RORC overexpression as a sign of Th17 lymphocytes accumulation in multiple myeloma bone marrow.

The role of the bone marrow microenvironment in supporting the proliferation and survival of the abnormal plasma cells in multiple myeloma (MM) is well established. Such microenvironment is rich of cytokines like IL-6, TGF-ß, IL-1 and IL-23 which are known to promote the differentiation of Th17 lymphocytes, a T helper subpopulation. Th17 cells secrete IL-17, a cytokine involved in the pathophysiology of several auto-immune diseases. Yet, its involvement in cancers remains unclear. Herein, we aimed to try to understand the role of Th17 lymphocytes in multiple myeloma. Bone marrow samples were prospectively collected from 29 MM patients and 23 healthy bone marrow donors for allograft. Mononuclear bone marrow cells were isolated by Ficoll-Hypaque gradient and CD138+ plasma cells were depleted using magnetic beads. The quantification of Th17 cells was performed by flow cytometry in the CD138 negative cells. The mRNA expression of IL17 and RORc was quantified using real time PCR in the same subset. The mRNA expression of IL17R was analyzed in plasma cells (CD138+ cells). Data obtained from patients and healthy donors were compared by both non-parametric Mann-Whitney U test and Spearman test. A significant increase of IL17 and RORC mRNA expression was found in the bone marrow microenvironment of MM patients compared to healthy donors. Th17 cells were also increased in the bone marrow of MM patients compared to healthy donors. Interestingly, the mRNA expression of IL17R was significantly decreased in MM patients. Yet, no correlation was found between the gene expression IL17, RORC and IL17R and the bone marrow infiltration or the stage of the disease. Collectively, our results suggest the involvement of Th17 cells in the pathophysiology of MM. Such data further support the use of anti-IL-17 antibodies as a therapeutic approach in MM.

Chronic myeloid leukemia patients in Tunisia: epidemiology and outcome in the imatinib era (a multicentric experience).

Data are limited in developing countries regarding the clinicopathologic features and response to therapy of chronic myeloid leukemia (CML) in the era of imatinib (IM). The objective of this study is to report on the clinicoepidemiologic features of CML in Tunisia, to evaluate the long-term outcome of patients in chronic (CP) or accelerated phase (AP) treated with IM 400 mg daily as frontline therapy, and to determine imatinib's efficacy and safety. From October 2002 to December 2014, 410 CML patients were treated with IM in six Tunisian departments of hematology. Response (hematologic, cytogenetic, and molecular responses) and outcome-overall survival (OS), event-free survival (EFS), and progression-free survival (PFS)-were evaluated. The following prognostic factors were analyzed for their impact on the European leukemia net (ELN) response, OS, EFS, and PFS at 5 years: age, sex, leukocyte count, Sokal score, European Treatment and Outcome Study (EUTOS) score, CML phase, time to starting IM, and impact of adverse events. The median age was 45 years (3-85 years). Two hundred ten (51.2%) patients were male. Splenomegaly was present in 322 of the 410 (79%). Additional cytogenetic abnormalities were encountered in 25 (6.3%) patients. At diagnosis, 379 (92.4%) patients were in CP, 31 (7.6%) were in AP. The Sokal risk was low in 87 (22.5%), intermediate in 138 (35.7%), and high in 164 patients (41.9%). The EUTOS risk was low in 217 (74%), and high in 77 (26%) patients. The rates of cumulative complete cytogenetic response (CCyR), major molecular response (MMR), and molecular response 4/5 log (MR4.5) in CP/AP-CML patients were 72, 68.4, and 46.4%, respectively. The median time to reach CCyR, MMR, and MR4.5 was 6 months (3-51), 18 months (3-72), and 24 months (3-100), respectively. According to the ELN criteria, optimal, suboptimal response, and failure were noted in 206 (51.8%), 61 (15.3%), and 125 (31.4%) patients, respectively. Five-year event-free survival (EFS), progression-free survival (PFS), and overall survival (OS) were 81, 90, and 90%, respectively. By multivariate analysis, AP, high EUTOS risk, and baseline WBC ≥ 150G/l remained independent predictive factors of non-optimal response to IM. The adverse events (AE) of IM were moderate and tolerable. With the caveats that the monitoring of the disease was not optimal, response rates were similar to those reported in previous studies. It is clear to us that improvements should be made in treatment of AP-CML and high Sokal risk group of CP-CML. The frontline use of second-generation tyrosine kinase inhibitor (TKI) is expected to improve the results of the first-line treatment of these high-risk Tunisian patients, but cost and accessibility of this therapy remain the problems in developing countries.

