bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunologic changes after transfusion of autologous or allogeneic buffy coat‐poor versus white cell‐reduced blood to patients undergoing arthroplasty. I. Proliferative T‐cell responses and the balance of helper and suppressor T cells
Authors: Innerhofer P. , Luz G. , Spötl L. , Hobisch‐Hagen P. , Schobersberger W. , Fischer M. , Nussbaumer W. , Lochs A. , Irschick E.
Published: March 2003

BACKGROUND: Donor white cells (WBCs) contained in red cell (RBC) transfusions are thought to provoke down‐regulation of T‐cell‐mediated immunity. This study investigated this topic in otherwise healthy patients receiving buffy coat‐depleted or WBC‐filtered RBCs and undergoing standardized perioperative management.

STUDY DESIGN AND METHODS: Patients undergoing elective orthopedic surgery (primary hip and knee replacement surgery) were enrolled in a prospective study. Perioperative changes in T‐cell proliferation (stimulation with phytohemagglutinin and mixed lymphocyte culture) and T‐cell balance (T‐lymphocytes, helper T cells, and suppressor T cells) were compared after random assignment to allogeneic buffy coat‐depleted (Group 2, n = 8) or WBC‐reduced RBC (Group 3, n = 11) transfusion regimens. Recipients of autologous buffy coat‐depleted RBC transfusions (n = 15) served as controls (Group 1).

RESULTS: Compared to that in autologous transfusion recipients, alloantigen‐induced T‐cell proliferation was significantly reduced in recipients of allogeneic WBC‐reduced RBCs (Day 3, p = 0.0274). After the transfusion of allogeneic buffy coat‐depleted RBCs, a weak trend toward decreased T‐cell proliferation was observed (p = 0.0933) and the numbers of CD4+ T cells were also significantly lower (Day 7, p = 0.0389). On Day 10, alloantigen‐induced T‐cell proliferation remained significantly below baseline after transfusion of WBC‐reduced RBCs (p = 0.05), the numbers of CD3+ cells decreased in allogeneic RBC recipients (Group 2, p = 0.078; Group 3, p = 0.05), and those of CD8+ cells decreased significantly after the transfusion of allogeneic buffy coat‐depleted RBCs (p = 0.0234) concomitant with an increased CD4:CD8 ratio (p = 0.0391).

CONCLUSION: Results of the present study confirm the hypothesis of impaired T‐cell‐mediated immunity after allogeneic transfusion.
Seroindeterminate patterns and seroconversions to human T‐lymphotropic virus type I positivity in blood donors from Martinique, French West Indies
Authors: Cèsaire R. , Bera O. , Maier H. , Lezin A. , Martial J. , Ouka M. , Kerob‐Bauchet B. , Amar A.K. Ould , Vernant J.C.
Published: March 2003

BACKGROUND: Screening for human T‐lymphotropic virus type I (HTLV‐I) antibodies in volunteer blood donors has been systematic in the French West Indies since 1989. Western blot‐indeterminate results are commonly obtained. The significance of these indeterminate serologic patterns in HTLV‐I‐endemic areas is still unclear.

STUDY DESIGN AND METHODS: During a 2‐year period, 9759 blood donors were tested for HTLV‐I antibodies. The epidemiologic features of HTLV‐I‐seropositive,‐seroindeterminate, and ‐seronegative donors were compared. A lookback investigation was performed for the HTLV‐I‐seropositive donors, and the HTLV‐I‐seroindeterminate individuals were followed up.

RESULTS: Thirty‐nine donors (0.4%) were HTLV‐I seropositive and 49 (0.5%) were seroindeterminate. The age and sex ratio characteristics of the seroindetermi‐nate donors are divergent from those of the HTLV‐I‐seropositive group and are more like those of the seronegative population. However, during the study period, three cases of seroconversion after an initial seroindeterminate profile were reported. Two cases were detected through follow‐up of 38 HTLV‐I‐ seroindetermi‐nate donors over a mean of 8 months (2‐24 months). The third seroconverter belonged to the HTLV‐I‐seropositive group and was identified through lookback investigation. This case is atypical, with p19 reactivity for several months before HTLV‐I seropositivity.

CONCLUSION: These findings indicate that, although HTLV‐I‐seroindeterminate donors mainly are HTLV‐I‐noninfected, the rate of seroconversion in a repeat blood donor population from an endemic region must be taken into consideration. Moreover, the case of delayed seroconversion observed in this study suggests the difficulty of counseling seroindeterminate blood donors in endemic regions.
Semi‐automated flow cytometric analysis of CD34‐expressing hematopoietic cells in peripheral blood progenitor cell apheresis products
Authors: Gutensohn K. , Carrero I. , Krueger W. , Kroeger N. , Schäfer P. , Luedemann K. , Kuehnl P.
Published: March 2003

BACKGROUND: The measurement of CD34+ cells is the most important step in the quality control of peripheral blood progenitor cell apheresis products. For this purpose, flow cytometry is applied. Recently, a new test kit has been introduced for the enumeration of CD34‐expressing cells, in combination with software support for semi‐automation of data acquisition and analysis.

STUDY DESIGN AND METHODS: This study evaluated the ProCOUNT kit. Ninety samples obtained from peripheral blood progenitor cell apheresis products from 39 patients with hemato‐oncologic diseases were analyzed. For data acquisition and analysis, ProCOUNT software was used. Data comparison was performed with parallel measurements according to the International Society for Hematotherapy and Graft Engineering (ISHAGE) guidelines and the German reference protocol for analysis of CD34‐expressing cells.

