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High production of reactive-oxygen species (ROS) by blood cells is involved in damage of the vascular endothelium and multiple organ dysfunction in sepsis. However, little is known about the intraplatelet ROS production in sepsis and its consequences on platelet reactivity. In this study, we evaluated whether the treatment of rats with lipopolysaccharide (LPS) affects platelet aggregation through intraplatelet ROS generation. Rats were injected with LPS (1 mg/kg, i.p.), and at 2 to 72 h thereafter, adenosine diphosphate (ADP) (3-10 µM) induced platelet aggregation was evaluated. Production of ROS in platelets was measured by flow cytometry using 2′,7′-dichlorofluorescein diacetate (DCFH-DA). Treatment of rats with LPS time-dependently inhibited ADP-induced platelet aggregation within 72 h. The inhibitory effect of LPS on platelet aggregation was further increased when the platelets were incubated with polyethylene glycol-superoxide dismutase (PEG-SOD; 30 U/mL), polyethylene glycol-catalase (PEG-CAT; 1000 U/mL) or the NADPH oxidase inhibitor diphenyleneiodonium (DPI; 10 µM). The ROS production in non-stimulated platelets did not differ between control and LPS-treated rats. However, in ADP-activated platelets, generation of ROS was increased by 3.0- and 7.0-fold, as evaluated at 8 and 48 h after LPS injection, respectively. This increased ROS production was significantly reduced when platelets were incubated in vitro with DPI, PEG-SOD or PEG-CAT. In contrast, treatment of rats with N-acetylcysteine (150 mg/kg, i.p.) significantly reduced the inhibitory effect of LPS on platelet aggregation, and prevented the increased ROS production by in vivo LPS. Our results indicate that the increased intraplatelet ROS production does not contribute to the inhibitory effect of LPS on platelet aggregation; however, the maintenance of redox balance in LPS-treated rats is fundamental to restore the normal platelet response in these animals. [ABSTRACT FROM AUTHOR]

We examined the influence of the glutathione S-transferase mu 1 ( GSTM1), theta 1 ( GSTT1), and pi 1 ( GSTP1) polymorphisms, which are involved in the metabolism of alkylating agents and anthracyclines, on the outcome of patients with Hodgkin lymphoma (HL) treated with conventional chemotherapy. Genomic DNA from 125 consecutive cases was analyzed by polymerase chain reaction and enzymatic digestion for polymorphism determination. The GSTM1 undeleted genotype was associated with more advanced tumor stage and worse disease-free survival. The GSTT1 undeleted genotype was associated with higher recurrence rate. In contrast, higher toxicity of chemotherapy was attributed to absence of the GSTT1 gene. Concerning overall survival, lower tumor stage ( p = 0.006) and International Prognostic Score ( p = 0.02), lower peripheral leukocyte count ( p = 0.0003), higher serum albumin level ( p = 0.08), and GSTT1 undeleted genotype ( p = 0.04) were predictive of a better outcome of patients. In multivariate analysis comparing staging and GST polymorphism, only tumor stage and GSTT1 genotype remained in the model. Our results suggest that the GSTT1 polymorphism influences the outcome of Brazilian patients with HL. However, studies of toxicity, pharmacokinetics, and protein function may clarify whether carriers of the distinct genotypes should receive different doses of chemotherapeutic agents. [ABSTRACT FROM AUTHOR]

Alterations in platelet aggregation in septic conditions are well established. However, little is known about the effects of lipopolysaccharide (LPS) on platelet adhesion. We have therefore investigated the effects of LPS in human platelet adhesion, using an in vitro model of platelet adhesion to fibrinogen-coated wells. Microtiter plates were coated with human fibrinogen, after which washed platelets (6 × 108 platelets/ml) were allowed to adhere. Adherent platelets were quantified through measurement of acid phosphatase activity. Calcium mobilization in Fura2-AM-loaded platelets was monitored with a spectrofluorimeter. Platelet flow cytometry in thrombin-stimulated platelets was performed using monoclonal mouse anti-platelet GPIIb/IIIa antibody (PAC-1). Prior incubation of washed platelets with LPS (0.01–300 µg/ml) for 5 to 60 min concentration- and time-dependently inhibited non-activated platelet adhesion. In thrombin-activated (50 mU/ml) platelets, LPS inhibited the adhesion to a significantly lesser extent than non-activated platelets. Cyclohexamide, superoxide dismutase polyethylene glycol (PEG-SOD) or catalase polyethylene glycol did not affect the LPS responses. No alterations in cyclic GMP levels were seen after platelet incubation with LPS, except with the highest concentration employed (300 µg/ml) where an increase of 36% ( P < 0.05) was observed. Thrombin increased by 7.5-fold the internal Ca2+ platelet levels, an effect markedly inhibited by LPS. Thrombin induced concentration-dependent platelet GPIIb/IIIa activation, but LPS failed to affect the activation state of this membrane glycoprotein. In conclusion, LPS inhibits human platelet adhesion to fibrinogen by mechanisms involving blockade of external Ca2+, independently of cGMP generation and activation of GPIIb/IIIa complex. [ABSTRACT FROM AUTHOR]