Inflammations mediators and circulating levels of matrix metalloproteinases: Biomarkers of diabetes in Tunisians metabolic syndrome patients.

AIMS: This study investigates the relationships between matrix metalloproteinases, inflammations mediators and type 2 diabetes mellitus in Tunisians metabolic syndrome (Mets) patients. METHODS: The study has included 239 MetS patients and 247 controls. Mets was defined according to the NCEP-ATPIII report. Mets patients were also divided into two categories: 29 MetS non-diabetics and 210 MetS diabetics. Dysglycemia markers, matrix metalloproteinase-9 (MMP-9), Tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2), tumor necrosis factor α (TNF-α), C-reactive protein (CRP) levels and White Blood Cells (WBC) counts were determined in patients and controls. RESULTS: In our study, the level of inflammatory markers WBC, TNF-α and matrix metalloproteinases (MMP-8 and MMP-9) were significantly higher in diabetic patients with MetS, as compared with non-diabetic MetS patients. Inflammation mediators and MMP-9 were significantly associated with many clinical characteristics of MetS. The use of ROC "Receiver Operating Characteristic" analysis revealed the impact of TNF alpha on diabetes patients with MetS. In fact TNF alpha was found as a sensitive parameter in these patients with a sensitivity of 85%. CONCLUSION: Inflammation, matrix metalloproteinases and dysglycemia markers are not expressed in isolation but rather concurrently and are continuously interacting with each other, in MetS and diabetics patients. These markers fit with an early stage of cardiovascular disease (CVD); and measuring them could improve the risk evaluation, an early diagnosis, and the prognosis of CVD.

Causes of vitamin B12 deficiency.

BACKGROUND: B12 Vitamin deficiency is common in adults (20% of general population of industrialized populations), especially in elderly patients (30-40%). The etiologies of Vitamin B12 deficiency have been dominated by the cobalamin syndrome nutrient and the Biermer disease, rarely by the intake or nutritional deficiency and bad absorptions. STUDY OBJECTIVE: Establish an etiology of vitamin B12 in a Tunisian population Methods: In a prospective study involving 100 patients with macrocytic anemia, a comprehensive assessment has been carried out of: B12 vitamin and folate intake, homocysteine, immunological assessment (antibodies, intrinsic anti-factor and anti-gastric parietal cells), an endoscopic exploration, and a dietary nutritional survey. RESULTS: The mean age of patients was 53.6 ± 17,6 years (13 - 88 years), the gender ratio (female/male) is 1.22. The clinical symptomatology shows a functional anemia syndrome in 89% of cases, a digestive syndrome in 88% of cases, and neurological disorders in 67% of cases. The intake of B12 vitamin was reduced (<180 pg/ml) in 99 patients, associated with a hyperhomocysteinemia in 81.63% of cases.The intrinsic anti-factor antibodies were positives in 32 patients, and the antibody anti-gastric parietal cells in 85 patients. Gastric biopsy was performed in 54 patients, showing a chronic atrophic gastritis of fundic localization in 44 patients, antral in 5 patients and pan- gastric in 3 patients. The diagnosis of Biermer anemia was held in 75% of patients, that of FCS in 16% of patients, and a lack of intake in 8% of patients. The etiology was undetermined in 1% of cases. CONCLUSION: Vitamin B12 deficiency is common in the general population, its causes and origins are multiple, we list them in order of occurrence: Biermer disease, the FCS, and the intake deficiency in our population.

Extracorporeal lithotripsy in patients with hemophilia: systematic review.