RESULTS: Correlation of the German and ISHAGE techniques was excellent (r2 = 0.99). The initial correlation coefficient of ProCOUNT analysis with the German protocol was r2 = 0.89. In 21 (23.3%) of 90 ProCOUNT analyses, a warning message was encountered from the ProCOUNT software. Following manual reevaluation of these data with CellQUEST software, a correlation of r2 = 0.96 with the German protocol and r2 = 0.97 with the ISHAGE analyses was obtained. ANOVA testing revealed significant differences between ProCOUNT and ISHAGE techniques (p<0.05) and between ProCOUNT and the German protocol (p<0.05). No statistically significant difference between ISHAGE and German protocol was observed (p = 0.19).

CONCLUSION: The ProCOUNT kit and software for semi‐automated data acquisition and analysis represents a further step toward standardization of CD34 cell quantitation in peripheral blood progenitor cell apheresis products. However, the occurrence of software warnings is high, and analysis or data reevaluation by experienced staff is still mandatory. Therefore, currently there is no definite advantage of the kit and software over the existing guidelines for CD34+ analysis in peripheral blood progenitor cell grafts.
Variations in the expression of granulocyte antigen NB1
Authors: Matsuo K. , Lin A. , Procter J.L. , Clement L. , Stroncek D.
Published: March 2003

BACKGROUND: Between 87 and 97 percent of whites express NB1 alloantigen on some but not all of their granulocytes. The expression of NB1 has not been compared among large groups of adults of different sexes, ages, and ethnic groups. Previous testing of whites suggests that the expression of NB1 is variable.

STUDY DESIGN AND METHODS: Serologic testing of granulocytes from 224 persons with two examples of MoAb to NB1 (1B5 and 7D8) was performed to distinguish phenotypic differences among age, sex, and ethnic groups and differences in reactivity to MoAbs. The donors were from 17 to 82 years of age, and 87 were female. They were from four ethnic backgrounds: 54 were African American (black), 10 were Asian, 9 Hispanic, and 152 white. Granulocytes were tested by flow cytometry. Parallel testing with MoAbs to CD16, CD11b, and CD45 served as controls. The size of the granulocyte population reacting with 1B5 and 7D8 and the respective mean, median, and peak cell fluorescence intensities were analyzed.

RESULTS: The expression of 7D8 and 1B5 was greater on granulocytes from female donors. The expression of 7D8 fell in older women but not in men. There were no differences among the four racial groups in either the frequency of NB1 as determined by 1B5 or 7D8 or in the size of the population of granulocytes reacting with either antibody. When the fluorescence intensities of the antibody reactions were compared among groups, there were no differences in reactivity with 1B5. However, reactions with 7D8 were all greater in blacks. Comparison of the size of the antigen‐positive granulocyte population, as determined by antibody reactivity, showed that only 30 donors differed by more than 10 percent. These discordant results were more likely to occur in whites than in blacks (18% vs. 4%, p<0.02).

CONCLUSIONS: NB1 is composed of at least two epitopes as determined by serologic studies. The expression of both antigens is greater in females. The 7D8‐reactive epitope appears to be more prevalent or more accessible on granulocytes of blacks. Variations in the expression of NB1 are more likely to occur in whites. The biochemical and molecular basis of these variations are not known.
Effects of three Hb‐based oxygen‐carrying solutions on neutrophil activation in vitro: quantitative measurement of the expression of adherence receptors
Authors: Toussaint Marie , Latger‐Cannard Veronique , Caron Alexis , Lecompte Thomas , Stoltz Jean‐Francois , Vigneron Claude , Menu Patrick
Published: March 2003

BACKGROUND: Hb‐based oxygen carriers (HbOCs) are currently under investigation as RBC substitutes. These solutions have vasoactive properties and may modify the behavior of the RBC in vitro. The effects of these agents on other blood cells have not been extensively investigated. The aim of this study was to evaluate the potential effect(s) of three differently modified HbOCs on the activation of PMNs.

STUDY DESIGN AND METHODS: Whole blood from humans was incubated with Hb dextran‐benzene‐tetracarboxylate, αα‐Hb, or o‐raffinose‐poly‐Hb for 15, 30, and 60 minutes. The expression of adherence receptors CD62L, CD18, and CD11b, which reflects the activation state of the neutrophils, was assessed in vitro by a direct immunofluorescence method using calibration beads measured by flow cytometry.

RESULTS: The observed expression of each of the three adherence receptors for each solution at each time point was similar to that noted with the RPMI control. Decreases in CD62L and increases in CD18 expression were noted at 15 minutes of incubation but, when those values were compared to the values obtained with TNFα as the positive control of PMN activation, the profile of expression of adherence receptors obtained with HbOC solutions did not conform to the expected activation profile of PMNs.

CONCLUSION: Hb solutions modified by a variety of methods, as obtained and employed in this study, did not appear to activate neutrophils in vitro.
First‐time blood donors: demographic trends
Authors: Wu Ying , Glynn Simone A. , Schreiber George B. , Wright David J. , Lo Annie , Murphy Edward L. , Kleinman Steven H. , Garratty George ,   for the Retrovirus Epidemiology Donor Study (REDS) Group
Published: March 2003

BACKGROUND: With changing demographics of the United States population and the continuous need to recruit new donors, it is important to monitor the demographic profile of first‐time donors and to evaluate changes in the donor pool to improve recruitment targeting.

STUDY DESIGN AND METHODS: First‐time whole blood (n = 901,862) donors at five United States blood centers between 1991 and 1996 were analyzed.

RESULTS: The total number of first‐time donors appears to be decreasing. Over the 6‐year period, there was an overall increase in the proportion of Hispanic and other minority first‐time donors and a concurrent decrease in the proportion of white donors at Retrovirus Epidemiology Donor Study centers. Other variables, including age, sex, and education, did not show a consistent trend.

