bluuebunny/crossref_metadata_2025 huggingface dataset:
Epitope Specificity of Blood‐Group‐A‐Reactive Murine Monoclonal Antibodies
Authors: Lubenko A. , Ivanyi J.
Published: March 2009

Abstract. Monoclonal antibodies (MABs) towards blood group A manifested three broad patterns of binding distinguished on the basis of affinities for A1 or A2B phenotype red blood cells and blood group substances. Competitive inhibition of binding of radiolabelled MABs by other unlabelled antibodies and absorption studies with synthetic haptens suggested that GpA specificity was expressed as two topographically related structures, one being the terminal GpA trisaccharide and the second probably involving part of the oligosaccharide backbone. The enhancement of binding of an extremely low‐affinity antibody by higher affinity antibodies recognising either epitope indicated a topographic, possibly conformational relationship between these epitopes. It is suggested that only those antibodies that recognise terminal GpA trisaccharides can agglutinate weak A2B cells.
Rh(D) Alloimmunization in the Absence of Exposure to Rh(D) Antigen
Authors: Biggins Kieran R. , Bowman John M.
Published: March 2009

Abstract. An anti‐D was detected in the serum of an Rh(D)‐negative primigravida. There is no history of previous pregnancies or blood transfusions. The husband and infant are both Rh(D)‐negative.
Effect of Sodium Phosphonoformate on Blood Group Antibody Detection
Authors: Hunt A.F. , Jones M. T.A. , Howkins G.J.
Published: March 2009

Abstract. A panel of 47 blood group antibodies covering a range of blood group antigens has been tested in the presence of 680 μmol concentration sodium phosphonoformate as a putative chemical additive to serum to reduce infectivity of ‘high risk’ HTLV‐III/HBsAg samples. No inhibitory effect could be demonstrated using the indirect antiglobulin test or prepapainized cell test.
Proposal for the Nomenclature of Human Plasminogen (PLG) Polymorphism
Authors: Skoda Ulrike , Bertrams J. , Dykes D. , Eiberg H. , Hobart M. , Hummel K. , Kühnl P. , Mauff G. , Nakamura S. , Nishimukai H. , Raum D. , Tokunaga K. , Weidinger S.
Published: March 2009

Abstract. Since its discovery, human plasminogen (PLG) polymorphism has received widespread acceptance in population genetics and forensic haematology. Due to the large number of variant alleles described, a PLG reference typing and Plasminogen Symposium was held, at which a nomenclature proposal was inaugurated. The technology of comparing PLG variants was based on isoelectric focusing and subsequent detection by caseinolytic overlay and ‘Western’ blotting. Typing results permitted comparison of so far described variant designations and resulted in a new nomenclature proposal for PLG polymorphism. It is recommended that the two most common alleles found in all investigated races be called: PLG*A (previously also PLG*1) and PLG*B (previously also PLG*2), the known variants with acidic pI: PLG*A1 to *A3, intermediate variants: PLG*M1 to *M5, PLG*M5 being functionally inactive, and basic variants: PLG*B1 to *B3. For future classification of newly discovered variants, samples should be compared at any of the laboratories participating in the reference typing.
Screening of Factor VIII:C Levels in Blood Donors
Authors: Carlebjörk G. , Blombäck M. , Blomstedt M. , Åkerblom O.
Published: March 2009

Abstract. A new chromogenic peptide substrate method, modified for assay with semi‐micro tubes, Cobas Bio centrifugal analyser and microplates, was used for screening of F VIII:C in blood donors. The precision of the assays is high and the costs are reasonable. The assay with microplates is especially suitable for selection of donors for a plasma programme and for quality control in blood banks.
Limitations of Polybrene to Detect ABO Incompatibility
Authors: Mintz Paul D. , Anderson Garth
Published: March 2009

Abstract. The ability of the manual Polybrene™ test to detect ABO incompatibility was assessed since the anti‐human globulin phase of the crossmatch is no longer mandated under many circumstances by the Standards of the American Association of Blood Banks, while the ability of the crossmatch to detect ABO incompatibility remains a requirement. 10,084 randomly selected patient samples (from approximately 6,000 different patients) were entered into a double‐blind study. Sera from 97 different patients gave unexpectedly negative reactions with A2 and/or B cells in the manual Polybrene test. 94 of these sera reacted as expected in saline (87 after an immediate‐spin examination). The manual Polybrene test does not provide adequate assurance of ABO compatibility.
Preservation of Red Blood Cells: Content of Microaggregates and Di‐2‐Ethylhexylphthalate (DEHP) in Red Blood Cells Stored in Saline‐Adenine‐Glucose‐Mannitol (SAGM) Medium
Authors: Gulliksson H. , Karlman G. , Segerlind A. , Gullbring B.
Published: March 2009

Abstract. The content of microaggregates was determined in leukocyte‐poor red blood cells (RBC) after 5 weeks of storage in citrate‐phosphate‐dextrose (CPD)‐adenine plasma and in saline‐adenine‐glucose‐mannitol (SAGM) medium. The size range was 3,600‐200,000 fl, corresponding to a particle diameter of 19–73 μm, presuming a spherical particle shape. In comparison with CPD‐adenine whole blood (100%), the total number of particles per unit of RBC was found to be 41.2% (CPD‐adenine anticoagulant) and 25.4% (SAGM medium). The corresponding data for total particle volumes were 43.3 and 30.3%. The significant differences between the two categories of RBC could not be explained on the basis of differences concerning pH or cell composition. A majority of particles (50–62%) was observed in the lowest size range (19–29 μm), corresponding to only 18–26% of the total particle volume. The accumulation of di‐2‐ethylhexylphthalate (DEHP) was determined in two variations of RBC stored in SAGM medium and in RBC stored in CPD‐adenine plasma. The quantities of DEHP per unit of RBC after 5 weeks were significantly smaller in SAGM medium (9.0 ±3.6 and 22.7 ± 6.1 mg, respectively) compared to CPD‐adenine plasma (30.1±9.5mg).
Serum ALT Levels in Patients with Primary Hypogammaglobulinemia Receiving Replacement Therapy with Intravenous Immunoglobulin or Fresh Frozen Plasma
Authors: Leen C.L.S. , Yap P.L. , Neill G. , McClelland D.B.L. , Westwood A.
Published: March 2009

