Search

Background and aim Sickle cell anemia (SCA) is characterized by variable patterns of clinical expression. Polymorphisms linked to different genes have been associated with specific complications of the disease. Herein, we focused on the study of the association of 4 polymorphisms of Insulin like Growth Factor 1 receptor ( IGF1R ) gene with infections, which are the major cause of death in SCA. Material and methods This study involved 116 sickle cell patients among whom 58 SS have the same confirmed infectious phenotype. Allele-Specific PCR was performed for the study of rs1319868, whereas the PCR/sequencing method was carried out for rs1567811, rs2872060 and rs8041224. Results The results showed that rs1319868 and rs1567811 were associated with a decreased risk of infection among SS patients ( p = 0.038, RR = 0.54; p = 0.044, RR = 0.56, respectively). Interestingly, the combination of different genotypes showed the association of the genotype GT of rs1319868 and the genotype CC of rs8041224 with further decreased infection risk in SCA ( p = 0.028, RR = 0.04). Conclusion These significant associations of IGF1R SNPs with infection suggest that this gene could play an important role in the immune function in SCA.

Resume La fibrillation atriale est une situation clinique qui peut exposer a un risque de complications thromboemboliques. Plusieurs marqueurs biologiques ont ete testes afin d’affiner la stratification de ce risque. Le but de cette etude est d’etudier l’interet du dosage des d -dimeres dans cette pathologie. L’etude etait prospective, interessant 50 malades et 19 temoins colliges au service de cardiologie de l’hopital Mongi Slim entre le 1er juillet–1er novembre 2012. Le dosage des d -dimeres a ete realise sur mini-VIDAS® et analyse en fonction des donnees cliniques, echographiques et biologiques. L’âge moyen des patients etait de 61,8 ± 10,6 ans. Le score CHA2DS2-VASc moyen etait de 2,58 ± 1,31. Le taux moyen des d -dimeres des patients etait de 590 ± 506 ng/mL et de 225,26 ± 112,95 ng/mL chez les temoins (p = 0,02). Il n’a pas ete retrouve de difference significative entre le taux des d -dimeres selon l’âge, le sexe, le type et l’etiologie de la fibrillation atriale, le score CHA2DS2-VASc, la surface de l’oreillette gauche et la presence d’un thrombus intra-auriculaire gauche. Le taux des d -dimeres etait significativement plus eleve chez les patients sous acenocoumarol ayant un INR d -dimeres (300 ng/mL) a ete determinee permettant de distinguer significativement les patients des temoins (p d -dimeres pourrait permettre d’identifier les patients en fibrillation atriale avec un etat d’hypercoagulabilite. Il pourrait etre complementaire au dosage de l’INR dans la surveillance du traitement anticoagulant et ceux en permettant d’affiner le risque thrombogene.

We report here the clinical, hematological and molecular data in a 50-year-old patient with β-thalassemia intermedia (β-TI) caused by a homozygous β+ mutation on the β-globin gene polyadenylation (polyA) signal (AATAAA>AAAAAA). β Haplotype analysis was accomplished by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Haplotype and framework analysis showed that this mutation is associated with the [- - - - + + +] β haplotype and framework 1 (CCGCT) (FW1). This mutation was previously reported in the heterozygous state in association with the codon 9 (+TA) mutation in a β-TI patient originating from Tunisia. To the best of our knowledge, this is the first report describing this mutation in the homozygous state. The case reported here, coinherited Gilbert's syndrome, which is characterized by hyperbilirubinemia. This conclusion was reached by the investigation of the promoter region [A(TA)nTAA] motif of the UGT1A1 gene, showing the (TA)6/(TA)7 genotype.

ResumeObjectif Evaluer le taux d’immunisation anti-erythrocytaire et anti-HLA dans les hemoglobinopathies. Patients et methodes Etude transversale (octobre 2009–mars 2010) concernant 83 patients suivis pour hemoglobinopathie. La recherche d’anticorps irreguliers est realisee par deux techniques : Coombs indirect et technique enzymatique sur carte gel. La recherche des anticorps anti-HLA de classe I est faite par la technique de lymphocytotoxicite complement dependante. Resultats L’âge moyen est de 30 ans (14–64 ans), le sex-ratio M/F est de 0,84. Il s’agit de 42 cas de syndromes drepanocytaires majeurs (29 drepanocytoses homozygotes et 13 drepano-thalassemies) et de 41 cas de syndromes thalassemiques (26 thalassemies majeures et 15 intermediaires). La frequence d’allo-immunisation anti-erythrocytaire est de 22,89 % sans difference entre les syndromes drepanocytaires et thalassemiques (7,31 %). La frequence d’auto-immunisation anti-erythrocytaire est de 24,8 %. Elle est plus elevee dans les syndromes thalassemiques (41,46 %) que dans les syndromes drepanocytaires (7,14 %) (p

