bluuebunny/crossref_metadata_2025 huggingface dataset:
Fragmentation of Therapeutic Human Immunoglobulin Preparations
Authors: Page M. , Ling C. , Dilger P. , Bentley M. , Forsey T. , Longstaff C. , Thorpe R.
Published: March 2009

Some commercial batches of human therapeutic immunoglobulins (Ig) have been found to show evidence of molecular fragmentation when examined by molecular sizing methodologies including sodium dodecyl sulphate polyacrylamide gel electrophoresis [SDS‐PAGE] and size exclusion high performance liquid chromatography (SE‐HPLC). These batches all demonstrated impaired immunobiological activity (efficacy) as assessed by Fc function measured using a rubella haemolytic assay and as such are likely to be subpotent for therapeutic use. Fragmented Igs were characterized by the presence of at least three protein bands and peaks additional to monomeric IgG. Incubation of Igs with blood enzymes (plasmin and kallikrein) reproduced the fragmentation patterns observed for intrinsically degraded batches, suggesting that fragmentation occurred by contamination with these proteases from the source material (human blood) during manufacture. Intravenous Igs (IVIG) were found to be more susceptible to proteolysis than intramuscular Igs, probably as a consequence of the post‐fractionation processing that some IVIGs receive which may induce molecular alterations, allowing enzyme access and fragmentation. Two of the products examined were found to be relatively resistant to proteolysis and both were formulated by processes that limit enzyme activity. These processes were inclusion of an enzyme inhibitor, α2‐macroglobulin, and formulation at acidic pH. Enzyme carry‐over into the final product is a likely cause of Ig fragmentation, and reduction in levels of such contamination should lead to improvements in product stability and efficacy.
A Comparative Study of Platelets Stored in Polyvinyl Chloride Containers Plasticised with Butyryl Trihexyl Citrate or Triethylhexyl Trimellitate
Authors: Turner V. S. , Mitchell S. G. , Kang S. K. , Hawker R.J.
Published: March 2009

Platelet concentrates (PCs), stored for 5 days in PL 2209, a new polyvinyl chloride (PVC) storage container plasticised with butyryl trihexyl citrate, were compared with those stored in PL 1240, a PVC platelet container plasticised with triethylhexyl trimellitate. In part 1 of the study, pooled platelet‐rich plasma (PRP) was aliquoted into each type of pack and pH, pCO2, pO2, hypotonic shock response, aggregation responses, lactate, glucose and ATP concentrations, and lactate dehydrogenase and β‐thromboglobulin release were compared at days 1, 3 and 5. In part 2, 12 volunteers gave a unit of blood on two separate occasions and PCs produced by the PRP method were stored in PL 2209 or PL 1240 for 5 days before autologous reinfusion of a 111In‐labelled sample. In vitro results demonstrated that PL 2209 was more gas permeable than PL 1240. In part 2 of the study, at day 5, pCO2 was 3.13±0.62 versus 5.14±0.69 (p<0.001), whilst pO2 was not significantly different for PL 2209 versus PL 1240, respectively. pH was better maintained in PL 2209 than in PL 1240 (7.38±0.13 vs. 7.24±0.10, respectively, p<0.01) after storage for 5 days. These results were confirmed by those from part 1. In vivo data were similar for PC stored in the two plastics with a multiple‐hit recovey of 40.9±12.1% for PL 2209 and 37.4±11.3% for PL 1240, and a multiple‐hit survival of 4.89±1.20 days and 5.28±2.06 days for PL 2209 and PL 1240, respectively. γ‐Camera imaging of volunteers showed similar biodistribution of radiolabeled platelets stored in each container. These results demonstrate that PL 2209 is a suitable container for storage of PCs for 5 days.
A Comparative Evaluation of the Sensitivity of Seven Anti‐Hepatitis C Virus Screening Tests
Authors: Couroucé Anne‐Marie , Barin Francis , Botté Christine , Lunel Françoise , Maisonneuve Pascale , Maniez Michèle , Trepo Christian
Published: March 2009

The sensitivity of the most recent generation of anti‐hepatitis C virus (anti‐HCV) screening tests from seven manufacturers was evaluated with a common panel of 530 specimens from 320 HCV‐infected subjects. This panel included 221 samples from 57 seroconverters (53 pre‐sero conversion negative specimens and 168 positive samples) and 309 selected specimens from 263 other HCV‐infected patients of which 19% exhibited NS3 or core reactivity alone with the presence of HCV‐RNA assessed by PCR. None of the seven screening tests detected all infectious and antibody‐positive specimens. However, important differences were observed between these assays in the number of false‐negative results, which seemed mainly due to nonreactivity of antibody to the NS3 antigen.
Impact of Allergy Screening for Blood Donors: Relationship to Nonhemolytic Transfusion Reactions
Authors: Wilhelm D. , Klüter H. , Klouche M. , Kirchner H.
Published: March 2009