Abstract. The intravenous immunoglobulin (IV. IgG) preparation used in this study is manufactured by the Scottish National Blood Transfusion Service (SNBTS) by the pH 4/mild pepsin method. Recent reports suggest that non‐A, non‐B hepatitis may be transmitted by certain intravenous immunoglobulin preparations. Serum ALT levels were therefore measured prospectively in 16 patients with primary hypogammaglobulinaemia who received an intravenous immunoglobulin replacement therapy (SNBTS IV IgG) over a period ranging from 6 to 25 months. Retrospective analysis of serum ALT levels was also carried out in 8 patients with primary hypogammaglobulinaemia who received fresh frozen plasma (FFP) for periods ranging from 8 months to 13 years. There was no evidence of non‐A, non‐B hepatitis transmission by either SNBTS IV IgG or by FTP in all the patients studied.
Epidemiology of Delta Agent Infection in Arabia: Geographical Distribution and Prevalence of Anti‐Delta
Authors: El‐Hazmi M.A. , Ramia S.
Published: March 2009

Abstract.The epidemiology of infection with the recently discovered delta (δ) agent was assessed in Saudi Arabia from the prevalence of antibody to delta (anti‐δ) in 488 HBsAg‐seropositive subjects from various regions of the country. In the Riyadh area, anti‐δ was prevalent in 22.2% of chronic hepatitis B patients compared to 7.9% and 6.7% in active hepatitis B patients and HBsAg carriers, respectively. Anti‐δ was also prevalent in HBsAg carriers from A1‐Hafouf (5.3%) and Najran (9.6%), but no anti‐δ was detected in 40 HBsAg‐positive polytransfused thalassemics and in 145 HBsAg carriers from Jaizan and Khaiber. It seems that δ infection is present in only certain regions of Saudi Arabia, and infection is possibly transmitted parenterally in these populations.
II. Relation between Cellular Morphology and Viability
Authors: Högman Claes F. , de Verdier Carl‐Henric , Borgström Lena
Published: March 2009

Abstract. Red cells collected in CPD, suspended in SAGM solution and stored for 42 days at +4°C, were tested before and after incubation for 1 h at 37°C with respect to autologous 24‐hour posttransfusion red cell survival, morphology, fluidity at low shear rate and adenylate content. Warming and to some extent chromium labelling of the cells resulted in improvement of the morphology and fluidity but in no increase in adenosine triphosphate, total adenylate concentration or posttransfusion survival. The conclusion is drawn that short‐term improvement of the morphology before transfusion does not result in increased viability.
Instability of Red Cell Shape Associated with the Absence of Membrane Glycophorin C1
Authors: Kuczmarski Candace A. , Orlina Armando R. , Delahanty Lauri K. , Kuczmarski Edward R.
Published: March 2009

Abstract. Erythocytes from 3 Gerbich‐negative siblings readily converted to echinocytes when subjected to changing concentrations of sodium or lithium chloride. When erythrocyte membrane proteins were assayed by immunoblotting with human anti‐Ge 1,2, normal membranes were found to contain a reactive component of an approximate molecular weight of 33,000 daltons which was absent from the Gerbich‐negative individuals. PAS staining and mobility on SDS gels identified this component as glycophorin C. These observations indicate that glyocophorin C (β‐sialoglycoprotein) is a component of the Gerbich antigen and is involved in normal cell shape maintenance.
Density Distribution of Hematopoietic Progenitor Cells and T Lymphocytes from Peripheral Blood: Heterogeneity of the Human Population1
Authors: Dooley Douglas C. , Alsop Patricia , Law Ping
Published: March 2009

Abstract. Density gradient centrifugation has been used in animal systems to purify stem cells and eliminate T lymphocytes prior to allogeneic transplantation. There is substantial disagreement whether the same approach can be used to purify hematopoietic stem cells obtained from human peripheral blood. The purpose of the present study was to resolve that issue by determining the density distribution of 4 classes of human leucocytes: total mononuclear cells, T lymphocytes, CFUc, and BFUe. To ensure a representative sampling, a large number of randomly selected donors were analyzed. The results show that most T lymphocytes band between 1.068 and 1.071 g/ml, with relatively little variation from individual to individual. In contrast, the density distributions of both CFUc and BFUe fluctuated markedly from donor to donor. As a consequence, there was significant variability in the degree of progenitor‐T cell separation. The implications of these results for clinical application of the density separation technique are discussed.
A Survey of Transfusion Practices among Anesthesiologists
Authors: Stehling L.C. , Ellison N. , Faust R.J. , Grotta A.W. , Moyers J.R.
Published: March 2009

Abstract. A survey of transfusion practices was conducted among anesthesiologists practicing in the United States to determine if significant variation exists and to identify those areas toward which research and educational efforts should be directed. Thirty‐seven percent (389) of 1,043 active members of the American Society of Anesthesiologists who received the survey responded. The indications for transfusion as well as the types of components administered were found to vary considerably. Among the areas which need to be addressed are arbitrary preoperative hemoglobin requirements, indications for fresh frozen plasma administration, preoperative blood ordering and autologous transfusion.
Sensitive Two‐Stage Papain Technique without Cell Washing
Authors: Scott Marion L. , Phillips P.K.
Published: March 2009