Resume En Tunisie, la transfusion de globules rouges s’inscrit dans un cadre reglementaire mais reste sujette a des defaillances en raison de la meconnaissance de la legislation et des regles de securite. L’objectif de notre travail est d’evaluer les connaissances des medecins en matiere de securite immunologique en transfusion erythrocytaire avant et apres une lecture d’un CD-ROM d’auto-enseignement. Il s’agit d’une evaluation d’une intervention. Quatre-vingts medecins hospitaliers ont repondu a un questionnaire anonyme, comportant sept questions a choix multiple (QCM) de 29 items. Le taux de bonnes reponses (TBR) calcule par question et par item a pris en compte l’impact du CD-ROM sur l’amelioration des reponses apres lecture. La note globale moyenne est de 2,9/7. Les reponses par questions sont variables (25 % a 76 %). Tous les participants ont repondu juste a plus de 50 % des items. Deux participants ont repondu correctement a tous les items. Parmi les participants, 31,3 % ont repondu correctement aux cinq items « importants » concernant les regles de compatibilite dans le systeme ABO et l’epreuve ultime au lit du malade. Quatre-vingt-trois pour cent ont visionne le CD-ROM et accepte de participer a l’evaluation finale. L’impact du CD-ROM a ete statistiquement significatif. La note globale a augmente de 2,9 a 5,8/7, 31,5 % ont repondu correctement a toutes les questions (vs 2 %) et 95,5 % ont repondu correctement aux cinq items importants (vs 31,3 %). Ce travail a revele une meconnaissance des medecins en matiere de securite immunologique erythrocytaire. Le CD-ROM a significativement ameliore les reponses. L’instauration d’une formation continue est imperative dans notre pays.

International audience; The pro-inflammatory context of sickle cell disease promotes the liberation of cytokines such as CCL5, encoded by a gene located on chromosome 17. Herein, the occurrence of three variations of CCL5 in sickle cell anemia (SCA) and their relations to two major complications - painful crisis and presence of infections - were investigated. 100 SCA Tunisian patients and 100 healthy subjects were included in the case control study. Then the sample of patients was divided into two groups according to the presence or absence of each complication. The polymorphisms, namely g.-403G>A, g.-28C>G and g.In1.+1T>C, were analyzed by PCR/sequencing. Our findings show the presence of eight genotypes, namely GG, GA and AA of g.-403G>A, CC, CG and GG of g.-28C>G, and TT and TC of g.In1.+1T>C. The frequencies of studied single nucleotide polymorphisms (SNPs) and haplotypes in SCA patients do not differ significantly from healthy control group results. There is also no significant association between the analyzed polymorphisms and complications as for painful crisis and presence of infections (p > 0.05). Altogether, our data support the conclusion that the three polymorphisms of CCL5, namely g.-403G>A, g.-28C>G and g.In1.+1T>C, do not seem to be involved in the clinical variability of SCA in Tunisia.

The C/EBPE gene, located in 14q11.2, encodes for a B/zip-type transcription factor. The C/EBPɛ is involved in terminal differentiation and functional maturity of granulocyte progenitor cells and in cell apoptosis during myeloid differentiation. A C/EBPE gene has recently been described as a candidate gene involved in clinical variability of β-thalassemia (β-thal). In this study, the C/EBPE gene was sequenced in 146 subjects divided into the severe type of β-thal major (β-TM) and moderate type of β-thal intermedia (β-TI), and a control group. The analysis identified the rs45496295 (C > T) polymorphism in the heterozygous state in 73.9% β-TI patients, which was not the case in the β-TM patients or in the control group. Thus, the T allele is consequently associated with the β-TI group (p = 10-3). According to the Human Splicing Finder (version 3.0, Marseille, France), the presence of the rs45496295 polymorphism leads the creation of a new intronic exotic splicing enhancer (ESE) site. Moreover, the T allele of rs45496295 is associated with a lower transfusion regimen (p = 10-3) and a higher pretransfusion hemoglobin (Hb) rate (p = .006). The comparison of several factors concerning T allele carriers and non-carriers showed that the T allele does not act on the Hb F rate. The T allele of rs45496295, associated with moderate type of β-thal, seems to modify the C/EBPɛ action, thereby preventing the hemolysis.

Resume Objectif La determination avec precision de la lignee cellulaire au cours de la leucemie aigue (LA) est une etape cruciale dans le diagnostic et la conduite therapeutique ulterieure. En Tunisie, faute de cytometrie de flux (CMF), certaines LA continuent d’etre traitees sur la base de la cytologie seule. Notre objectif est de confronter les resultats de la cytologie a ceux de l’immunophenotypage et d’analyser les degres de discordance. Patients et methodes L’etude concerne 100 cas de LA. Elle consiste a effectuer une deuxieme lecture a posteriori en double insu de lames de moelle de LA deja diagnostiquees par deux cytologistes differents et de confronter aux resultats de l’immunophenotypage. Resultats Deux cas classes cytologiquement en LAM ont ete reclasses a l’immunophenotypage en LAL T et leucemie aigue biphenotypique. Trois cas classes cytologiquement LAL ont ete reclasses leucemies aigues indifferenciees (2 cas) et leucemie aigue biphenotypique (1 cas). Quatre cas non identifies a la cytologie ont ete classes LAL B (3 cas), leucemie aigue biphenotypique (1 cas). Dans les deux cas de discordance entre les deux cytologistes, la cytometrie de flux a permis de retenir le diagnostic de leucemie aigue biphenotypique dans un cas et de LAM dans l’autre. Conclusion L’etude cytologique demeure insuffisante dans le diagnostic de lignee meme avec des cytologistes experimentes. L’immunophenotypage est un examen incontournable pour confirmer ou redresser le diagnostic de LA.

Auteur(s) : Wijden El Borgi1, Emna Gouider1, Nour Louati1, Nawel Ben Salah1, Halima Senana2, Ramzi Jeddi3, Ali Saad2, Raouf Hafsia1 1Service d’hematologie biologique, Hopital Aziza Othmana, Tunis 2Service de cytogenetique, Hopital Farhat Hached, Sousse 3Service d’hematologie clinique, Hopital Aziza Othmana, Tunis, Tunisie L’hemophagocytose par les cellules blastiques est rarement observee au cours des leucemies aigues (LA) myeloides, elle est surtout rapportee dans les [...]