There has been some discussion whether the atopic disposition of a blood donor is associated with a potentially higher incidence of hypersensitivity nonhemolytic transfusion reactions (NHTRs). Serum samples from patients who had suffered from NHTRs and samples from the platelet concentrates (PCs) responsible for the reactions were examined for total and specific IgE as diagnostic markers for allergic events. In addition, the allergy prevalence among 1,088 blood donors was determined to analyze the allergy prevalence among our blood donors. Our results indicate that in 90% of cases, allergic NHTRs were associated with specific IgE antibodies in the recipient's serum, indicating the allergic disposition of the patient. In contrast, specific IgE antibodies were detected in only 22% in the transfused PCs. However, among all investigated NHTRs, there was not a single case in which specific IgE antibodies were detected exclusively in the PC. The allergy prevalence among our blood donors was about 26%. In our opinion, the few cases in which the allergic disposition of blood donors in combination with the allergic disposition of the recipients was associated with NHTRs reflects the allergy prevalence among our blood donors in general (26%). On the basis of these findings, we conclude that allergy diagnosis for blood donors is only of minor value in the prevention and prediction of NHTRs, whereas allergy diagnosis for patients who require multiple PC transfusion might be helpful.
Alloimmunization against ly, a Low‐Frequency Antigen on Platelet Glycoprotein Ib/IX as a Cause of Severe Neonatal Alloimmune Thrombocytopenic Purpura
Authors: Kiefel V. , Vicariot M. , Giovangrandi Y. , Kroll H. , Böhringer M. , Greinacher A. , Breitfeld C. , Santoso S. , Mueller‐Eckhardt C.
Published: March 2009

Neonatal alloimmune thrombocytopenia (NAIT) is usually induced by platelet‐specific antibodies against HPA‐la (Zwa) or HPA‐5b (Bra). Recently, low‐frequency alloantigens on the platelet glycoprotein (GP) IIb/IIIa complex have been discovered as a cause for NAIT. In this report, a new low‐frequency platelet‐specific alloantigen, ly, is described which induced severe NAIT. The corresponding antigen was detected in 1/249 unrelated German blood donors. Antibody binding assays with trypsin‐digested platelets (ELISA, immunoprecipitation with biotin‐labelled platelets) indicate that the antigen is not localized on the glycocalicin moiety of GP Ibα, but may be situated on the remnant moiety of GP Ibα, GP IX or GPIbβ. Apparently, ly is not related to the HPA‐2 (Ko) antigen system.
Prediction of the Severity of Haemolytic Disease of the Newborn: Quantitative IgG Anti‐D Subclass Determinations Explain the Correlation with Functional Assay Results
Authors: Garner S.F. , Gorick B.D. , Lai W.Y.Y. , Brown D. , Taverner J. , Hughes‐Jones N.C. , Contreras M. , Lubenko A.
Published: March 2009

Sera containing anti‐D, taken from 44 RhD‐negative women with RhD‐positive infants, were tested in antibody‐dependent cellular cytotoxicity (ADCC) and monocyte monolayer assays (MMA) which used similar target and effector cell populations. In addition, the anti‐D concentration was measured in the Auto Analyzer and the number of IgG1 and IgG3 anti‐D molecules bound to the target red cells was measured by flow cytometry. The results of the functional assays and AutoAnalyzer quantitation were examined for correlation with IgG subclass quantitation and all results were compared for their ability to predict the severity of haemolytic disease of the newborn (HDN). ADCC correctly predicted HDN in 39/44 (88.6%) cases, AutoAnalyzer quantitation in 35/44 (79.5%) and the MMA in 32/44 (72.7%). For all three assays, the number of correct predictions was highest when the maternal serum contained both IgG1 and IgG3 anti‐D. ADCC activity and HDN were correlated with the number of cell‐bound IgG1 molecules (r≥0.58), but MMA activity was most closely correlated with the number of cell‐bound IgG3 molecules (r = 0.68). Hence the superior predictive value of ADCC is due to its ability to reflect the IgG1 component of maternal anti‐D, which has a better correlation than IgG3 anti‐D with the severity of HDN.
A ‘New’ Low‐Incidence Red Cell Antigen, WARR: Unique to Native Americans?
Authors: Coghlan Gail , Crow Martha , Spruell Peggy , Moulds Marilyn , Zelinski Teresa
Published: March 2009

Investigation of a mild case of hemolytic disease of the newborn has led to recognition of a ‘new’ low‐incidence red cell antigen, WARR (ISBT No. 700.55). Data gleaned from two kindreds, both with Native American heritage, exclude WARR from the MNS, Lutheran, Lewis, Duffy, Kidd, Xg, Chido/Rodgers, Kx and Gerbich blood group systems. Serologic or genetic evidence suggests it is not part of the Kell or Yt systems.
Haemolytic Disease of the Newborn due to Anti‐M
Authors: Duguid J.K.M. , Bromilow I.M. , Entwistle G.D. , Wilkinson R.
Published: March 2009

A patient with haemolytic disease of the newborn (HDN) due to anti‐M which required exchange transfusion is described. Anti‐M antibodies are usually assumed to be naturally occurring and to consist of immunoglobin M (IgM); many however have an immunoglobin G (IgG) component. In view of this and the described occurrence of HDN, recommendations are made regarding the management of a pregnancy in which anti‐M antibodies are detected.
Studies in Red Blood Cell Preservation 10. 51Cr Recovery of Red Cells after Liquid Storage in a Glycerol‐Containing Additive Solution
Authors: Greenwalt T.J. , Dumaswala U.J. , Rugg N.
Published: March 2009

The purpose of the present study was to compare the 24‐hour recovery of red blood cells stored for 9 weeks in a hypoosmolar additive solution containing 150 mM glycerol to cells stored in Adsol®. Seven units of packed red cells were split into 2 aliquots. To one sample, 100 ml of the experimental additive solution (EAS 25) was added, and to the other, 50 ml of Adsol. At the end of the storage period the cells were labeled with 51Cr. A double chromium technique was used to make it possible to perform comparative autologous studies in the same donor. The 24‐hour 51Cr recovery value for EAS 25 was 73.03 ± (SD) 4.2% and for Adsol 60.9 ± 7.1. At 9 weeks the adenosine triphosphate levels were not significantly better compared to Adsol but the other in vitro measurements were better. New approaches to the study of red cell preservation are suggested.
Evaluation of Storage Conditions of Platelet Concentrates Prepared from Pooled Buffy Coats
Authors: de Wildt‐Eggen J. , Bins M. , van Prooijen H.C.
Published: March 2009

