Early T precursor acute lymphoblastic leukaemia/lymphoma shows differential immunophenotypic characteristics including frequent CD33 expression and in vitro response to targeted CD33 therapy.

The differential immunophenotypic characteristics of early T precursor (ETP) acute lymphoblastic leukaemia/lymphoma (ALL) remain incompletely characterized. The study group (n = 142) included 106 (74·7%) men and 36 (25·3%) women with a median age of 34·9 years (range, 2-79) at diagnosis. Patients were subtyped by flow cytometry immunophenotyping as follows: 33 (23·2%) ETP; 32 (22·5%) early non-ETP; 60 (42·2%) thymic; and 17 (12·1%) mature. Excepting definitional markers, there was a significant differential expression of the markers CD2, CD10, CD33 and TdT between ETP-ALL and non-ETP-ALL. Positive CD33 expression (≥20% of leukaemic blasts) was detected in 21/33 (63%) ETP-ALL compared with 17/95 (17·9%) non-ETP-ALL (P < 0·001). Notably, targeted anti-CD33 therapy with IMGN779 resulted in significant growth inhibition and increased apoptosis in ETP-ALL cells in vitro. An 11-marker T-ALL immunophenotype score discriminated reliably between ETP and non-ETP ALL. Longitudinal analysis of ETP-ALL cases in this study demonstrated that the immunophenotype may be occasionally dynamic but is largely stable over the disease course. In summary, identification of ETP-ALL might be enhanced by using an 11-marker T-ALL immunophenotype score. CD33 expression is frequent in ETP-ALL, and in vitro data suggest that exploring anti-CD33 therapy in ETP-ALL is warranted.

Clinicopathologic and molecular features in hairy cell leukemia-variant: single institutional experience.

Hairy cell leukemia-variant is rare. Only a small number of cases have been reported in the literature with little cytogenetic or molecular data available. In this study, we describe the clinicopathologic and genetic features of 23 patients with hairy cell leukemia-variant (16 men and 7 women) with a median age of 70 years. Most patients had splenomegaly (90%), leukocytosis (77%), and lymphocytosis (82%); no patients had monocytopenia. Histologically, the bone marrow biopsy specimens showed a mixed pattern of predominantly interstitial and lesser intrasinusoidal infiltration by leukemic cells. In bone marrow aspirate smears most cells had villous cytoplasmic features and a small nucleolus. We describe unusual sites of hairy cell leukemia-variant involvement in 4 patients, including brain, omentum, terminal ileum, and skin at the time of initial presentation. Immunophenotyping showed monotypic B-cells positive for pan B-cell antigens, CD11c, and CD103, and negative for CD25 and annexin A1. Conventional cytogenetic or fluorescence in situ hybridization analysis showed deletions of 17p13/TP53 and 11q22/ATM gene in 5/12 (42%) and 2/9 (22%) cases, respectively. Sequencing of the variable region of IGVH showed mutations (>2% deviation from germline) in 40% of the cases assessed. MAP2K1 mutation (p.C121S) was seen in 1 of 14 (7%) patients tested. No BRAF V600E mutations were detected. The patients were treated in a heterogeneous manner, but most often with therapies designed for classical hairy cell leukemia and the 5-year overall survival was 84%. In summary, hairy cell leukemia-variant exhibits a heterogeneous spectrum of clinical, morphologic, immunophenotypic, and genetic features that may overlap with classic hairy cell leukemia and other hairy cell-like B-cell neoplasms. A subset of patients can have an aggressive clinical course. In our experience MAP2K1 mutations are uncommon in this disease.

Plasma asymmetric dimethylarginine levels in healthy people.