Von Willebrand's disease (VWD) is the most commonly inherited bleeding disorder. It is characterized by clinical, biological and molecular heterogeneity. In certain types of the disease, diagnosis can be difficult.We report the clinico-biological characteristics of VWD's patients and analyze diagnosis difficulties.33 cases were diagnosed in the laboratoryfrom February to May, 2011. Screening hemostasis included the measuring ofC, VWF: Ag and VWF: RCo. Blood cell count and blood group were performed in all cases.Mean age at diagnosis is 13 years [10 months -43 years]. The sex ratio M/F is 0.5. The patients are classified type 3 VWD in 52% of the cases, type 2 VWD in 30 % of the cases and type 1 VWD in 18 % of the cases. The diagnosis of type 2B VWD suspected in combination of the ratio VWF:RCo / VWF: Ag0,7 and thrombocytopenia in one case. Required tests for positive diagnosis and distinction between the primary categories of VWD are available. Specialized tests will allow a best characterization variants type 2 VWD for a better therapeutic approach.

International audience; Aims: Fetal hemoglobin (HbF) modulates the phenotype of sickle cell anemia (SCA) by inhibiting deoxy sickle hemoglobin (HbS) polymerization. HbF genes are genetically regulated, and the level of HbF and its distribution among sickle erythrocytes is highly variable. Herein, we aimed to determine whether two functional polymorphisms of BCL11A are implicated in the variation of HbF and clinical events in SCA Tunisian patients. Material and methods: The studied population consisted of 148 SCA patients with SS phenotype. The group of patients was divided into two subgroups according to the threshold point of %HbF which is 15%. Genotyping of rs11886868 and rs4671393 was performed using PCR/Sequencing. To test for trait association with the candidate SNPs, genotype and allele frequencies between `group who had %HbF < 15' and `group who had %HbF > 15' (controls) were compared using Pearson's chi-square test (compare 2, version 1.02). The association of each genotype and the combined genotype with complications was performed by logistic regression test. Results: Our findings showed that the majority of patients carried genotype CT of rs11886868 and genotypes AG and GG of rs4671393 present HbF level < 15%. RR = 0.08, RR = 0.176, and RR = 0.189, respectively. The results showed a significant association between the alleles T of rs11886868 and G of rs4671393 and %HbF < 15% with P = 0.016; RR = 0.39 and P = 8.9 x 10(-3): RR = 0.567, respectively. Interestingly, the C allele of the rs11886868 and the A allele of the rs46713939 were associated with an ameliorated phenotype in patient's SCA. The combination of the genotypes GG and CT explains more phenotypic variance than the sum of the two BCL11A SNPs taken individually.

International audience; Hypereosinophilic syndromes (HES) are a heterogenous group of rare disorders characterized by sustained and otherwise unexplained overproduction of eosinophils with organ involvement and consecutive dysfunction. Detection of the FIP1L1-PDGFRA fusion gene or the corresponding cryptic 4q12 deletion in HES supports the diagnosis of chronic eosinophilic leukemia (CEL) and provides a molecular explanation for the pathogenesis of this disorder. We screened seven Tunisian patients fulfilling the WHO criteria of HES for the presence of the FIP1L1-PDGFRA fusion gene using nested reverse transcription polymerase chain reaction on peripheral blood samples. Four of the seven patients were positive for this fusion gene. Sequence analysis revealed a substantial heterogeneity of the fusion transcripts due to the involvement of several FIP1L1 exons. All patients were male. The median age at diagnosis was 24 years (range, 18-50); one patient had a history of hypereosinophilia of more than 10 years. Two patients had clinically important and symptomatic eosinophilic endomyocardial disease with thrombotic events. Splenomegaly was constant in FIP1L1-PDGFRA positive CEL but not in the other HES patients (only 1/3).; Les hyperéosinophilies essentielles sont caractérisées par une hyperéosinophilie supérieure à 1500 éléments/mm 3 persistante au-delà de sixmois, après exclusion d ’ une éosinophilie réactionnelle. Différentes altérations viscérales peuvent s’y associer leur pronostic est souvent redou-table et jusqu’à une période récente, il n’existait pas de traitement spécifique ou réellement efficace. En 2003 un gène de fusion :FIP1L1-PDGFR alpha a été retrouvée de façon récurrente dans ces syndromes, ce gène de fusion est généré à la suite d’une délétion interstitielle en 4q12 qui est cryptique à l’ analyse cytogénétique standard, la protéine chimérique résultante a une activité tyrosine kinase constitutive qui répond au traitement par l’ imatinib utilisé par ailleurs pour le traitement de la leucémie myéloïde chronique. Nous rapportons quatre cas d’hyperéosinophilie associéesà ce remaniement génétique

As a result of chronic hemolysis, hyperbilirubinemia is often observed, leading to the formation of pigment cholelithiasis which could be busted by the presence of uridine diphosphoglucuronosyltransferase 1A1 defects.Herein, we investigated the effect of glibert mutation on the occurrence of pigment cholelithiasis in Tunisian patients with beta (β) hemoglobinopathy including sickle cell anemia and β thalassemia (minor).Our study included 151 subjects divided in 75 SCA patients and 76 β thalassemia patients. Both groups of patients were divided into two sub-groups according to the presence or absence of cholelithiasis. The relationship between A(TA)nTAA variation of UGT1A1 gene, the serum bilirubin level and the occurrence of cholilithiasis was investigated.Our results show a significant association between genotypes carrying variant (TA)7 and hyperbilirubinemia (p0.05). Furthermore, we demonstrated a significant association between (TA)6/(TA)7 and (TA)7/(TA)7 genotypes with cholelithiasis among sickle cell anemia and thalassemia patients (p0.05).Altogether, our data provide evidence that genotypes (TA)6/(TA)7 and (TA)7/(TA)7 and (TA)7 variant present a risk factor of developing gallstone among β hemoglobinopathy Tunisian patients.