Five days storage of pooled platelet concentrates (PCs) with high yields often results in a pH fall and poor platelet morphology despite the use of specific containers. In this study we evaluated two techniques for prolonged storage of PCs with high platelet counts, by measuring pH and platelet swirl. In routine procedures, 90 PCs, prepared from five buffy coats, were stored in a single 1‐litre PL 732 container. After 5 days storage, 51 PCs with platelet counts below 3.1 times 1011/U showed a pH above 6.8 and optimal swirling scores. 29 of 39 PCs (74%) with yields above 3.1 times 1011/U showed pH values below 6.8 and poor swirling scores. These poor results can be prevented by shortening the duration of storage. PCs with high yields can be stored for 5 days either by dividing the concentrate into two containers or by adjusting the platelet count to 3.1 times 1011/U. In the latter procedure, we reduced the volume of the concentrate and found that of 102 PCs, only 9 showed poor values for pH and platelet swirl. The data clearly indicate the importance of measuring the prestorage platelet count to select the optimal procedure for concentrate storage.
The Effect of Cysteine Modification and Proteinases on the Major Antigens (D, C, c, E and e) of the Rh Blood Group System
Authors: Schmitz G. , Sonneborn H.H. , Ernst M. , Blanchard D. , Gielen W. , Dahr W.
Published: March 2009

We have confirmed and extended previous observations showing that the (Rh) D antigen of erythrocyte membranes is destroyed by various reagents that modify cysteine (Cys) residues (Res.) and by trypsin as well as chymotrypsin, using thirty examples of monoclonal or polyclonal anti‐D in heamglutination inhibition assays. We have also shown that most C, c, E, e and BS58 epitopes are inactivated or weakened by most Cys reagents and by these proteinases, using monoclonal and polyclonal antibodies. Inactivation by 5,5‐dithiobis‐(2‐nitrobenzoic acid) was always fully reversible after subsequent dithioerythritol treatment. The essential Cys Res. appear to be buried in the membrane in view of the inability of some reagents to inactivate (iodoacetamide, iodoacetic acid) or reactivate (reduced glutathione) the antigens. Data obtained with N‐ethylmaleimide indicate that inactivation of the C and c antigens is, at least in part, attributable to (a) Cys Res. that is (are) different from that (those) involved in the E and e antigens. Data obtained with the Cys reagents and the proteinases suggest that more than one peptide loop of the Rh proteins is involved in the major Rh antigens.
CD8+ Lymphocyte Decrease in HIV Disease: Association with Anti‐CD4+ but Not with Anti‐CD8+ Lymphocyte Autoantibodies
Authors: Daniel V. , Süsal C. , Weimer R. , Zipperle S. , Kröpelin M. , Zimmermann R. , Huth‐Kühne A. , Gerhard I. , Maier H. , Opelz G.
Published: March 2009

HIV+ patients form autoantibodies against CD4+ and CD8+ lymphocytes. It was shown that anti‐CD4+ lymphocyte autoantibodies are associated with the depletion of CD4+ cells. In the present study we analyzed the relationship of anti‐CD4+ and anti‐CDS+ autoantibodies with the CD8+ lymphocyte decrease commonly observed during HIV disease. IgM and IgG antibodies as well as complement fragments were determined on the surface of CD4+ and CD8+ lymphocytes using double fluorescence flow cytometry. Anti‐CD8+ lymphocyte autoantibodies were found more often in HIV+ hemophilia patients (75/105 = 71%) than HIV‐ hemophilia patients (13/37 = 35%; p<0.0001), patients with pharyngeal carcinoma (20/44 = 45%; p = 0.002), habitual abortions (3/13 = 23%; p = 0.0009) or healthy individuals (93/223 = 42%; p< 0.0001). Anti‐CD8+ antibodies, mostly of the IgM type, occurred significantly more frequently than anti‐CD4+ antibodies in healthy controls (p< 0.0001), patients with pharyngeal carcinoma (p = 0.0001), or HIV‐ patients (p = 0.01). In HIV+ patients, however, anti‐CD4+ autoantibodies were found more often than anti‐CD8+ antibodies (85 vs 71%; p = 0.02). 70 of 104 (67%) HIV+ patients had autoantibodies on both CD4+ and CD8+ lymphocytes and the IgG/IgM/C3d autoantibody pattern was identical in 31 (44%) of the patients. Interestingly, peripheral blood CD8+ cell counts were significantly associated with anti‐CD4+ (p = 0.01) but not with anti‐CD8+ lymphocyte autoantibodies. It is hypothesized that the inhibition and depletion of CD4+ cells by anti‐CD4+ autoantibodies is associated with a loss of regulatory functions that leads to a depletion of antiviral cytotoxic CD8+ lymphocytes.
Spectrotype Analysis of Human ABO Antibodies: Evidence for Different Clonal Heterogeneity of IgM, IgG, and IgA Antibody Populations
Authors: Rieben Robert , Frauenfelder Agnes , Nydegger Urs E.
Published: March 2009

