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The second example of anti-Lub was found in a Milwaukee woman who had a mild atypical hemolytic transfusion reaction. The family study revealed an amazing pedigree. The parents of the propositus represent a mating with an expected frequency of 0.0002. The properties of this anti-Lub and five examples of anti-Lua will be discussed. The results of a large population survey utilising these anti-sera will be presented. Eight examples of the rare genotype LuaLua have been found. Representative family studies will be included in the discussion.

Background and Objectives Red blood cell (RBC) storage systems are licensed based on their ability to prevent haemolysis and maintain RBC 24-h in vivo recovery. Preclinical testing includes measurement of RBC ATP as a surrogate for recovery, 2,3-diphosphoglycerate (DPG) as a surrogate for oxygen affinity, and free haemoglobin, which is indicative of red cell lysis. The reproducibility of RBC ATP, DPG and haemolysis measurements between centres was investigated. Materials and Methods Five, 4-day-old leucoreduced AS-1 RBC units were pooled, aliquotted and shipped on ice to 14 laboratories in the USA and European Union (EU). Each laboratory was to sample the bag twice on day 7 and measure RBC ATP, DPG, haemoglobin and haemolysis levels in triplicate on each sample. The variability of results was assessed by using coefficients of variation (CV) and analysis of variance. Results Measurements were highly reproducible at the individual sites. Between sites, the CV was 16% for ATP, 35% for DPG, 2% for total haemoglobin and 54% for haemolysis. For ATP and total haemoglobin, 94 and 80% of the variance in measurements was contributed by differences between sites, and more than 80% of the variance for DPG and haemolysis measurements came from markedly discordant results from three sites and one site, respectively. In descending order, mathematical errors, unvalidated analytical methods, a lack of shared standards and fluid handling errors contributed to the variability in measurements from different sites. Conclusions While the methods used by laboratories engaged in RBC storage system clinical trials demonstrated good precision, differences in results between laboratories may hinder comparative analysis. Efforts to improve performance should focus on developing robust methods, especially for measuring RBC ATP.

Serum specimens submitted for confirmation of hepatitis B antigen (HBsAg) reactivity were tested by counterelectrophoresis (CEP) and by three modifications of the solid phase radioimmunoassay "sandwich" technique (SPRIA). In addition, the specificity of each reaction was determined for two of the SPRIA test methods. One hundred seventy-five of the 688 specimens were reactive by all techniques and a further 60 ere specifically reactive by two of modifications of SPRIA. An additional five specimens were positive by a third SPRIA modification. The primary SPRIA technique, using a homologous guinea pig antibody sandwich, detected 184 nonspecific reactive samples. With the same test done at 45C with a reduced incubation time, there were only 37 nonspecific reactive samples. None of the nonspecific samples was detected when the SPRIA technique used a heterologous guinea pig-human antibody sandwich. The results provide a basis for estimating the effectiveness of the SPRIA technique for HBsAg testing.

The purpose of this study was to determine if the removal of most of the leukocytes and platelets would affect the in vitro characteristics of stored red blood cells (RBC). Fresh RBC concentrates prepared by removing platelet-rich plasma and filtration through Imugard IG 500 filters were compared with unmanipulated units after storage for up to 56 days. The filtered units were significantly better after 56 days storage for supernatant K+ (p = 0.001), hemolysis (p = 0.05), total vesicle membrane protein shed (p = 0.03), and RBC morphology score (p = 0.04). These differences occurred even though ATP levels were well maintained in both groups. The measurements that did not differ significantly were pH, hematocrit, ATP, 2,3-DPG, glucose and supernatant Na+. It is suggested that enzymes, leukotrienes, catecholamines and eicosanoids released by degenerating leukocytes and platelets may be inimical to RBC. Some may act as agonists on alpha-adrenergic and cholinergic muscarinic receptors present on RBC membranes.

Red blood cells (RBCs) must be stored in polyvinyl chloride (PVC) bags plasticized with di-2-ethylhexyl phthalate or a similar plasticizer to achieve their full storage life with conventional storage solutions. Improved storage solutions might remove this requirement and allow blood storage in other plastics. Experimental Additive Solution-61 (EAS-61), which maintains RBCs for 9 weeks with reduced haemolysis and satisfactory 51Cr 24-h recovery, is an appropriate candidate improved RBC storage solution.Twenty-four units of packed RBCs were pooled in groups of four units, each pool was realiquoted into four units and stored, six pooled units per arm, in one of the following: 100 ml of EAS-61 in PVC; 200 ml of EAS-61 in PVC; 100 ml of EAS-61 in polyolefin (PO); and 200 ml of EAS-61 in PO. Haemolysis, RBC morphology indices, RBC ATP concentrations, and other measures of RBC metabolism and function were measured weekly.RBC haemolysis exceeded 1% by 7 weeks in PO bags containing 100 ml or 200 ml of EAS-61. In PVC bags, haemolysis was less than 1% at 11 weeks. RBC ATP concentrations were 1 mol/g of haemoglobin (Hb) higher at 2 weeks in the PVC-stored units.RBCs stored in PVC had markedly less haemolysis and higher RBC ATP concentrations than those stored in PO. Haemolysis would limit RBC storage in PO bags to a duration of 6 weeks, even with EAS-61.