α‐Thalassemia (thal) is one of the most common genetic disorders in man. It results from defects in the α‐globin genes, and is characterized by absent or decreased rate of α‐globin synthesis in fet...

Thiamine-responsive megaloblastic anaemia (TRMA) syndrome with diabetes and deafness was found in two patients from a Tunisian kindred. The proband was homozygous for a novel mutation, 287delG, in the high-affinity thiamine transporter gene, SLC19A2. We demonstrated that fibroblasts from this patient exhibited defective thiamine transport. These data confirm that the SLC19A2 gene is the high-affinity thiamine carrier and that this novel mutation is responsible for TRMA syndrome.

We report two cases of atypical defibrination syndromes in patients with respectively acute monoblastic leukemia (chronic myeloid leukemia initially) and acute lymphoblastic leukemia. Hemostasis studies show low fibrinogen level, elevated D-dimers, decreased alpha 2 antiplasmin and factor V, normal antithrombin III values. Plasminogen is below the normal range in one patient. Soluble complexes, which are an important argument for diagnosis of intravascular coagulation disease, are not detected in both patients. Primary or secondary hyperfibrinolysis seems also excluded since euglobulin clot lysis time was normal. Enzymatic proteolysis of fibrinogen (or fibrin) by the blast cells has been reported by some authors; this mechanism could account for the hemostasis abnormalities observed in these two patients.

Les leucemies aigues lymphoblastiques (LAL) sont des hemopathies malignes se definissant par l'envahissement de la moelle par une population lymphoide immature et monoclonale. L'etude de la maladie residuelle (MRD) au cours de ces hemopathies permet d'evaluer la reponse therapeutique. L'objectif de notre travail est l'evaluation de la masse tumorale residuelle a la fin du traitement d'induction (J35). Cette etude porte sur 59 patients suivis au service d'hematologie de l'Hopital Aziza Othmana. A J35, l'estimation d'une remission complete est basee sur une etude cytologique. Elle est definie par un taux de blastes < 5 %. L'etude immunophenotypique a objective 42 cas de LAL B et 17 cas de LAL T. La MRD est < 10 -4 dans 29 % des cas seulement. Ce taux reste inferieur a celui rapporte dans les differentes series de la litterature (60-70%). L'immunophenotypage permet de quantifier les blastes non detectables en cytologie, d'evaluer la masse tumorale residuelle et d'adapter les protocoles therapeutiques. La cytometrie en flux est une technique simple, rapide, tres sensible et moins couteuse que la biologie moleculaire dans l'evaluation de la maladie residuelle et permet ainsi un suivi therapeutique rigoureux des patients.

Hypereosinophilic syndromes (HES) are a heterogeneous group of disorders characterized by marked peripheral blood and tissue eosinophilia resulting in organ damage. Recent advances in molecular biology have led to the identification of a FIP1L1-PDGFRA fusion gene as a recurrent abnormality in some patients with HES. This fusion gene results from a cryptic 4q12 interstitial deletion involving an 800 kb region. Recent reports indicate that this subtype of HES is imatinib responsive with rapid and complete haematological remissions. Here we report two patients successfully treated with imatinib.

L’approvisionnement de notre hopital en concentres globulaires se fait a partir du centre national de transfusion sanguine. Les produits sanguins sont demandes a l’avance avant la date de transfusion pour eviter un delai d’attente aux polytransfuses. L’objectif de ce travail est d’evaluer le taux de satisfaction des patients inclus dans une etude transversale de type enquete-evaluation (fevrier/mi-mars 2015). L’enquete est basee sur un questionnaire anonyme portant sur la joignablite du centre par telephone, l’ecoute, la prise en compte de la demande, l’accueil, le respect du nombre d’unites delivrees, et de la date prevue. Le degre de satisfaction est evalue a l’aide d’une cotation (tres satisfaisant [++], satisfaisant [+], peu satisfait [-], non satisfait [--], sans opinion). Cinquante-trois patients parmi 111 patients inclus dans le programme ont repondu a notre questionnaire (48 %). La moyenne d’âge est de 41 ans ; 63,5 % sont suivis pour hemoglobinopathies, 21,1 % pour syndromes myelodysplasiques et 15,4 % pour d’autres pathologies. Tous les malades recoivent des concentres de globules rouges (CGR) phenotypes, 70 % recoivent du sang filtre et 12 % du sang lave ; 87,5 % des malades rapportent une amelioration apres inclusion dans le programme. Soixante-neuf pour cent sont satisfaits de la facilite a joindre la banque du sang, 55,3 % des malades sont satisfaits du respect de la date prevue. Bien que ce programme transfusionnel ait permis d’ameliorer la qualite du service rendu, plusieurs mesures restent a entreprendre afin de paliers a certaines insuffisances.

