Romélia Pinheiro Gonçalves Lemes

The Effect of a Selective Inhibitor of Phosphodiesterase-9 on Oxidative Stress, Inflammation and Cytotoxicity in Neutrophils from Patients with Sickle Cell Anaemia

The aim of the study was to investigate the possible anti‐inflammatory and antioxidant effects of BAY 73‐6691 on neutrophils from SCA patients. This study included 35 patients with a molecular diagnosis of SCA, whose neutrophils were isolated and treated with BAY 73‐6691 at the concentrations 100, 10, 1.0 and 0.1 μg/mL. LDH release and MTT assays were performed to verify cell viability. To evaluate oxidative stress, the following parameters were determined by spectrophotometric assays: NO and malondialdehyde (MDA) levels and activity of catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPx). As inflammatory markers, myeloperoxidase (MPO) levels were evaluated by colorimetric assay and TNF‐α by enzyme immunoassay. The results showed that neutrophils from SCA patients not treated with hydroxyurea (HU) had significantly lower NO levels and catalase and SOD activity, as well as significantly higher MDA, MPO and TNF‐α levels when compared with neutrophils from SCA patients treated with HU and neutrophils from control group. Treatment of SCA neutrophils with BAY 73‐6691 resulted in 94%, 200% and 168% increase in NOx levels, SOD and catalase activity, respectively. In addition, there was a reduction of approximately 46% and 45% in TNF‐α and MPO levels, respectively. In SCAHU neutrophils, there was a 30% and 44% increase in NOx levels and SOD activity, respectively, and a 28% and 37% decrease in TNF‐α and MPO levels, respectively. However, these effects were observed at cytotoxic doses only. The results of this study are original and demonstrate that inhibition of phosphodiesterase‐9 in neutrophils from SCA patients with BAY 73‐6691 was able to increase the NO bioavailability and attenuate oxidative stress and inflammation in neutrophils from patients not treated with HU.

Genotoxicity associated with the use of tyrosine kinase inhibitors in patients with chronic myeloid leukemia

Genotoxicity AssociatedWith the Use of Tyrosine Kinase Inhibitors in Patients With Chronic Myeloid Leukemia Pedro Aurio Maia Filho ,* Tarcı sio Paulo de Almeida Filho, Caroline de F atima Aquino Moreina-Nunes, Rommel Rodriguez Burbano, Jos e Alexandre Rodrigues de Lemos , Edivaldo Herculano Correa de Oliveira, Bruno Coêlho Cavalcanti, Jamilly Florêncio Pereira,Maritza Cavalcante Barbosa, Fernando Barroso Duarte, Marilena Facundo de Castro, Acy Telles de Souza Quixad a, and Rom elia Pinheiro Gonçalves Lemes Faculty of Pharmacy, Federal University of Cear a, Fortaleza, Brazil Nucleus of Research and Development of Medicines, Federal University of Cear a, Fortaleza, Brazil Biological Science Institute, Universidade Federal do Par a, Bel em, Brazil Hematology Hospital at Walter Cantı ıdio University Hospital, Fortaleza, Cear a

Bone marrow fibrosis at diagnosis is associated with TP53 overexpression and adverse prognosis in low‐risk myelodysplastic syndrome