Clonal characteristics of ABO antibodies in 18 paired samples of serum and breast milk were analyzed by isoelectric focusing and affinity immunoblotting. Anti‐A and/or ‐B (anti‐A/B) IgM showed a uniform, polyclonal spectrotype with more than 20 bands and only minimal interindividual differences. In contrast, IgG spectrotypes were oligoclonal (< 12 bands) and individually distinct. IgA of serum as well as of breast milk showed oligoclonal motifs of up to 15 bands, but with more interindividual variance in intensity than IgM. The bands did not appear at an identical pH in milk and serum samples. The uniformity of IgM spectrotypes could be due to an absence of somatic mutations and thus reflect the use of unmutated germline genes. The greater clonal heterogeneity of IgG as compared to IgA indicates a difference in isotype‐switch regulation. Somatic hypermutation may be less active during the switch from IgM to IgA than during the switch to IgG, or the latter switch may be accompanied by a repertoire shift. Alternatively, the anti‐A/B IgA and IgG antibody populations could be derived from different clonal precursors.
The Rh Antigen D: Partial D Antigens and Associated Low Incidence Antigens
Authors: Tippett P. , Lomas‐Francis C. , Wallace M.
Published: March 2009

The expression of the Rh antigen D varies quantitatively and qualitatively (partial D); published information and 15 years' work studying D variants are discussed in this review. D epitopes correspond to the reaction patterns of monoclonal anti‐D with partial D antigens. Partial D antigens can be reported in terms of their D epitopes but the epitope profile of cells with a quantitative variant of D (weak D) is difficult to determine reliably by haemagglutination tests. Nine partial D antigens, categories II‐VII, DFR and two not previously reported, are identified by their epitope profiles and by association with low incidence antigens. Monoclonal anti‐D recognize 16 D epitopes and more epitopes are anticipated. The specificities of polyclonal anti‐D made by people with partial D antigens are considered in terms of possible D epitope specificities: recognized epitope specificities, or combination thereof, were not able to account for all observed reaction patterns of anti‐D made by immunized individuals with partial D pheno‐types. An attempt is made to understand partial D antigens and their associated low incidence antigens in terms of the molecular genetic information available.
HCV‐Infection in Blood Donors: Association between Anti‐HCV Core IgM Antibodies and Serum HCV RNA
Authors: Ounanian‐Paraz A. , Morel‐Baccard C. , Barlet V. , Gueddah N. , Schweizer B. , Bensa J.C. , Zarski J.P. , Seigneurin J.M.
Published: March 2009

Among 47 blood donors tested positive with HCV EIA 2.0 Abbott, 27 (57.4%) also reacted with four ‘third‐generation’ EIAs. The presence of anti‐HCV antibodies was confirmed with 3 different immunoblot assays in 16 of 27 sera (34.0%) while 10 samples (21.3%) had indeterminate profile with antibodies usually directed against structural core antigen. Anti‐HCV core IgM response was found in 12 of 47 sera (25.5%) and HCV viremia detected by the polymerase chain reaction (PCR) procedure was observed in 15 samples (31.9%). A comparative study of the different markers confirmed a good correlation between a strong antibody response in EIAs and immunoblot assays and the presence of HCV RNA in the serum; only 2 immunoblot indeterminate samples were PCR positive. An association was observed between IgM antibodies against ‘core’ epitopes and HCV RNA carriage: all IgM‐positive sera were found positive by PCR. However, the direct detection of viral genome remains the best method for identifiying HCV carriers in the blood donor population.
False Negative Anti‐HIV‐1/HlV‐2 ELlSAs in Acute HIV‐2 Infection
Authors: Christiansen Claus Bohn , Jessen Torben E. , Nielsen Claus , Staun‐Ohen Poul
Published: March 2009

Since HIV‐2 was isolated in 1986, only 1 case of acute HIV‐2 infection has been reported. We have identified another patient with primary HIV‐2 infection. Follow‐up samples were requested from the patient due to discrepant results. The HIV‐2 infection was confirmed with HIV‐2‐specific proviral DNA amplification by PCR. The HIV‐2 seroconversion panel obtained was used to evaluate the sensitivity of both combined and specific ELISAs currently in use in Europe, and to investigate the Western‐blot patterns on both HIV‐1‐ and HIV‐2‐specific Western blots. The window period was determined to be less than 37 days with the most sensitive assays. A remarkable difference in sensitivity to HIV‐2 antibodies in acute HIV‐2 infection was found in combined HIV‐l/HIV‐2 ELISAs. Three out of the 4 combined sandwich ELISAs appeared to be less sensitive than the indirect ELISAs in HIV‐2 seroconversion, leading to a prolonged window period. One HIV‐2‐specific ELISA was also negative on the first sample, but positive on the second sample. In the HIV‐2 Western blot, early reaction with HIV‐2‐specific env and gag proteins was seen, whereas the HIV‐1 Western blot on the first sample revealed gag (p24, p55) reactivity only.
A New Microplate Red Blood Cell Monolayer Technique for Screening and Identifying Red Blood Cell Antibodies
Authors: Llopis F. , Carbonell‐Uberos F. , Planelles M.D. , Montero M. , Plasencia I. , Carrillo C.
Published: March 2009

A new method has been developed to immobilize red blood cells in wells of microplates using a cell fixation buffer. This method has been employed for detecting and identifying red blood cell antibodies with greater sensitivity than haemagglutination antiglobulin tests, without loss of specificity. This method decreases the amount of test erythrocytes and anti‐human globulin reagents employed per test, consequently lowering the cost.
Effect of High‐ versus Intermediate‐Purity Blood Coagulation Factor Concentrates on HIV‐1 Replication
Authors: Schögl Daniela , Zimmermann Klaus , Turecek Peter L. , Schwarz Hans Peter , Eibl Martha M. , Mannhalter Josef W.
Published: March 2009