We tested in this study whether the polymorphisms of the glutathione S-transferase Mu1 (GSTM1), glutathione S-transferase Theta 1 (GSTT1) and glutathione S-transferase Pi 1 (GSTP1), involved in metabolism of chemical agents, cell proliferation and cell survival, alter the risk for Hodgkin lymphoma (HL). Genomic DNA from 110 consecutive patients with HL and 226 controls was analysed by polymerase chain reaction and restriction digestion for the polymorphism analyses. Similar frequencies of the GSTM1 and GSTT1 genotypes were seen in patients and controls. In contrast, the frequency of the GSTP1 wild genotype (59.1%versus 36.3%, P = 0.004) was higher in patients than in controls. Individuals with the wild genotype had a 2.68 (95%CI: 1.38-5.21)-fold increased risk for the disease than others. An excess of the GSTP1 wild genotype was also observed in patients with tumors of stages III + IV when compared with those with tumors of stages I + II (39.1%versus 20.0%, P = 0.03). These results suggest that the wild allele of the GSTP1 gene is linked to an increased risk and high aggressiveness of the HL in our cases but they should be confirmed by further studies with larger cohorts of patients and controls. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented in response to the article regarding chronic myeloid neoplasm with t(5;8)(q27;p11).

The role of apoptosis in the pathobiology of myelodysplastic syndromes (MDS) remains controversial. We studied the expression of CD95 and CD95L in CD34 + cells of patients with newly diagnosed MDS by flow cytometry, and examined the relation between this expression and FAB and WHO types, total number of CD34 + bone marrow (BM) cells and the degree of peripheral cytopenias. The patients with refractory anemia (RA) and sideroblastic anemia showed CD34 + cells in numbers comparable to normal donors, but had a higher percentage of CD95/CD34 and CD95L/CD34 positive cells. An inverse correlation was found between these cells and the total CD34 + cells. This was also observed when only patients with RA were analyzed. In RAEB, however, CD95/CD95L expression correlated with CD34 + cells but not with the percentage of BM blasts. After classification by the WHO proposal, the patients with refractory cytopenias with multilineage dysplasia showed features intermediary between RA and RAEB. No significant correlation was seen between the expression of CD95/CD95L and the peripheral blood counts. These results are in keeping with the hypothesis that, as the number of MDS precursors increases during disease progression they become less succeptible to apoptosis. [ABSTRACT FROM AUTHOR]

Inactivation of tumor suppressor genes, whose products exert an inhibitory influence on cell cycle progression, can lead to neoplastic transformation. In acute myeloid leukemia (AML), the frequency of p53 gene mutations ranges from 4 to 15% in populations from USA and Europe. In an attempt to investigate the frequency of point mutations in the p53 gene in AML Brazilian patients, DNA samples of 35 patients were studied using PCR-SSCP techniques, screening exons 4-10. Mutations were identified in bone marrow DNA in 5 of the 35 AML patients (14.3%), a frequency similar to those reported for Northern American and European populations. The overall survival of patients with mutations in the p53 gene was significantly shorter than for patients without mutations. [ABSTRACT FROM AUTHOR]

Fourteen pediatric cases of myelodysplastic syndrome according to French-American-British Cooperative Group (FAB) criteria were identified in a retrospective review of all cases of hematologic malignancies referred to the Pediatric Oncology division of the Cancer Hospital A. C. Camargo over a 12-year period: 1 case of refractory anemia, 8 cases of refractory anemia with excess of blasts, and 1 case of refractory anemia with excess of blasts in transformation. Three children had features consistent with chronic myelomonocytic leukemia, and one child was diagnosed with secondary myelodysplastic syndrome. The median age was 3.5 years (1 month-11 years). In 11/14 cases the disease evolved to acute leukemia. In 3 patients blasts had morphological and cytochemical features of lymphoblasts. Two of these patients had a good response to acute lymphoblastic leukemia chemotherapy protocol. The time of progression to leukemia in these cases was shorter than in those who evolved to acute myeloid leukemia. The authors believe this to be the first series of pediatric myelodysplastic syndrome documented in Brazil. Cases were characterized by aggressive FAB type, conspicuous cell atypias in all 3 hemopioetic cell lines, and a high rate of evolution to acute leukemia. [ABSTRACT FROM AUTHOR]