HAEMATOLOGY


Diagnostic Efficacy and Safety of Computed Tomography-Guided Transthoracic Needle Biopsy in Patients with Hematologic Malignancies

M.H. Kallenberg, F.M. Marty, R.H. Rubin, R.R. Gill, F.L. Jacobson, F.E. Factor, J.M. Bryar

Brigham and Women’s Hospital

Introduction: The role of transthoracic needle biopsy (TTNB) in patients with hematologic malignancies, particularly discriminating between malignant and benign etiologies, has not been well studied. Hence, a retrospective analysis to evaluate the diagnostic efficacy and safety of computed tomography (CT)-guided TTNB in this population was performed.

Material and Methods: Records of 54 patients with hematologic malignancies, who underwent TTNB from August 1, 1999 to July 31, 2007, were retrospectively reviewed and analyzed. Particular consideration was given to changes in antifungal therapy based on results of TTNB.

Results: The most common underlying hematologic malignancy was non-Hodgkin lymphoma in 21 patients (38.9%). Of lesions biopsied, the most common lesion location was the left upper lobe in 16 cases (29.6%); 33 lesions were pleural-based (62.3%), and 9 had cavitation (16.7%). Samples were considered cytopathologically adequate in 24 cases (48%). TTNB established a specific diagnosis in 24 patients (44.4%): malignancy was found in 12 (22.2%), infection in 10 (18.5%), and a non-specific benign diagnosis in 2 (3.7%). Sensitivity for detecting malignancy was 47.8%, and sensitivity for detection of a specific infection was 38.5%. Complications occurred in 9 patients (16.7%), pneumothorax in 8 cases (14.8%) and self-limited small volume haemoptysis in 1 case (1.9%). One patient (1.9%) required chest tube placement. The results of TTNB led to a change in antifungal therapy in 14 of the 24 patients with a specific diagnosis (58.3%).

Conclusion: Optimized TTNB in this population is a safe and efficacious diagnostic test, and has decreased morbidity and mortality compared with surgical procedures
Conclusion:

  1. Patients with anterior MI usually have worse outcome and prognosis compared with those with inferior (posterior) MI.
  2. Men with AMI develop complications more often than in woman (ratio 2:1).
  3. Appearance of complications in patients with AMI is connected with systolic dysfunction of the left ventricle.

Variations of Iron Metabolic Indexes in Megalobalstic Anemia

A. Kameli, M. Zare, F. Bagheri, D. Maleki, M. Mousavi, A. Asem, Z. Sabahi

Urmia Medical University

Introduction: Iron metabolic indexes, including serum iron (SI), total iron binding capacity (TIBC) and serum ferritin change in hematologic disorders. TIBC increases in iron deficiency or pregnancy, and decreases in hemolytic anemia and anemia of chronic diseases .also ferritin increases in hemolytic anemia, leukemia and inflammatory disease and decreases in iron deficiency or pregnancy. We studied the change of these indexes in megalobalstic anemia patients.

Material and Methods: By reviewing the files of the patients admitted in UEKEH, with diagnosis of M.A and we charted the range of S.I, serum ferritin and TIBC.

Results: Out of 135 patients with M.A- 46% female and 54% male in the range of 15-95 years old with the average of 52.9- we found mentioned lab tests in 104 of them and the average of 110 for SI, 251 for TIBC and 245.4 for serum ferritin.

Conclusion: This study indicates that the change of iron metabolism indexes in M.A is similar to their range in inflammatory diseases, therefore, in patients with these range of iron indexes and without any significant signs or symptoms of inflammation – specially for the cases without macrocytosis such as minor thalassemia – we have to think of M.A beside hemolytic anemia as a differential diagnose.

Cardio Toxic Effects of Anthracycline Therapy in Children with Acute Lymphocyte Leukemia

A. K. Kato

Ohrid

Introduction: Over the last 25 years of clinical trials, a significant rise in the rate of complete remissions as well as an increase in long-term survival of children with acute lymphoblastic leukemia has been achieved. Therefore, growing attention is now focused on the toxic effects of chemotherapy.
Cardio toxicity is well known side effect of chemotherapy with antracyclins.

The aim of our study was to evaluate the cardio toxic effects of chemotherapy based on the EF (Ejection fraction), FS (Shortening fraction) and tachycardia.

It is retrospective study of 60 patient records with ALL in the period July 2001-June 2007. This study is a part of a bigger study that involves all toxic effects of the ALL-BFM-95 protocol.