Abstract. A two‐stage papain technique is described in which cell washing after papain treatment is replaced by the addition of a specific papain inhibitor. This technique permits optimal enzyme treatment of red cells while digestion of immunoglobulin following the addition of serum is avoided. The technique therefore combines the design and consequent sensitivity advantage of two‐stage tests with the convenience of one‐stage tests, rendering it suitable for use in compatibility testing.
Alloimmunization to Red Cell Antigens in Thalassemia: Comparative Study of Usual versus Better‐Match Transfusion Programmes
Authors: Michail‐Merianou Vasiliki , Pamphili‐Panousopoulou Lena , Piperi‐Lowes Louli , Pelegrinis Elias , Karaklis Aris
Published: March 2009

Abstract. Transfusion records and serological data concerning 120 regularly transfused thalassemic children were followed up, beginning with their first transfusion, for a 5‐year period. All patients had been phenotyped for 18 red cell antigens before their first transfusion. The usual‐match (UM) group consisted of 64 children who received blood compatible with their ABO and Rho (D) antigens. The better‐match (BM) group comprised 56 children who received blood compatible with ABO, CcDEe and K antigens. It was found that a statistically significant difference does not exist in the overall frequency of alloimmunization between the UM (23.43%) and BM group (14.28%) and also between the children that started transfusion therapy before they were 12 months old regarding UM (9%) and BM policies (5.2%). However, a large numerical difference, which might become statistically significant with a larger number of patients was observed in the group of children who were started on transfusions after they were 12 months old, between the UM (38.7%) and BM (18.9%) policy. Finally, a statistically significant difference (<0.005) was found only between the children that started transfusions early (7.69%) and those that started them later in life (27.9%), irrespective of the transfusion policy observed. We report, in this study, the mean transfusion numbers of immunized and nonimmunized patients and the frequency, specificity and clinical significance of the identified antibodies are analyzed. Our conclusion is that a BM policy in transfusion programmes, including at least the Rhesus and Kell antigens, is recommended for all thalassemic children that start transfusion therapy after they are 12 months of age; if they start earlier, observation of the BM policy is not necessary.
Chronic Hepatitis B Carriers Found at Blood Donation: Do They Need Regular Follow‐Up?
Authors: Howdle P.D. , Wood G.M. , Ramskill S.G.A. , Losowsky M.S.
Published: March 2009

Abstract. It is recommended by the Department of Health and Social Security that hepatitis B virus (HBV) carriers found at blood donation should be followed up by specialist liver units. We attempted to do this in 74 such patients found by the Yorkshire Regional Blood Transfusion Service during a 4.5‐year period. Only 32 of the patients eventually attended for further investigation and follow‐up. However, 43 chronic HBV carriers found at blood donation have been followed for varying periods from 2 to 14 years, and the prognosis has been good. Although this raises the question as to whether such carriers need to be seen routinely in a specialist unit, we suggest that there are good reasons to continue to advocate this policy.
Human Immunodeficiency Virus Antibody Screening in Blood Donors from India, Nigeria and Thailand
Authors: Kühnl P. , Seidl S. , Ray V. , Kulkarni A.G. , Mba E.C. , Chandanayingyong D.
Published: March 2009

Abstract. Screening for human immunodeficiency virus (HIV) (LAV/HTLV‐III) antibodies in 3 blood donor populations from India (n=1,000), Nigeria (n=500) and Thailand (n=650; sampling in 1982) with a sensitive enzyme immunoassay (EIA; Abbott) yielded seropositivity rates of 0.5, 2.2 and 1.7%, respectively. Two EIAs with control antigens prepared from uninfected cell cultures (‘ELAVIA’, VIRGO), a recombinant Escherichia coli DNA EIA (‘ENV/CORE’), Western blot, an immunofluorescence assay and a radio‐immunoprecipitation assay confirmed none of the EIA‐reactive specimens as truly positive. The lack of specificity of the screening test was also attributable to monochromatic evaluation of the test trays at 492 nm only, and to reactivities against determinants of H9 cells used to grow HIV (HLA antibodies).
Human Erythrocyte Antigens
Authors: Telen Marilyn J. , Scearce Richard M. , Haynes Barton F.
Published: March 2009

Abstract. Human erythrocyte membrane proteins express antigens which serve as markers for erythroid differentiation as well as targets for human blood group alloantibodies. We have produced and characterized a new panel of five monoclonal antibodies to erythrocyte membrane proteins. Three monoclonal antibodies (E3, E4, E5) were specific for erythrocyte glycophorins. One antibody (E3) identified the sialoglycoprotein α and β homologous regions proximal to the plasma membrane, a second antibody (E4) was specific for sialoglycoprotein α, while a third (E5) was a sialogylcoprotein‐β‐specific antibody. Two antibodies (E6 and TE10) to the 65,000‐dalton chymotrypsin cleavage product of band 3 were also produced. These antibodies constitute a new panel of probes for investigation of normal erythroid differentiation, erythroleukemia, and the expression of normal and anomalous blood group antigens.
Endotoxin‐Polymyxin Complexes in an Improved Enzyme‐Linked Immunosorbent Assay for IgG Antibodies in Blood Donor Sera to Gram‐Negative Endotoxin Core Glycolipids
Authors: Scott Boyd B. , Barclay G. Robin
Published: March 2009