Stroke is a devastating and potentially fatal complication to sickle cell Anemia. Strokes are difficult to explain on the basis of the central pathological process in SCA, namely the occlusion of small vessels by deformed sickled cells. We examined whether Single Nucleotide Polymorphism (SNP) variants in the MCP-1 or CCR2 genes independently or in combination are associated with occurrence of Cerebrovascular Accidents (AVC) in SCA Tunisian patients. Material and methods: 100 SCA patients among whom 19 have AVC were enrolled in this study. Clinical diagnosis of stroke was performed by the use of Transcranial Doppler ultrasonography (TCD). The genotyping of rs1026611 in the MCP-1 gene and V64I of CCR2 was performed using PCR/RFLP. Results: Our findings showed no association of the polymorphisms studied with occurrence of AVC in SCA Tunisian patients.

Immunophenotyping is a major tool for the diagnosis of the chronic lymphoid leukaemia (CLL). Its interest remains limited in the classification of the other B chronic lymphoproliferative syndromes (B-CLPS). We evaluate the place of the flow cytometry (CMF) in the diagnosis and classification of the non CLL B-CLPS. The cases with Matutes score of 4 or more are excluded. A confrontation of the results to the histology is made. 28 cases of non CLL B-CLPS are diagnosed. CMF shows a κ monoclonal population in 15 cases and λ in 13 cases. A co-expression CD19+CD5 + is found in 11 cases concording with an atypic CLL or a mantel cell lymphoma in 6 cases with Matutes score of 3. In 5 cases, we concluded to non CLL B-CLPS (Matutes

International audience; AIMS: The skeletal manifestations of sickle cell disease are the result of changes in bone and bone marrow caused by chronic tissue hypoxia that is exacerbated by episodic occlusion of the microcirculation by the abnormal sickle cells. Furthermore, the occurrence of osteonecrosis is under the control of some modifier gene. BMP6 (Bone morphogenetic protein) has been reported as associated with osteonecrosis in sickle cell anemia (SCA). Herein, we intend to study the impact of rs267196, rs267201, rs408505 and rs449853 of BMP6 gene in the occurrence of osteonecrosis among sickle cell patients in Tunisia. METHODS: Our study involved 100 SCA patients among whom 19 have osteonecrosis of the head of the femur. The latter polymorphisms of BMP6 gene were analyzed for all subjects by PCR/sequencing. To test for trait association with the candidate SNPs, genotype and allele frequencies between cases (osteonecrosis group) and controls (non-osteonecrosis group) were compared using Pearson's chi_square test with a significance threshold of P

Immunophenotyping is an essential step in the diagnosis of acute myeloid leukemia. Its prognostic value remains controversial with contradictory results.To assess prognostic impact of the immunophenotyping in AML.Our study is retrospective (October, 2005 - July, 2007) concerning 56 cases of AML (AML3 excluded) of the adult from 18 to 55 years old diagnosed and treated in Tunis Aziza Othmana Hospital. The immunophenotyping was performed by flow cytometry (Beckman Coulter EPICS XL MCL®). We studied clinical and biological characteristic, immunophenotypic expressions, and parameters of the response to the treatment: complete remission (CR), overall survival (OS), relapse free survival (RFS) and relapse in a delay of 1 year and 2 years. SPSS software was used to perform the statistical analysis.The median age of the patients is of 37,7±11.8 years. Sex ratio (M/F) is 1.33. Among individual antigenic expressions, only CD7 is associated to lower CR rates (p=0.044). We did not find any statistically significant association between immunophenotypic expressions and OS nor with relapse or RFS.The impact of immunophenotyping in AML remains controversial because of contradictory results. The research of molecular changes would be an interesting alternative in our context.

We screened 423 patients referred to our laboratory after hemolysis triggered by fava beans ingestion, neonatal jaundice or drug hemolysis. Others were asymptomatic but belonged to a family with a history of G6PD deficiency. The determination of enzymatic activity using spectrophotometric method, revealed 293 deficient (143 males and 150 females). The molecular analysis was performed by a combination of PCR-RFLP and DNA sequencing to characterize the mutations causing G6PD deficiency. 14 different genotypes have been identified : G6PD A(-) (376A>G;202G>A) (46.07%) and G6PD Med (33.10%) were the most common variants followed by G6PD Santamaria (5.80%), G6PD Kaiping (3.75%), the association [c.1311T and IVS11 93c] (3.75%), G6PD Chatham (2.04%), G6PD Aures (1.70%), G6PD A(-) Betica (0.68%), the association [ 376G;c.1311T;IVS11 93c] (0.68%), G6PD Malaga, G6PD Canton and G6PD Abeno respectively (0.34%). Two novel missense mutations were identified (c.920A>C: p.307Gln>Pro and c.968T>C: p.323 Leu>Pro). We designated these two class III variants as G6PD Tunis and G6PD Nefza. A mechanism which could account for the defective activity is discussed.

In Tunisia, the ATRA era began in 1998 with the use, consecutively, of two regimens combining ATRA and an anthracycline with cytarabine (APL93), and without cytarabine (LPA99). From 2004, 51 patients with confirmed APL either by t(15;17) or PML/RARA were treated according to the PETHEMA LPA 99 trial. Forty three patients achieved CR (86%). The remaining seven patients had early death (one died before treatment onset): four caused by differentiation syndrome (DS) and three died from central nervous system hemorrhage. Multivariate analysis revealed that female gender (P=0.045), baseline WBC> 10 G/L ( P=0.041) and serum creatinine > 1.4mg/dl ( P=0.021) were predictive of mortality during induction. DS was observed in 16 patients (32%) after a median onset time of 15 days from treatment onset (range, 2–29). Body mass index ≥ 30 (P=0.01) was the only independent predictor of DS. Occurrence of hypertensive peaks significantly predicted occurrence of DS (P=0.011) and was significantly associated with high BMI (p=0.003). With a median follow-up of 50 months, 5 year cumulative incidence of relapse, event free and overall survival were 4.7%, 74% and 78%, respectively.