CONCLUSION: The demographic profile of first‐time donors is changing. These data highlight the importance for blood centers to continuously monitor the donor population. A better understanding of the donor population may help blood centers adjust their donor outreach, recruitment, and retention programs. New recruitment efforts appear needed to counter general apathy toward donating blood, and minority groups appear to be receptive to becoming blood donors.
RBC transfusion in sickle cell anemia (HbSS):experience from the Jamaican Cohort Study
Authors: Thame Jennifer R. , Hambleton Ian R. , Serjeant Graham R.
Published: March 2003

BACKGROUND: RBC transfusion is widely advocated in the management of sickle cell anemia (SCA), but it carries potentially serious risks, especially in the setting of chronic transfusion. The Jamaican Sickle Cell Clinic is conservative in its use of transfusion, and this experience is presented to allow comparison with other centers in defining the role for transfusion in SCA.

STUDY DESIGN AND METHODS : Reported here is a retrospective, descriptive study of all RBC transfusions given to 311 subjects with SCA who were followed in a cohort study from birth and are, at this writing, 16.3 to 24.7 years old.

RESULTS: There were 520 transfusion episodes in 197 (63.3%) of the 311 subjects; 1 transfusion in 80 (41%) of those who received transfusion(s), 2 transfusions in 54 (27%), 3 transfusions in 21 (11%), 4 in 17 (9%), 5 in 9 (5%), and 6 or more in 16 (8%). Single transfusions were usual for acute anemia of aplastic crises or acute splenic sequestration, and multiple transfusions (up to 21 episodes) were usual in the prophylaxis of recurrent stroke. Indications were aplastic crisis (102), acute chest syndrome (90), acute splenic sequestration (75), stroke (62), septicemia (46), hypoplasia (40), hypersplenism (34), surgery (31), gastroenteritis (10), and miscellaneous (30).

CONCLUSION : Despite conservative transfusion use, 70.6 percent of patients had received at least one transfusion by 20 years of age in the Jamaican Sickle Cell Clinic.
Incidence rates of viral infections among repeat donors:are frequent donors safer?
Authors: Schreiber George B. , Glynn Simone A. , Busch Michael P. , Sharma Usha K. , Wright David J. , Kleinman Steven H. ,   for the Retrovirus Epidemiology Donor Study
Published: March 2003

BACKGROUND: Incidence rates (IRs) for viral infections may vary with the frequency of donation among repeat, community, whole‐blood (WB) donors, with IRs thought to be lower among donors with higher frequency of donation.

STUDY DESIGN AND METHODS: IRs for HIV, HTLV, HCV, and HBV infection were stratified by frequency of donation among 868,403 repeat WB donors who gave approximately 4 million donations at five United States blood centers from 1991 through 96. All donors had given at least 2 donations during those years, with the first donation being nonreactive on confirmatory testing. Frequency of donation was measured in three ways: by the number of donations per year; at the time of donation, by the number of donations given within the preceding 2‐year period; and by the number of donations given from 1991 through 1993.

RESULTS: The IRs for HIV, HCV, and HBV infection did not appear to differ among donors with lower or higher numbers of donation per year. However, the IR for HTLV infection decreased as the number of donations per year increased (p = 0.0004). IRs for all viral markers remained stable, regardless of the number of donations given within the 2‐year period before the donation. Although IRs for HIV, HTLV, and HCV infection did not vary by the number of donations given in 1991 through 1993, the IR for HBV infection appeared to be lower in donors who gave more donations in that period (p = 0.01).

CONCLUSION: These findings do not provide evidence of a lower IR for transfusion‐transmissible viral infections among repeat WB donors who give more frequently. Abbreviated screening histories for frequent repeat donors might not be advisable.
Effect of prophylactic transfusion of stored RBCs on oxygen reserve in response to acute isovolemic hemorrhage in a rodent model
Authors: D'Almeida Mark S. , Gray Daryl , Martin Claudio , Ellis Christopher G. , Chin‐Yee Ian H.
Published: March 2003

BACKGROUND: The storage of RBCs results in a time‐related decline in 2,3 DPG that may reduce the ability to unload oxygen (O2) to tissue. The objective of this study was to compare the effect that transfusion of stored 2,3 DPG‐depleted rat blood (7 days in CPDA‐1) had on the O2 reserve in conscious rats, with that of the transfusion of fresh blood (<2‐hour storage).

STUDY DESIGN AND METHODS: Anemic rats (Hb, 80 g/L) received either fresh packed RBCs or stored RBCs to raise Hb levels to 140 g per L. They then underwent isovolemic hemorrhage mimicking surgical blood loss to the point of O2 supply dependency (OSD). Critical O2 delivery (DO2crit), Hb concentration, and O2 extraction at OSD were measured in a metabolic chamber.

RESULTS: After transfusion, RBC DPG decreased by 50 percent in the stored‐blood group, and the p50 value decreased by 5 mmHg (32.1 ± 2.5 mmHg vs. 37.5 ± 3.0). DO2crit was similar in the two groups (fresh blood: 2.79 ± 0.44 mL/min × g–1; stored blood, 2.99 ± 0.76 mL/min × g–1). The critical Hb concentration at DO2crit was higher in the stored‐blood group (44 ± 4 g/L) than in the fresh‐blood group (38 ± 5 g/L); the cardiac index and O2 extraction ratio in the two groups were not different. Under conditions of severe normovolemic anemia in rats, depletion of DPG and a decrease in p50 had only minor effects on the O2 reserve. At OSD, under these conditions, O2 consumption is not limited by diffusion.

CONCLUSION: The physiologic impact of DPG depletion in transfused stored blood on oxygen availability in normal rats appears to be small and may be clinically inconsequential.
Risk behavior in Norwegian blood donors
Authors: Stigum Hein , Bosnes Vidar , Ørjasæter Harald , Heier Hans Erik , Magnus Per
Published: March 2003

BACKGROUND: Blood banks ensure the safety of blood components by testing them for a set of known infectious agents and by careful selection of donors based on a self‐administered questionnaire and an interview. The purpose of this study is to describe the risk behavior for sexually transmitted diseases in Norwegian blood donors.