Myelodysplastic syndrome (MDS) is a heterogeneous group of stem cell clonal alterations, culminating in a high risk of progression to acute myeloid leukaemia (AML) (Greenberg et al, 2012). The prognosis of these patients is commonly determined by the International Prognostic Score System (IPSS), which considers the number of cytopenias, cytogenetic alterations and number of blasts in the bone marrow at diagnosis. Patients with a lower risk of progression to AML are classified as low risk; however, a subgroup of these patients develops a disease with an aggressive course and a lower survival rate (Mittelman et al, 2010). This clinical heterogeneity reinforces the need to identify auxiliary markers for prognostic stratification systems. Bone marrow fibrosis is a histological finding in approximately 10–20% of patients with de novo MDS; however, MDS with fibrosis is not yet recognized as a distinct entity and its prognostic role is still under debate (Ramos et al, 2016). The immunohistochemical expression of TP53 has been used as a predictive marker for mutations in the TP53 gene in several haematological diseases due to the low-cost and easy to perform technique (Kulasekararaj et al, 2013). Moreover, studies have demonstrated that TP53 expression is an independent adverse prognostic factor in patients with highrisk MDS, being associated with complex karyotype and therapy-related MDS (Kulasekararaj et al, 2013). Therefore, the present study investigated the impact of bone marrow fibrosis and TP53 expression at diagnosis on the clinical characteristics and overall survival of patients with low-risk MDS. Seventy-three low-risk MDS patients treated at a reference University Hospital in the northeast of Brazil participated in the study. Patients were stratified according to the IPSS and revised IPSS (IPSS-R) criteria (Greenberg et al, 2012). Clinical data at diagnosis, such as karyotype, complete blood count, number of blasts, presence of fibrosis and evolution to death or AML, were collected from medical records. TP53 expression was determined by immunohistochemistry, according to Shah et al (2012). Briefly, 5-lm sections of bone marrow were incubated with the p53-DO7 monoclonal antibody (Dako, Santa Clara, CA, USA). TP53 protein was identified by the ABC – peroxidase technique, staining with 3,30-Diaminobenzidine (DAB; Dako), and counterstaining with haematoxylin. The percentage and intensity of TP53 staining was assessed based on a total manual count of 1000 granulocytic precursors. TP53 expression was defined as positive for a strong nuclear staining pattern, according to the Modified Quick Score (Kulasekararaj et al, 2013) in at least 1% of the cells analysed (Saft et al, 2014). Patients were stratified for the presence of bone marrow fibrosis at diagnosis according to the criteria used by the European Myelofibrosis Network (Thiele et al, 2005). The study was approved by the Research Ethics Committee of the Federal University of Cear a (protocol #129/12) and all patients agreed to participate. Statistical analysis was performed using the GraphPad Prism 5.0 software (GraphPad Software Inc., La Jolla, CA, USA), using chi-square and Student’s t or Mann–Whitney test, according data normality, verified by Kolmogorov–Smirnov test. The Kaplan–Meier curve was performed to verify differences in overall survival. Significance was set at P < 0 05. The median age of the study patients was 65 years, with a predominance of females (64 3% females and 35 7% males). Most of the patients (49 32%) were classified as single-lineage dysplasia, followed by multi-lineage dysplasia (41 09%), MDS with ringed sideroblasts (6 85%) and MDS with isolated del(5q) (2 74%). Patients with bone marrow fibrosis (MF-2 and MF-3) had significantly lower haemoglobin and haematocrit values at diagnosis when compared to patients without fibrosis (MF-0 and MF-1) (P = 0 001). High TP53 expression was associated with the presence of bone marrow fibrosis (P = 0 008). There was no difference between the other analysed parameters (Table I). Patients with bone marrow fibrosis [P = 0 0007, hazard ratio (HR) = 0 02, 95% confidence interval (CI) = 0 002– 0 20] and high TP53 expression (P = 0 0039, HR = 14 34, 95% CI = 2 0–102 8) had lower overall survival when compared to patients without these findings. Overall survival in patients with bone marrow fibrosis (MF-2 and MF-3) did not demonstrate a significant difference regarding TP53 expression (P = 0 3794, HR = 2 62, 95% CI = 0 34–17 40) (Fig 1). Some studies have reported on the importance of bone marrow fibrosis in prognostic characteristics and survival of patients with de novo MDS; however, the role of these Correspondence

Presence of new mutations in the TP53 gene in patients with low-risk myelodysplastic syndrome: two case reports

BackgroundMyelodysplastic syndromes are heterogeneous disorders. Patients with myelodysplastic syndrome disease often have ineffective hematopoiesis, cytopenias, blood cell dysplasia in one or more cell types, and are at high risk for developing acute myeloid leukemia. In myelodysplastic syndrome, mutations of TP53 gene are usually associated with complex karyotype and confer a worse prognosis. In the present study, two mutations in this gene are presented and discussed with the clinical evolution of the patients.Case presentationThe first case is a 77-year-old Brazilian woman diagnosed as having multiple lineage dysplasia myelodysplastic syndrome according to World Health Organization 2016 and classified as very low-risk by Revised International Prognostic Scoring. The second case is an 80-year-old Brazilian man also diagnosed as having multiple lineage dysplasia myelodysplastic syndrome and classified as low risk. The mutation described in the first case was already identified in some neoplasias and it is associated with a poor prognosis, but it had never been reported before in myelodysplastic syndrome. The second mutation has never been described.ConclusionsThis is a novel report for the scientific community and may be very helpful as we can better understand the disease and the impact of mutations through the follow-up of these patients and others in the future. Both patients are in a good clinical condition, suggesting that these mutations may not alter the clinical course of the disease or may be associated with a good prognosis, but their role in the disease must be investigated more deeply in a larger population.

Comments on the clinical and laboratory characteristics of patients with dengue hemorrhagic fever manifestations and their transfusion profile

See paper by Fujimoto DE et al. on pages 115-20. *Corresponding author at: Hemocentro do Estado do Ceara, Hospital Universitario Walter Cantidio, Rua Capitao Francisco Pedro, 1210, Rodolfo Teofilo, 60430-370, Fortaleza, CE, Brazil. E-mail address: romelia.pinheiro@pg.cnpq.br (R. P. G. Lemes). 1516-8484/

Clinical events and their relation to the tumor necrosis factor-alpha and interleukin-10 genotypes in Sickle-Cell- Anemia patients

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