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Patients with chronic liver disease (CLD) and cirrhosis present a rebalanced hemostatic system in the three phases of haemostasis. This balance is however unstable and can easily tip towards bleeding or thrombosis. Management of both spontaneous bleeding and bleeding during invasive procedures remains a challenge in this patient population. Transfusion of blood products can result in circulatory overload and thereby worsen portal hypertension. As an alternative to fresh frozen plasma (FFP), prothrombin complex concentrates (PCC) may have merit in patients with liver disease because of their low volume. The impact of PCC in in-vitro spiking experiments of cirrhotic plasma is promising, but also warrants cautious use in light of thromboembolic risk. The majority of existing studies carried-out in CLD patients are retrospective or do not have an adequate control arm. A prospective study (the PROTON trial) was set up in 2013 to investigate the utility of PCC in patients undergoing liver transplantation. However, the study has never recruited the planned number of patients. Robust data on PCC safety in CLD is also required. The limited existing evidence does not seem to indicate an excessive thromboembolic risk. Currently, the utilisation of PCC in CLD cannot be routinely recommended but can provide an option for carefully selected cases in which other measures were not sufficient to control bleeding and after delicately weighing risks and benefits.

In this unprecedented crisis of severe acute respiratory syndrome coronavirus 2 and its associated coronavirus disease 2019 (COVID-19), polymerase chain reaction and then serological testing platforms have been massively developed to face the important screening demand. Polymerase chain reaction and serological testing platforms are not the only actors impacted by the crisis, transfusion services are facing important difficulties. A positive direct antiglobulin test is frequently observed for patients encountering COVID-19. Patients with severe symptoms may develop anaemia and become good candidates for blood transfusions. The interpretation of a positive direct antiglobulin test for patients recently transfused and suffering from COVID-19 is complex. The differentiation between COVID-19 induced antibodies and possible associated transfusion alloantibodies is therefore crucial. In this context, the elution technique incorporated in an appropriate decision-making process plays its full role. This intricate topic is presented through a case report followed by literature review and finally decision-making process for COVID-19 patients necessitating red blood cells administration.

While transfusion is a common and safe therapeutic procedure in health care facilities, transfusion reactions can occur, whether acute or delayed, mild or life-threatening. In face of these reactions, the biological analysis laboratory plays a central role in their diagnosis. The objective of this article is to develop decisional algorithms for laboratory tests to be performed according to the clinical symptoms developed by the patient during or after transfusion.Based on the information collected by reviewing the literature and the procedures used in our hospital, we then developed biological investigation algorithms according to the symptoms presented by the patient, rather than the presumed reaction.We have developed symptom-based algorithms for acute transfusion reactions management that streamline laboratory testing and simplify the differential diagnosis.

BACKGROUND: The major causes of death of combat casualties in austere environments are related to hemorrhage and occur early after injury. The implementation of a walking blood bank may overcome the logistical issues raised using blood component therapy. Nonetheless, it is important to ensure that this buddy transfusion is not going to compromise the mission success by altering the donor's performance. The results available so far cannot rule out this issue with certainty. Therefore, this study aimed at investigating the immediate effect of a 450-ml blood donation on the performances of elite soldiers in laboratory and field environments. STUDY DESIGN AND METHODS: This double-blind, randomized controlled study included two experiments. For both experiments, subjects were randomly assigned either to a control group (n1 = n2 = 7) or to a 450-ml-blood-bag donation group (n1 = 7 and n2 = 8). All participants underwent before and after a potential blood donation a multifactorial assessment including adapted physical tasks, hematological variables, vigilance parameters, and subjective assessments. RESULTS: No significant results were evidenced in this study. There was no impact of blood donation on the participants' performances in both the hospital and the combat-like environments. CONCLUSION: From a donor's point of view, a 450-ml blood donation has no impact on the required abilities of our elite soldiers to fulfill a demanding tactical mission. Thus, the results of this study support the fact that buddy transfusions could be part of the operational clinical armamentarium in austere environments for elite soldiers when no blood components are available.

Background and Objectives Belgian health authorities launched a national platform in 2011 to improve the quality of transfusion practices and blood use in Belgian hospitals. No data were available about the quality of hospital transfusion practice at the national level. Materials and Methods Three consecutive national surveys (2012, 2014 and 2016) were performed in all 111 Belgian hospitals to assess the degree of implementation of standards in four process domains related to red blood cell (RBC) transfusion: general quality aspects, ordering of RBC, electronic traceability and reporting of adverse events. The surveys were part of a methodology based on informing, feedback and benchmarking. Responses to questions were analysed semi-quantitatively, and hospitals could score 10 points on each of the domains. Results The proportion of hospitals scoring below 5 per domain decreased from 16%, 70%, 14% and 11% (2012) to 2%, 17%, 1% and 1% (2016), respectively. Similarly, scores above 7.5 increased from 25%, 1%, 23% and 36% (2012) to 64%, 30%, 68% and 81% (2016), respectively. In 2016, overall quality of transfusion practices, including the four pre-specified domains, improved continuously with an average total score (max = 40) increasing from 24.2 to 30.5 (p = 0.0005). In addition, there was a decrease in the number of distributed and transfused RBC per 1000 population between 2011 and 2019 from 47.0 to 36.5 and 43.5 to 36.1, respectively. Conclusion These data show that the applied methodology was a powerful tool to improve quality of transfusion practices and to optimize utilization of RBC at the national level.

