Flow cytometry and morphology analysis of bone marrow in a child with brucellosis and hematologic manifestations.

Constitutional symptoms and pancytopenia are occasionally the initial presentation of pediatric brucellosis. Therefore, in endemic areas, in children with pancytopenia, both brucellosis and malignancy should be included in the deferential diagnosis. We report here a child with pancytopenia and hepatosplenomegaly as manifestations of brucellosis in whom bone marrow morphology and flow cytometry data revealed hemophagocytosis, left shift in myeloid cells and activation changes in antigenic properties of T and B lymphocytes and monocytes. The patient had an uneventful and complete recovery after appropriate antibiotic therapy. Our report demonstrates that bone marrow and flow cytometry findings in children with brucellosis may include significant reactive changes in hematopoiesis.

Moderate to severe thrombocytopenia during pregnancy.

OBJECTIVE: The objective was to investigate obstetric risk factors, complications, and outcomes of pregnancies complicated by moderate to severe thrombocytopenia. MATERIALS AND METHODS: A retrospective case-control study comparing 199 pregnant women with moderate to severe thrombocytopenia (platelet count below 100x10(9)/l) with 201 pregnant women without thrombocytopenia, who delivered between January 2003 to April 2004. Stratified analysis, using the Mantel-Haenszel procedure was performed in order to control for confounders. RESULTS: The main causes of thrombocytopenia were gestational thrombocytopenia (GT) (59.3%), immune thrombocytopenic purpura (ITP) (11.05%), preeclampsia (10.05%), and HELLP (Hemolysis, elevated liver enzymes and low platelet count) syndrome (12.06%). Women with thrombocytopenia were significantly older (30.7+/-5.9 versus 28.7+/-5.7; p=0.001) compared with patients without thrombocytopenia, and had higher rates of labor induction (OR=4.0, 95% CI=2.2-7.6, p<0.001) and preterm deliveries (OR=3.5, 95% CI=1.9-6.5, p<0.001). Even after controlling for labor induction, using the Mantel-Haenszel technique, thrombocytopenia was significantly associated with preterm delivery (weighted OR=3.14, 95% CI=1.7-6.0, p<0.001). Higher rates of placental abruption were found in pregnant women with thrombocytopenia (OR=6.2, 95% CI=1.7-33.2, p=0.001). In a comparison of perinatal outcomes, higher rates of Apgar scores <7 at 5 min were noted in infants of mothers with thrombocytopenia (OR=6.3, 95% CI=1.8-33.8, p=0.001), intrauterine growth restriction (IUGR; OR=4.6, 95% CI=1.5-19.1, p=0.003), and stillbirth (65/1000 versus 0 p<0.001). These adverse perinatal outcomes were found in rare causes of thrombocytopenia such as disseminated intravascular coagulation (DIC), familial thrombotic thrombocytopenic purpura (TTP), anti-phospholipid antibodies (APLA) syndrome, and myeloproliferative disease, and not among patients with GT. CONCLUSIONS: Moderate to severe maternal thrombocytopenia points to a higher degree of severity of the primary disease, which increases perinatal complications. However, the adverse outcome is specifically attributed to preeclampsia, HELLP syndrome, and rare causes, while the perinatal outcome of GT and ITP is basically favorable. Special attention should be given to patients with thrombocytopenia due to preeclampsia, HELLP syndrome, and rarer causes during pregnancy.

Expression of CD24 on CD19- CD79a+ early B-cell progenitors in human bone marrow.

CD24 is a surface marker expressed in immature and mature B cells and involved in cellular adhesion and apoptosis. There are no data, which delineate the stage in early development of human B cells, which marks the expression of CD24. We studied lymphopoiesis in normal pediatric bone marrow (BM) and found that 1.5+/-0.2% of WBC were CD24(+) lymphocytes which did not express CD19. A significant fraction of these cells expressed low levels of CD45 (CD19- CD24+ CD45low cells). Small numbers of CD19- CD24+ CD45low cells were found in the regenerating BM of children with acute lymphoblastic leukemia after the completion of chemotherapy and in normal adult BM. Flow cytometric analyses have shown that CD19- CD24+ CD45low lymphocytes express CD10, CD34, CD79a, CD179a (VpreB), and TdT markers, i.e., displayed antigenic properties of early B-cell progenitors. Our data indicate that CD19- early B-cell progenitors in human BM express CD24, and that the expression of CD24 in human B-cell development precedes the expression of CD19.

Conversion of childhood acute lymphocytic leukemia (L2) with a double t(12;21) to juvenile myelomonocytic leukemia with a novel t(4;11)(p12;q23): a cytogenetic, morphologic, and immunophenotypic study.