Controversy exists as to whether treatment of HIV‐1‐seropositive hemophiliacs with blood coagulation products of high purity might help prevent the decline of CD4‐positive lymphocytes and thus delay progression toward AIDS. As viral load has recently been shown to be associated with disease progression in HIV‐1 infection, we tested for a possible direct interference of high‐ or intermediate‐purity blood coagulation products with replication of HIV‐1. The data obtained revealed comparable replication of HIV‐1 in the presence and absence of all blood coagulation products tested (assessed by PCR‐based quantitation of proviral HIV‐1 DNA in infected cells after a 10‐day incubation period). These data suggest that coagulation factor concentrates per se will also have no direct effect on HIV‐1 replication in vivo.
A Safety Study of Albumex®5, a Human Albumin Solution Produced by Ion Exchange Chromatography
Authors: Wolf M. , Kronenberg H. , Dodds A. , Miach P. , Isbister J. , Levidiotis M. , Dear K.
Published: March 2009

Human serum albumin solutions have been used as plasma replacement fluids for many years and adverse events associated with their use are rare. Albumex®5 is a new albumin solution manufactured by a combination of the Cohn fractionation process and a chromatographic technique. A multicentre, randomised study was conducted to compare the safety of Albumex®5 and 5% normal serum albumin (5% NSA) in patients undergoing large volume therapeutic plasmapheresis for a variety of disorders. Up to six exchanges were performed on each patient. A total of 208 evaluable exchanges were performed on 40 patients, 109 5% NSA and 99 Albumex®5. There were 9 adverse events (reaction rate of 8.3%) in 6 patients with 5% NSA and 12 events (reaction rate 12.1%) in 8 patients with Albumex®5. The difference is not statistically significant (p = 0.37, Fisher's exact test). None of the adverse events were considered serious. Only six reactions were thought to be possibly related to the product and three exchanges were modified as a result of an adverse event; one with Albumex®5 was interrupted and two with 5% NSA were slowed. Albumex®5 can be considered to be a safe product with a low adverse event rate.
Liquid Pasteurization of an Immunoglobulin Preparation without Stabilizer: Effects on its Biological and Biochemical Properties
Authors: Bridonneau P. , Marcilly H. , Vernois‐Martin M. , Goigoux P. , Bourdel V. , Laulan A. , Deramoudt F.X. , Desmadril M. , Sitbon M. , Basuyaux B. , Steinbuch M. , Schmitthaeusler R.
Published: March 2009

Intravenous immunoglobulins (IVIg) purified by cold ethanol fractionation have a very good safety record with regard to the transmission of many viruses. However, a few cases of non‐A‐non‐B hepatitis have been described after intravenous injection of some immunoglobulin preparations. To ensure even higher safety for our IVIg, an additional virus inactivation step, based on pasteurization, was developed. The heating of aqueous IVIg was performed without stabilizer, and at a very low salt concentration (< 1 mM) at acidic pH. No generation of polymer was detected after pasteurization and a significant decrease in the proportion of dimers was observed. Analysis of the secondary structure by circular dichroism showed a very slight change in the secondary structure. The biological properties of the Fc region as well as the Fab region were not affected by the pasteurization. Our method has several advantages: (1) improvement of viral safety; (2) there is no need to add stabilizer which may stabilize viral particles, and (3) the absence of any hypotensive effect and low anticomplementary activity indicates a good clinical tolerance of IgG preparation.
Autologous Blood Donation for Elective Surgery in Children Weighing 8–25 kg
Authors: Mayer M.N. , de Montalembert M. , Audat F. , Brusset M.C. , Houfani B. , Merckx J. , Barrier G. , Gazengel C.
Published: March 2009

To determine whether autologous blood donation can be used safely and efficiently in children weighing 8–25 kg, we studied children whose perioperative blood losses were expected to exceed 25% of total blood volume. Blood donations were performed in pediatric units, under the direction of an anesthesiologist and a blood bank physician experienced in pediatric care. Twenty‐four children, median age 6 years (1–13), were included. They underwent surgery mainly for digestive or urological disorders, and for orthopedic defects. Forty blood collections were performed of the 46 prescribed. Phlebotomies could not be performed in 1 child because of the mother's apprehension, and in 5 cases because of venous access problems. All phlebotomies were hemodynamically well tolerated. Hemodilution was also performed in 17 children, and cell saver used in 2. Allogeneic blood transfusion was avoided in 21/24 children.
Multicenter Evaluation of the 3% Paraformaldehyde Method for White Cell Counting in Leukocyte‐Reduced Red Blood Cells
Authors: Prati Daniele , Brandwein Harvey , Capelli Carmen , Dzik Walter H. , Masse Maurice , Myllylä Gunnar , Stromberg Robert R. , Takahashi Tsuneo , Vyas Girish N. , Wenz Barry
Published: March 2009

Background/Aim: The 3% paraformaldehyde (PFA) method is a simple technique for counting residual white blood cells (WBC) in leukocyte‐depleted red blood cells (RBC). Preliminary data suggested that its sensitivity is at least equal to PCR and flow cytometry. We report the results of a multicenter study conducted by the BEST Working Party to determine precision and accuracy of the 3% PFA method. Study Design: In the 7 participating laboratories, 5 sets of samples containing nominal concentrations of 200, 100, 50, and 10 WBC/ml were prepared by diluting whole blood into ‘WBC‐free’ RBC. Ten milliliters of each sample were processed using the 3% PFA method, which is based on erythrocyte lysis and WBC concentration into 5% of the original sample volume; a Nageotte chamber is used to count concentrated WBC. Results: The precision of the technique varied according to the nominal concentration, ranging from a CV of 12% at 200 WBC/ml to 57% at 10 WBC/ml. The technique measured fewer than the nominal WBC concentrations (mean of all laboratories, ‐12.4%); underestimation was probably due to cell loss during sample manipulation. Overall accuracy was however acceptable, because statistical considerations establish that the actual WBC concentration would unlikely exceed 2 times the estimated count. Conclusions: The 3% PFA method is suitable for the enumeration of residual WBC at concentrations ≥50/ml. It represents a useful tool for evaluation of high performance filters by reference laboratories.
IN VITRO (EX VIVO) EXPANSION OF MURINE HEMATOPOIETIC STEM CELLS Running Title: In Vitro Expansion of Stem Cells
Authors: OGAWA MAKIO , YONEMURA YUJI , KU HSUN
Published: March 2009