Geographical distribution of ABO and RHD antigens is important for blood transfusion services and population genetics studies. There are few data on this topic in Burkina Faso, a multi-ethnic country. Our study aims at reporting phenotypic and allelic frequencies of ABO and RHD blood groups among voluntary blood donors from various ethnical regions of Burkina Faso. We conducted a cross-sectional study including 81,486 blood donors. ABO allelic frequencies were determined using the Bernstein method. Differences in phenotypic distribution of blood groups were assessed using the chi-square test; a p value0.05 being considered as statistically significant. We noticed that O+B+A+AB+O-B-A-AB- in our population. Phenotypic frequencies of blood groups A, B, O and AB were respectively 22.54%, 28.56%, 43.30% and 5.60%. RHD+was 92.24%. The allelic frequencies of A, B, O and D were respectively 0.1524; 0.1887; 0.6590 and 0.7214. We noticed statistical differences (p 0.05) between these administrative regions which corresponded roughly to some natural ethnic areas. Indeed, the phenotype O was more frequent in the Central-west, Central and East regions corresponding to "Mossi," "Gourounsi," "Gourmantché" areas while the phenotype A and AB were more reported in "Boucle du mouhoun" and "Hauts-Bassins" regions where we have "Bwaba" and "Bobo." The phenotype O negative was less frequent in "Bwaba." Our study provides interesting information to blood services that will allow them to better refine their donor recruitment strategies.

Objectives Recently, daratumumab has been included in the therapeutic strategies for myeloma patients. This molecule is an antibody directed against CD38, strongly expressed on plasma cells. Nevertheless, as CD38 is also present on erythrocyte membrane, daratumumab interferes with immunohaematological tests, complicating the selection of compatible blood. Methods A total of 14 patients treated by daratumumab have been followed in our transfusion laboratory. Among them, 11 have been transfused. Dithiotreitol (DTT) has been used to inhibit the daratumumab's interference, in the pre-transfusion tests (irregular antibody screening and cross-match). Results The red blood cell treatment with DTT has been very efficacious to inhibit the daratumumab's interference in 13 patients out of 14. Some precautionary measures had to be taken into account, especially the pH and the storage conditions. An extended pheno/genotype was an additional security element in the selection of compatible blood. To simplify and to optimize the laboratory practices, a decisional flow chart has been written. Conclusion DTT red blood cell treatment is very useful and efficacious in the pre-transfusion tests of patients treated with daratumumab. It allows to avoid the selection of blood bags only on the basis of an extended pheno/genotype, what is more secure and more ethical with respect to other at higher risk patients. A clear decisional flow chart allows a quality assurance gait. Collaboration with physicians is essential.

Le test de Coombs direct est utilise en premiere ligne pour le diagnostic de l'anemie hemolytique auto-immune (AHAI). Il permet de detecter la presence d'anticorps ou de fragments du complement a la surface d'erythrocytes. La positivite du test peut atteindre 0,1 % dans une population de donneurs de sang et jusqu'a 15 % chez des personnes malades hospitalisees. Vu le nombre important de publications associant la COVID-19 et l'AHAI, le taux de positivite du test de Coombs direct a ete evaluee aux Cliniques Universitaires Saint-Luc (Bruxelles). Une large cohorte de patient hospitalises dans notre hopital (n = 225) a ete selectionnee sur base d'une PCR recemment positive pour le SARS-CoV-2. 44 % des echantillons recoltes ont donne un resultat positif. Une difference statistiquement significative a pu etre mise en evidence entre le taux de positivite des patients hospitalises dans une unite de soins classique et celui des patients aux soins intensifs. Ceci plaide en faveur du fait qu'un test de Coombs positif pourrait former un critere de gravite de la maladie. Le plasma de patients SARS-CoV-2-positifs ayant un Coombs direct a ete incube avec des globules rouges de patients COVID-19 ne presentant pas de test de Coombs direct positif. Une agglutination systematique est observee lorsque ces globules sont testes pour un Coombs direct. Le plasma des patients COVID a aussi ete incube avec des globules de poches sanguines, generant egalement une agglutination. Avec le plasma de patients non-COVID, ces derniers globules n'agglutinaient pas. Le Coombs direct semble donc « transmis » d'un globule rouge a l'autre par l'intermediaire du plasma des patients COVID ayant un Coombs direct positif. (French) [ABSTRACT FROM AUTHOR] Copyright of Transfusion Clinique et Biologique is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Resume Introduction Le Centre national de transfusion sanguine, operateur unique de la transfusion sanguine au Burkina Faso est engage dans la demarche qualite suivant la norme ISO 9001. Aussi, l’ecoute-client constitue-t-il un fondement majeur de son systeme. Notre etude est construite sur le concept de la « satisfaction client » comme fonction de la qualite du service percue suivant le modele SERVQUAL. Objectifs Identifier les facteurs associes a la satisfaction des prescripteurs de produits sanguins afin de donner aux decideurs des elements d’aide a la decision pour l’amelioration continue des services. Materiel et methodes Nous avons conduit une enquete transversale aupres des prescripteurs de la ville de Ouagadougou entre le 27 fevrier et le 30 avril 2015. Nous avons utilise un questionnaire anonyme, auto-administre comprenant 13 items associes aux 5 dimensions du SERVQUAL. Les differents gaps ou index de satisfaction (ISC) ont ete calcules et une regression lineaire a permis de determiner les associations statistiques avec un seuil significatif a 5 %. Resultats Sur 256 questionnaires distribues, le taux de retour etait de 94,5 %. Environ 30 % des enquetes etaient globalement satisfaits a tres satisfaits. L’index global de satisfaction des clients etait de −5,74. Le delai de livraison des produits, l’efficacite et l’innocuite des produits, le conseil transfusionnel, la pro-activite de la communication, la gestion des demandes de reservation de produits et la couverture des besoins etaient les facteurs associes a la satisfaction des prescripteurs. Conclusion Cette etude, premiere du genre en transfusion sanguine dans notre contexte, a permis d’evaluer la satisfaction des clients et d’identifier les facteurs sur lesquels cibler les actions a mener dans le but d’utiliser efficacement les ressources disponibles.

