groups. The average time spent with each mother was approximately 20 minutes. In serologic testing, one indeterminate anti‐HCV seroconversion (c22) was detected. Collection of the baby's postnatal history reported one case of congenital urinary malformation not known at delivery, one of protein C deficiency, one of phenylketonuria, one of mucoviscidosis, and one of 10q‐ chromosomal abnormality. The cord blood components from all these births were discarded.

CONCLUSION: These data support the feasibility of a routine 6‐month program of evaluating mothers and babies giving cord blood at a cord blood bank. Such programs may increase the quality of components stored for transplantation.
Peripheral blood progenitor cell collection after epirubicin, paclitaxel, and cisplatin combination chemotherapy using EPO‐based cytokine regimens: a randomized comparison of G–CSF and sequential GM–/G–CSF
Authors: Perillo Alessandro , Pierelli Luca , Scambia Giovanni , Serafini Riccardo , Paladini Umberto , Salerno Maria Giovanna , Bonanno Giuseppina , Fattorossi Andrea , Leone Giuseppe , Mancuso Salvatore , Menichella Giacomo
Published: March 2003

BACKGROUND: The peripheral blood progenitor cell (PBPC) mobilization capacity of EPO in association with either G–CSF or sequential GM–CSF/G–CSF was compared in a randomized fashion after epirubicin, paclitaxel, and cisplatin (ETP) chemotherapy.

STUDY DESIGN AND METHODS: Forty patients with stage IIIB, IIIC, or IV ovarian carcinoma were enrolled in this randomized comparison of mobilizing capacity and myelopoietic effects of G–CSF + EPO and GM–/G–CSF + EPO following the first ETP chemotherapy treatment. After ETP chemotherapy (Day 1), 20 patients received G–CSF 5 μg per kg per day from Day 2 to Day 13 and 20 patients received GM–CSF 5 μg per kg per day from Day 2 to Day 6 followed by G–CSF 5 μg per kg per day from Day 7 to Day 13. EPO (150 IU per kg) was given every other day from Day 2 to Day 13 to all patients in both arms of the study. Apheresis (two blood volumes) was performed during hematologic recovery.

RESULTS: The magnitude of CD34+ cell mobilization and the abrogation of patients' myelosuppression were comparable in both study arms; however, GM–/G–CSF + EPO patients had significantly higher CD34+ yields because of a higher CD34+ cell collection efficiency (57.5% for GM–/G–CSF + EPO and 46.3% for G–CSF + EPO patients; p = 0.0009). Identical doses of PBPCs mobilized by GM–/G–CSF + EPO and G–CSF + EPO drove comparable hematopoietic recovery after reinfusion in patients treated with identical high‐dose chemotherapy.

CONCLUSION: The sequential administration of GM–CSF and G–CSF in combination with EPO is feasible and improves the PBPC collection efficiency after platinum‐based intensive polichemotherapy, associating high PBPC mobilization to high collection efficiency during apheresis.
The expression of NA antigens in people withunusual Fcγ receptor III genotypes
Authors: Matsuo Kazuhiko , Procter Jo L. , Chanock Stephen , Stroncek David F.
Published: March 2003

BACKGROUND: The Fcγ receptor IIIb (FcγRIIIB) genes that encode neutrophil‐specific antigens NA1 and NA2 differ at 5 nucleotides (nts); in 4, the result is an amino acid (AA) difference between the two alleles. The role of each of these differences in antigen expression is not known. Persons with FcγRIIIB genes that differ from NA1‐FcγRIIIB and NA2‐FcγRIIIB by 1 nt have been described. This study compared NA1 and NA2 expression on granulocytes in persons with variant FcγRIIIB genes and in healthy blood donors.

STUDY DESIGN AND METHODS: Reactions of NA1‐ and NA2‐specific MoAbs and alloantibodies with granulocytes were assessed by flow cytometry in 74 healthy blood donors and 6 persons with known variant FcγRIIIB genes. The granulocytes were tested with 1 NA1‐specific MoAb, 1 NA2‐specific MoAb, 4 NA1‐specific alloantibodies, and 4 NA2‐specific alloantibodies.