Sickle cell disease is an autosomal, recessive hemoglobinopathy characterized by hemolytic anemia. Red blood cell transfusions are uncommon therapeutic mainstay in sickle cell disease and repeated transfusions can result in iron overload. The predicted risks of iron overload and organ failure increase with both the duration of disease requiring transfusion therapy and the number of transfusions.To assess the state of iron overload in patients with sickle ce anemia according to their number of transfusions.The medical records of 94 patients with sickle cell anemia (46 had homozygous sickle cell disease, 41 had sickle-ß thalassemia, 7 had compound heterozygous hemoglobin: 4 SC and 3 SOArab) were retrospectively reviewed for the following: clinical exam, serum ferritin level, liver function tests, abdominal ultrasound exam and heart Doppler.61% of our patients are from the Northern- west of the country. The average age is 18.29 years (2 to 62 years) and the sexratio is 0.62. In addition to parental consanguinity which is found in 28.72% of the cases. The average level of ferritin is 660.35 ng/ml. 41.5% of the patients have a high status of ferritin witch ranged from 521.4 to 3360 ng/ml. There is not a significant difference of ferritin level according to age, sex and a phenotype of sickle cell anemia. However, it is higher among the transfused patients with a same phenotype (p0.05). We found a correlation between serum ferritin levels and the number of transfusions (r =+0.74). Splenectomy has a preventive role because it allowed stopping the transfusion in 65% of the cases. The evaluation of organ dysfunction has found a hepatomegaly in 29% of the cases, half of witch were have a high status of serumferritin (1000 ng/ml). Left ventricular hypertrophy associated to valvulopathy was classified in 10 % of the cases.Iron overload in sickle cell anemia, though relying on transfusion, remains moderate. The repetitive assessment of serum ferritin level is considered as the best test though it does not evaluate an organic dysfunction. To evaluate them better, other tests are requiring: magnetic resonance imaging and Tc-Squid biosusceptometers.

Thalassemia intermedia empasses a mild clinical and biological spectrum. The aim is to report the clinical and biological features and treatment of this disease.It is a retrospective study about 36 thalassemia intermedia patients (17 males, 19 females). Epidemiological, haematological aspects and treatment were reportedThe diagnosis was carried out at a relatively old age 15 years (1-72).The thalassemia intermedia was characterized by mild facial deformities, splenomegaly and moderate anemia ( Hb = 9.1 g/dl). The mean serum ferritin was 518 ng/ml (25-1800). Three phenotypes are caracterised: heterozygosis beta thalassemia, beta degrees thalassemia and beta + thalassemia. Clinical complications were hypersplenism, extra medullary hematopoiesis, leg ulcers, thrombosis and pulmonary hypertension. Treatment was based on occasionally transfusion and splenectomy on event of hypersplenious (47%). Evolution of this disease was generally good with a long lifespan at 31 years (6-83).Thalassemia intermedia is well tolerated. Transfusions and splenectomy were indicated in case of hypersplenious.

In inflammatory diseases, classical parameters of iron status (serum iron, serum ferritin, total iron-binding capacity of transferrin and transferrin saturation) are not very reliable.The purpose of this study is to investigate soluble transferrin receptor, its index and classical parameters of iron status [serum iron, serum ferritin, total iron-binding capacity of transferrin and transferrin saturation] during iron-deficiency anemia and combined iron deficiency and inflammatory anemia.Our study concerned 24 patients: 18 patients with iron-deficiency anemia and 6 patients with combined iron-deficiency and inflammatory anemia. 55 healthy subjects were included as controls. Both groups underwent classical parameters of iron status [serum iron, serum ferritin, total iron-binding capacity of transferrin and transferrin saturation] and measurement of soluble transferrin receptor with its index.In iron-deficiency anemia, total iron-binding capacity of transferrin, soluble transferrin receptor and its index were enhanced, whereas serum iron, ferritinemia and transferrin saturation were low compared to controls. Compared to patients with iron-deficiency anemia, those with combined iron-deficiency and inflammatory anemia showed higher levels of serum iron and ferritinemia. In contrast, soluble transferrin receptor and its index did not vary significantly between both groups.Our findings show the interest of soluble transferrin receptor and its index in the detection of iron deficiency during anemia of inflammatory states.

Hairy cell leukemia is a rare lymphoproliferative disorder.with cytological and immunophenotypic features.We report 6 cases of hairy cell leukemia diagnosed in the Biological Department of Hematology at the Aziza Othmana Hospital of Tunis.Hairy cells was observed in blood smears of 5 cases. Flow cytometry analysis shown monoclonal a monoclonal population B while gatinting on the expression of the CD19 and SSC signal. The positivity of the CD103 is noted in 5 cases and the CD11c signal is intense in all the cases.Immunophenotype is of great interest in the diagnosis of hairy cell leukemia.