Objectives Our study aimed to update the seroprevalence and factors associated with anti-dengue virus (DENV) antibody positivity among blood donors and to discuss their implications for blood supply. Background Questions on the potential transmission of DENV by transfusion increased after the documentation of the risk of transmission of the West Nile virus. This risk was estimated after transfusion of DENV RNA-positive blood units of up to 37.5%. In Burkina Faso, very few studies on DENV in blood donors have been conducted. As a result, there were no reliable data on DENV to allow the implementation of appropriate measures to control the risk of transmission of the dengue virus by blood transfusion. Methods We conducted a 4-week cross-sectional study from December 4 to 30, 2016. Blood donors of both genders, aged 18-60 years, accepted for blood donation after medical selection were consecutively enrolled. Results Our study included a total of 1007 blood donors, in which donors living in urban areas represented 78.2%. The mean age was 26.1 ± 8.1 years. After adjustment in a multiple regression logistic model, the odds of having IgG anti-DENV increased as age increased. The odds of DENV was 53% lower in rural areas (OR = 0.47; P = .000) compared to urban settings and 42% lower in mobile sites (OR = 0.58; P = .03) compared to fixed ones. Conclusion Our study provides new and useful insights for future research on the risk of TT-DENV throughout blood transfusion.

Aim: This study aims to describe the characteristics of the red cell concentrates (RCCs) produced by whole blood settling in Burkina Faso. Methods: We conducted a cross-sectional study in the blood centers of Ouagadougou and Koudougou between 12th February and 15th March 2015. 427 blood units selected at Koudougou and divided into three groups were processed by settling method over three durations (methods I = 36–48 h, II = 48–72 h and III = 72–96 h). Other 139 units selected at Ouagadougou were processed by centrifugation (method IV). The RCCs units' characteristics were described by the mean of their hematocrit, volume, hemoglobin level and content. The residual plasma level, the hematocrit yield and the hematocrit concentration ability (ability of method to raise hematocrit in comparison with whole blood) have been used to assess methods efficacy. Results: The volume and hemoglobin content of overall units met standard requirements. 2.1%, 14.2% and 27.7% of units respectively issued from methods I, II, III had hematocrit over 50%. The average volume (mL) was 372.88, 357.91, 350.30 and 332.26 respectively for method I to IV (p

Traceability is an essential tool for haemovigilance and transfusion safety. In Burkina Faso, the implementation of haemovigilance has been achieved as part of a pilot project from 2005 to 2009. Our study aims to evaluate the traceability of blood transfusions and reporting of adverse reactions over the 6-year postpilot phase. A cross-sectional study including all blood units ordered between 2010 and 2015 has been conducted in public and private health care facilities supplied with blood products by the transfusion center of Bobo-Dioulasso. The complete traceability was possible for 83.5% of blood units delivered. Adverse reactions were reported in 107 cases representing 2.1/1,000 blood units per annum. Transfusions of wrong blood to wrong patient were reported in 13 cases. Our study shows that the haemovigilance system in Burkina Faso must be improved. Healthcare workers have to be sensitized on how traceability and haemovigilance could impact the quality of care provided to patients.

Peripheral blood progenitor cells (PBPC) infusion allows rapid haematological recovery after high dose chemotherapy. Efficient PBPC collection is therefore essential as rescue therapy for transplantation. In order to validate a new equipment (ComTec®, Fresenius Kabi), we compared the efficiency of three cell separators for PBPC collection in patients with haematological malignant diseases. From June 2014 to December 2015, 83 PBPC were collected in 48 patients. Three aphaeresis machines were used: Cobe Spectra® (Terumo BCT, 11), Amicus® (Fenwall, 30), and ComTec® (Fresenius Kabi, 42). The median collection efficiency was similar between the three separators. The evaluation of cell contamination in the final product revealed a lower red cell contamination with Spectra® and ComTec®, whereas the platelet contamination was lower with Amicus®. The new equipment has been validated and can be further used in routine, with a total running cost that turned out to be quite lower. Each separator has its own characteristics and advantages. Further study is needed to suggest that the choice of separator could be guided following the patient's blood characteristics.

To estimate the seroprevalence of Borrelia burgdorferi (Bb), Anaplasma phagocytophilum (Ap), and Francisella tularensis (Ft) in Belgium, we tested sera from three population-based samples in which exposure to pathogen is assumed to vary: 148 samples from workers professionally exposed, 209 samples from rural blood donors, and 193 samples from urban blood donors. Sera were tested using ELISA or the immunofluorescence assay test. The seroprevalence of Bb was 5.4% in workers professionally exposed, 2.9% in rural blood donors, and 2.6% in urban blood donors, which is similar to other studies. The fraction of negative results decreases significantly from urban blood donors and rural blood donors to workers. Regarding the seroprevalence of Ap, the cutoff titer of 1:64 recommended by the manufacturer may be set too low and produces artificially high seroprevalence rates. Using a cutoff titer of 1:128, the seroprevalence of Ap was estimated at 8.1% for workers professionally exposed, 6.2% for rural blood donors, and 5.7% for urban blood donors. Tularemia sera confirmed the presence of the pathogen in Belgium at 2.0% for workers and 0.5% for rural and urban blood donors. Our study is one of the few providing an estimation of the seroprevalences of Bb, Ap, and Ft in three different populations in Belgium, filling the gap in seroprevalence data among those groups. Our findings provide evidence that the entire Belgian population is exposed to Bb, Ap, and Ft infections, but a higher exposure is noticed for professionals at risk. Education on the risk factors for tick bites and preventive measures for both professionals exposed and the general population is needed.