RESULTS: Analysis of granulocytes from persons with variant NA genotypes found that single‐base substitutions in FcγRIIIB at 141 and at 349 are important in NA1 expression and those at 227 and 277 are important in NA2 expression. Among blood donors, neither age, sex, nor race affected the expression of NA1 or NA2. The NA2‐specific MoAb reacted more intensely with granulocytes from NA2‐double‐dose cells than with those from NA‐single‐dose cells, but this was not true for the NA2‐specific alloantibodies. There was no difference in the reactions of the NA1‐specific MoAbs and alloantibodies with donor samples of known NA1‐double‐dose or NA‐single‐dose cells. The intensity of reactions of both the NA1‐ and NA2‐specific MoAbs and alloantibodies were strongly correlated on double‐dose cells but not on single‐dose cells. In fact, granulocytes from 7 healthy blood donors, phenotyped as NA‐single‐dose with the MoAbs, were phenotyped as NA2‐double‐dose with the alloantibodies. Variations in FcγRIIIB are common in blacks, but 5 of the 6 donors were white. These results suggest that FcγRIIIB variations may be common in both whites and blacks.

CONCLUSIONS : NA2 expression is affected by polymorphisms in FcγRIIIB 227 and FcγRIIIB 277, both of which are involved in an FcγRIIIb N‐glycosylation site. Polymorphisms in FcγRIIIB at 141 and 349 appear more important to NA1 expression.
The effect of two additive solutions on the postthaw storage of RBCs
Authors: Hess John R. , Hill Heather R. , Oliver Cindy K. , Lippert Lloyd E. , Greenwalt Tibor J.
Published: March 2003

BACKGROUND: Sterile systems for freezing and for washing thawed blood will allow the storage of RBCs for more than 24 hours after removal of the cryoprotectant glycerol. This study assessed the effect of two ASs in maintaining deglycerolized RBCs.

STUDY DESIGN AND METHODS: Twenty‐four RBC units were stored for 6 days, pooled in groups of 4, realiquoted, sterilely glycerolized, and frozen. One month later, the units were thawed, sterilely deglycerolized by using an automated system (H215; Haemonetics), and stored for 5 weeks in either 100 or 200 mL of AS‐3 or an experimental AS (EAS‐61). Sterile samples were taken weekly for chemical and morphometric analysis.

RESULTS: The glycerolization and deglycerolization process produced highly comparable RBC units, but it caused a marked reduction of RBC pH, to about 6.4 at the beginning of storage. The addition of acidic AS‐3 further reduced the pH, which in turn reduced glucose consumption, lactate formation, and RBC ATP concentrations. Alkaline EAS‐61 increased these measures. Hypotonic EAS‐61 caused increased cell swelling and hemolysis, despite better RBC morphology.

CONCLUSIONS: Automation of sterile glycerolization and deglycerolization with the H215 works well, but the solutions should be reformulated for extended postthaw storage. This would best be accomplished by raising the pH of the wash solutions by the addition of disodium phosphate or sodium bicarbonate or both, by using alkaline ASs, and by matching the osmolality of the wash solution and ASs.
Causality assessment of suspected virustransmission by human plasma products
Authors: Schosser Rudolf , Keller‐Stanislawski Brigitte , Nübling C. Micha , Löwer Johannes
Published: March 2003

BACKGROUND: Causality assessment of reports on suspected virus transmission is crucial for early detection of infectious plasma products. Commonly used algorithms, such as the WHO criteria, do not meet the specific requirements for causality assessment of suspected virus transmission.

STUDY DESIGN AND METHODS: A special algorithm, based on nucleic acid amplification and gene sequencing technology, effectiveness of validated virus‐inactivation methods, empirical data concerning the safety record of the product, and information on batch‐related infection clusters, was developed. The algorithm is focused on laboratory test results or otherwise standardized data, with few clinical data being required. To facilitate practical application, the algorithm has been converted into a graphical decision tree.

RESULTS: The feasibility of the algorithm is shown by causality assessment of sample cases. Three cases are presented with the details of each case used in the 12‐question checklist. The answers provided by the checklist led to the causality classification.