STUDY DESIGN AND METHODS: A survey of the sexual habits of 5,859 blood donors in the capital of Norway was performed by using anonymous questionnaires. The results were compared with a previous survey of 10,000 randomly selected Norwegian subjects aged 18 to 60 years. The response rates were 70.3 percent and 48.4 percent, respectively.

RESULTS: Blood donors had considerably more education than the general population. Their general sexual behavior was similar to that of the rest of the population, although the blood donors had later sexual debut, fewer new partners per year, and a lower frequency of intercourse. In addition, homosexual experience among males was much lower in the donor group. Blood donors were less likely to engage in risk behavior for sexually transmitted diseases than were the general population. Nevertheless, 1.5 percent of the donors reported behavior that would have led to deferral had the behavior been disclosed at the predonation interview. Deferrable donors were more likely to be male and young and to have had many partners.

CONCLUSION: Anonymous questionnaires reveal information that is not given at the time of blood donation.
Transfusion‐transmitted bacterial infectionin the United States, 1998 through 2000
Authors: Kuehnert Matthew J. , Roth Virginia R. , Haley N. Rebecca , Gregory Kay R. , Elder Kathy V. , Schreiber George B. , Arduino Matthew J. , Holt Stacey C. , Carson Loretta A. , Banerjee Shailen N. , Jarvis William R.
Published: March 2003

BACKGROUND: Bacterial contamination of blood components can result in transfusion‐transmitted infection, but the risk is not established.

STUDY DESIGN AND METHODS: Suspected cases of transfusion‐transmitted bacteremia were reported to the CDC by participating blood collection facilities and transfusion services affiliated with the American Red Cross, AABB, or Department of Defense blood programs from 1998 through 2000. A case was defined as any transfusion reaction meeting clinical criteria in which the same organism species was cultured from a blood component and from recipient blood, with the organism pair confirmed as identical by molecular typing.

RESULTS: There were 34 cases and 9 deaths. The rate of transfusion‐transmitted bacteremia (in events/million units) was 9.98 for single‐donor platelets, 10.64 for pooled platelets, and 0.21 for RBC units; for fatal reactions, the rates were 1.94, 2.22, and 0.13, respectively. Patients at greatest risk for death received components containing gram‐negative organisms (OR, 7.5; 95% CI, 1.3‐64.2; p = 0.009).

CONCLUSION: Bacterial contamination of blood is an important cause of transfusion‐transmitted infection; infection risk from platelet transfusion is higher compared with that from RBCs, and, overall, the risk of infection from bacterial contamination now may exceed that from viral agents. Recipients of components containing gram‐negative organisms are at highest risk for transfusion‐related death. The results of this study may help direct efforts to improve transfusion‐related patient safety.
Lack of transmission of TT virus through immunoglobulins
Authors: Azzari Chiara , Resti Massimo , Moriondo Maria , Gambineri Eleonora , Rossi Maria Elisabetta , Novembre Elio , Vierucci Alberto
Published: March 2003

BACKGROUND: A high prevalence of TT virus (TTV) infection has been found in patients who received blood or blood components. Viral DNA was demonstrated in commercial preparations of FVIII and F IX, but very few data have been reported on immunoglobulins. The risk of TTV infection associated with intramuscular or IV immunoglobulin administration is unclear.

STUDY DESIGN AND METHODS: The prevalence of TTV infection in a group of patients undergoing lifelong therapy because of congenital immunodeficiency has been evaluated in a long term follow‐up (median, 6 years). Seventeen patients with congenital immunodeficiency receiving monthly administration of IVIG were included in the study. TTV DNA was repeatedly evaluated by PCR in serum samples from each patient during the follow‐up. Research of antibodies against TTV was not applicable, as the patients studied were unable to produce antibodies. The presence of TTV was also evaluated in 15 IVIG lots.

RESULTS: The total amount of immunoglobulin administered was 18,773 g. TTV infection was not found in any patients included in the study. None of the 15 immunoglobulin preparations analyzed was found positive for TTV DNA.

CONCLUSION: Despite the high prevalence of TTV in blood donors, commercial immunoglobulins are safe and unable to transmit TTV.
Processing of peripheral blood progenitor cell components in improved clean areas does not reduce the rate of microbial contamination
Authors: Cassens Uwe , Ahlke Christoph , Garritsen Henk , Krakowitzky Petra , Wüllenweber Jörg , Fischer Rudolf‐Josef , Peters Georg , Sibrowski Walter
Published: March 2003

BACKGROUND: Microbial contamination of peripheral blood progenitor cell components (PBPCs) may cause severe complications in immunosuppressed recipients. Therefore, principles of Good Manufacturing Practice (GMP) are applicable for processing of PBPC components to reduce potential risks of contamination.

STUDY DESIGN AND METHODS: It was investigated in a retrospective study whether the microbial contamination of PBPC components could be reduced after processing in improved clean areas according to the “Manufacture of Sterile Medicinal Products.” Starting in 1994, a total of 1478 autologous and allogeneic PBPC components have been collected and processed into 3149 cryopreservation bags at the Department of Transfusion Medicine. Sterility testing was performed for all bags. Until December 1998, 783 PBPC components were processed at a clean bench only (group I). Thereafter, 695 PBPC components have been processed at a clean bench located in a clean area with an airlock system for personnel and equipment (group II).

RESULTS: In group I, 16 of 1555 bags (1.03%) showed positive results in the first sterility testing. In group II, 21 of 1594 bags (1.32%) were positive (p = NS). The clinical follow‐up was inconspicuous.