Abstract. Common or cross‐reactive epitopes of Gram‐negative endotoxins are found in the inner‐core glycolipid region of the outer membrane lipopolysaccharides (LPS). Hydrophobic LPS from rough mutants of Gram‐negative bacteria, lacking serotype polysaccharide O‐antigen chains, did not bind satisfactorily to polystyrene microplates for ELISA detection of cross‐reactive IgG anti‐endotoxin antibodies. When these LPS molecules were reacted with the cationic polypeptide polymyxin B, complexes were formed which were stable when coated on microplates. LPS‐polymyxin complexes allowed optimisation of conditions for an ELISA for IgG antibodies to the core glycolipid region of endotoxins which could be used for screening large numbers of blood donor sera.
5‐Day Storage of Human Platelet Concentrates in 30 ml of Plasma or Artificial Medium1
Authors: Adams George A. , Swenson Sonia D. , Rock G.
Published: March 2009

Abstract. Optimal conditions for the storage of platelet concentrates were studied by changing 5 environmental parameters: bag composition (PL146 vs. PL732), volume of plasma (60 vs. 30 ml), anticoagulant (CPDA‐1 vs. heparin), nutrient (glucose vs. fructose) and medium (plasma vs. artificial medium). A full bilevel factorial study was conducted to evaluate each variable alone and in combination with the other variables for their effects on platelet aggregation and release in response to single and pairs of stimuli. Serotonin uptake, pCO2, platelet count, lactate, glucose, pO2, pH and white blood cell concentration were also measured after 3 and 5 days of storage. Platelets that were stored in PL 146 bags had reduced responses to stimulation by 3 days and markedly impaired responses after 5 days relative to platelets that were stored in PL732 bags. There was a large drop in pH and platelet responsiveness when platelets were stored in a volume of 30 ml in PL146 bags; these were not found when platelets were stored in 30 ml in PL732 bags. Replacing plasma with an artificial medium or adding fructose or heparin and calcium to plasma yielded platelets that were equally functional as routine controls in CPD‐A1 plasma. It was concluded that replacement of plasma with 60 ml of artificial medium or a reduction of plasma volume with storage in PL732 bags are two possible mechanisms of obtaining more plasma from blood donations without compromising maximum platelet storage life.
Opsonic and Physicochemical Characteristics of Intravenous Immunoglobulin Preparations
Authors: van Furth R. , Braat A.G.P. , Leijh P.C.J. , Gardi A.
Published: March 2009

Abstract. The composition and opsonizing activity of five commercially available immunoglobulin preparations for intravenous use (Venoglobulin I, Venilon, Gammagard, Polyglobin, and Sandoglobulin) were studied. The composition of these preparations does not differ very much as far as total protein, immunoglobulin class and IgG subclass concentrations are concerned. The only exceptions were that Veniglobulin I, Gammagard and Sandoglobulin contain IgA, which might cause side effects in patients with anti‐IgA antibodies, Gammagard contains very little IgG4, and Venilon and Polyglobin contain no and almost no IgG3, respectively, which might explain their very low opsonic activity. It was found that Venilon and Gammagard activate complement in the ready‐for‐infusion state. The opsonic activity of Venoglobulin I, Sandoglobulin and Gammagard is about equal to that of inactivated serum: Staphylococcus aureus, Escherichia coli with K antigen, Streptococcus pyogenes and Streptococcus group B are well opsonized and E. coli without K antigen and Streptococcus pneumoniae are poorly opsonized.
Evaluation of a New Microporous Filtration Membrane System for Therapeutic Plasma Exchange
Authors: Kurtz Sanford R. , Carey Patricia M. , McGill Manley , Pineda Alvaro A. , Zaroulis Charles G. , Case Marvin T.
Published: March 2009

Abstract. A new therapeutic plasma exchange device developed by Sams Inc./3M was evaluated in plasmapheresis of 20 healthy volunteers and in a multicenter clinical study of therapeutic plasma exchange that included 49 patients. Safety and efficacy of plasma separation from whole blood were assessed for a module that contains Durapore® microporous surfactant‐free polyvinylidene fluoride membrane (Millipore Corp., Bedford, Mass., USA). The extra‐corporeal volume was 80 ml. Citrate and heparin anticoagulants were utilized. Mean plasma separation efficiency was 62% with unhindered passage of plasma proteins through the membrane pores and no hemolysis or activation of complement as measured by total hemolytic complement (CH50) and C3 conversion. Mean decrease in platelet count after procedures was 10%. No severe reactions occurred, and citrate effects (13%) were comparable to values reported with centrifugal instruments. The Sarns Inc./3M Therapore® device is a rapid, safe and efficient system for plasma exchange and potentially for source plasma collection. The principal benefits are small extracorporeal volume and cell‐free filtrate.
Recognition of a Polymorphic Monocyte Antigen
Authors: Menzel Gerd Rainer , Rose Michael
Published: March 2009

Abstract. 138 sera from renal transplant recipients were screened for the presence of monocyte‐specific antibodies. Most of the sera contained antibodies against monocytes, T‐ and/or B‐lymphocytes. One serum was identified which defined a monocyte‐specific antigen, MOLL This serum was investigated in intensive population and family studies for the estimation of the formal genetic criteria of this ‘new’ monocyte antigen. A gene frequency of 0.0614 was obtained by population analysis. Family investigations conveyed the information that the gene coding for MOLI was transmitted in linkage with HLA genes. A positive linkage disequilibrium of MOLI and HLA‐B17 was found.
HLA‐Cw4 Association with Acute Lymphoblastic Leukaemia in Sicilian Patients
Authors: Caruso Calogero , Cammarata Giovanna , Sireci Guido , Modica Maria Assunta
Published: March 2009

Abstract. The HLA frequencies of 50 Sicilian patients affected with acute lymphoblastic leukaemia (ALL) were examined. The frequency of Cw4 antigen was significantly increased in patients. Thus results obtained in our homogeneous population confirm in part previous reports suggesting that Cw4‐related genetic factors might be involved in the susceptibility to aetiological or pathogenetic mechanisms which play a role in some haematological malignancies.
Evaluation of Adenosine Triphosphate and 2,3‐Diphosphoglycerate after Twenty‐Four Hours in Red Cells Washed for Neonatal Transfusion
Authors: Wrotnowski Ute , Gray Lloyd S. , Anderson Garth , Samsell James E. , Mintz Paul D.
Published: March 2009