Material and methods: Evaluation of ECG and cardiac echosonography in children who manifested cardio toxicity during the chemotherapy. We evaluated the ones made before starting the protocol and further ECG (made weekly) and echo sonograms made after indication of cardiologist. We evaluated Ef (Ejection fraction), Fs (Shortening fraction) and changes in the rhythm.

Results: From the total number of 60 patients, 21 manifested cardiotoxic effects. Fifteen were female and six male (2,5:1), and the average age was 7 years, (1.5 – 12 years). The changes that were registrated were: tachycardia in 19 cases (90%), frontal pericardial separation in 2 cases, problems with repolarization in 5 patients (23.8%), 3 cases of initial secondary cardiomyopathy (14.3%) and 1 case of cardiac hypertrophy(4.76%). All of the changes disappeared after the end of the chemotherapy, except the tachycardia. Tachycardia manifested 10 patients in protocol I, after the cumulative dosage of 120 mg/m² of anthracyclines, in protocol M manifested 4 patients, and 5 patients in protocol II after cumulative dosage of 240 mg/m². Patients were treated with Digoxin and at the end of the treatment they were recommended to continue Digoxin therapy and to do regular checkup at the cardiology department. The EF was in the referent values of 65-83 in all of the patients. It was similar with the FS; there were no significant changes in the values.

Conclusion: The major problem was sinus tachycardia, a disorder of the rhythm in 90% of the children. Several children manifested reversible changes in echocardiogram. None of them had significant changes in EF and FS. Further follow-up of these patients is necessary to detect eventual late cardio toxic effects of chemotherapy.

Correlation Between Cerebrospinal Fluid Tau Protein and Cognitive Functioning in Children with Acute Lymphoblastic Leukemia

P.T. Protas, A. Grabowska, A. Panasiuk, K. Muszynska- Roslan, A. Holownia, M. Krawczuk- Rybak, J.J. Braszko

Bialystok

Introduction: Event free survival in childhood acute lymphoblastic leukaemia (ALL) rose above 70%, allowing us to observe a variety of long-term complications, including neurological disorders. After replacing central nervous system (CNS) radiotherapy and systemic plus intrathecal chemotherapy with high-dose of methotrexate, number of neurological disorders has decreased. However decline in neurocognitive functions can occur even years after the treatment. The aim of the study was to assess whether cerebrospinal fluid tau protein is associated with cognitive changes in children with ALL.

Material and methods: We have examined 38 patients (22 boys, mean age at the diagnosis 7.59 years) with ALL from Department of Pediatric Oncology and HAEMATOLOGY of Medical University in Bialystok, Poland. Cerebrospinal fluid (CSF) samples were collected from children during lumbar punctures performed at diagnosis (point 1), after induction treatment [protocol I (point 2)], during consolidation [protocol M (points 3, 4, 5, 6)] and before maintenance therapy [protocol II or protocol III (point 7)]. The reference group consisted of 22 children (12 boys, mean age 8.23), diagnosed with the clinical symptoms of cerebrospinal meningitis. The Human Total Tau Enzyme-Linked Immunosorbent Assay Kit (BIOSOURCE) was used to determine tau protein levels. Cognitive functioning was established in 19 patients, on average 3.7 years after diagnosis, using Wechsler’ Intelligence Scale for Children-Revised (WISC-R). Data were analysed with statistical package (Statistica 6.0).

Results: The total protein level in the cerebrospinal fluid was not elevated in any of the samples. The mean value of tau protein at diagnosis was 286.81+/-121.26 pg/ml in the study group and 297.63+/-96.81 pg/ml in the reference group. Age and gender did not influence the level of tau protein in CSF of both group. Examination revealed a statistically significant increase in tau protein on the 59-th day of the treatment and at two points during consolidation phase. The level of tau protein at point 7 (before maintenance therapy) was negatively correlated with verbal intelligence quotient measured on an intellectual scale (r=-0.65 p<0

Effect of the Selective Serotonin Reuptake Inhibitor Paroxetine on Platelet Function is Modified by a SLC6A4 Serotonin Transporter

N. Abdelmalik, H.G. Ruhé

Academic Medical Center

Introduction: Selective serotonin reuptake inhibitors (SSRIs) have been associated with an increased bleeding tendency. Polymorphisms of the serotonin transporter (SERT) gene (SLC6A4) promoter region (5-HTTLPR) are associated with the transcriptional activity of the SERT-gene and the rate of serotonin uptake. The objective of this study was to prospectively quantify the dose-response effects of paroxetine and the influence of the 5-HTTLPR polymorphism on platelet function.