BACKGROUND: The management of urolithiasis in patients with haemophilia poses a real challenge to the urologist. AIM: We conducted a systematic literature review to assess the safety and efficacy of extracorporeal shock wave lithotripsy (ESWL) in the treatment of urolithiasis in hemophiliacs. METHODS: A systematic review was conducted by using the National Library of Medicine (PubMed) search engine between January 1985 and June 2013. We've used these key words: "haemophilia" and "extracorporeal shock wave lithotripsy". All articles dealing with the treatment of nephrolithiasis by ESWL in patients with hemophilia were included. Two independent reviewers extracted the data from each article. The data was included into a systematic review and analyzed. RESULTS: A total of 12 medical articles were selected with a total of 25 patients. The stone size varies from 6 to 21 mm. The substitution of the deficient clotting factor started the day before the ESWL. ESWL was effective in all patients except one after 1-6 sessions / patient. An ultrasound was performed after the procedure to look for potential bleeding complications. The judgment of the substitution therapy depends on the patient's condition, the presence of hematuria and the absence of signs of bleeding. Major bleeding complications were observed in 4 patients. CONCLUSIONS: With effective substitution of deficient clotting factors, ESWL is a safe and low morbidity method in the treatment of urinary calculi in hemophiliacs.

[The JAK2 mutation in myeloproliferative disorders: A predictive factor of thrombosis]. / La mutation JAK2 dans les syndromes myéloprolifératifs BCR-ABL négatifs: facteur prédictif de thrombose.

BACKGROUND: BCR-ABL negative myeloproliferative neoplasms (MPN) include polycythemia Vera (PV), essential thrombocythemia (ET) and primitive myelofibrosis (PMF). the JAK2 V617F mutation has been introduced since 2008 as a major diagnostic criterion on the one hand and on the other hand, it would be linked to increased risk of thrombotic complications. AIM: This study aimed to evaluate the association of JAK2 mutation and thrombotic events in MPN. METHODS: A retrospective study concerning 45 BCR-ABL negative MPN patients (mean age=53 old years, sex ratio=0.8) was conducted. RESULTS: They were classified as PV (22 patients), ET (17 patients), PMF (3 patients) and atypical MPN (3 patients). The JAK2 mutation was found in 64.4% of patients: 72.7% of PV patients, 47% of ET patients and 66.7% of PMF patients. Thrombotic events were recorded in 11 patients (24.4%). Cerebral arteries and portal vein were the most frequent localizations. The JAK2 mutation was an independent risk factor of thrombotic events. CONCLUSION: Consequently, it seems that screening for JAK2 mutation in BCR-ABL negative MPN could play a role in identifying patients at high risk of vascular complications.

[Adult lymphocytosis etiology by flow cytometry]. / Classification par cryométrie en flux de 104 cas d'hyperlymphocytose de l'adulte.

BACKGROUND: Positive and differential diagnosis of chronic lymphocytic leukemia (CLL) is based on immunophenotyping analysis. CLL is searched whenever a persistent lymphocytosis is found. AIM: To evaluate the performance of flow cytometry in etiologic diagnosis of lymphocytosis. Could it allow us to distinguish CLL from other causes of lymphocytosis? METHODS: Blood samples from 104 adult patients having a rate of lymphocytes> 5000 élé/mm3 persisting more than three months were analyzed using a large panel of monoclonal antibodies in three colors and Cell Quest software. results: Lymphoproliferative B disorder was retained in 83 cases, including 50 cases of typical CLL with Matutes score≥ 4 and 12 cases of atypical CLL with Matutes score = 3 . Diagnosis of hairy cell leukemia and follicular lymphoma were guided by the respective specific antigen expression CD103 and CD10. Large granular T lymphoma (LGL-T) was the most common etiology of lymphoid T proliferation. Unusual cases of Natural Killer (NK) and NK/T proliferations were found. CONCLUSION: The Flow cytometry is a powerful tool to establish lymphocytosis etiological diagnosis; it avoids invasive investigations in a large number of cases.

[Transfusion safety: state of the arts (ART)]. / Sécurité transfusionnelle: etat des lieux.