Abstract. Adenosine triphosphate and 2,3‐diphosphoglycerate concentrations in 14 U of CPDA‐1 stored red cells (PRBC) and washed red cells (WRBC) were measured to assess indirectly the quality of WRBC for neonatal transfusion after 24 h. The results indicate that there is no difference in red cell ATP and 2,3‐DPG concentrations between PRBC and WRBC after a 24‐hour period.
Efficacy of Heat Treatment of Factor VIII Concentrate
Authors: Cuthbert R.J.G. , Ludlam C.A. , Brookes E. , McClelland D.B.L.
Published: March 2009

Abstract. Two batches of heat‐treated factor VIII concentrate were found to contain anti‐HIV‐positive plasma donations. The batches were dry‐heat‐treated at 68 °C for 2 and 24 h, respectively. No HIV seroconversions occurred in 13 susceptible haemophiliacs receiving a total of 540 bottles of these factor VIII preparations.
Plasminogen Hemizygosity
Authors: Skoda Ulrike , Goldmann S.F. , Händler Christa , Hummel K. , Lechler E. , Lübcke Ingeborg , Mauff G. , Meyer‐Börnecke Dorothee , Pesch Stefanie , Pulverer G.
Published: March 2009

Abstract. The accumulation of the homozygous plasminogen (PLG) variant A3 in 4 siblings of a family led to the detection of 5 cases of apparent inverse homozygosity of PLG phenotypes which seemed to exclude paternity. Determination of 22 blood group markers and HLA typing, but under exclusion of PLG phenotypes, confirmed paternity in all cases (biostatistical probability of paternity >99.9985%). Comparing the results of ‘Western blots’ with functional‐caseinolytic phenotyping, the existence of inactive plasmin, as described earlier, could be excluded. Besides inverse homozygosity the assumption of a silent allele was confirmed by reduction of PLG antigenic levels and functional activities to approximately 50% of normal range. The PLG phenotype A in 1 individual with anamnestic thrombosis, reduced values of PLG antigen, and reduced functional activity, although in accordance with Mendelian inheritance, was also considered as indicative for PLG hemizygosity.
Comparison between a Solid‐Phase Low‐Ionic‐Strength Solution Antiglobulin Test and Conventional Low‐Ionic‐Strength Antiglobulin Test: Assessment for the Screening of Antierythrocyte Antibodies
Authors: Guignier F. , Domy M. , Angue M. , Richaud P. , Chatelain P.
Published: March 2009

Abstract. A solid‐phase low‐ionic strength salt antiglobulin test (LISS‐SPAT) has been developed using a micro‐plate coated with dried sera as a solid phase. Before coating, the in vitro C3d fragment generation was activated by adding heat‐aggregated immunoglobulin. The LISS‐SPAT was compared with low‐ionic strength conventional antiglobulin test (LISS‐AGT) and also with a test using polybrene or papain microplates. When detecting the IgG and IgM antierythrocyte antibodies the reaction was developed in the same way in LISS‐SPAT and LISS‐AGT. In routine work, the LISS‐SPAT provides a fast, reliable, handy and inexpensive screening of antibodies. This method appears to be an additional method to the papain and polybrene tests in microplates.
A Fluorescent Microsphere Method for the Investigation of Erythrocyte Chimaerism after Allogeneic Bone Marrow Transplantation Using Antigenic Differences
Authors: de Man A.J.M. , Foolen W.J.G. , van Dijk B.A. , Kunst V.A.J.M. , de Witte T.M.
Published: March 2009

Abstract. A method for the investigation of erythrocyte chimaerism in patients after bone marrow transplantation (BMT) has been developed using fluorescent microspheres coated with anti‐human IgG. Blood group antigens different in patient and donor were used as marker. The specificity and reliability of this method was evaluated measuring artificial mixtures of blood group positive and negative red cells. Red cell antigens tested were D, c, K, Fya, Fyb, Jka, Jkb, S and s. Mean linear regression lines for mixtures with percentages positive cells ranging from 0.01 to 1% and 1 to 100% were 0.91 x‐0.03 (r=0.99) and 1.00X‐0.05 (r=0.99), respectively. The sensitivity level of the assay was one positive cell per 10,000 blood group negative cells. In negative control samples (n= 15) a mean percentage positive cells was found of 0.002 ± 0.004 (SD). Intra‐ and inter‐assay coefficients of variation were 4.9 and 5.3% for mixtures of 10%, and were 11.1 and 24.6% for mixtures of 0.1% blood group positive cells. It is concluded that the described method can be used both to establish the take of donor marrow in an early phase after transplantation and to investigate mixed erythrocyte chimaerism long after BMT.
Irradiated Blood Platelet Concentrates Stored for Five Days ‐Evaluation by in vitro Tests
Authors: Espersen Geert T. , Ernst Erik , Christiansen Ole Bjarne , Jersild Casper , Grunnet Niels
Published: March 2009

Abstract. Platelet concentrates, irradiated with 15 Gy and stored for 5 days at room temperature under standardized conditions, were evaluated by in vitro tests and electron microscopy, in a paired study with nonirradiated platelets from the same concentrates, to investigate their usefulness for transfusion. The results showed no significant differences between the two groups in pH, pO2, PCO2, in vitro platelet aggregation, LDH, β‐thromboglobulin and thromboxane‐B2. Examination by electron microscopy showed a higher degree of degranulation in the 5‐day‐old platelets but no certain difference between irradiated versus nonirradiated platelets. On the basis of satisfactory in vitro storage properties, platelet concentrates can be stored for 5 days in PL‐1240 bags after irradiation. However, we recommend irradiation just before transfusion whenever possible.
Three Epitopes on the Human Rh Antigen D Recognized by 125I‐Labelled Human Monoclonal IgG Antibodies
Authors: Gorick B.D. , Thompson K.M. , Melamed M.D. , Hughes‐Jones N.C.
Published: March 2009