Currently there is significant interest in in vitro (ex vivo) expansion of human hematopoietic stem and progenitor cells in hematology and oncology. Transplantation of umbilical cord blood stem cells promises to be a new and exciting mode of bone marrow/stem cell transplantation since it is inexpensive and appears to be associated with milder graft vs. host disease (GVHD) than transplantation of adult cells. A potential problem of cord blood transplantation is the limited number of stem cells that may be harvested. The majority of cases of cord blood transplantation, therefore, have been attempted in children. If it were possible to expand the population of stem cells in umbilical cord blood and transplant them to adult recipients, it would significantly enlarge the scope of this form of transplantation. Already many preclinical studies have shown that it is possible to expand the population of total cells and progenitors in culture in the presence of combinations of cytokines. Whether or not it is possible to expand the population of hematopoietic stem cells which have long‐term reconstitution capabilities remains to be clarified (1–3). The cytokines used in the majority of attempts of in vitro expansion were selected from a group of early‐acting cytokines such as steel factor (SF, kit ligand), flt3 ligand (FL), interleukin (IL)‐3, IL‐6, IL‐11, granulocyte/ macrophage‐colony stimulating factor (GM‐CSF), and granulocyte‐CSF (G‐CSF). The rationale for combining these cytokines has been explained in a recent review (4). Almost all protocols for in vitro expansion include IL‐3 and many contain IL‐1 because of their known myelopoietic effects.
PRESENT AND FUTURE PROBLEMS OF TRANSFUSION ‐ TRANSMITTED HIV INFECTION
Authors: BEAL R.W.
Published: March 2009

To an external viewer from a developed country, there would appear to be few problems associated with transfusion‐transmitted HIV infection (TTHIV) in 1996. There has been virtually no TTHIV recorded in most developed countries since effective screening tests were introduced in the mid‐1980s. Such a conclusion would be simplistic for a developed country, and inappropriate for a developing country. Both settings require effort and vigilance if TTHIV is to be reduced and minimised, and where it is already of low prevalence, to avoid complacency and a reduction in safety.

Blood transfusion was (and regretfully still is in some countries) the most efficient and, in theory, the most easily preventable means of HIV transmission. The infectivity of a unit of blood is related to the large volume of virus transmitted, the intravenous route of exposure and, sometimes, the debilitated state of the recipient. In those countries where universal screening was introduced early, prevention has been effective, if not total. This situation does not apply to many countries in the world, where health funding does not allow universal screening.
PRACTICE OF QUALITY ASSURANCE IN BLOOD CENTERS — THE SYSTEM AND DAILY WORK
Authors: NEVALAINEN DAVID E.
Published: March 2009

Using process documentation is not an occasional activity that gets done, but rather must be woven into the very fabric of an organization's activities and values. Management must take active leadership roles in the quality process. If management gets involved in the implementation of the quality program, and makes quality a strategic part of their business plan, it will result in a more focused organization, that can more effectively meets its financial requirements and be increasingly competitive in the ever more competitive environment of health care in the 21st century. To do that will require that blood banks learn how to consistently apply the use of quality records, audits, training, and quality metrics. The table and graph demonstrate the areas where blood bank management can work more effectively to lead their organizations beyond GMP and into the quality systems world of the future.
Detection of Sclerotinia sclerotiorum in Planta by a Real‐time PCR Assay
Authors: Yin Yanni , Ding Laisong , Liu Xin , Yang Jinghui , Ma Zhonghua
Published: March 2009

Stem rot caused by Sclerotinia sclerotiorum is a very serious disease on oilseed rape worldwide. In this study, a pair of polymerase chain reaction (PCR) primers was designed based on the nucleotide sequence of a DNA region amplified by a microsatellite primer M13. The primer pair amplified a 252‐bp fragment from all S. sclerotiorum isolates collected from oilseed rapes at different locations in different years, but not from any other fungus tested. Using this pair of primers, a real‐time PCR assay was developed to rapidly detect early infection of S. sclerotiorum on petals of oilseed rape. The real‐time PCR assay developed in this study could help growers make a timely decision on fungicide application.
Assessing the ecological status of the North‐western Adriatic Sea within the European Water Framework Directive: a comparison of Bentix, AMBI and M‐AMBI methods
Authors: Simonini Roberto , Grandi Valentina , Massamba‐N’Siala Gloria , Iotti Mirko , Montanari Giuseppe , Prevedelli Daniela
Published: March 2009