Resume But de l’etude Evaluer les besoins de formation du personnel dans les centres d’Afrique subsaharienne francophone. Methodologie Cette etude preliminaire a analyse le nombre, le niveau de formation de tout le personnel de quatre centres de transfusions, leurs domaines de formation, leurs durees de formation et les etablissements formateurs. Resultats Les besoins restent eleves dans tous les domaines et sont cruciaux en administration des services de transfusion, en maintenance des equipements et transfusion clinique.

Background and Objectives Photochemical treatment (PCT) based on amotosalen and ultraviolet A light (UVA) demonstrated a wide range of pathogen inactivation. However, coagulation proteins are affected by this treatment. The aim of this study was to evaluate the coagulation parameters in apheresis plasma units after thawing and processing by PCT. Materials and Methods Thirty apheresis plasma units were rapidly frozen at ≤ –30°C after collection. Plasma units were thawed after 7 days for PCT with amotosalen and UVA light. Treated apheresis units were refrozen and stored at ≤ –30°C for 1 month. Samples were collected for each plasma units at several times of process. Coagulation times (prothrombin time, activated partial thromboplastin time), coagulation factors (fibrinogen, Factor [F] II, FV, FVII, FVIII, FIX, FX, FXI), prothrombin fragments 1 and 2, antithrombotic proteins (protein C, protein S, antithrombin) and total protein content were measured. Functionality of ADAMTS-13 was also tested. Results After thawing, coagulation times were slightly increased and a decrease of FV, FVIII and protein C activity was found. The mean recovery for all proteins, except one, ranged from 81% to 97% of the baseline activity in plasma units after thawing and PCT. FVIII was more affected with a mean recovery of 69 ± 8%. ADAMTS-13 function was also preserved after the whole process. The effect of an additional 1-month frozen storage on coagulation parameters was minimum. Conclusion Coagulation protein levels after thawing and processing of plasma by PCT with amotosalen and UVA were preserved well in the physiological ranges.

Chromosomal translocations represent an important prognostic indicator in B-cell chronic lymphocytic leukemia (B-CLL). However, their value had been neither determined in homogeneously treated patients nor compared to that of IgV(H) mutational status. Sixty-five B-CLL patients were investigated using cytogenetics, interphase fluorescence in situ hybridization (FISH), analysis of IgV(H) and of TP53 mutational status before treatment with 2-chloro-2'-deoxyadenosine (CdA). Translocations (n=45) were detected in 42% of the patients, including both balanced (n=12) and unbalanced (n=33) types. IgV(H) was mutated in 43% of the patients. Patients with translocations were more heavily pretreated (P=0.05), presented with more complex karyotypes (P

Activation of the coagulation process is frequently encountered in patients with venous vascular malformations and results in the local formation of clots and the consumption of components of the coagulation process (platelets, clotting factors). This activation accounts for multiple biological abnormalities such as the elevation of the D-dimers, reduction of fibrinogen and platelets count and less frequently a local intravascular coagulation (LIC). This process seems to be responsible for the local painful symptoms. In case of LIC, bleeding complications may be observed. Therapeutically, antithrombotic treatment with low-molecular weight heparin can improve the local coagulopathy with beneficial effects on blood tests disturbances, pain and bleeding tendency.

Objectives: Follicular lymphoma (FL) is the most common adult non-Hodgkin's lymphoma. Diagnosis is based on morphology and can be confirmed by cytogenetic, flow cytometry (FCM) or molecular studies. Despite all these complementary approaches, diagnosis sometimes remains difficult. The purpose of the present work was to characterise the expression of new specific follicular cells markers which allows us to target specifically the abnormal FL cell population in FCM. Methods: A total of 153 samples from healthy subjects and from patients with chronic B-cell lymphoproliferative disorders were analysed by FCM in the same conditions for purpose of comparison. Results: We showed that CD44 is weakly expressed in FL cells compared with peripheral blood mononuclear cell from normal blood donors and others cells from B lymphoproliferative diseases. We nevertheless observed bone marrow samples where some immature B-cell population express CD44 with lower fluorescence intensity. Therefore, we developed a double antibody combination, using CD44 and CD38, which allowed us to separate the normal immature cells from the pathological population using FCM. Conclusion: This new phenotypic approach offers an accurate (sensitivity and specificity of 93% and 96%, respectively), fast and low sample consuming method for the diagnosis of FL.

This work aims to further characterize the newly described leukemic plasmacytoid dendritic cells (LPDC), for which we had previously demonstrated their normal, PDC-like ability to produce IFN-alpha. In addition, LPDC also express the specific antigens BDCA-2 and BDCA-4. Importantly, they become fully competent antigen-presenting cells (APC) after a short maturation induced by IL-3 + CD40L or virus, exhibiting a characteristic APC phenotype (high expression of CD83 and of the costimulatory molecules CD40, CD80, CD86). Whereas IL-3 + CD40L-activated LPDC prime naive CD4(+) T cells towards a Th2 pathway (IL-4-secreting T cells), virus-activated LPDC drive a Th1 profile (IFN-gamma-secreting T cells). Moreover, we show in one case that LPDC are able to capture, process and present exogenous antigens, leading to the activation of both CD4(+) and CD8(+) T cell clones in an antigen-specific manner. This study further characterizes the phenotype and immunological functions of LPDC.