Abstract. Seven purified monoclonal antibodies specific for the D antigen of the human Rh blood group system were examined for the characteristics of their reactions with red cells (phenotype CcDEe). The average number of sites available for binding to the antibodies ranged from 8,900 to 26,000/cell. In mutual inhibition studies, all the antibodies inhibited each other but the extent of inhibition varied in that antibodies recognizing a lower number of sites only partially inhibited those recognizing a higher number of sites. It was concluded from the evidence that these six monoclonals recognize at least three different epitopes on the D polypeptide. In order to explain the variation in the number of epitopes on each polypeptide, it is suggested that there is heterogeneity in the placement of the molecule in the red cell membrane resulting in variation in access to the epitopes.
An Example of Anti‐Yta Demonstrating a Change in Its Clinical Significance1
Authors: AuBuchon J.P. , Brightman A. , Anderson H.J. , Kim B.
Published: March 2009

Abstract. The clinical significance of some red cell alloantibodies remains in doubt and can best be studied with long‐term 51Cr survival studies. We report a patient whose IgG anti‐Yta was initially shown not to shorten the lifespan of 51Cr‐labeled Yt(a+) red cells. At the time of this study, the subclass of the antibody could not be determined. Twelve weeks after transfusion with 4 units of Yt(a+) red cells, the alloantibody for the first time was demonstrable as IgG1; a repeat radiolabeled red cell survival demonstrated significant shortening of the lifespan of Yt(a+) red cells when they were followed for 7 days. These cells had a marked ‘two‐component’ survival curve. Because the patient also demonstrated autoantibody coating his red cells, the clinical effect of this autoantibody was followed with autologous red cells labeled with 111In; the survival of autologous red cells was normal throughout these studies. Evaluation of the clinical significance of an alloantibody in a patient may require long‐term 51Cr red cell survival studies and repetition of these studies after exposure to large quantities of the antigen.
Demonstration of Seven Epitopes on the Rh Antigen D Using Human Monoclonal Anti‐D Antibodies and Red Cells from D Categories
Authors: Lomas C. , Tippett P. , Thompson K. M. , Melamed M. D. , Hughes‐Jones N. C.
Published: March 2009

Abstract. The agglutination patterns have been established for the reaction between 29 monoclonal antibodies with specificity for the Rh antigen D and red cells of D categories IIIa, IIIc, IVa, IVb, Va, Vc, VI and VII, which are known to lack certain epitopes on the D polypeptide. Six different agglutination patterns were recognized and interpreted to indicate the recognition of seven different epitopes. These epitopes are termed epDl through to epD7. The separate existence of epD6 and epD7 is deduced from previous observations in inhibition studies using purified 128I‐labelled antibodies: they cannot yet be distinguished in agglutination tests. The number of epitopes lacking from cells of each category varied between two and five. As all the antibodies agglutinated cells of categories IIIa, IIIc and VII and cells of categories II, IIIb and Vb were not available, it is probable that there are epitopes other than the seven presently recognized. Eighteen out of the 29 antibodies which were examined recognized epitopes epD6/7 and it is suggested either that antibodies recognizing these epitopes predominate in polyclonal anti‐D sera, or that the lymphocytes producing these antibodies are preferentially selected during establishment of cell lines.
Low AIDS Attack Rate Among Dutch Haemophiliacs Compared to Homosexual Men: A Correlate of HIV Antigenaemia Frequencies
Authors: Wolfs Tom F. W. , de Wolf Frank , Breederveld Cees , Sjamsjoedin‐Visser Liesbeth J. M. , Roos Marijke , Bakker Margreet , Goudsmit Jaap
Published: March 2009

Abstract. A cohort of 180 haemophiliacs followed between 1983 and 1986 and a cohort of 961 homosexual men followed between 1984 and 1986 were compared for the prevalence and incidence of HIV‐1 antibody (HIV‐1‐Ab) seropositivity, the incidence of AIDS‐related complex (ARC) and AIDS and the prevalence and incidence of serological and immunological markers for HIV‐related disease progression. Among the haemophiliacs 23 (12.8%) patients were HIV‐1‐Ab seropositive at entry and 20 (12.7%) of the remaining 157 seroconverted for HIV‐1‐Ab during follow‐up. Of the homosexual men 238 (24.8%) were HIV‐1‐Ab seropositive at entry and 68 (9.4%) of the 723 at entry seronegatives seroconverted during follow‐up. Clinical follow‐up of the HIV‐1‐Ab seropositive and seroconverted men was 59 months in the haemophiliac cohort and 60 months in the homosexual cohort. Among the HIV‐1‐Ab seropositive and seroconverted haemophiliacs and homosexual men the cumulative ARC/AIDS incidence was 2 and 18%, respectively. Occurrence of HIV‐1‐antigenaemia was more frequent among seropositive and seroconverted homosexual men (28%) than among haemophiliacs (7%) (p = 0.001). The groups did not differ significantly for the absence or loss of anti‐HIV core antibodies or the occurrence of low CD4+ cell numbers. These data indicate a slower progression of HIV‐related disease in seropositive haemophiliacs compared to seropositive homosexual men.
Follow‐Up of Anti‐IgA Antibodies in Primary Immunodeficient Patients Treated with γ‐Globulin
Authors: Ferreira A. , Rodriguez M.C. García , Fontán G.
Published: March 2009