UNLABELLED: Asymmetric dimethylarginine (ADMA) is an endogenous competitive inhibitor of the endothelial nitric oxide synthase (eNOs). ADMA is believed to be implicated in angiogenesis because it regulates the nitric oxide biosynthesis; any pathological abnormalities in ADMA play a crucial role in the pathogenesis and progression of atherosclerosis. The AIM of the present study was to determine the reference range for plasma concentration of ADMA in a sample of Bulgarian population. PATIENTS AND METHODS: To establish the reference interval of ADMA plasma levels and study the impact of sex and age we recruited 150 healthy subjects of Bulgarian nationality aged between 18 and 65 years. The selection criteria for the reference group were made to comply with the generally approved recommendations of the International Federation of Clinical Chemistry (IFCC). Plasma concentrations of ADMA were determined using ELISA assay (DLD, Diagnostics, Hamburg, Germany). RESULTS: The reference ranges for ADMA, given as 95% of the measured values, were from 0.22 to 0.69 micromol/1. We found no sex-related differences in ADMA concentration (P > 0.05), which obviates the need for separate reference intervals for men and women. Single-factor dispersion analysis found no age dependency ofADMA (P > 0.05, F = 1.038) in the studied reference group in the age range 18-65 which makes unnecessary establishment of reference intervals for lower age ranges in this age group. CONCLUSION: The reference values for ADMA plasma concentrations calculated according to the type of distribution of results can be used as baseline criteria in clinical laboratory studies and for clinical purposes.

Gamna-Gandy bodies in fine-needle aspiration from abdominal splenosis: A clue to underlying portal hypertension.

A 40-year-old woman presented with abdominal pain and jaundice. Past medical history was significant only for splenectomy following a motor vehicle accident. Owing to presence of multiple peritoneal nodules on computerized tomography (CT) and elevated serum CA-125, ovarian peritoneal carcinomatosis was suspected. Ultrasound-guided fine-needle aspiration (FNA) revealed presence of abundant hemosiderin, leukocytes, endothelial cells, and fungal hypha-like structures. No evidence of neoplasia was found. Findings were consistent with Gamna-Gandy bodies (GGBS) within splenic tissue. Based on history of splenectomy and FNA findings, a diagnosis of abdominal splenosis with presence of GGBS was made. Workup for hepatic cirrhosis and portal hypertension was recommended. Liver biopsy confirmed presence of cirrhosis. To our knowledge, this is the first report of GGBS identified within abdominal splenosis. It is important for pathologists to be able to recognize GGBS and to be aware of their relationship to portal hypertension and other conditions associated with severe vascular congestion or hemorrhage. History and pathogenesis of GGBS, their diagnostic morphologic features and a review of cases of GGBS diagnosed via cytology are given.

IgM plasma cell myeloma in the era of novel therapy: a clinicopathological study of 17 cases.

IgM plasma cell myeloma (PCM) is a rare subtype of myeloma, and its response to novel therapies has not been fully characterized. We describe clinicopathological features and outcome of 17 patients with IgM PCM (11 men and 6 women) with a median age of 63 years. Patients presented with serum hyperviscosity (77%), bone lesions (71%), anemia (65%), renal dysfunction (53%), and hypercalcemia (35%). Median serum IgM level was 6.4 g/dL (0.7-12.1 g/dL). Bone marrow plasma cells (median, 80%; range, 20%-90%) were frequently of lymphoplasmacytic type (8/17; 47%). Immunophenotypically, the myeloma cells were positive for CD38, CD138, CD20 (5/16; 31%), CD56 (4/16; 25%), and CD117 (2/12; 17%); negative for CD19; and decreased or absent CD27 and CD81 in all cases. Seven (41%) patients had a complex karyotype, and fluorescence in situ hybridization showed CCND1-IGH (13/16; 81%), and deletions of 17p13/TP53 (29%) and 13q14/RB1 (38%). No MYD88 L265P mutation was detected. Most patients (94%) received proteasome inhibitor with or without immunomodulatory drug, 62% of patients required multiple regimens because of refractory disease, and 11 (65%) of 17 patients underwent autologous stem cell transplant (ASCT). The median OS was 67 months. After a median follow-up of 38 months (range, 3-106 months), only 5 patients achieved complete remission, 5 had persistent disease, and 7 died (2 progressed to plasma cell leukemia and 1 to blastic variant). In summary, IgM PCM is highly associated with t(11;14) and lymphoplasmacytic morphology. Patients are refractory to novel therapy and progression to high-risk myeloma is common, suggesting a need for alternative novel therapies.

Associations between serum selenium and total plasma homocysteine during the acute phase of ischaemic stroke.