CONCLUSION: Microbial contamination of PBPC components could not be reduced by installation of improved clean area conditions.
Volume of RBCs, 24‐ and 48‐hour posttransfusion survivals, and the lifespan of 51Cr and biotin‐X‐N‐hydroxysuccinimide (NHS)‐labeled autologous baboon RBCs: effect of the anticoagulant and blood pH on 51Cr and biotin‐X‐NHS elution in vivo
Authors: Valeri C. Robert , Pivacek Linda E. , Cassidy George P. , Ragno Gina
Published: March 2003

BACKGROUND: The RBC injury that occurs during collection of the first few milliliters of blood into the pH 5.0 ACD (NIH, Formula A) is referred to as the lesion of collection. The RBC injury was evaluated by labeling the ACD RBCs with 51Cr and measuring the 24‐hour posttransfusion survival. The effect of the acidification of ACD blood on the in vivo elution of 51Cr from the RBC has not been reported.

STUDY DESIGN AND METHODS: Baboon blood was collected in heparin and in ACD and CP2D at different ratios of blood to anticoagulant. ACD blood with a pH of 5.7 to 6.9 was labeled with 51Cr. Heparinized blood with a pH of 7.4 was labeled with biotin‐X‐N‐hydroxysuccinimide (NHS). ACD blood with a pH of 5.9 was labeled with both 51Cr and biotin‐X‐NHS. The RBC volumes, 24‐ and 48‐hour posttransfusion survivals, and lifespans (T50) were measured.

RESULTS: The RBC volume of ACD blood with a pH ranging from 5.7 to 6.9 was not affected by 51Cr labeling. 51Cr‐labeled ACD blood with a pH of 6.9 had an RBC volume that was significantly greater than that seen in heparinized blood with a pH of 7.4 labeled with biotin‐X‐NHS. In vivo elution of 51Cr from the RBCs prepared from the ACD blood with a pH of 5.7 to 6.9 and in vivo elution of biotin‐X‐NHS from RBCs prepared from ACD blood with a pH of 5.9 were associated with reductions in 24‐ and 48‐hour posttransfusion survivals and T50.

CONCLUSIONS:The anticoagulant and the pH of the medium in which the RBCs were labeled with 51Cr or biotin‐X‐NHS affected in vivo elution of the label from the RBCs and may reduce posttransfusion survival values.
Evaluation of a new generation of culture bottle using an automated bacterial culture system for detecting nine common contaminating organisms found in platelet components
Authors: Brecher M.E. , Heath D.G. , Hay S.N. , Rothenberg S.J. , Stutzman L.C.
Published: March 2003

BACKGROUND : An automated bacterial culture system (BacT/ALERT 3D, bioMérieux) has been previously validated with a variety of bacteria in platelets. The recovery of bacteria in platelets using a new generation of culture bottles that do not require venting and that use a liquid emulsion sensor was studied.

STUDY DESIGN AND METHODS
:
Bacillus cereus, Enterobacter cloacae, Escherichia coli, Klebsiella oxytoca, Staphylococcus aureus, Staphylococcus epidermidis, Serratia marcescens, Streptococcus viridans,
and
Propionibacterium acnes isolates were inoculated into Day 2 platelets to concentrations of 10 and 100 CFU per mL. Samples were then studied with current and new aerobic, anaerobic, and pediatric bottles.

RESULTS
: All organisms, except
P. acnes,
were detected in a mean time of 9.2 to 20.4 (10 CFU/mL) or 8.7 to 18.6 (100 CFU/mL) hours.
P. acnes was detected in a mean time of 69.2 (10 CFU/mL) or 66.0 (100 CFU/mL) hours. The 10‐fold increase in inoculum was associated with a mean 9.2 percent difference in detection time. The aerobic, anaerobic, and pediatric bottles had a mean difference in detection time (hours) between the current and new bottles of 0.10 (p = 0.61), 0.4 (p = 0.38), and 1.0 (p < 0.001), respectively.

CONCLUSION : No difference in detection time between the current and new aerobic and anaerobic bottles was demonstrated. The new pediatric bottles had a small but significant delay in detection.
The band III ligand dipyridamole protects human RBCs during photodynamic treatment while extracellular virus inactivation is not affected
Authors: Besselink G.A.J. , Van Engelenburg F.A.C. , Korsten H.G. , Blokker D.E. , Lagerberg J.W.A. , Brand A. , De Korte D. , Verhoeven A.J.
Published: March 2003

BACKGROUND : Recently, the potential usefulness of dipyridamole (DIP) in protecting RBCs against the harmful side effects of photodynamic sterilization was demonstrated. In the present study, the use of DIP for selective protection of RBCs was investigated under conditions more relevant for blood bank practice.

STUDY DESIGN AND METHODS : WBC‐reduced RBC suspensions (30% Hct) were treated with 1,9‐dimethylmethylene blue and red light, and the influence of the inclusion of DIP on photohemolysis was assessed as a function of sensitizer concentration, light dose, and storage time. Furthermore, the possible interference of DIP with inactivation of extracellular virus by use of a panel of different viruses (HIV‐1, pseudorabies virus [PRV], bovine viral diarrhea virus [BVDV], VSV, encephalomyocarditis, and canine parvovirus) was investigated.

RESULTS : In WBC‐reduced RBC suspensions (30% Hct), DIP exerted a clear protective effect against photohemolysis. Part of this protection was achieved with concentrations near the dissociation constant for band III binding. Importantly, efficiency of inactivation of extracellular HIV‐1, PRV, BVDV, and VSV was not significantly impaired by the inclusion of DIP. Phototreatment conditions, resulting in a 4 to 5 log inactivation of extracellular HIV‐1 and PRV, resulted in a high level of hemolysis after 28 days of storage. This long‐term hemolysis could be decreased, but not completely prevented, by the inclusion of DIP.