Abstract. The levels of anti‐IgA antibodies and the appearance of adverse reactions following γ‐globulin administration in 41 patients affected by primary antibody defects treated with intramuscular (IMGG) or intravenous γ‐globulin (IVGG), and 3 patients with the Wiskott‐Aldrich syndrome (WAS) have been studied during a 31‐month period. Anti‐IgA antibodies were restricted to patients with circulating B lymphocytes and measurable amounts of IgA. The incidence of anti‐IgA antibodies in the immunodeficient patients studied was 22.7%, and 2 of the 3 WAS patients also had high levels of anti‐IgA antibodies. The presence of moderate levels of anti‐IgA antibodies (up to 1/1,600) was not associated with adverse reactions. Our results indicate a significant relationship <0.02) between persistence of anti‐IgA antibodies and IMGG administration.
Influence of Blood Storage Time and Plasma Histamine Levels on the Pattern of Transfusion Reactions
Authors: Frewin D.B. , Jonsson J.R. , Frewin C.R. , Russell W.J. , Russell M.W. , Davis K.G. , Beal R.W.
Published: March 2009

Abstract. The data from 359 transfusion reactions reported over a 3‐year period have been compared with those obtained from 359 matched units given at the same time as the former but not implicated in a reaction. It was found that patients who developed reactions which were not accompanied by a change in body temperature received units with a longer storage time than patients experiencing febrile reactions. In addition it was noted that reactions which included a skin rash also tended to be associated with units stored for longer. In a further prospective assessment plasma histamine levels were measured in a group of 71 patients experiencing ‘anaphylactoid’, febrile and mixed transfusion reactions as well as in individuals in whom blood transfusions did not cause a reaction. The ‘anaphylactoid’ reaction group demonstrated a mean plasma histamine level in excess of 1 ng/ml, whereas mean concentrations below 0.50 ng/ml were recordeα in the other 3 groups. Our data also suggested that in general, more and ‘older’ blood was given to the ‘anaphylactoid’ reactors. The high plasma histamine concentrations could readily contribute to the rash, wheeze and flushing recorded in these individuals. Thus, if a long shelf‐life for stored blood is required, strategies which remove histamine from the unit may be worth considering.
Analysis of Natural Killer Cells in Patients with Idiopathic Autoimmune Hemolytic Anemia
Authors: Conte Roberto , Dinota Angelo , Tazzari Pier Luigi , Belletti Daniela , Sermasi Giorgio
Published: March 2009

Abstract. A functional and phenotypic analysis of the circulating natural killer (NK) cell population was carried out in 9 patients with idiopathic autoimmune hemolytic anemia (IAHA). The NK‐cell activity, assessed by a sensitive method based on the inhibition of target clone growth in plasma semisolid medium, was markedly decreased in all patients as compared with normal controls and was not restored by stimulation of the cells with recombinant α interferon (α‐IFN). Analysis of peripheral blood phenotypic markers showed that cells bearing Leu7 and CD16 antigens numbered in the normal range. These findings suggest that IAHA patients exhibit a functional impairment of the NK compartment.
New Platelet Antigen, Siba, Involved in Platelet Transfusion Refractoriness in a Japanese Man
Authors: Saji Hiroh , Maruya Etsuko , Fujii Hiroshi , Maekawa Taira , Akiyama Yuichi , Matsuura Takao , Hosoi Takemitsu
Published: March 2009

Abstract. Siba, a new platelet‐specific alloantigen involved in a case of platelet transfusion refractoriness is reported. The IgG platelet alloantibody was detected in a multiply transfused patient of Japanese extraction (Sib), by the presence of HLA antibodies. After transfusion of HLA‐compatible platelets, the patient suffered from refractoriness. Adsorption studies with pooled lymphocytes showed that the serum contained anti‐platelet activity. Family studies indicate that Siba is inherited as an autosomal codominant trait and separate from HLA and Baka. As of this report, segregation from Zw (PlA) and Yuk (Pen) antigen systems have not yet been determined. The gene frequency of Siba in the Japanese population is estimated to be 0.136.
Comparison of a Conventional Quadruple‐Bag System with a ‘Top‐and‐Bottom’ System for Blood Processing
Authors: Pietersz R. N. I. , Dekker W. J. A. , Reesink H. W.
Published: March 2009

‘Top‐and‐bottom’ bags have an outlet at the top and at the bottom of the collecting bag, allowing simultaneous expression of plasma and red cells, whereas the buffy coat remains in the collecting bag. The composition of blood components was investigated following manual separation of whole blood in a conventional 4‐bag system (A) or automated separation in a ‘top‐and‐bottom’ system (B). To overcome inter‐donor differences, two units of whole blood were pooled and redistributed into the original bags (A and B) prior to centrifugation. Leukocyte‐poor platelet concentrates (LPPC) were manufactured from both types of buffy coat (A and B). The volumes of plasma, red cell concentrates (RCC) and buffy coat were similar in both methods. However, the residual leukocytes and platelets in the RCC from the top‐and‐bottom system were significantly lower than in the conventional system, 140 ± 59 times 106 (mean ± SD) versus 762 ± 228 times 106, respectively (p<0.01). Both types of LPPC contained less than 107 leukocytes and could be stored for 7 days maintaining a pH above 6.5. We conclude that the top‐and‐bottom system enables automated and standardized preparation of RCC and plasma with a significantly better buffy‐coat removal than with manual processing.
Biochemical and Physical Properties of a Solvent‐Detergent‐Treated Fibrin Glue
Authors: Burnouf‐Radosevich M. , Burnouf T. , Huart J. J.
Published: March 2009