BACKGROUND/AIMS: Risk of ischaemic stroke (IS) was associated with total homocysteine (tHCY). On the other hand, serum selenium (Se) exhibited anti-aging and cardiopreventive effects. Se and tHCY showed relationships in animals but these were contradictory or inconclusive in humans; therefore, we searched for such associations in acute IS. METHODS: Ninety-four participants aged around 47 years were identified and 39 patients versus 46 healthy controls were analysed. Clinical, laboratory (blinded) and risk factor questionnaire methods were used. Comparison, correlation and multifactorial regression analyses were applied. RESULTS: IS patients were similar to controls concerning age and gender. IS was prevalent in the carotid system (76.9%); 82.1% had a subacute onset. IS patients expressed higher tHCY (14.65 +/- 9.79 micromol/l) and lower Se levels (1.3 +/- 0.5 micromol/l). Twice as many IS patients (23%) had optimal Se levels of <1.01 mumol/l. Subjects with hyperhomocysteinaemia (tHCY > or =15 micromol/l) showed lower Se levels during IS; Se accounted for 15.4% of tHCY variations (R = -0.393; p = 0.015) with unit change increasing tHCY by 8.25 units. Se remained predictive of tHCY levels after adjustments (vitamin B6, fibrinogen, triglycerides). CONCLUSIONS: Lower Se was observed during acute IS, being inversely associated with and predicting increased tHCY levels. Of note, there were more IS patients with suboptimal Se than controls.

Total plasma homocysteine in Bulgarian population measured by high-performance liquid chromatography with fluorescence detection--comparison with fluorescence polarization immunoassay.

Over the last decade, evidence has accumulated that elevated total homocysteine (tHcy) is an independent risk factor for vascular disease. Due to the variety of Hcy determinants (age, gender, ethnicity and lifestyle), it is now recommended that the distribution of plasma Hcy concentrations should be established for different populations. Therefore the objective of our study was to evaluate a modified HPLC with fluorescence detection procedure for reliable quantification of tHcy and to demonstrate its successful application to determine the distribution of tHcy levels in healthy Bulgarians. The presented method showed good analytical performance (intra- and interassay CVs were <3.9% and <6.7%, respectively; inaccuracy was <6.5%, and analytical recovery 95%-98%, the detection limit was 0.3 micromol/l) and no drug interference was registered. Comparison between HPLC-FD and FPIA using Passing-Bablok regression analysis (r=0.9906) showed good agreement. We describe the distribution of plasma tHcy in a group of 162 healthy Bulgarian adults and examined its relation with age and gender. Our results indicate that higher Hcy concentrations were associated with male sex and increasing age. The higher plasma Hcy observed in our population compared to the rest of Europe corresponds to the high prevalence and mortality of cardiovascular disease in Bulgaria.

A study of plasma total homocysteine levels in healthy people.

UNLABELLED: Elevated plasma levels of homocysteine have been identified as an independent risk factor for atherosclerosis. AIM: The aim of this study was to determine the reference limits of plasma total homocysteine for Bulgarian population. MATERIALS AND METHODS: We investigated 153 healthy individuals without vitamin deficiency aged from 18 to 65 years. The reference group consisted of 74 males and 79 females with mean age respectively 37.80 +/- 1.36 and 39.32 +/- 1.33 years. Plasma total homocysteine was determined by high performance liquid-chromatography (HPLC) modified and validated in our laboratory. RESULTS: The reference intervals were 7.4-18.5 micromol/l for males and 5.5-14.5 micromol/ 1 for females. The mean levels of plasma homocysteine were significantly higher in males in comparison with females (11.86 +/- 0.33 micromol/l vs. 9.88 +/- 0.27 micromol/l; P < 0.001), without considerable correlation with age. Comparing the values of total homocysteine between the two groups of age - < or = 49 and > or = 50 years showed that the investigated individuals > or = 50 years had higher plasma concentration, and the difference was significant only for the group of females. Hyperhomocysteinemia according to ECAP cut-off value (> 12.1 micromol/l) was registered in 30.7% of healthy volunteers. CONCLUSIONS: The results of our study demonstrated that homocysteine levels depend on sex and, to a lesser degree, on age. We have determined plasma total homocysteine reference intervals for the Bulgarian population. This will help the interpretation of the results and contribute to adequate and efficient prevention of blood vessel diseases.