Immunophenotyping is a major tool to assign acute leukemia blast cells to the myeloid lineage. However, because of the large heterogeneity of myeloid-related lineages, no clinically relevant immunological classification of acute myeloblastic leukemia (AML) has been devised so far. To attempt at formulating such a classification, we analyzed the pattern of expression of selected antigens, on blast cells collected at AML diagnosis. Patients were eligible if they had a first diagnosis of de novo AML and a sufficient number of blast cells for proper immunophenotyping. The relative expression of CD7, CD13, CD14, CD15, CD33, CD34, CD35, CD36, CD65, CD117, and HLA-DR were analyzed by cytometry in a test series of 176 consecutive AML cases. Statistical tools of clusterization allowed to remove antigens with overlapping distribution, leading us to propose an AML classification that was validated in a second AML cohort of 733 patients. We identified five AML subsets (MA to ME) based on the expression of seven antigens within four groups (CD13/CD33/DC117, CD7, CD3!5/CD36, CD15).-MA and MB-AML have exclusively myeloid features with seldom extramedullary disease and rare expression of lymphoid antigens. No cases of acute promyelocytic leukemia (APL) were observed within MB AML. MC AML have either myeloid or erythroblastic features. MD AML have more frequently high WBC counts than other subsets, which were related to the expression of CD35/CD36 and CD14 and to monoblastic differentiation. ME AML lack CD13, CD33, and CD117 but display signs of terminal myeloid differentiation. Specific independent prognostic factors were related to poor overall survival in each immunological subset: CD34+ (P

En Belgique, c’est en 2011 que le service publique federal (SPF) sante publique, securite de la chaine alimentaire et environnement a cree un Comite national ayant pour mission l’accompagnement des hopitaux pour la fonction d’hemovigilance. Ensuite, en 2014, l’optimisation de la consommation en composants sanguins est venue s’ajouter a cette mission. C’est la Plate-forme nationale de politique transfusionnelle (PNPT) qui a alors vu le jour et a ete rapidement rebaptisee « BeQuinT » pour Belgian Quality in Transfusion. Pour assurer ses differentes missions, BeQuinT a constitue des groupes de travail parmi lesquels un plus specialement dedie a la « formation et education en transfusion ». Le groupe s’est donne comme objectif de mettre a la disposition des soignants du materiel didactique dans les deux langues nationales (diaporamas, videos, e-learning) reprenant les notions essentielles et des Quiz permettant d’objectiver l’acquis de connaissances. La transfusion a ete decomposee en 13 modules couvrant en ± 300 diapositives la chaine transfusionnelle depuis l’organisation de la transfusion en Belgique et le recrutement des donneurs de sang jusqu’a la mise en route d’une transfusion, en passant par la preparation, la conservation et le transport des composants sanguins, la prescription, le prelevement pretransfusionnel, les transfusions urgentes, les reactions transfusionnelles (immediates et retardees), la transfusion neonatale et pediatrique et l’hemovigilance. C’est sur le site officiel de l’Institut national d’assurance maladie et invalidite (INAMI) que les apprenants ont acces aux modules dont la consultation est prise en compte pour l’accreditation professionnelle.

Introduction BeQuinT (Belgian Quality in Transfusion), une plateforme mise en place par le service public federal de sante publique a organise 3 enquetes nationales (2012–2014–2016) ayant pour but de dresser une carte et determiner l’evolution dans le temps de la qualite de la politique transfusionnelle dans les hopitaux belges relatives a differents champs d’application : la « Transfusion en general », la « Prescription », la « Tracabilite » et le « Rapportage ». Methodologie Tous les hopitaux belges avec un comite de transfusion ont ete invites a remplir l’enquete en ligne ; c’etait la condition sine qua non du financement de la fonction d’hemovigilance. La distribution et les tendances des scores par champ d’application et pour la totalite de l’enquete ont ete examines et un benchmarking anonymise pour les 4 champs d’application a ete calcule. En juin 2013, 2015, 2017, les resultats ont ete presentes aux presidents des comites de transfusion, ainsi que les points a ameliorer, pour aider les comites de transfusion a orienter leur politique transfusionnelle et pour etablir les priorites de la politique de BeQuinT. Resultats Entre la 1 re et la 3 e enquete, les scores medians sont plus eleves dans tous les champs d’application ; l’augmentation des scores est significative ( p re et la 3 e enquetes est significative ( p = 0,005). Nous pouvons donc conclure a une amelioration significative de la qualite transfusionnelle entre 2012 et 2016.

Le Daratumumab est un anticorps anti-CD38 utilise dans le traitement du myelome multiple, avec des resultats tres encourageants. Comme la molecule CD38 est exprimee sur les erythrocytes, ce medicament peut interferer avec les tests d’immunohematologie, en particulier les reactions utilisant de l’antiglobuline et parfois, le phenotypage erythrocytaire. Cette interference peut se prolonger jusqu’a 6 mois apres l’arret du traitement. A ce jour, dans le cadre de protocoles therapeutiques, 14 patients ont beneficie de cette molecule au CHU UCL NAMUR et 10 d’entre eux ont du etre transfuses. Dans tous les cas, la RAI et le test d’identification pre-transfusionnels etaient positifs. Un traitement des globules rouges par le dithiotreitol a ete valide et utilise afin de contrecarrer l’interference de l’anti-CD38. Des poches compatibles ont pu ainsi etre selectionnees et les transfusions ont ete efficaces. Un logigramme decisionnel de prise en charge a ete redige. Un bilan immunohematologique avant l’instauration du traitement et de toute transfusion est hautement recommandee, comportant un phenotypage erythrocytaire etendu. Des tests complementaires habituellement non systematiques doivent parfois etre realises. Afin de prevenir toute difficulte en cas de transfusion, une information claire, pertinente et adaptee doit etre mise a disposition des prescripteurs, des patients eux-memes et des banques de sang hospitalieres. Une collaboration entre les laboratoires d’immunohematologie est souhaitable et, dans tous les cas, une communication efficiente entre toutes les parties concernees par le traitement est indispensable.