The purpose of these studies was to evaluate the functional properties of blood components collected with an automated collection system.Single-donor platelets (n = 44) and packed red cell (RBC) units (n = 10) were collected. In vitro and in vivo assays were used to assess the function of single-donor platelet components stored for 5 days and of packed RBC units after storage for 42 days at 4 degrees C.Adverse events observed in the 44 study subjects were minor. The mean 24-hour recovery value for the packed RBC units stored for 42 days was 83.6 +/- 5.4 percent, with a mean percentage of hemolysis on Day 42 at 0.46 +/- 0.19 percent. The 25 patients receiving platelet components achieved a mean corrected count increment of 15.1 +/- 10.4 x 10(3). All platelet concentrates had less than 1 x 10(6) total white cells.Both in vitro and in vivo testing for the packed RBCs collected and stored for 42 days met the standards for both hemolysis and percentage of 51Cr 24-hour RBC recovery. The in vitro results and transfusion data on white cell-reduced platelet components transfused to thrombocytopenic patients were comparable to those on available platelet components.

The purpose of this presentation was to demonstrate that the red blood cell is not a hapless sac of hemoglobin and that much research is still needed for better understanding of its complexities. After a brief historical introduction the following subjects are presented: 1). Phosphofructokinase is the rate limiting step in the anaerobic glycolytic pathway. Ribose-5-phosphate, a metabolite of the oxidative pentose phosphate pathway is essential for the generation of phosphoribosylpyrophosphate which in turn is needed for the synthesis of adenosine monophosphate from adenine by the action of adenine phosphoribosyl transferase. 2). There are at least 17 blood group systems with more than 400 epitopes expressed on the red cell membrane. The Rh null and the McLeod phenotypes associated with abnormally shaped red cells and hemolytic anemia are briefly described as is the present understanding of the nature of the Rh complex. 3). The structure of the cytoskeleton and the composition and behavior of the lipid bilayer are presented with some discussion of the MN and Ss sialoglycoproteins and the Leach phenotype. 4). Touched upon is the role of phosphoinositides with some emphasis on recent discoveries relating to the glycophosphoinositide protein anchor. 5). The intricacies of the many faceted transport mechanisms are introduced. Briefly mentioned are the mechanisms activated when regulatory volume adjustments occur in fine tuning red cell volume after exposure respectively to hypotonic or hypertonic stress. Sufficient evidence is presented to convince that a cell doesn't have to have a nucleus to be respected even though it is just a corpuscle.

Studies were conducted to examine whether an experimental additive solution (EAS-2) containing, in mM: 20 NH4Cl, 30 Na2HPO4, 2 adenine, 110 dextrose, 55 mannitol, pH 7.15, would be useful to extend the storage shelf life of human RBCs. With 6 pairs of split units, ATP concentrations were better maintained for 12 weeks with EAS-2 than with Adsol (1.8 vs. 1.1 mumol/g Hb, respectively, p = 0.002). Autologous 24-hours 51Cr viability values for split units in the same donors were: on 6 paired units at 8 weeks, EAS-2 87.0 +/- 4.5%, Adsol 72.6 +/- 2.3%, p = 0.004; on 11 paired units at 9 weeks, EAS-2 79.5 +/- 7.1%, Adsol 68.2 +/- 10.1%, p = 0.0003. The data suggest that packed RBCs stored for 9 weeks with EAS-2 will be suitable for transfusion following the removal of supernatant with a single washing step.

The purpose of this study was to examine whether vesiculation of RBC plays a significant role in their rejuvenation. Outdated units of Adsol blood, were divided into two aliquots and incubated with equal volumes of a solution of 100 mM pyruvate and inosine, 103 mM phosphate and 5 mM adenine (PIPA) or 0.9% saline. Following 1 h incubation, vesicles were isolated from the supernatants and quantitated for hemoglobin content. Restoration of RBC ATP, 2,3-DPG, morphology, and osmotic fragility after rejuvenation was satisfactory. The postrejuvenation mean corpuscular volumes (88.2 +/- 6.9 fl) were significantly lower (p less than 0.001) than the prerejuvenation (94.6 +/- 6.8 fl) and control (104.0 +/- 7.3 fl) volumes. The hemoglobin shed in vesicles during rejuvenation was significantly greater than in the saline controls (0.44 +/- 0.31 vs. 0.18 +/- 0.10 mg/dl RBCs; p = 0.026). These data suggest that the decreased MCV following rejuvenation is in part due to membrane loss in exocytic vesiculation.