CONCLUSION : Photohemolysis in RBC concentrates can be reduced substantially by the application of DIP, while the efficacy of inactivation of HIV‐1 and other viruses remains unchanged.
The role of the paternal health history in cord blood banking
Authors: Askari Sabeen , Miller John , Clay Mary , Moran Sheila , Chrysler Gayl , McCullough Jeffrey
Published: March 2003

BACKGROUND : Umbilical cord blood (UCB) transplantation is becoming more widely used, yet ethical and policy issues regarding consent and health history persist. Whereas most UCB banks do not require paternal consent or paternal health history (PHH), both are obtained at this institution whenever possible. This study assessed the value of PHH in making UCB safer.

STUDY DESIGN AND METHODS : A retrospective review was performed of all cord blood units (CBUs) collected by this bank between November 1999 and October 2000. All discarded CBUs were studied to identify those deferred based exclusively on PHH provided by the father in the PHH questionnaire.

RESULTS : PHH was obtained for 301 of 655 (46%) CBUs collected. Of the 339 CBUs banked, 269 (79%) had PHH available. Three of the 301 CBUs in which PHH was available were discarded based solely on PHH, since maternal medical history and infectious disease testing were negative. Paternal high‐risk factors in those three cases were: gave money or drugs for sex; traveled to an HIV high‐risk area; and did not answer high‐risk questions.

CONCLUSION : Considerable time and effort is expended in the process and follow‐up of obtaining PHH with an overall indistinct and unconvincing role in minimizing infectious disease transmission risk in UCB banking.
A phased consent policy for cord blood donation
Authors: Vawter Dorothy E. , Rogers‐Chrysler Gayl , Clay Mary , Pittelko Larrie , Therkelsen Dave , Kim Debora , McCullough Jeffrey
Published: March 2003

This article focuses on ethical and policy questions concerning when consent may be sought for the collection and donation of cord blood. It reviews the advantages and disadvantages of alternative times for securing consent, challenges common objections to seeking consent during labor or after collection, and describes a phased consent process—a process that permits consent during early labor to the ex utero collection of cord blood followed by after‐consent collection to donation. The phased consent policy attends to the unique characteristics of cord blood collection and donation, respects donors and their families, maximizes the number and diversity of cord blood units collected, preserves the relationship between providers and patients, and preserves public trust in cord blood and other types of tissue banking.
Improved efficiency of national HIV, HCV, and HTLV antibody testing algorithms based on sequential screening immunoassays
Authors: Seed Clive R. , Margaritis Angelo R. , Bolton Wayne V. , Kiely Philip , Parker Susan , Piscitelli Lisa ,   Virology Subcommittee of the National Donor and Product Safety Committee, Australian Red Cross Blood Service
Published: March 2003

BACKGROUND : Traditional strategies for clarifying the antibody status of donors giving repeatedly reactive (RR) results on primary screening immunoassays (IA1) have usually involved direct testing by immunoblot. However, such strategies can generate nonspecific in determinate results. The aim of this report is to present the results of an alternative strategy based on the use of sequential immunoassays (SI) before immunoblot testing.

STUDY DESIGN AND METHODS : The efficiency of traditional and SI strategies was compared in terms of the number of IA1 RR samples requiring immunoblot testing and the percentage of immunoblot tests giving indeterminate results. In addition, the biologic false‐ reactive overlap between the PRISM assays selected as IA1 and candidate secondary screening immuno‐ assays (IA2) was calculated to determine the most efficient IA1/IA2 combinations.

RESULTS
: There was a significant decrease in the proportion of IA1 RR samples requiring immunoblot testing under the SI strategy when compared with existing site‐specific strategies for HIV (0.49 vs. 0.08, p
<
0.05), HCV (0.85 vs. 0.42, p
<
0.05), and HTLV (0.69 vs. 0.05, p
< 0.05) algorithms. In addition, there was a significant decrease in the percentage of immunoblot tests giving indeterminate results for HIV and HTLV under the SI strategy. However, there was no significant difference in the proportion of confirmed positive results for HIV, HCV, or HTLV before and after national SI algorithm implementation. For the anti‐HIV IA2s, there was considerable variation of biologic false‐reactive overlap with the PRISM HIV O plus chemiluminescent immunoassay (range, 1.6‐15.6%).

CONCLUSIONS : The results presented in this report demonstrate that the sequential use of screening immunoassays before immunoblot testing can significantly reduce both the number of immunoblot tests and proportion of indeterminate results, without impacting sensitivity, thereby improving algorithm efficiency and simplifying donor management.
Understanding and Preventing the Edge Effect
Authors: CHENEAU EDOUARD , WOLFRAM ROSWITHA , LEBORGNE LAURENT , WAKSMAN RON
Published: March 2003

Edge stenosis, combining neointimal proliferation and negative remodeling, remains a serious limitation of vascular brachytherapy. This review comprehensively presents terminology, definitions, mechanisms, and treatment strategies to better understand the complexities of edge narrowing. The major contributors to this phenomenon are known; understanding the practical solutions will enable us to further minimize the problem of the edge effect. (J Interven Cardiol 2003;16:1–7)
Transport of the sulfated, amidated bile acid, sulfolithocholyltaurine, into rat hepatocytes is mediated by Oatp1 and Oatp2
Authors: Meng Ling-Jie , Wang Pijun , Wolkoff Allan W. , Kim Richard B. , Tirona Rommel G. , Hofmann Alan F. , Pang Sandy K.
Published: February 2003