R. N. I. Pietersz, H. W. Reesink, S. Panzer, S. Oknaian, S. Kuperman, C. Gabriel, A. Rapaille, M. Lambermont, V. Deneys,D. Sondag, S. Ramirez-Arcos, M. Goldman, G. Delage, F. Bernier, M. Germain, T. Vuk, J. Georgsen, P. Morel, C. Naegelen,L. Bardiaux, J.-P. Cazenave, J. Dreier, T. Vollmer, C. Knabbe, E. Seifried, K. Hourfar, C. K. Lin, M. Spreafico, L. Raffaele,A. Berzuini, D. Prati, M. Satake, D. de Korte, P. F. van der Meer, J. L. Kerkhoffs, L. Blanco, J. Kjeldsen-Kragh,A.-M. Svard-Nilsson, C. P. McDonald, I. Symonds, R. Moule, S. Brailsford, R. Yomtovian & M. R. JacobsSeptic reactions after transfusion, particularly of plateletconcentrates, still occur and belong to the most serioustransfusion reactions. From a previous InternationalForum [1] on the subject, it could be concluded that inpart of the countries that participated in the forum, plate-let concentrates (PCs) were tested for bacterial contamina-tion and that culture-based methods, particularly theBacT/Alert system, were used.In recent years, several rapid bacterial detection meth-ods, such as surrogate measurements of the pH or glu-cose, the detection of bacteria with a scan system orPCR tests that detect bacterial RNA, have been devel-oped. These tests can either be performed immediatelyprior to transfusion of the PC or at a variety of testmoments at which culture and release tests are com-bined.Pathogen inactivation (PI) methods also affect bacterialcontamination of PCs. In 2007 [1], in some countries, theIntercept method of PI of PCs was implemented insteadof bacterial screening.It seemed of interest to evaluate the present state ofthe art of this subject. In order to obtain the desiredinformation, the following questions were sent to expertsin the field.Question 1: How long do you store PC and is there adifference between whole-blood-derived PC and apheresisPC? Which method of preparation do you use for whole-blood-derived PC? Are PCs leuco-reduced?Question 2: Do you use a culture method to detect bac-terial contamination of PC? If so

BACKGROUND AND OBJECTIVES: Immunophenotyping has become a useful tool for the differential diagnosis of chronic B-cell lymphoproliferative disorders. The aim of this work was to determine reference values of normal B-cell subpopulations. MATERIAL AND METHODS: Blood samples from 38 healthy volunteers were analyzed by multidimensional flow cytometry, using a panel of directly conjugated antibodies. Results were expressed as percent of positive B cells and as median fluorescence intensity, an indirect assessment of the expression level. RESULTS: CD20, CD22, CD24, CD40, CD79a, CD79b, FMC7, CD11a, CD18, CD44 were positive in the whole B cell population, whereas CD10, CD86, CD103, CD154 and FasL were almost absent from the B-lymphocyte population. 75% were IgD positive. The kappa/lambda ratio was 1.5. CD5, CD23, CD25, CD38, CD43, CD54, CD62L, CD80 and CD95 were positive in different B-cell subpopulations. The utility of all these markers in the differential diagnosis of chronic B-cell lymphoproliferative disorders is discussed. CONCLUSION: In order to interpret a pathological immunophenotype, it is necessary to refer to quantitative and qualitative values of normal B-cell subpopulations.

Umbilical cord blood (CB) transplantations are associated with a lower risk of severe graft-versus-host disease (GVHD) compared to BMT. GVHD is an immune reaction that involves interaction between cell surface molecules resulting in cell activation and release of many cytokines. Monocytes are known to be an important source of cell adhesion (CAM) and co-stimulatory molecules which play a crucial role in the efficient activation of T and B cells. We analyzed the phenotype of CB monocytes in the presence or absence of an inflammatory signal (rIFN-gamma) and compared them to adult blood (AB); the expression of HLA-DR and 17 different markers (CD11a, CD11b, CD11c, CD18, CD29, CD40, CD44, CD49a, CD49d, CD49e, CD49f, CD54, CD58, CD62L, CD80, CD86 and CD102) was measured by flow cytometry. Statistical analysis showed that, compared to AB, CB monocytes did not express CD11b, CD11c, CD49d and after stimulation with rIFNgamma, they lost the expression of CD58 and CD102, whereas CD80 and CD86 expression was induced. The analysis of fluorescence intensity (MFI) revealed that CB monocytes expressed some CAM (CD29, CD54, CD102) with a lower intensity than AB monocytes except CD44. In conclusion, absence and reduced expression of some markers argue for a different phenotypic profile of CB monocytes compared to AB monocytes, which might partly contribute to their impaired immune response and to the low incidence of GVHD observed after CB transplantations. However, CB monocytes expressed CD80 and CD86 co-stimulatory molecules, but this expression did not prove a normal co-stimulatory function.

Translocation t(12;21)(p13;q22) is the most frequent cytogenetic abnormality in childhood acute lymphoblastic leukemia (ALL) and is generally associated with favorable prognosis. In this report, we assessed the value of dual-color interphase fluorescence in situ hybridization (FISH) for the detection of t(12;21). Fifty-three patients were screened for ETV6/CBFA2 fusion by means of FISH, using two cosmid probes mapped on ETV6 and on CBFA2, respectively. The cut-off value (mean + three standard deviations) for positivity established on control patients was 9.3%. A comparison between FISH and molecular methods [reverse-transcriptase polymerase chain reaction/Southern blot (RT-PCR/SB)] was possible in 52 patients: 34 of 52 (65.4%) showed negative results with both approaches, and 13 of 52 (25%) were positive; 5 of 52 (9.6%) showed discrepancies: four patients who were positive using RT-PCR/SB were negative using FISH. Conversely, one patient negative when using RT-PCR/SB was positive with FISH. Further investigations on this patients, cytogenetically characterized by add(12p), showed an atypical breakpoint on ETV6, located 5' to the common breakpoint. Compared with RT-PCR and SB, dual-color interphase FISH with the cosmid probe set proved to be highly specific but showed limited sensitivity.