Sera and questionnaires were evaluated retrospectively from 128 volunteer blood donors whose blood had been implicated in cases of clinically recognized post-transfusion hepatitis in recipients of one- or two-unit blood transfusion between 1971 and 1977. Serologic markers of hepatitis B virus (HBV) infection were found in 23 percent, compared to 9.7 percent of 3,230 prospective blood donors. The prevalence of antibody to hepatitis A virus was similar among implicated donors (44%), prospective donors (58%), and among those implicated donors with (41%) and without (44%) HBV markers. Among implicated donors, none had a history at the time of donation of having had clinically recognizable hepatitis, 93 percent had no history of prior blood transfusion, and 80 percent had normal hepatic enzymes. Data from this study confirm that non-A, non-B hepatitis has been a common form of posttransfusion hepatitis in recent years, since 77 percent of these implicated donors had no HBV serologic markers. In addition, these donors could not be distinguished by age, race, sex, history of clinical hepatitis or of prior blood transfusion, or in most cases by hepatic enzyme levels.

Sixteen patients who received Konyne for bleeding episodes or for hemostasis during surgical procedures were evaluated for signs of posttransfusion hepatitis. Four patients died from their primary disease. Of the remaining 12 patients at risk, six patients developed posttransfusion hepatitis. Sera from four of the six patients with hepatitis were positive for the hepatitis B antigen (HBAg). Aliquots of Konyne from the specific lot (K4797) implicated in three cases of HBAg-positive hepatitis were negative for HBAg by immunodiffusion, immunoelectroosmophoresis, and radioimmunoassay. The high incidence of hepatitis associated with Konyne considerably limits the use for this specialized coagulation factor concentrate. HBAg screening by available technics does not necessarily detect Konyne preparations capable of transmitting HBAg-positive hepatitis.

Resume Anti-Jka etait la cause d'une forme legere de la maladie hemolytique du nouveau-ne. Zusammenfassung Anti-Jka-Antikorper haben eine leichte Form des Morbus haemolyticus neonatorum hervorgerufen.

The uptake and incorporation of P32O4 by the cord blood erythrocytes of normal and erythroblastotic infants and the erythrocytes of adults was studied by incubating specimens in ACD at 37 C. in air. The uptake of p32 from the plasma was slower in the infants than in the adults. It was possible to demonstrate an initial "fast" phase followed by a "slow" phase in all the plasma p32 uptake experiments. The half-value time and the percentage turnover per minute indicated greater activity in the adults compared to the infants. The balance of movement of phosphorus was from red cells to plasma in all the experiments. No striking differences in phosphate metabolism were demonstrated between the normal and erythroblastotic newborns, except for the large turnover in µM of P per minute by the red cells of the latter during the first 20 minutes of incubation. Labeling of intraerythrocytic inorganic phosphorus (Pi), adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) proceeded more slowly in the infants than in the adults. The labeling of the erythrocyte 2,3-DPG of all the infants lagged far behind Pi and ATP whereas in the adults the r.s.a. of 2,3-DPG had equalled or exceeded the r.s.a. of Pi and ATP in four hours. It is suggested that there is decreased synthesis of phosphoglycerates by the erythrocytes of the newborn which may be due to a lower level of glyceraldehyde-3-phosphate dehydrogenase activity.

SummaryStudies on three patients with platelet antibodies associated with thrombocytopenia but with different clinical backgrounds are presented. The first patient had chronic lymphocytic leukemia without known contact with an outside source of platelet antigen. Platelet antibodies were detected before a platelet transfusion was given as a measure of platelet survival. The second patient developed a drug (Diuril) induced thrombocytopathy and thrombocytopenia. The platelets appearing in the circulation immediately following the thrombocytopenia exhibited abnormal qualitative and quantitative ultrastructure and abnormal function of platelet factor 3. This constituents the first drug induced thrombocytopathy observed. The third patient had post-hepatitic cirrhosis with functional and histologic impairment. Possible explanations of antibody production were suggested.The antibodies were demonstrated by the platelet agglutination method of Tullis using normal stored and normal stored irradiated platelets as the source of antigens. Coagulation factors were studied together with the platelet antibodies. Qualitative (platelet thromboplastic function) and quantitative (thrombocytopenia) changes were observed in one patient.Patient’s circulating platelets and agglutinated normal stored and normal stored irradiated platelets were studied under the electron microscope. Morphological changes were observed and described.The sera of the patients and normal controls were centrifuged at 126,000 g for 6 hours and platelet agglutination tests were performed with the top and bottom layers. Increased agglutination in the top layers and decreased agglutination in the bottom layers occurred in the patient’s sera without any changes in the normal sera.