The uptake of the sulfated bile acid sulfolithocholyltaurine (SLCT) was investigated in isolated rat hepatocytes and in HeLa cells transfected with complementary DNAs (cDNAs) of organic anion transporting polypeptides (Oatps) 1 and 2 cloned from rat liver. In hepatocytes, transport of SLCT was greatly reduced by bromosulfophthalein (BSP), estrone sulfate, the precursor bile acids cholyltaurine and lithocholyltaurine, and 4,4'-diisothiocyanostilbene-2-2'-disulfonic acid (DIDS). However, SLCT transport was insensitive to 4-methylumbelliferyl sulfate, harmol sulfate, digoxin, fexofenadine, and lack of sodium ion. Because the estimation of kinetic constants was enhanced with use of inhibitors, BSP (1-50 μmol/L) was added to isolated rat hepatocytes to assess the various transport components for SLCT uptake. The resulting data showed a nonsaturable pathway and at least 2 pathways of different Michaelis-Menten constants (Km
) (70 and 6 μmol/L) and similar maximum velocities (Vmax
) (1.73 and 1.2 nmol/min/mg protein) and inhibition constants of 0.63 and 10.3 μmol/L for BSP. In expression systems, SLCT was taken up by Oatp1 and Oatp2 expressed in HeLa cells with similar Km
values (12.6 ± 6.2 and 14.6 ± 1.9 μmol/L). These Km
values were comparable to that observed for the high-affinity pathway in rat hepatocytes. In conclusion, the results suggest that transport of SLCT into rat liver is mediated in part by Oatp1 and Oatp2, high-affinity pathways, a lower-affinity pathway of unknown origin, and a nonsaturable pathway that is compatible with a transport system of high Km
and/or passive diffusion.
Aspirin and NS-398 inhibit hepatocyte growth factor-induced invasiveness of human hepatoma cells
Authors: Abiru Seigou , Nakao Kazuhiko , Ichikawa Tatsuki , Migita Kiyoshi , Shigeno Masaya , Sakamoto Miyuki , Ishikawa Hiroki , Hamasaki Keisuke , Nakata Keisuke , Eguchi Katsumi
Published: February 2003

Nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit cyclooxygenase (COX) activity and are considered to exert antitumor actions in a variety of cancer cells, although the effects are unlikely entirely due to COX inhibition. Because clinical observations suggest that hepatocyte growth factor (HGF) can promote metastasis of hepatoma cells while stimulating tumor invasiveness, we investigated the effect of aspirin and NS-398, a selective COX-2 inhibitor, on HGF-mediated invasiveness of HepG2 human hepatoma cells. HGF stimulated the invasiveness of HepG2 cells in Matrigel cell invasion assay, together with increased expression of matrix metalloproteinase (MMP) 9. Addition of aspirin or NS-398, similar to PD98059, which acts as a specific inhibitor of mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK), an upstream kinase regulating extracellular signal-regulated kinase (ERK)1/2, abrogated such actions of HGF without affecting cell viability. Aspirin and NS-398, in contrast to PD98059, did not suppress ERK1/2 phosphorylation induced by HGF. However, both agents inhibited the kinase activity of ERK1/2 induced by HGF and repressed HGF-induced phosphorylation of 90-kd ribosomal S6 kinase (RSK) and Elk-1, key downstream substrates of ERK1/2, resulting in the suppression of transcriptional activity of Elk-1 as well as nuclear factor κB (NF-κB) and AP-1, which are involved in MMP-9 gene regulation. In conclusion, our results suggest that aspirin and NS-398 inhibit HGF-induced invasiveness of HepG2 human hepatoma cells through ERK1/2.
Naked DNA injection for liver metastases treatment in rats
Authors: Baqué Patrick , Pierrefite-Carle Valérie , Gavelli Adolfo , Brossette Nicole , Benchimol Daniel , Bourgeon André , Staccini Pascal , Saint-Paul Marie-Christine , Rossi Bernard
Published: February 2003

The cytosine deaminase (CD) gene converts the nontoxic prodrug, 5-fluorocytosine (5-FC), into 5-fluorouracil (5-FU). We previously showed that injection of CD-bearing cancer cells followed by 5-FC treatment can act as an autologous tumor vaccine in a syngenic liver metastasis model in rats. In the present work, we analyzed the antitumor efficiency of a direct intratumoral injection of a CD-expressing plasmid. In rats bearing microscopic or macroscopic metastases in right and left liver lobes, an injection of a CD-expressing plasmid was performed in the left lobe tumor, followed by 5-FC treatment of the animals. A significant regression of the DNA-injected tumor was observed in 5-FC-treated rats, both in microscopic (P = .007) or advanced (P < .0001) tumor models. Moreover, this treatment also induced a potent distant bystander effect on untreated controlateral liver tumors and extrahepatic metastases, resulting in an increased survival compared with control animals in both tumor models (P < .05). In conclusion, these data suggest that direct intratumoral injection of a CD-expressing plasmid, associated to 5-FC administration, can constitute a powerful and innocuous alternative treatment for unresectable liver metastases from colon carcinoma.
Characterization of two hepatitis B virus populations isolated from a hepatitis B surface antigen-negative patient
Authors: Jeantet Damien , Chemin Isabelle , Mandrand Bernard , Zoulim Fabien , Trepo Christian , Kay Alan
Published: February 2003

In a study of surface antigen-negative, but weakly hepatitis B virus (HBV) DNA-positive, patients, we were able to amplify and clone whole HBV genomes from the serum of a cirrhotic patient. Sequencing showed that the patient harbored two different HBV populations, one of genotype A and the other of genotype D, with the genotype D genome apparently predominating. The surface antigen of the genotype A virus is heavily mutated, especially in the extracellular ≪ determinant a ≫ region, with several mutations that have not been previously described. The genotype D virus is a precore mutant. Both genomes possess the common A1762T-G1764A double mutation of the basal core promoter (BCP), and the genotype D virus is also mutated in the ≪ TATA box ≫ of the large surface antigen promoter. Biological characterization showed that the genotype A genome was fully replication-competent, whereas the genotype D genome replicated poorly. The small surface antigen of the genotype A virus was only very weakly recognized by commercial tests. The small surface antigen of the genotype D virus could be recognized by the tests, but it was mainly retained within transfected cells, probably because of an excess of large surface antigen. In conclusion, the cryptic nature of this double HBV infection is characterized by the predominance of the replication-deficient genotype D virus over the replication-competent genotype A virus.
Ursodeoxycholate and tauroursodeoxycholate inhibit cholangiocyte growth and secretion of BDL rats through activation of PKC alpha
Authors: Alpini Gianfranco , Baiocchi Leonardo , Glaser Shannon , Ueno Yoshiyuki , Marzioni Marco , Francis Heather , Phinizy Jo Lynne , Angelico Mario , LeSage Gene
Published: February 2003