BACKGROUND AND OBJECTIVES: Freezing is a practical approach for cell preservation for retrospective studies. The aim of this work was to check the cryopreservation impact on B cell chronic lymphocytic leukaemia phenotype. MATERIAL AND METHODS: Blood samples from 15 CLL patients were analyzed freshly and after freezing at -196 degrees C, without separation, and thawing. Results were compared by Student's paired t-test. RESULTS: The phenotype of fresh CLL cells was as follows: CD19+, CD5+, faint CD20, CD23+/-, weak CD22 and sIg, CD37+, HLA-DR+, FMC7-. After cryopreservation, the percentage of CD5 and CD23 positive cells decreased, whereas HLA-DR positive cells increased moderately. The CLL Matutes's score was modified in 6 cases out of 15 (40%). CONCLUSION: Cryopreservation modifies B cell chronic lymphocytic leukaemia phenotype, by decreasing CD5 and CD23 expression.

Because 2-chloro-2'-deoxyadenosine (CdA) is active in B-chronic lymphocytic leukemia (B-CLL), and may interfere with DNA repair, we investigated the potentiating effect of CdA on the cytotoxicity induced in vitro in B-CLL lymphocytes by cyclophosphamide (CP) derivatives, which induce DNA damage by DNA cross-linking. Exposure to CdA at clinically achievable concentrations for 2 h, followed by mafosfamide (MAF) or 4-hydroxycyclophosphamide (4HC) for 22 h, resulted in synergistic cytotoxicity in the majority of B-CLL samples tested. Synergy between CdA and MAF was observed in cell samples of sensitive/untreated patients, as well as in cells of resistant/pretreated patients, particularly at the highest concentrations of MAF. In the cells treated with CdA and MAF, we observed loss in ATP and hallmarks of apoptosis, as evidenced by cellular morphology and high molecular weight DNA fragmentation. The synergy could be explained neither by an influence of MAF on the phosphorylation of CdA, nor by an increase in the incorporation of CdA into DNA in the presence of MAF. The in vitro synergy between CdA and CP derivatives provides a rationale for the use of this association in B-CLL patients.

We report the case of a 3-year-old boy with severe haemophilia A presenting with recurrent haemarthroses despite daily infusions of factor VIII delivered through a central venous access device (CVAD). Regular rinsing of the CVAD with heparin, according to a standard protocol, resulted in systemic anticoagulation, as demonstrated by prolonged thrombin time and therapeutic anti-Xa levels. The bleeding symptoms resolved after replacing heparin with a normal saline solution. This case illustrates that heparin administered to maintain CVAD patency should be used with caution in young haemophiliacs. Prolonged thrombin time should alert the physician to this possible CVAD complication.

It is widely believed that multiple sclerosis is a T-cell mediated autoimmune disease associated with abnormalities in immunoregulation. This large, prospective study evaluated the lymphocyte immunophenotypic profile of 246 MS patients, divided clinically into a remitting/relapsing group (n = 176) and a progressive group (n = 70), and compared their results to those of 117 healthy controls. All patients were found to have significantly elevated percentage and absolute numbers of IL2R(+)CD3(+) cells as well as depressed percentages of CD45RA(+)CD4(+) cells. However, when the factor of treatment with cyclophosphamide (CY) versus no treatment or treatment with other agents was used to group patients, dramatic declines in both percentages and absolute numbers of CD45RA(+) CD4(+) cells were discovered. These declines were associated specifically with CY and could be explained by this factor independent of the clinical state of the patient. The effects were seen in patients undergoing current treatment or in those exposed to CY in the near or remote past. These findings highlight the confounding effect of specific treatments on the immune profile of MS patient groups and suggest that there may be important implications for cellular function and clinical outcome in these and other patient groups.

Reduction of leucocytes in blood components, achieved by filtration, may reduce the risk of HLA alloimmunization, virus transmission, and febrile reactions. Nevertheless, white blood cell (WBC) concentrations obtained in this way can be too low to be accurately counted with automated and manual techniques. We describe here a rapid, reliable, and very sensitive method for counting low leucocyte numbers using flow cytometry. WBC nuclei are labelled by propidium iodide. A known amount of fluorescein- and phycoerythrin-conjugated beads (Standard Brite, Coulter) is added as an indicator of the examined volume. The time required for analyzing one tube is approximately 5 min. This method was first validated by comparing WBC counts (120-2,570/microliters) assessed on a haemocytometer (Bürker) and by flow cytometry. The second step was to determine the sensitivity of the method: we diluted normal platelet concentrates with phosphate-buffered saline. The comparison of expected with observed values showed a very good correlation up to a concentration of 1 WBC/microliter. This technique is an accurate method that can be applied in blood bank quality controls and in clinical studies.