BACKGROUND: Blood transfusion is a high risk activity. AIM: To evaluate transfusion safety in planned cardiac surgery. METHODS: This study was conducted in the blood bank of the Rabta Hospital in two phases: a phase to observe transfusion acts followed by corrective actions and a phase to evaluate the impacts of these corrections on the transfusion practices. Characteristics of the potentially transfused patients, the eventually prescribed, dispensed and transfused blood products and transfusion practices were studied. RESULTS: During the observation phase, 70 patients were enrolled, 51 potentially transfused. Weaknesses concerned the mention of phenotype and transfusion history when ordering blood components as well as the double ABO/D group typing, the phenotype and the cross match performing. Final bedside controls were done in a wrong way. The distribution and the blood administration were established respectively for 208 and 232 blood products. The traceability was established for 86 blood products. During the evaluation phase, 30 patients were enrolled, 15 potentially transfused. Improvement was achieved in the transfusion history notification, phenotype and antibodies screen performing and cross matching. CONCLUSION: Optimisation of blood transfusion can be conceived only with collaboration between the different transfusion structures.

[Protein S, C and antithrombin deficiency: association with myocardial infarction and thromboembolism in the young]. / Déficits en protéine S, C et antithrombine: Association à l'infarctus du myocarde et aux évènements thrombo-emboliques chez le sujet jeune.

BACKGROUND: The pathogenesis of myocardial infarction (MI) in young involves new factors including constitutional or acquired thrombophilia. AIM: To determine in patients ² 50 years, the association between coagulation factors deficiency, myocardial infarction and cardiovascular events during follow-up. METHODS: Protein C (PC), PS and antithrombin (AT) were screened in 50 patients admitted for acute MI and in a healthy control group. Univariate and multivariate analysis were performed using SPSS 11.5 version. RESULTS: PS and PC deficiency were associated to MI (respectively 24% vs 0%, p=0.001 and 14% vs 0%, p=0.016), independently for PC. No AT deficiency was detected in both groups. During followup, PS and C deficiency were predictive for venous thrombosis (p<0.05) and PS deficiency for pulmonary embolism. CONCLUSION: Protein C and S deficiency may play an important role in MI in young and also in thromboembolic complications during follow-up. Nevertheless, therapeutic implications remain controversial.

[Erythrocyte abnormalities in thyroid dysfunction]. / Les anomalies ertyrocytaires au cours des dysthyroïdies.

BACKGROUND: Erythrocyte abnormalities are frequently associated with thyroid dysfunction. However, they are rarely investigated and related to the thyroid. AIM: This study was aimed to determine the nature and frequency of erythrocyte abnormalities in thyroid disease and look for their evolution after thyroid function restoration. METHODS: This retrospective study included 412 patients with peripheral thyroid disease; hyperthyroidism (n=235) or hypothyroidism (n=177). Hyperthyroidism was considered for TSH<0.10 ÌUI/ml and hypothyroidism for TSH>5.0 ÌUI/ml. Anemia was defined by hemoglobin level<13 g/dl in men and <12 g/dl in women, microcytosis by mean corpuscular volume (MCV)<80 fl, macrocytosis by MCV>98 fl, and hypochromia by mean corpuscular hemoglobin (MCH)<25 pg. Restoration of euthyroid state was considered in patients with normal TSH levels for at least 3 months. RESULTS: Anaemia was observed in 40.9% of patients with hyperthyroidism and 57.1% of patients with hypothyroidism. Among these, normocytic or macrocytic anaemia was present in 46.3% of cases. Whereas, microcytosis, with or without anaemia, was noted in 87.7% of patients with hyperthyroidism. FT4 was positively correlated with the number of red blood cells and haemoglobin, and inversely correlated with MCV and MCH. After restoration of euthyroid state, most erythrocyte abnormalities were corrected. CONCLUSION: Thyroid diseases are frequently associated with erythrocyte abnormalities, including normocytic anaemia in hypothyroidism and microcytosis in hyperthyroidism. These abnormalities should be investigated and corrected. Their presence could steer towards subclinical thyroid dysfunction, allowing its early management.

The hypercoagulable state in multiple myeloma: The contribution of thrombin generation test.