Material and Methods: Nineteen drug-free psychiatric outpatients with anxiety disorder or depression (44.5 +/-10.8 years), were tested before and after 6 weeks of paroxetine treatment (20mg/day). Based on clinical symptoms, paroxetine dosages were increased (40-50mg/day) for 6 more weeks in 11 patients. Parameters related to platelet function were assessed by bleeding time, platelet function analyzer (PFA), platelet serotonin, platelet factor 4 (PF4), beta-thromboglobulin (beta-TG), and aggregation tests.
Results: Paroxetine 20mg/day increased mean bleeding time by 1.2 minutes (95% confidence interval (95%CI) -0.2-2.7) and reduced median platelet serotonin level (463 ng/10(9) platelets; Inter Quartile Range (IQR) 361-666), and platelet ss-TG concentration (3.1 IU/10(6) platelets; IQR 0.3-6.0). Other platelet parameters did not change significantly. Serial platelet aggregation tests did not become abnormal. Paroxetine dose-escalation did not further influence platelet function. However, 5-HTTLPR polymorphisms modified these effects: in L(A)/L(A)-carriers, bleeding times did not change (-0.2 minutes; (95%CI -0.6 to 0.9)), while bleeding times significantly increased in /=1 L(A)-allele carriers (457 ng/10(9) platelets; (IQR 392 to 598); p=0.035). PFA closure time and PF4 increased significantly in patients without L(A)-alleles.

Conclusion: Paroxetine 20mg/day does not increase overall bleeding time, but impairs platelet function by decreasing the levels of platelet serotonin and platelet ss-TG. These paroxetine effects appear to be mediated by 5-HTTLPR, with most pronounced effects in patients without L(A)-alleles.

Characterization of a Novel Fusion Gene Possibly Involved in the Development of T-cell Acute Lymphoblastic Leukemia

I. Elens1, I. Lahortiga1, C. Graux2, N. Mentens1, J. Cools1

1Center for Human Genetics
2Cliniques Universitaires Université Catholique de Louvain

Introduction: T-ALL is a T-cell malignancy affecting children and adolescents, caused by cooperation of mutations affecting proliferation, survival, cell cycle, and differentiation.

To detect novel genomic rearrangements, we performed 1 Mb resolution array CGH with extra probes covering candidate oncogenes, as the RTKs (Receptor Tyrosine Kinase). A new fusion gene involving an RTK which functions by kinase activity and nuclear translocation, was detected in one cell line. The other gene involved is a transcription factor expressed in lymphocytes that regulates development and differentiation. Specific screening for this fusion transcript showed it’s presence in 31% in a higher number of T-ALL cell lines.

Materials and Methods: constructions: we generated the sequence of the different fusion variants found in the cell lines by PCR followed by clonation into the MSCV-puromycin vector. Transfection of Hek293T cells was done with Genejuice reagent and the expression and activity level of the RTK was analyzed by Western Blotting. Screening in patients: Samples from 25 patients with T-ALL were screened for the presence of the fusion transcript at RNA level. Cell culture: T-ALL cell lines were cultured in RPMI-1640 supplemented with 20% FCS. We cultured BaF3 cells in RPMI-1640 medium supplemented with 10 % FBS and 1 ng/ml mouse IL3.By retroviral transduction and subsequent puromycin selection we made BaF3 cells express the different constructs. To test IL3-independency, stable BaF3 cell lines were washed three times in PBS and cultured in IL3-free medium. The number of viable cells was counted with a Vi-cell XR cell viability analyzer (Beckman Coulter, Fullerton, CA).
Results: The new fusion transcript was detected in 16% (4/25) of the patients.

Transduction of the constructs in BaF3 cells revealed that none had transforming capacities. Overexpression of the wild type RTK could make the cells IL3 independent after stimulation of the kinase by its ligand.

Conclusion: The presence of a fusion transcript in 31% of the cell lines and 16% of patients with T-ALL suggests a possible role in the development of leukemia. The observation that the addition of the ligand to cells overexpressing the RTK made them IL3 independent, confirms that this kinase can transform thymocytes. Functional studies are still in progress.