CONCLUSION: The algorithm is a tool for evaluating reports of suspected virus transmission in a standardized manner. It thus has the potential to improve early signal detection in pharmacovigilance of plasma products by confirmation or exclusion of suspected infectivity in most cases.
The effect of hot beverages, cold beverages, and chewing gum on oral temperature
Authors: Newman Bruce H. , Martin Christin A.
Published: March 2003

BACKGROUND: A prospective blood donor's oral temperature is affected by external stimuli such as drinking a hot or cold beverage or chewing gum. These stimuli can cause a deferral or an inappropriate acceptance for a certain unknown time.

STUDY DESIGN AND METHODS: Ten volunteer subjects were exposed to each of the three stimuli in separate studies. Oral and tympanic membrane temperatures were taken before exposure, immediately after exposure, and at various times until the subject's temperature returned to baseline.

RESULTS: The drinking of a hot beverage caused an immediate mean temperature elevation of 2.6°F which would lead to deferral for 7 of 10 subjects. All subjects were acceptable for blood donation (temperature <99.6°F) by 5 minutes. A cold beverage lowered the temperature; the temperature in 6 subjects returned to baseline by 10 minutes and that in 4 did so by 30 minutes. Gum chewing caused minimal temperature elevations; only 1 donor's temperature reached 99.6°F, and it was at acceptable levels for blood donation by 5 minutes.

CONCLUSION: Drinking a hot beverage and, to a much lesser degree, gum chewing can lead to a deferral for elevated temperature, but subjects were acceptable by 5 minutes. Cold beverages are of more concern, because they suppress the temperature for a short time and could theoretically lead to acceptance of a febrile, possibly infectious donor.
Preclinical ex vivo expansion of cord blood hematopoietic stem and progenitor cells: duration of culture; the media, serum supplements, and growth factors used; and engraftment in NOD/SCID mice
Authors: Lam Audrey C. , Li Karen , Zhang Xiao B. , Li Chi K. , Fok Tai F. , Chang Allan M.Z. , James Anthony E. , Tsang Kam S. , Yuen Patrick M.P.
Published: March 2003

BACKGROUND: Ex vivo expansion of cord blood (CB) hematopoietic stem and progenitor cells increases cell dose and may reduce the severity and duration of neutropenia and thrombocytopenia after transplantation. This study's purpose was to establish a clinically applicable culture system by investigating the use of cytokines, serum‐free media, and autologous plasma for the expansion of CB cells and the engraftment of expanded product in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice.

STUDY DESIGN AND METHODS: Enriched CB CD34+ cells were cultured in four media (Iscove's modified Dul‐becco's medium with FCS, Gibco; X‐Vivo‐10, BioWhittaker; QBSF‐60, Quality Biological; and StemSpan SFEM, Stem Cell Technologies) with four cytokine combinations (thrombopoietin [TPO], SCF, Flt‐3 ligand [FL] with and without G–CSF, and/or IL‐6). The effect of autologous CB plasma was also investigated. The read‐out measures were evaluated on Days 8 and 12. After expansion at the optimized condition, cultured cells were transplanted into sublethally irradiated NOD/SCID mice. The engraftment of human CD45+ cells and subsets in the bone marrow, spleen, and peripheral blood was determined.

RESULTS: QBSF‐60 or StemSpan SFEM supported high yields of early progenitors (CD34+ cells, < 64.8‐fold; CD34+CD38– cells, 330‐fold; CFU–granulocyte erythroid macrophage megakaryocyte [GEMM], 248‐fold) and CFUs of the myeloid (CFU–GM, 407‐fold) and erythroid (BFU/CFU–E, 144‐fold) lineages. The expansion of the megakaryocytic lineage was consistently higher in X‐Vivo‐10 (CFU–megakaryocyte, 684‐fold). Autologous plasma promoted colony formation but reduced CD34+ cells and CFU–GEMM. The addition of G–CSF or IL‐6 improved cell yields; G–CSF was more effective for committed progenitors. Expansion products from cultures in QBSF‐60 with the cytokines engrafted and differentiated into the myeloid and lymphoid lineages in NOD/SCID mice.

CONCLUSION: The data supported the strategy of expansion. The optimized condition may be applicable to clinical expansion for the abrogation or reduction of posttransplant cytopenia.
A rabbit model for monitoring in vivo viability of human platelet concentrates using flow cytometry
Authors: Leytin Valery , Allen David J. , Mody Meera , Rand Margaret L. , Hannach Barbara , Garvey Bernadette , Freedman John
Published: March 2003

BACKGROUND : Viability in vivo of novel platelet components cannot be readily determined in human transfusions. Elaboration of valid animal models may be useful for this purpose.

