Routine diagnostic procedures of myelodysplastic syndromes: value of a structural blood cell parameter (NEUT-X) determined by the Sysmex XE-2100™.

INTRODUCTION: Diagnostic features of myelodysplastic syndromes (MDS) are often polymorphic and nonspecific including anemia in most cases. Standard parameters provided by an automated analyzer seldom bring any argument for this diagnosis. The aim of this study was to investigate whether some structural parameters, not routinely provided by Sysmex™ XE 2100 analyzer, could help diagnose MDS in a simple way, adapted to routine practice. METHODS: Blood samples from 184 MDS fully annotated cases and 3545 normal blood count controls were performed with XE 2100 Sysmex™ analyzer. Quantitative and structural parameters were considered. RESULTS: We found that the structural neutrophil parameter, NEUT-X, converted into a semi-quantitative parameter, the granularity index (GI), could be used as a flag for MDS in front of anemia. Negative GI and anemia were able to make otherwise unrecognized MDS stand out in routine practice, increasing the number of slides addressed to review from 67% to 96%, without leading to a large excess of unfounded slide review among non-MDS. CONCLUSION: Including the GI index in the routine parameters provided by the Sysmex analyzer could be of major help for nonspecialized routine laboratories in detecting MDS.

Recurrent thromboembolism in a patient with beta-thalassemia major associated with double heterozygosity for factor V R506Q and prothrombin G20210A mutations.

Double heterozygosity for factor V R506Q and prothrombin G20210A mutations was identified in a 24-year-old man with beta-thalassemia major. The patient experienced a first thrombotic event at the age of 19 years and three recurrent thromboses in a short time interval, the third occurring while the patient was receiving long-term anticoagulant treatment. This case suggests that patients with major thalassemia and congenital thrombophilic mutations need intensive and long-lasting anticoagulant treatment. Thus, even if thrombotic events could be explained by a hypercoagulable state observed in patients with major thalassemia, after a first thrombotic event has occurred these patients should be screened for acquired and congenital thrombophilia.

Fibrinogen Saint-Germain I: a case of the heterozygous Aalpha GLY 12 --> VAL fibrinogen variant.

A fibrinogen variant was suspected based on the results of routine coagulation tests in a 2-year-old asymptomatic child. Coagulation studies showed marked prolongation of both the thrombin and reptilase times, and discrepancy was noted between the level of plasma fibrinogen as measured by a kinetic versus immunological determination. Family studies revealed that the father beared the same abnormality. Studies of purified fibrinogen revealed an impaired release of both fibrinopeptides by thrombin. Fibrin monomer polymerization and fibrin stabilization were normal. DNA sequencing revealed a heterozygous G --> T point mutation in exon 2 of the gene coding for the Aalpha chain, which substituted a Gly for Val at position 12. Although the mutation is the same as in fibrinogen Rouen, fibrinogen Saint-Germain I shows a different fibrinopeptide release pattern and a mild factor V deficiency.

Three new cases of dysfibrinogenemia: Poissy III, Saint-Germain I and Tahiti.

In order to identify unknown mutations, the FAMA method was used to rapidly screen the fibrinogen chain genes in individuals with dysfibrinogenemias. Chemical cleavage at mismatches on heteroduplexes DNA end-labeled with strand-specific fluorescent dyes reliably detects sequence changes in DNA fragments of up to 1.5 kb and locates them precisely. This method was successfully used for the detection of three new dysfibrinogenemias: Poissy III, Tahiti (heterozygous Aalpha Arg16His) and Saint-Germain I (heterozygous AalphaGly12Val). The mutations were confirmed by dideoxy sequencing.

Lymph node infection by Trichomonas tenax: report of a case with co-infection by Mycobacterium tuberculosis.

In an 82-year-old woman, presenting with fever and asthenia, cervical adenopathy was noted. Clinical and radiological investigations were fruitless. Laboratory examinations detected a refractory anemia. The lymph node was excised and showed numerous trichomonads on touch preparations. Histologically, the node showed caseous necrosis and macrophagic reaction. Diagnosis of lymph node infection by Trichomonas tenax was made. Three weeks later, culture of the node showed Mycobacterium tuberculosis and let us conclude co-infection. T tenax is usually regarded as a harmless saprophyte of the oral cavity. This exceptional observation shows for the first time an invasive potential of T tenax. It raises questions about links with tuberculosis and refractory anemia.

[Activated C protein resistance manifested by cutaneous necrosis after interferon alpha injection: case report]. / Nécrose cutanée après injection d'interféron alpha révélant une résistance à la protéine C activée: rapport d'un cas.

INTRODUCTION: Cutaneous necrosis occurring in the course of treatment by alpha interferon is an uncommon side-effect. Its physiopathologic mechanism remains obscure. A local thrombotic action of interferon has been suggested to explain its occurrence. EXEGESIS: A 64-year-old male patient with human immunodeficiency virus-related cutaneous Kaposi's sarcoma presented cutaneous necrosis after a 9-month treatment by interferon alpha, while his resistance to activated protein C had already been demonstrated. To our knowledge, this is the first case ever described regarding the association of interferon-induced cutaneous necrosis with activated protein C resistance. CONCLUSION: This suggests that in case of interferon treatment-induced cutaneous necrosis coagulation disorders should be investigated and questions the existence of a particular "pro-coagulant profile" facilitating this side effect.

[Pre-leukemic granulocytic sarcoma of the uterus. Report of a case]. / Sarcome granulocytaire pré-leucémique de l'utérus. A propos d'un cas.