Hemoglobin O Arab is a rare abnormal hemoglobin.We report the Clinical and biological features of this disease20 patients.:16 were compound hétérozygous Hb O Arab/Béta thalassemia and 4 homozygous Hemoglobin O Arab. Patients are 7 men and 13 women.Most of them are originated from the North West of Tunisia with a age average of 39.7 years. Diagnosis was carried out at a relatively old age (26.9 years old). The homozygous form was not very symptomatic. The compound heterozygous form was more severe and characterized by a mild form of thalassemia with a moderate microcytic hypochromic anaemia (Hb = 8.8 g/dl). It was often complicated of thrombopenia due to hypersplenism in 40% of the cases. Treatment was based on occasionally transfusion and splenectomy on event of hypersplenism. Evolution of this disease was generally good with a long lifespan of patients.Haemoglobin O Arab is an abnormal hemoglobin well tolerated except for heterozygous category which requires iterative transfusions. Spelenectomy is indicated in case of hypersplenious. The evolution is generally good with a long survival.

was to provide the clinical and biological patterns hemoglobine disease in Tunisia.This retrospective study collected to 16 cases of hemoglobin C disease : 6 homozygotic Hb C and 10 heterozygotic Hb C/beta-thalassemia.The clinical profile is characterized by mild hemolytic anemia (Hb = 11.7 g/dl) associated with splenomegaly and hypersplenism. Contrary to homozygous state, the Hb C/beta-thalassemia is associated with microcytosis and pseudopolycythemia. The diagnosis is based on target cells, specific intraerythrocytic Hb C crystals in blood smear and Hb C level at 100%.The Hb C disease must be considered as a benign hemoglobinopathy which is associated with a long survival without major complications.

Acute myeloid leukemia (AML)'s diagnosis is clinical and biological. We report here 80 AML with cytology and immunophenotype features to establish correlations. 21 AML1, 23 AML2, 12 AML3, 2 AML4, 18 AML5 and 3 AML6 were diagnosed by cytology. Only one case of AML0 was diagnosed by immunophenotype. Myelogysplasia is present in 29.8% cases. CD19 and CD56 expression was significantly associated to AML +t(8;21). Additionally, concomitant negativity of CD34 and HLA-DR was discrimininatif to AML3 diagnosis. Prognostic value to expression some CD needs time backwards.

La transfusion erythrocytaire est une therapeutique substitutive souvent indiquee en milieu hematologique. Elle expose a des complications notamment immunologiques. Nous etudions dans ce travail les resultats de recherche d’anticorps irreguliers (RAI) pratiques dans notre service. Il s’agit d’une etude retrospective (janvier 2007–mars 2014). La RAI associe la technique de Coombs indirect a basse force ionique et la technique enzymatique sur cartes gel (Biorad®). Au total, 3672 RAI sont realisees ; 5,9 % sont positives correspondant a 95 patients. Les anticorps sont de specificites RH (1, 2, 3, 4, 5) KEL1 dans 48,42 % des cas, reconnaissant au moins un antigene des autres systemes dans 51,58 % des cas. Une association d’anticorps de specificites RH (1, 2, 3, 4, 5) KEL1 et autres specificites est retrouvee dans 24,21 % des cas. Les patients presentant une RAI positive sont suivis pour syndrome drepanocytaire majeur dans 61 % des cas et syndrome myelodysplasique dans 16,5 % des cas. Les specificites des anticorps les plus frequentes dans les systemes autres que RH KEL1 sont : FY1 (27,1 %), MNS3 (18,6 %), MNS1 (18,6 %) et JK1 (16,9 %). L’anti- RH3 et l’anti-KEL1 sont les plus frequemment associes a ces anticorps. Les anti-FY1 et anti-MNS1 sont le plus souvent identifies en association a d’autres allo-anticorps (p = 0,001 ; p = 0,016 respectivement). L’apparition d’anticorps anti-erythrocytaires en milieu hematologique alourdit la recherche de globules rouges compatibles surtout en presence d’associations. Une adaptation des protocoles transfusionnels chez les patients a risque devrait tenir compte du cout et de la disponibilite du produit.

Les medecins anesthesistes-reanimateurs (MAR) representent des prescripteurs reguliers des transfusions sanguines. Le but de ce travail est d’evaluer les pratiques transfusionnelles des MAR exercant dans le secteur prive. Il s’agit d’une etude d’evaluation qui a cible toutes les polycliniques du Grand Tunis (23 cliniques). Le moyen d’evaluation est un questionnaire anonyme de 10 questions a choix multiples (QCM) de 5 items chacune. Nous calculons la note moyenne par copie, et le taux de bonne reponse (TBR) par QCM et par item. Certains items sont classes « dangereux ». Il s’agit d’items concernant l’incompatibilite ABO et la pratique du test ultime au lit du malade. Parmi 96 MAR, 75 % ont accepte de participer a l’etude. Soixante-huit pour cent ont deja exerce dans le secteur public. L’anciennete moyenne d’exercice est de 9,4 ans (0,5 a 26 ans). Le nombre moyen d’annees d’experience est de 17 ans (1 a 43 ans). Aucun participant n’a repondu correctement a toutes les questions. Dix-huit pour cent ont repondu correctement a 5 QCM ou plus. La note moyenne est de 3,1/10 (0 a 9). Le TBR par question varie de 4,2 % a 87,2 %. Il varie de 21,7 % a 100 % pour les items. Le TBR aux items classes dangereux varie de 29 % a 100 %. La carriere hospitaliere anterieure influence positivement les TBR avec une difference significative. Une anciennete d’exercice de moins de 7 ans est statistiquement associee a de meilleurs TBR. Ce travail a revele des lacunes dans les pratiques transfusionnelles. Un programme de formation continue doit etre entrepris afin de pallier a ces insuffisances.