The true incidence of bedside transfusion errors, i.e. those happening when blood products have left the blood bank, is underestimated because published figures rely on reporting of clinically relevant events or on indirect methods. The SAnGUIS project assessing blood practice in a prospective and randomized fashion for 6 elective surgical procedures gave the opportunity to trace all transfused units and to identify steps at risk during blood delivery in surgery. We considered transfusion of a wrong unit as a major error and poor execution or documentation as a recording error. Over 15 months, 808 patients out of 1,448 were transfused with 3,485 units. A total of 165 errors were found after blood products had left the blood banks. Seven were misidentifications (0.74% of patients, 0.2% of units). Eight other major errors occurred in 4 (0.5%) patients. Major errors occurred during nonemergency situations, in wards or intensive care units. The remaining ('recording') 150 errors consisted of misrecordings (61), mislabellings (6), or failures to document transfusions in the medical records (83). All errors were uneventful except one misidentification which induced a transient, yet unreported, reaction. The 'descending' inquiry method used for this study showed that most errors pass unnoticed and are therefore not reported. Measurement of error rates may constitute an important quality indicator. Retrospective information of this survey to the concerned staff people provided an impetus to take adequate measures to reduce these bedside errors.

A reference range for lymphocyte populations, with particular emphasis on T lymphocyte subsets, was obtained for normal individuals covering age cohorts from birth through adulthood. This report confirms and extends findings from a developmental reference range published earlier (1). Absolute numbers of WBC, lymphocytes, and T, B, and NK subsets decline significantly during childhood. However, differences in the rate of decline of certain lymphocyte subsets leads to discordance between absolute numbers and percentages. Those lymphocyte subsets which decline less rapidly with age than the total lymphocyte count will show an increase in percentage, whereas those which decline more rapidly will show further declines in percentage values. T cell percentages were seen to increase over time whereas B cell percentages decline. Markers of immaturity such as CD45RA on CD4 cells and CD38 on CD8 cells declined in both percentages and absolute numbers. Activation markers, such as HLA-DR on CD8 cells and IL2-R on CD3 cells, increased in percentages with time but changed inconsistently in cell number from infancy to adulthood. These findings extend the lymphocyte reference range to markers thought to be informative in various disease states, including HIV infection.

H. W. Reesink, S. Panzer, C. A. Gonzalez, N. Lena, P. Muntaabski, S. Gimbatti, E. Wood, M. Lambermont, V. Deneys, D. Sondag, T. Alport, D. Towns, D. Devine, P. Turek, M.-K. Auvinen, T. Koski, C. K. Lin, C. K. Lee, W. C. Tsoi, E. Lawlor, G. Grazzini, V. Piccinini, L. Catalano, S. Pupella, H. Kato, S. Takamoto, H. Okazaki, I. Hamaguchi, J. C. Wiersum-Osselton, A. J. W. van Tilborgh, P. Y. Zijlker-Jansen, K. M. Mangundap, M. R. Schipperus, D. Dinesh, P. Flanagan, O. Flesland, C. T. Steinsvag, A. Espinosa, M. Letowska, A. Rosiek, J. Antoniewicz-Papis, E. Lachert, M. B. C. Koh, R. Alcantara, M. Corral Alonso & E. Muniz-Diaz

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BACKGROUND: The study of lineage markers by real-time quantitative polymerase chain reaction (RT-qPCR) at diagnosis enables differentiation between acute myeloblastic leukemia, B- or T-lineage acute lymphoblastic leukemia, without cell sorting. Our objective was to assess the relationship between protein expression and the amount of lineage marker mRNA in acute leukemia samples and to determine whether four lineage markers could be used to differentiate between normal and acute leukemia bone marrow (BM) without cell sorting. METHODS: Quantification of the mRNA of CD19, CD79a, CD3e, and myeloperoxidase was performed by RT-qPCR on 130 acute leukemia BM samples at diagnosis and on 20 BM samples from healthy donors, without cell sorting. Immunophenotyping of leukemia samples was performed after manual gating around the blastic population. RESULTS: Reference values for the four lineage markers were established by RT-qPCR for normal BM. The mRNA expression levels of these four lineage markers allowed the distinction between normal samples and 100% of acute leukemia samples. CONCLUSIONS: With 92% congruence for protein expression and amount of mRNA in acute leukemias, these four lineage markers, essential for diagnosis and subclassification of acute leukemias by flow cytometry, also represent excellent candidate genes when using RT-qPCR technology as a diagnostic tool for molecular cancer class prediction.

OBJECTIVE: To determine which venous malformations (VMs) are at risk for coagulopathy. Venous malformations are slow-flow vascular malformations present at birth, and localized intravascular coagulopathy (LIC) causes pain and thrombosis within a lesion and severe bleeding during surgical procedures. DESIGN: Prospective convenience sample accrued from 2 multidisciplinary sites in Brussels, Belgium, and Caen, France. PARTICIPANTS: The study population comprised 140 patients with clinical data and coagulation parameters. Magnetic resonance imaging was performed for 110 patients. MAIN OUTCOME MEASURE: Measurement of D-dimer levels. RESULTS: Of the 140 participants, 59 (42%) showed high D-dimer levels, 36 (61%) of whom had levels higher than 1.0 microg/mL. Six of the participants had low fibrinogen levels. In univariate analysis, large surface, presence of palpable phleboliths, and truncal localization were associated with high D-dimer levels. In the multivariate analysis, only large surface area and presence of phleboliths remained independently associated with high D-dimer levels. Severe LIC, characterized by concomitant low fibrinogen level, was associated with extensive venous malformations of the extremities. CONCLUSIONS: Localized intravascular coagulopathy is statistically significantly associated with large and/or deep venous malformations that affect any location, which can have a palpable phlebolith. These patients are at risk of local pain due to thrombosis. Lesions with elevated D-dimer levels associated with low fibrinogen levels (severe LIC) commonly affect an extremity and have a high risk of hemorrhage. Low-molecular-weight heparin can be used both to treat the pain caused by LIC and to prevent decompensation of severe LIC to disseminated intravascular coagulopathy.