Red cell ghosts prepared by conventional methods were mixed with an equal volume of 0.6% γ globulin and converted to an insoluble “slurry” with glutaraldehyde. A 50% suspension of the washed “slurry” in phosphate buffered saline using 0.1% NaN3 as preservative was used for preparing hemoglobin-free eluates of anti-D and other blood group antibodies suitable for radiolabeling; it remained active during storage at room temperature for longer than 3 months. The procedure can be used for preparing immunoadsor-bents containing red cell stroma coupled to other substances.

A. The action of EDTA on the coagulation mechanism was studied. EDTA was found: 1. To exert no direct effect on prothrombin or thrombin. 2. To have an inhibitory action on labile factor. 3. To increase the speed of conversion of prothrombinogen to prothrombin. B. The effect of incubation or aeration of plasma on thrombin time was studied. It was found that aeration or incubation increases the thrombin time of plasma. The following observations lead one to attribute this prolongation to a heparinoid activity: 1. The property of incubated plasma to prolong the thrombin time of fresh plasma when added to it. 2. The ability of protamine to shorten the thrombin time of incubated or aerated plasma. 3. The sensitivity of the thrombin time of incubated or aerated plasma to pH changes and calcium concentration. C. The incidental observation was made that cooling reverses the prolonged thrombin time produced by incubation.

Summary The eighth child of a woman who had received five pints of blood in the past was born with mild erythroblastosis foetalis due to anti-Duffy(anti-Fya). The specificity of the antibodies on the infant's erythrocytes was established by elution. The maternal antibody was shown to be a gamma globulin by neutralization studies. Seven additional cases of erythroblastosis due to anti-Fya were culled from the files of other laboratories making a total of eleven known to the authors. It is suggested that anti-Fya is a more frequent cause of haemolytic disease of the newborn than was suspected. Reume: Le huitieme enfant d'une femme qui avait regu, de par le passe, 5 ampoules de sang, presenta a la naissance une maladie hemolytique peu grave due a un anticorps anti-Fya. Ce dernier put etre elue des hematies de l'enfant, et l'etude du serum maternel montra, par des tests de neutralisation, qu'il s'agissait d'une gamma globuline. Sept autres cas d'erythroblastose due a l'anti-Fya ont ete recueillis dans les listes d'autres laboratoires, ce qui fait un total de onze cas connus des auteurs. Ceux-ci sont par consequent d'opinion que l'anti-Fya est une cause plus frequente de maladie hemolytique du nouveau-ne qu'on ne le pensait jusqu'a maintenant. Zusammenfassung Das achte Kind einer Frau, welche fruher 5 Vollblutkonserven erhalten hatte, wurde mit einem Anti-Fya-bedingten leichten Morbus haemolyticus neonatorum geboren. Mit den kindlichen Erythrozyten wurde ein Elutionsversuch durchgefuhrt, wobei die Spezifitatsbestimmungder Antikorper bestatigt werden konnte. Die mutterlichen Antikorper zeigten die Antigenstruktur von γ-Globulinen. Auser dem erwahnten Fall wurden aus dem Untersuchungsgut anderer Laboratorien 7 weitere Falle ermittelt. Die Autoren kennen heute 11 Falle von Anti-Fya-bedingtem Morbus haemolyticus neonatorum. Es ist deswegen naheliegend, anzunehmen, das Anti-Fya-bedingte Morbus haemolyticus-Falle haufiger vorkommen, als bisher vermutet wurde.

Summary Anti-N was found in the serum of a healthy blood donor of group MN. The behavior of the donor's blood group antigens and the anti-N appeared to be straightforward. The only explanation suggested is that the antibody was of heterologous origin. Resume Un anti-N a ete trouve dans le serum d'un donneur de sang en bonne sante de groupe MN. Le comportement des antigenes de groupe sanguin de ce donneur et l'anti-N est tout â fait net. La seule explication possible semble etre que cet anticorps soit d'origine heterologue. Zusammenfassung Im Serum eines Spenders mit den Gruppen MN wurde ein Anti-N gefunden. Die MN-Antigene wie auch der Antikorper entsprechen der Norm. Moglicherweise beruht der Antikorper auf einem heterologen Antigenstimulus.