In this paper we looked for the most suitable method for benthic ecological quality status (EcoQS) assessment of the North‐western Adriatic Sea. Taking into account the historical and present information, we inter‐compared our expectations in terms of EcoQS, with the Bentix, AMBI and M‐AMBI classification metrics results, and investigated the sensitivity of these indices to the spatial, seasonal and inter‐annual variability of benthic communities. The sampling sites were located at increasing distances with respect to the Po River mouth, along an (inverse) gradient of eutrophication. Moreover, a 2‐year monthly monitoring plan (October 2004–September 2006) was carried out at the CEBEB site, located in a central position with respect to the investigated area. The results highlighted that the three indices differed in the sensitivity to spatial and temporal variability of the benthic communities. The strong seasonal and inter‐annual changes in the benthic communities had a significant effect on the Bentix, whereas no relevant modifications were observed on the EcoQS assignation obtained using the M‐AMBI and AMBI methods. The environmental quality ratios (EQR) values and EcoQS classification obtained by the Bentix and AMBI did not match expectations, as they were not inversely correlated with the site distances with respect to the Po mouth, whereas the M‐AMBI clearly discriminated the most and the least eutrophicated sites. It thus appears that the M‐AMBI could be considered to be the most valuable method for the ecological‐status assessment of the North‐western Adriatic Sea benthic ecosystem.
Primary central nervous system lymphoma initially mimicking lymphomatosis cerebri: An autopsy case report
Authors: Sugie Masayuki , Ishihara Kenji , Kato Hirotaka , Nakano Imaharu , Kawamura Mitsuru
Published: March 2009

A 59‐year‐old immunocompetent man was admitted to our hospital because of progressive dementia with concomitant bilateral uveitis. The first brain MRI revealed diffuse hyperintense lesions in the cerebral white matter of both hemispheres on a T2‐weighted image and fluid‐attenuated inversion recovery image. However, another MRI taken more than 1 month later revealed enhanced cohesive mass lesions in the bilateral thalami, in addition to the white matter lesions. The white matter lesions were slightly hyperintense on a diffusion‐weighted image and apparent diffusion coefficient map image, suggesting vasogenic edema. One year after the onset of uveitis, he died of respiratory failure. Pathological diagnosis was diffuse large B‐cell lymphoma with perivascular proliferation and diffuse scattered infiltration in the cerebrum and brainstem. Microscopically, cohesive mass lesions in the bilateral thalami were a massive cluster of lymphoma cells. This is a case of primary CNS lymphoma (PCNSL) mimicking ‘lymphomatosis cerebri (LC)’ at first but later exhibiting typical mass lesions, giving rise to the possibility that cases of LC might unmask features of regular lymphomas in their later course more often than believed thus far.
The first Japanese patient with variant Creutzfeldt‐Jakob disease (vCJD)
Authors: Shinde Akiyo , Kunieda Takenobu , Kinoshita Yoshimi , Wate Reika , Nakano Satoshi , Ito Hidefumi , Yamada Masahito , Kitamoto Tetsuyuki , Nakamura Yosikazu , Matsumoto Sadayuki , Kusaka Hirofumi
Published: March 2009

Eleven years after a brief visit to some European countries, a 48‐year‐old Japanese man developed writing difficulty, irritability and general fatigue. Then he complained of dysesthetic pains in his legs, for which benzodiazepines were prescribed. However, at the time pulvinar sign was retrospectively confirmed on brain MRI. Eighteen months after the onset, his gait became ataxic with rapid deterioration of mental status over the following several months. Thirty‐one months after the onset, he became akinetic and mute with periodic synchronous discharges on EEG, and died at the age of 51. The total clinical course was approximately 43 months. Pathological examination revealed the characteristic alterations of spongiform encephalopathy, severe in the thalamus, moderate but widely spread in the cerebral cortices, and moderate in the cerebellum. Abundant amyloid plaques were easily identified in the cerebral cortex and the cerebellum on HE staining. Immunohistochemistry for abnormal prion protein (PrPsc) confirmed amyloid plaques in several forms, such as florid, uni‐ and multi‐centric plaques as well as perineuronal and periaxonal deposits in the basal ganglia and synaptic patterns in the thalami. A Western blotting study identified type 2B protease‐resistant PrP. This is the first Japanese patient who was definitely diagnosed as variant Creutzfeldt‐Jakob disease (vCJD). The pathological findings were similar to those of previous reports of vCJD in the UK. However, the changes were much more severe both in degree and distribution, probably due to a longer duration of the illness than those in the UK.
Prion protein oligomers in Creutzfeldt‐Jakob disease detected by gel‐filtration centrifuge columns
Authors: Minaki Haruhiko , Sasaki Kensuke , Honda Hiroyuki , Iwaki Toru
Published: March 2009

Prion diseases are diagnosed by the detection of accumulation of abnormal prion protein (PrP) using immunohistochemistry or the detection of protease‐resistant abnormal PrP (PrPres). Although the abnormal PrP is neurotoxic by forming aggregates, recent studies suggest that the most infectious units are smaller than the amyloid fibrils. In the present study, we developed a simplified method by applying size‐exclusion gel‐filtration chromatography to examine PrP oligomers without proteinase K digestion in Creutzfeldt‐Jakob disease (CJD) samples, and evaluated the correlation between disease severity and the polymerization degree of PrP. Brain homogenates of human CJD and non‐CJD cases were applied to the gel‐filtration spin columns, and fractionated PrP molecules in each fraction were detected by western blot. We observed that PrP oligomers could be detected by the simple gel‐filtration method and distinctly separated from monomeric cellular PrP (PrPc). PrP oligomers were increased according to the disease severity, accompanied by the depletion of PrPc. The separated PrP oligomers were already protease‐resistant in the case with short disease duration. In the cases with quite severe pathology the oligomeric PrP reached a plateau, which may indicate that PrP molecules could mostly develop into amyloid fibrils in the advanced stages. The increase of PrP oligomers correlated with the degree of histopathological changes such as spongiosis and gliosis. The decrease of monomeric PrPc was unexpectedly obvious in the diseased cases. Dynamic changes of both oligomerization of the human PrP and depletion of normal PrPc require further elucidation to develop a greater understanding of the pathogenesis of human prion diseases.
Intraventricular metaplastic meningioma in a child: case report and review of the literature
Authors: Dulai Mohanpal Singh , Khan Abdul‐Majid , Edwards Michael S. B. , Vogel Hannes
Published: March 2009