STUDY DESIGN AND METHODS
: Viability of platelet concentrates (PCs) WBC reduced before storage was determined by flow cytometry in rabbits whose reticuloendothelial system was inhibited by ethyl palmitate administration. PCs stored at 22°C for 2 and 5 days (D2‐ and D5‐PCs) or refrigerated PCs (3‐6 days at 22°C plus 1‐4 days at 4°C, RF‐PCs) were transfused into rabbits. Five parameters of PC viability in vivo were calculated from human platelet survival curves: survival time, recovery 0.5 and 24 hours after transfusion (R
0.5
, R
24
), maximal recovery (R
max
), and total recovery for 0 to 24 hours (R
Σ ).

RESULTS
: No differences in viability of D2‐ and D5‐PCs were found. In contrast, viability of RF‐PCs was significantly lower than that of D2‐PCs, as was revealed with diverse sensitivity by four parameters: R
Σ
> R
24
> R
0.5 = survival time (p < 0.001, p < 0.01, and p < 0.05, respectively).

CONCLUSION : The rabbit model elaborated is sufficiently sensitive to reveal differences in human platelet viability in vivo between conventional and cold‐damaged PCs. It may be useful for comparing viability of different platelet components that cannot be readily tested in human transfusions.
Use of percutaneous radial artery catheter for peripheral blood progenitor cell collection in pediatric patients
Authors: Bambi Franco , Fontanazza Silvia , Messeri Andrea , Lippi Alma , Tucci Fabio , Tamburini Angela , Tintori Veronica , Casini Tommaso , Lacitignola Laura , Tondo Annalisa , Veltroni Marinella , Bernini Gabriella , Faulkner Lawrence B.
Published: March 2003

BACKGROUND : Leukapheresis procedures require adequate flow rates, which in children may frequently involve invasive vascular access placement.

STUDY DESIGN AND METHODS : A minimally invasive peripheral radial artery catheter was used for drawing blood in 85 leukapheresis procedures performed in 33 pediatric patients. Blood return to the patients was provided by either a central Broviac‐type catheter or a peripheral venous access. The patients' age range was 1 to 18 years (median, 9.5) and the weight range was 9 to 73 kg (median, 29 kg). Vasocan Braunüle Luer Lock IV cannulas (22 gauge in 78 and 20 gauge in 7) were placed percutaneously under local anesthesia, and in 8 patients, catheter placement was carried out during general anesthesia for other procedures. A continuous flow cell separator was used in all cases (Fresenius AS104 in 23 and AS204 in 62).

RESULTS : Flow rates ranged from 18 to 45 mL per minute, the mean number of total blood volumes processed was 2.07 (range, 0.51‐2.51), and the mean duration of the procedures was 150 minutes (range, 90‐260). The 22‐gauge cannulas provided adequate flow rates independently of patient age and weight. No significant thrombotic, embolic, hemorrhagic, ischemic, or infectious complications were observed.

CONCLUSION : Peripheral radial artery catheters are safe, are minimally invasive, and provide steady, high‐flow rates, and they should be considered for patients requiring leukapheresis and lacking a suitable vascular access for drawing blood.
Sophisticated Architecture is Required for the Sinoatrial Node to Perform Its Normal Pacemaker Function
Authors: BOYETT MARK R. , DOBRZYNSKI HALINA , LANCASTER MATTHEW K. , JONES SANDRA A. , HONJO HARUO , KODAMA ITSUO
Published: March 2003

Structure‐Function Relationships of the SA Node. The heart's pacemaker, the sinoatrial node, does not consist of a group of uniform sinoatrial node cells embedded in atrial muscle. Instead, it is a heterogeneous tissue with multiple cell types and a complex structure. Evidence suggests that from the periphery to the center of the sinoatrial node, there is a gradient in action potential shape, pacemaking, ionic current densities, connexin expression, Ca2+ handling, myofilament density, and cell size. This complexity may be necessary for the sinoatrial node to pacemake under diverse conditions, drive the more hyperpolarized atrial muscle, and resist proarrhythmic perturbations.