Familiarity with the phenomenon of red cell polyagglutinability, while of prime importance to laboratory personnel, can be useful to the clinician as well. The presence of T-polyagglutinability, for instance, may be a clue to infection. Thrombocytopenia, leukopenia, or even hemolytic anemia can be associated with Tn-polyagglutinability. The Cad type of polyagglutinability is inherited.

Platelet concentrates prepared from freshly drawn blood were infused into patients with thrombocytopenic complicating disease. All exhibited a prolonged bleeding time, no clot retraction of a whole blood clot, platelet counts between 20,000 to 100,000 per cu. mm., usually, abnormal platelet ultrastructure and prothrombin consumption times under 15 seconds. Detection of platelet antibodies was attempted and the ultrastructure with a differential platelet count under the electron microscope of the patient's own platelets was documented. The platelet concentrates were given and it was possible to follow the fate of the donor's platelets by ultrastructural changes. Parallel studies of the coagulation mechanisms of the patients were followed serially. We found that some platelets survived up to seven days, although in some patients, platelets circulated but did not survive. Therefore immediate circulation of platelets cannot be equated with survival. In one patient with hypersplenism and a positive platelet agglutinin test no transfused platelets appeared to circulate. The ultrastructure of transfused platelets is greatly altered in bleeding patients. In the two patients with aplastic anemia and one with leukemia surviving platelets circulated which did not enter into the mechanisms of prothrombin activation, but which continued to circulate for some days with no improvement in the prothrombin consumption time over the pretransfusion level. Ultra-structural studies of these platelets showed that the circulating platelets possessed their granules which contain platelet factor 3 activity. The differential platelet count under the electron miscroscope becomes abnormal as the donor platelets are stored. Our work suggests that the ultrastructure and differential platelet count of a platelet concentrate must be essentially normal for survival to be achieved. We confirmed the work of others indicating that donor platelets do not survive if stored or, if the recipient has detectable platelet antibodies. Donor platelets did not survive in some patients for reasons we could not explain. This may be dependent on utilization of platelets in hemostatic plugs, in thrombin production or for the endothelial supporting function but to date we cannot estimate to what extent or in what fashion a thrombocytopenic patient utilizes donor platelets.

Blood group chimaerism was detected in a healthy fertile woman, not known to be a twin. Her peripheral lymphocytes had a male karyotype (46/XY); fibroblasts cultured from her skin had a female karyotype (46/XX). The mechanism of chimaerism could not be established.

Blood collected by 11 Red Cross Regional Blood Centers were screened by the method of agar gel diffusion (AGD) for the presence of hepatitis-B antibody. Of the 185,134 units tested, 114 were found to be positive for HBAb by the Regional Centers and were forwarded to National Special Projects Laboratory for confirmation. Only five out of 114 samples revealed lines of identity with a control anti-HBAg when reacted with a pool of plasma positive for hepatitis-B antigen that was different from that which was provided to the Centers. Apparently, the precipitation reactions observed by the Centers were largely due to the antithrombin antibodies in the donors' sera reacting with the residual thrombin used by the manufacturer to convert HBAg positive plasma to serum. We conclude that the incidence of hepatitis-B antibody as measured by agar gel diffusion in the Red Cross Blood donor population was extremely low.

With blood given exclusively by volunteer donors, the American National Red Cross (ANRC) Blood Program aims to supply patients needing transfusion with blood products of the highest quality. The use of blood from volunteers, with its established greater safety, combined with laboratory testing to detect carriers of hepatitis B surface antigen (HB-SAg) and modern computer technology, creates an effective system to reduce the risk of post-transfusion hepatitis. The ANRC has devised a national Donor Deferral System, which is designed to minimize the transmission of hepatitis by blood and blood products.

The results of hepatitis B surface antigen (HBs-Ag) testing in a large volunteer blood donor population are described. Counterelectrophoresis and three versions of solid-phase radioimmunoassay technic are compared and evaluated. Initial results suggested that the radioimmunoassay technic are compared and evaluated. Initial results suggested that the radioimmunoassay technic detected more than five times as many reactive donors as did counterelectrophoresis. The specificity of the radioimmunoassay technic has been increased by successive modifications, and recent results show that the technic detects 73 percent more reactive donors than does counterelectrophoresis. Not all of these reactions are specific, and it is estimated that the true gain in detection of HBsAg carriers is 49 percent of the value found by counterelectroesis. The incidence of HBsAg carriers in the America Red Cross donor population is about 1.25 per 1,000.