Childhood meningiomas are rare and display important differences from adult forms. We report the first case of an intraventricular metaplastic meningioma arising in a child. A 7‐year‐old female underwent resection of an enhancing tumor arising within the left lateral ventricle. It was composed of monomorphic cells embedded within an abundant myxoid stroma. The cells demonstrated epithelial membrane antigen and vimentin immunoreactivity. Ultrastructural analysis demonstrated intermediate filaments, complex intercellular interdigitations and desmosomes, and a diagnosis of myxoid (metaplastic) meningioma was rendered. This case reflects the higher incidence of intraventricular meningiomas in childhood and greater incidence of intraventricular meningiomas in the left lateral ventricle. Recognition of the grade I myxoid meningioma in this case is paramount since chordoid meningiomas, which share similar histologic features, are of a higher grade and worse prognosis.
Mental health nurses establishing psychosocial interventions within acute inpatient settings
Authors: Mullen Antony
Published: March 2009

Acute inpatient units provide care for the most acutely unwell people experiencing a mental illness. As a result, the focus for care is on the containment of difficult behaviour and the management of those considered to be ‘at high risk’ of harm. Subsequently, recovery‐based philosophies are being eroded, and psychosocial interventions are not being provided. Despite the pivotal role that mental health nurses play in the treatment process in the acute inpatient setting, a review of the literature indicates that mental health nursing practice is too custodial, and essentially operates within an observational framework without actively providing psychosocial interventions. This paper will discuss the problems with mental health nursing practice in acute inpatient units highlighted in the current literature. It will then put forward the argument for routine use of psychosocial interventions as a means of addressing some of these problems.
Vulnerable populations and multicentred research
Authors: McCauley‐Elsom Kay , Gurvich Caroline , Lee Susan , Elsom Stephen , O'Connor Margaret , Kulkarni Jayashri
Published: March 2009

There are many challenges facing researchers with projects requiring multicentred ethics approval. Achieving ethical approval at multiple sites, whether statewide or nationally, is a complex and time‐consuming experience, compounded by the research process itself, as well as the recruitment of clinical sites and participants. Human ethics and research committees act as research gatekeepers and, as many research activities involve multiple applications and multiple approvals, can considerably delay the commencement of a project. A delay in ethics approval results in delays recruiting staff and participants, delays in the utilization of funding, and in delays regarding the progress and completion of projects. Such problems are additional problems for researchers of vulnerable populations, such as those in mental health or palliative care, where multicentred research is necessary to ensure the validity of the project itself. A current example of work that has required multicentred human ethics and research approval from around Australia is the establishment of the National Register of Antipsychotic Medication in Pregnancy. The guidelines for embarking on such a project with the requirement of multicentred ethical approval are described. Some of the issues, recommendations, and guidelines presented by the authors are taken from their experiences in establishing multicentred research projects.
Testing and using Goldberg's General Health Questionnaire: Mental health in relation to home nursing, home help, and family care among older, care‐dependent individuals
Authors: Dale Bjørg , Sævareid Hans Inge , Söderhamn Olle
Published: March 2009

The aims of this study were to test the Norwegian version of Goldberg's 30‐item General Health Questionnaire (GHQ‐30) in a group of older, care‐dependent individuals living at home; to describe self‐reported mental health; and to relate mental health to receiving home nursing, home help, and family care. A sample of 234 home nursing patients in Norway aged 75 years and older was interviewed. Mental state was assessed using the GHQ‐30. Reliability and validity were calculated with Spearman's rank correlations, Cronbach's alpha coefficient, and Mann–WhitneyU‐test. The factor analysis was performed using the principal components analysis with varimax rotation and Kaiser normalization. Demographic characteristics and amounts of formal and family care were recorded, and descriptive statistics and stepwise multiple regression were used in the analyses. Cronbach's alpha coefficient for the GHQ was 0.92. The item–total correlations were generally acceptable. For items concerning depression and anxiety, the item–total correlations ranged from rs=0.60 to 0.77. The factors extracted in the factor analysis explained 70% of the variance in the group. Females<85 years of age living in urban areas were associated with reduced mental health. There were no associations between general mental health and the amounts of formal and family care provided.
Systematic screening for metabolic syndrome in consumers with severe mental illness
Authors: Brunero Scott , Lamont Scott
Published: March 2009

The high prevalence of metabolic syndrome (MetS) in people with a mental illness has been reported recently in the literature. Gaps have emerged in the widespread use of systematic screening methods that identify this collection of critical risk factors for cardiac and metabolic disorders in people with severe mental illness. A sample (n = 103) of consumers with severe mental illness was screened for MetS using the Metabolic Syndrome Screening Tool and compared to a sample (n = 72) of consumers who were not receiving a systematic approach to screening for MetS. The results demonstrated ad hoc screening of consumers for MetS in the comparison group, potentially leaving patients at risk of cardiac and metabolic disorders being untreated. Mental health nurses are well placed to show leadership in the screening, treatment, and ongoing management of MetS in people with severe mental illness. A potential new speciality role entitled the ‘cardiometabolic mental health nurse’ is proposed as a means leading to improved outcomes for consumers who have both the complication of physical health problems and a severe mental illness.