BACKGROUND: Multiple myeloma is a hematologic malignancy which confers a high venous thromboembolic risk. This risk is linked to patient-related factors, disease-specific mechanisms, and antimyeloma therapy, especially immunomodulatory drugs. Some studies have suggested that the thrombin generation assay may be a predictive marker of thrombosis. This study aimed to assess the hypercoagulable state in patients with multiple myeloma at diagnosis and after myeloma therapy. METHODS: Thirty-one patients with multiple myeloma were included in a prospective study and were compared with 31 matched controls with age and gender. Thrombin generation assay was performed in patients at diagnosis prior to treatment initiation and at the end of myeloma therapy, and in controls. Parameters of lag time, peak thrombin concentration, time to peak, endogenous thrombin potential, and velocity index were analyzed. RESULTS: Median age of patients at diagnosis was 58 years (11 men and 20 women). Twenty-three patients (74%) were classified as high vascular risk and received thromboprophylaxis. No thromboembolic events have been reported during follow-up, except a symptomatic pulmonary embolism in one patient which occurred at diagnosis. At baseline, patients with myeloma had significantly elevated velocity index as compared to controls (178 vs 128 nmol/L/min; P = .013). High-risk patients showed an elevation of plasma thrombin generation as compared to low-risk patients (endogenous thrombin potential = 1244 vs 1052 nmol/L/min; P = .043). Myeloma therapy did not significantly change the thrombin generation parameters. CONCLUSION: Thrombin generation appears to be higher in patients with myeloma compared with controls, especially in high-risk patients, and does not change significantly after treatment completion.

Heparin-like anticoagulant associated with multiple myeloma and neutralized with protamine sulfate.

A 55-year-old man with multiple myeloma developed sustained bleeding after bone marrow aspiration and cutaneous bleeding. Routine coagulation studies revealed a prolonged activated partial thromboplastin time and thrombin time (> 60 s) with a normal reptilase time. Further evaluation showed failure of the activated partial thromboplastin time to correct completely in a 1: 1 mixture with normal plasma. Treatment of the patient's plasma in vitro with protamine sulfate normalized the thrombin time. The presence of a heparin-like anticoagulant was suspected. The plasma heparin level was 0.73 IU/ml. Intravenous infusion of protamine sulfate appeared to neutralize the anticoagulant activity and stop the bleeding. The cancer cells themselves or the invasive nature of this type of cancer might result in a massive release of a heparinoid. Such coagulopathy appears to be a rare mechanism of bleeding and it is an important entity to consider since it is potentially reversible with protamine sulfate.

Differentiation of Fanconi anemia and aplastic anemia using mitomycin C test in Tunisia.

Fanconi anemia (FA) is a recessive chromosomal instability syndrome that is clinically characterized by multiple symptoms. Chromosome breakage hypersensitivity to alkylating agents is the gold standard test for FA diagnosis. In this study, we provide a detailed laboratory protocol for accurate assessment of FA diagnosis based on mitomycin C (MMC) test. Induced chromosomal breakage study was successful in 171 out of 205 aplastic anemia (AA) patients. According to the sensitivity of MMC at 50 ng/ml, 38 patients (22.22%) were diagnosed as affected and 132 patients (77.17%) as unaffected. Somatic mosaicism was suspected in an 11-year-old patient with a FA phenotype. Twenty-six siblings of FA patients were also evaluated and five of them (19.23%) were diagnosed as FA. From this study, a standard protocol for diagnosis of FA was developed. It is routinely used as a diagnostic test of FA in Tunisia.

Hemarthrosis revealing congenital factor XI deficiency.

Congenital factor XI deficiency (also known as the Rosenthal syndrome or hemophilia C) manifests as minor bleeding, usually after trauma or surgery. We report a case in which bilateral knee hemarthrosis was the first manifestation. The patient presented at 32 years of age with a 2-year history of mechanical pain and intermittent swelling in both knees. Knee aspiration recovered blood-tinged fluid. The laboratory workup showed severe factor XI deficiency. Replacement therapy with fresh frozen plasma was effective. Tests in the family showed factor XI deficiency in the patient's sister.