L’immunisation anti-erythrocytaire dans les syndromes drepanocytaires majeurs (SDM) alourdit la prise en charge transfusionnelle. L’objectif de l’etude est de determiner le taux d’immunisation anti-erythrocytaire dans les SDM et d’analyser les facteurs pouvant influencer son apparition. Il s’agit d’une etude retrospective (janvier 2013–juillet 2014) concernant 169 SDM. La recherche d’anticorps irreguliers associe : le test de Coombs indirect et la technique enzymatique sur cartes gel (Biorad ® ). Le test de Coombs direct (TCD) est realise sur carte gel (Biorad ® ) a distance des transfusions. Les concentres de globules rouges (CGR) transfuses sont compatibilises phenotypes. Un phenotypage elargi est indique en cas d’immunisation. Les patients sont repartis en : 96 cas de drepanocytose homozygote, 65 cas de S-β thalassemie et 8 cas d’heterozygote composite. L’âge median etait de 32 ans [5–70 ans]. Le sex-ratio M/F etait de 0,6. Une transfusion etait indiquee chez 77 % des patients. L’âge median a la premiere transfusion etait de 11 ans [1–40 ans]. Le nombre de CGR total transfuses etait de 2245 (une moyenne de 18,5/patient). Au moins, un allo-anticorps etait identifie chez 18,5 % des patients transfuses. Une allo-immunisation multiple etait notee dans la moitie des cas. Le TCD est positif chez 6,5 % de patients tous transfuses (IgG : 63,6 % des cas ; C3d et IgG+ C3d dans 18,2 % des cas respectivement). L’augmentation du risque d’immunisation est significativement associee aux besoins transfusionnels eleves. Le taux d’immunisation est eleve dans notre serie. Le phenotypage elargi des le diagnostic est recommande mais il reste confronte au cout.

The present work focuses on the therapeutic efficacy and the toxicity of alpha interferon in patients younger than age 18 years. 5 patients younger than 18 years were treated and followed up between 1990 and 1999 at the department of haematology (Aziza Othmana Hospital) Hydroxyurea was given as initial treatment to all patients. After a median period of 8 months, these patients received alpha interferon (5 millions units/m2 once). Six months after the beginning of the alpha interferon a complete hematologic response was obtained in all patients. The median overall survival was of 66 months: 3 patients are still alive (2 patients in an advanced stage and one patient in chronic phase) and 2 patients died after transformation. The most common reported side effects of alpha interferon were asthenia, weight loss, fever, myalgia, chills and headaches--these toxic manifestations were mild and were noticed in all our patients. Myelosuppression was noted in two patients. Interferon is well tolerated in patients younger than age years 18 old, with CML. It may offer an alternative to bone marrow transplantation in children in the chronic phase of CML without histocompatible donor. The role of new agents such as STI 571 needs to be evaluated as well.

Through a national retrospective study, the authors report the clinical and hematological characteristics of 124 acute lymphoblastic leukemia of the adult diagnosed during 5 years (1993-1997). The national prevalence is of 0.28/100.000 inhabitants/year. The sex-ratio is of 1.3. Sixty six per cent of patients were 16-35 years of age, and only 10% of them were more than 60 years of age. A tumoral syndrome was present at 71% of the cases with peripheral adenopathies in 55%, splenomegaly in 40%, hepatomegaly in 19% and a mediastinal tumor in 18% of the cases. The bone pain were rarely signaled (10%) and neuro-meningeal affection was found in only 3% of cases. There was no testicular lesions. The white blood cells count was less than 30.000/mm3 in 60% whereas an important hyperleucocytosis superior than 100.103/mm3 was observed in 20% of the cases. Anemia and thrombopenia were noted in 94% and 90% of the cases respectively. Acute lymphoblastic leukemia typing by cytological study of Bone marrow according to the Fransh-American-Britain criteria (FAB) had found 43%, 48% and 4% for type 1,2 and 3 respectively. In 5% of the cases the type of the acute lymphoblastic leukemia was not precised (diagnosis based on the Bone biopsy).

Background: Hemophilia is a hereditary bleeding disorder characterized by a deficiency of factor VIII (hemophilia A) or factor IX (hemophilia B). It is established that hemophilia is a ubiquitous condition that affects one birth in 10,000, regardless of race or geographic region. We propose in this work to assess the incidence of hemophilia in the North of Tunisia with a literature review. The hemophilia treatment center at the hospital Aziza Othmana supports patients in northern Tunisia, which represents 60% of all tunisians cases. The estimated number of inhabitants is 5.19 million (50% of the country population). Methods: Assays of factors VIII and IX are performed by STA Compact, Stago®. Confirmation of the diagnosis ismade on two different samples. Results: Twenty cases of hemophilia were diagnosed between January 2010 and December 2011 in our center. The average of age was 3.37 years (2 months-28 years). The most frequent circumstances of diagnosis are hematomas and hemarthrosis following minor trauma (12 cases). Family history of hemophilia is found in 6 cases. For all patients, an isolated prolonged APTT with a correction to the mixture test was found. We note 15 cases of hemophilia A and 5 cases of hemophilia B. The distribution of cases according to severity is: severe (9 cases), moderate (8 cases) and minor (3 cases). Comment: The incidence of hemophilia on the north of Tunisia is 1.1cases per 10 000 births per year. The incidence found in our series corresponds to the incidence reported in the literature. However, the prevalence of hemophilia diagnosed in Tunisia remains low only 50% of the theoretical number of hemophilia is diagnosed.