A case of pre-leukemic granulocytic sarcoma (GS) of the uterus was found in a 73-year old woman. The diagnosis was suggested by vaginal cytology and the green color of the gross lesions, then confirmed by naphthol AS-D chloro acetate esterase stain and immunohistochemistry on fixed tissue with the anti-lysozyme, anti-myeloperoxidase, CD 43 and CD15 antibodies. At the time of GS discovery, the patient presented no evident leukemia but myelogram contained 20% of blast cells. She developed acute myeloid leukemia 2 months later. Cytogenetic study of the bone marrow revealed chromosome 21 trisomy. GS is frequently mistaken for malignant lymphoma since it expresses some of the leukocytic antigens (leukocyte common antigen, CD 45 RO (UCHL1), MB2). Therefore, the use of a large panel of antibodies, including anti-myeloperoxidase, anti-lysozyme and CD15, is recommended. Precise diagnosis is essential because all GS must be treated as acute myeloid leukemias.

Variation in fetal hemoglobin parameters and predicted hemoglobin S polymerization in sickle cell children in the first two years of life: Parisian Prospective Study on Sickle Cell Disease.

Intracellular hemoglobin S (HbS) polymerization is most likely to be the primary determinant of the clinical and biologic manifestations of sickle cell disease (SCD). Fetal hemoglobin (HbF) does not enter the HbS polymer and its intracellular expression in sickle erythrocytes inhibits polymerization. HbF levels, high at birth but decreasing thereafter, protect the newborn from the clinical manifestations of this hemoglobinopathy. We have measured the sequential changes in HbF, F reticulocytes, and F cells in the first 2 years of life in 25 children with SCD and compared the results with those obtained in 30 normal children (AA). We have also calculated HbF per F cell (F/F cell), the preferential survival of F cells versus non-F cells, as measured by the ratio F cells versus F reticulocytes (FC/FR) and polymer tendency at 40% and 70% oxygen saturation. HbF levels decreased from about 80.4% +/- 4.0% at birth to 9.2% +/- 2.9% at 24 months. During this time, we observed a regular decrease of the F reticulocytes and the F cells. The kinetics of the decline of F/F cell was comparable with the decline of HbF, rapid from birth (mean, 27.0 +/- 3.6 pg) to 12 months of age (mean, 8.5 +/- 1.5 pg) and then slower from 12 to 24 months of age (mean, 6.2 +/- 1.0 pg) in the SCD children. In the AA children, the decrease in HbF, due to changes in both numbers of F cells and F/F cell, was more precipitous, reaching steady-state levels by 10 months of age. Calculated values for mean polymer tendency in the F-cell population showed that polymerization should begin to occur at 40% oxygen saturation at about 3 months and increase progressively with age, whereas polymerization at 70% oxygen saturation would not occur until about 24 months. These values correspond to HbF levels of 50.8% +/- 10.8% and 9.2% +/- 2.9%, respectively, and F/F cell levels of 15.6 +/- 4.5 pg and 6.2 +/- 1.0 pg, respectively. In the non--F-cell population, polymerization was expected at birth at both oxygen saturation values. Three individuals had significantly greater predicted polymerization tendency than the remainder of the group because of early decreases in HbF. These individuals in particular, the remainder of the cohort, as well as other recruited newborns, will be studied prospectively to ascertain the relationship among hematologic parameters, which determine polymerization tendency and the various clinical manifestations of SCD.

[Alpha-thalassemia]. / Les alpha-thalassémies.

alpha-Thalassaemias are probably the most common genetic disorder worldwide. alpha-Thalassaemias are haemolytic anaemias resulting from inherited deficient synthesis of alpha-globin chains. In this paper, the classification and nomenclature of alpha-thalassaemias are developed. Procedures to ensure the laboratory diagnosis are explained. The heterozygous carrier states for these disorders are, in most cases, not associated with any easily discernible change in the haemoglobin pattern, except, sometimes, a reduced MCV in the blood picture. Heterozygotes for alpha-thalassaemia deletions are now detectable by the accurate PCR method. Because of the high prevalence of these disorders in large segments of the world population, alpha-thalassemia and haemoglobinopathies often occur in the same individual. The laboratory features of these interactions and, particularly, the role of alpha-thalassaemia as a potential modulator of sickling haemoglobinopathies are discussed.

Donor for BMT with haemoglobin H disease.

We report a successful allogeneic BMT for the treatment of juvenile chronic myelogenous leukemia (JCML) in a 9-month-old Laotian boy using an HLA-matched sibling donor with HbH disease (--SEA/aaCS). In addition, before BMT the recipient had a complex haemoglobinopathy associating heterozygous state AE along with HbH disease (--SEA/-a3,7) without haemoglobin Constant Spring (HbCS). Because various haemoglobinopathies are frequently encountered in southeast Asia, when BMT is performed in Asian families the results may be evaluated by the differing haemoglobin characteristics of recipient and donor. However, there is also a significant risk of transmitting a new haemoglobinopathy to the recipient. Because transplantation from HLA-identical siblings offers the only chance of cure for JCML, the presence of HbH disease with mild clinical expression in the donor should not be taken as a contra-indication to BMT.

Frequency and prognostic importance of thrombocytopenia in symptom-free HIV-infected individuals: a 5-year prospective study.

The exact frequency of HIV-associated thrombocytopenia (TCP), defined as platelet count less than 150 x 10(9)/1, was studied in 435 symptom-free HIV-seropositive individuals. At the baseline control, 23 (5.5%) had TCP. TCP individuals had a significantly lower mean CD4 lymphocyte count than the non-TCP individuals. During a mean follow-up of 30 months, 79 out of the 435 individuals (18%) had TCP at least once. During the study period, only 1% of our patients had a platelet count less than 50 x 10(9)/l. TCP was more frequent in intravenous drug users than in other risk groups. A spontaneous normalization of platelet count was observed in more than 50% of TCP individuals.