Abstract. A fibrin glue preparation has been obtained from pooled human plasma using a procedure which includes a solvent‐detergent (SD) treatment to inactivate lipid‐enveloped viruses. The SD treatment inactivated ≥ 5.5 log10 of HIV in less than 45 min, and ≥5 log10 and ≥6.5 log10 of VSV and Sindbis virus, respectively, in less than 2 h. The product was found to contain high quantities of fibrinogen (116±2.49 g/l; n = 12), factor XIII (35±2.88 U/ml) and von Willebrand factor (23±1.9 U/ml ristocetin cofactor activity), and relatively low levels of fibronectin (5.9±0.51 g/l). Plasminogen, the precursor of plasmin, which may play a negative role by decreasing the resistance of the fibrin clot, was at only 0.03 g/l. Cellulose acetate electrophoresis showed 95% gamma‐proteins and 5% alpha‐2‐beta proteins. Sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions detected three main protein bands with apparent molecular weights of 65, 56 and 47 kilodaltons, probably corresponding to the alpha, beta, and gamma fibrinogen subunits. Other characteristics of the product included (1) high clottability of fibrinogen (over 85%); (2) absence of low molecular weight fibrin degradation products; (3) rapid solubilization at room temperature (less than 10 min); (4) high tensile strength (202±27 g/cm2 after 2 h of application), and (5) high elasticity of the fibrin clot. In addition, scanning electron microscopy revealed a highly organized structure showing tridimensional arrangement of the fibrin fibers. SD treated fibrin glue should efficiently replace autologous fibrinogen or cryoprecipitate preparations for surgical application.
Studies in Red Blood Cell Preservation 3. A Phosphate‐Ammonium‐Adenine Additive Solution1
Authors: Greenwalt T.J. , McGuinness C.G. , Dumaswala U.J. , Carter H.W.
Published: March 2009

Abstract. The maintenance of adenosine triphosphate (ATP) in red blood cells (RBC) during storage is largely dependent on the integrity of glycolytic metabolism in the Embden‐Meyerhof pathway. Meryman et al. [Transfusion 1986;26:500–505] hypothesized that a solution that increased the surface tension of the corpuscles through hypotonic swelling might retard the development of echinocytosis and membrane loss by the shedding of exocytic vesicles. Unexpectedly, maintenance of good ATP levels and satisfactory RBC survivals were found for as long as 18 weeks. The purpose of our study was to test their observations and to explore the possible mechanisms. Equal parts of units of packed RBC were stored in the experimental preservative and, for comparison, in ADSOL®. The most notable findings were ATP values at 4 weeks averaging 5.2 μmol/g Hb (130% of initial) and at 12 weeks 2.9 μmol/g Hb (73% of initial), whereas these values declined as expected in ADSOL. Mean RBC diameters and surface areas by morphometric analysis were not significantly different in the two preservatives indicating the absence of any hypotonic swelling. The morphology scores of the RBC were significantly better throughout (p<0.05) than in ADSOL. The shedding of exocytic hemoglobin‐containing vesicles was essentially the same in both preservatives. Our data confirm the observation that ATP levels are well maintained for at least 12 weeks, but do not show any evidence that hypotonic swelling was a part of the mechanism.
Manual Quantitative Method for the Study of Red Cell Agglutination Using Light Diffraction by Suspended Particles
Authors: Rasia R. J. , Valverde‐Rasia J. , Rosasco M. García
Published: March 2009

Abstract. A new manual technique, called the diffractometric technique, for quantitation of hemagglutination is described in this paper. After the equilibrium has been attained in the reaction, the whole product is resuspended in a dense transparent medium in order to obtain a stable suspension. The relative optical extinction produced by this suspension depends on the number and sizes of the suspended particles and on the photodetector geometry. Hence the agglutination percentage of the reaction can be monitored by using a visible light transmittance photometer. The results of this technique compare very well with the ones obtained by applying two other methods, the Dybkjaer and the Ropars methods.
Report on the Fourth ISBT/ICSH Platelet Serology Workshop (London 1988)
Authors: Metcalfe P. , Waters A. H.
Published: March 2009

Abstract. The fourth ISBT/ICSH platelet serology workshop took place in 1988. It consisted of a wet workshop carried out by the 17 participating laboratories during March 1988, followed by a discussion of the results at a meeting of the ISBT/ICSH Expert Panel on Platelet Serology at the ISBT/BBTS Congress in London in July 1988. Sixteen samples, including 1 negative control serum, were analysed by the participants using methods of their choice. There was general agreement on 10/16 samples. The results indicated progress in the following areas: (i) antibody identification; (ii) differentiation of platelet‐specific and HLA antibodies; (iii) glycoprotein localisation of platelet specific antigens; (iv) identification of new platelet antigen systems.
Agglutination of Red Blood Cells from Patients with Diabetes mellitus by a Polyclonal Human Antibody Specific for D‐Glucose
Authors: Ellisor S. S. , Toy P. T. C. Y. , Reid M. E. , Avoy D. R.
Published: March 2009

Abstract. We have previously described an antibody which will agglutinate red blood cells which had been incubated in vitro in D‐glucose. This antibody is specific for the ring form of glucose, β‐D‐glucopyranose. The current report demonstrates that without prior in vitro incubation with glucose, red blood cells from 36 of 38 patients with diabetes mellitus, and 7 of 70 patients not diagnosed as diabetic were agglutinated by this antibody. Strength of agglutination of red blood cells from diabetic patients correlated with both glucose (r = 0.61; p<0.001) and hemoglobin A1c levels (r = 0.50; p<0.01) in simultaneously obtained samples. This reactivity could be reversed by incubating red blood cells from diabetics for several hours in saline. This report suggests that red blood cells from diabetic patients have membrane‐bound glucose that can be detected immunologically. Reversibility of the reaction and rapidity of in vitro glycosylation suggests short‐term binding of glucose. To our knowledge, this is the first report documenting immunological detection of in vivo short‐term reversible binding of glucose to cellular membranes.