Here we describe a cytogenetic and flow-cytometric study of a case in which a conversion of childhood acute lymphocytic leukemia (ALL) into juvenile myelomonocytic leukemia (JMML) occurred. A 3-year-old boy diagnosed CALLA+, pre-B-ALL with double t(12;21) (by fluorescence in situ hybridization analysis), was treated as per the BFM protocol. A cytogenetic analysis performed at 17 months into treatment showed no t(12;21) in bone marrow (BM) cells; however, a novel translocation, namely, t(4;11), involving the p12 locus on chromosome 4 and the MLL gene at 11q23 was detected in monocytes. No cytogenetic abnormalities were found either in Epstein-Barr virus-transformed B cells or in phytohemagglutinin-stimulated T-lymphoid cells. Flow-cytometric analysis demonstrated an asynchronous expression of the antigenic determinants in populations of granulocytic and monocytoid cells: 60% of monocytes expressed low levels of CD14, an unusually high level of CD15, and no CD13 or HLA-DR antigens; 74% of myeloid cells expressed no CD13. Our results indicate that the transformation from B-cell ALL to JMML in this case occurred most probably in the granulocyte-erythroid-macrophage-megakaryocyte progenitor cells without involving the lymphoid cell line. To date, the child is 10 months off therapy and asymptomatic, with t(4;11) in only 3% of the cells.

Urinary excretion of retinol in patients with multiple myeloma: a preliminary study.

Urinary excretion of vitamin A was studied in patients with multiple myeloma (MM). Eight of the 12 patients studied excreted retinol in urine; only one of them had elevated serum creatinine (115-150 micromol/L). There was a highly significant correlation between urinary retinol and serum creatinine (P < 0.0004). Urinary retinol correlated also with urine protein (P < 0.0001) and albumin (P = 0.001), but not with urinary immunoglobulin light chains. Urinary retinol excretion may be an early manifestation of renal dysfunction in MM patients. The effect of urinary retinol excretion on vitamin A homeostasis in MM deserves further study.

Thalassemia intermedia and cavernous transformation of portal vein thrombosis in pregnancy.

We report a rare case of a cavernous transformation of portal vein (CTPV) thrombosis accompanied by Thalassemia and thrombophilia during pregnancy that was successfully treated by low molecular weight heparin. The clinical presentation, diagnosis and the treatment are discussed.

Acute myelogenous leukemia with splenic infarcts presenting as fulminant multi-organ failure.

A 60-year-old male was admitted with leukopenia, thrombocytopenia, splenic infarcts and a normal peripheral smear. Within few hours he rapidly deteriorated with fatal multi-organ failure. Autopsy revealed massive infiltration of leukemic cells in several organs. Acute myelogenous leukemia should be considered in a patient presenting with unexplained multiorgan failure.

Cytology of middle ear fluid during acute otitis media.

BACKGROUND: Limited information is available on the cellular characteristics of the middle ear fluid (MEF) during acute otitis media (AOM). OBJECTIVES: To determine the white blood cell (WBC) composition of the MEF in AOM before and during antibiotic therapy. MATERIALS AND METHODS: Total WBC and differential counts were determined in the MEF of 96 infants and children (ages 2 weeks to 3 years) with AOM who were receiving antibiotics. WBC counts were reported as number of WBC/mg MEF (mean +/- sd). RESULTS: One hundred forty-five MEF samples were obtained by tympanocentesis at enrollment (Day 1), and 36 samples were collected on Days 4 to 5 after initiation of antibiotic therapy. Sixty-one percent of the patients were <1 year of age, and 38% were receiving antibiotic therapy at enrollment. Twenty-eight MEF samples were paired (same ear, Day 1 and Days 4 to 5). One hundred twelve pathogens were isolated from 95 of 145 (66%) culture-positive samples obtained on Day 1: 67 Haemophilus influenzae, 40 Streptococcus pneumoniae and 5 others. MEF WBC counts were lower on Day 1 in patients who had received previous antibiotic therapy than in those who had not (432.4+/- 412.8 vs. 590.5 +/- 436.8, P = 0.03). WBC counts were higher on Day 1 in culture-positive than in culture-negative samples (603.9 +/- 504.9 vs.421.4 +/- 373.4, P = 0.02). WBC counts were higher on Day 1 in MEF samples positive for S. pneumoniae than in those positive for H. influenzae (799.2 +/- 641.5 vs.506.4 +/- 401.9, P = 0.04). There were no differences in the number of neutrophil WBC present in the samples obtained on Day 1 vs. Days 4 to 5 or between samples positive vs.samples negative for bacterial pathogens. CONCLUSIONS: WBC counts were higher in the MEF of patients with culture-positive AOM than in those with culture-negative AOM and in those with AOM caused by S. pneumoniae.