Accumulating bile acids (BA) trigger cholangiocyte proliferation in chronic cholestasis. The aim of this study was to determine if ursodeoxycholate (UDCA) or tauroursodeoxycholate (TUDCA) chronic feeding prevents the increased cholangiocyte growth and secretion in bile duct-ligated (BDL) rats, if UDCA and TUDCA effects are associated with increased cholangiocyte apoptosis, and to determine if this inhibition is dependent on increased intracellular Ca2+ ([Ca2+]i) and activation of protein kinase C (PKC) alpha. Immediately after BDL, rats were fed UDCA or TUDCA (both 275 μmol/d) for 1 week. We determined the number of bile ducts in liver sections, cholangiocyte proliferation (by measurement of H3 histone and proliferating cellular nuclear antigen in isolated cholangiocytes), and ductal secretion. In purified cholangiocytes from 1-week BDL rats, we evaluated if UDCA and TUDCA directly inhibit cholangiocyte proliferation and secretin-stimulated adenosine 3', 5'-monophosphate levels. We determined if UDCA and TUDCA activate PKC, increase [Ca2+]i, and alter the apical BA transporter (ABAT) expression in cholangiocytes. UDCA and TUDCA inhibited in vivo the cholangiocyte proliferation, secretion, and ABAT expression. In vitro UDCA and TUDCA inhibition of cholangiocyte growth and secretion required increased [Ca2+]i and PKC alpha. In conclusion, activation of Ca2+-dependent PKC alpha is required for UDCA and TUDCA inhibition of cholangiocyte growth and secretion. Reduced cholangiocyte ABAT may decrease endogenous BA stimulation of cholangiocyte growth and secretion.
Effects of Sho-saiko-to on hepatocarcinogenesis and 8-hydroxy-2'-deoxyguanosine formation
Authors: Shiota Goshi , Maeta Yoshiko , Mukoyama Tomoyuki , Yanagidani Atsushi , Udagawa Akihide , Oyama Kenji , Yashima Kazuo , Kishimoto Yosuke , Nakai Yoichiro , Miura Tetsuo , Ito Hisao , Murawaki Yoshikazu , Kawasaki Hironaka
Published: February 2003

Oxidative stress plays an important role in hepatocarcinogenesis. Although Sho-saiko-to (TJ-9), a Japanese herbal medicine which has been recently administered to patients with chronic liver disease in Japan, prevents hepatocarcinogenesis, the mechanism by which TJ-9 protects against cancer development is not fully understood. 8-Hydroxy-2'-deoxyguanosine (8-OHdG), a DNA adduct by reactive oxygen species, is known as a parameter of genetic risk for hepatocarcinogenesis. To clarify whether the preventive effect on hepatocarcinogenesis by TJ-9 is dependent on 8-OHdG, the effect on 8-OHdG levels by TJ-9 was examined by using high-performance liquid chromatography-mass spectrometry (LC-MS) in a diethylnitrosamine (DEN)-induced hepatocarcinogenesis model of male Fisher rats. TJ-9 reduced the number of preneoplastic cells, detected as the glutathione S transferase P (GST-P)-positive hepatocytes, and inhibited the development of liver tumors. TJ-9 also significantly decreased the formation of 8-OHdG, as indicated by LC-MS and immunohistochemical analysis. In addition, ornithine decarboxylase (ODC) activity and the number of proliferating cell nuclear antigen (PCNA)-positive cells were not altered. An electron paramagnetic resonance spin-trapping technique showed that TJ-9 scavenges hydroxyl radicals in a dose-dependent manner. In conclusion, the results of the present study suggest that TJ-9 prevents hepatocarcinogenesis in association with inhibition of 8-OHdG formation.
Severity and correlates of liver disease in hepatitis C virus-infected injection drug users
Authors: Rai Rudra , Wilson Lucy E. , Astemborski Jacquie , Anania Frank , Torbenson Michael , Spoler Charles , Vlahov David , Strathdee Steffanie A. , Boitnott John , Nelson Kenrad E. , Thomas David L.
Published: February 2003

Between May 1996 and June 1998, 210 members of a cohort of 1,667 hepatitis C virus (HCV)-infected injection drug users (IDUs) were selected for liver biopsy procedure after stratification based on 2 consecutive serum alanine transaminase (ALT) levels. Liver histology, which could be fully evaluated for 207 subjects, was classified by using the modified Ishak scores. At the time of biopsy, the median age of subjects was 41.3 years and the median estimated duration of HCV infection was 20.7 years; 94% were African American; 78% men; 31% were human immunodeficiency virus (HIV) seropositive; and 76% had HCV genotype 1a or 1b. Total modified histologic activity index (MHAI) scores ranged from 0 to 9, and 26.6% had a total MHAI score of 5 or greater. Persons with a total MHAI score of 5 or greater were more likely to be HIV infected (P = .04). Higher fibrosis, indicated by Ishak modified fibrosis scores of 3 to 6, was present in 10.1% of subjects and was found more often in those older than 46 years of age (the highest quartile) (P < .01). Both fibrosis scores of 3 or greater and total scores of 5 or greater were associated with elevated ALT, aspartate transaminase (AST), and γ-glutamyl transpeptidase (GGT) levels (P < .01). When serial values were considered, the results of liver enzyme testing could reduce the probability of an IDU having a fibrosis score of 3 or greater from 10% to 3%. In conclusion, these data indicate that severe liver disease is uncommon in this urban, HCV-infected IDU cohort, especially in younger persons and those with repeatedly normal liver enzymes.