André Fattori

Increased soluble guanylate cyclase activity in the red blood cells of sickle cell patients

Activation of soluble guanylate cyclase (sGC) has been reported to up‐regulate γ‐globin gene transcription in erythroid cell lines and primary erythroblasts. sGC is activated by nitric oxide (NO), subsequently catalysing the conversion of guanosine triphosphate to cyclic guanosine monophosphate (cGMP), which mediates various physiological responses. To study the importance of this mechanism in the erythroid cells of sickle cell patients, cGMP levels were measured in the red blood cells (RBC) of normal individuals, steady‐state sickle cell patients (SS) and SS patients on hydroxyurea (HU) therapy (SS + HU). cGMP levels were found to be significantly higher in RBC of SS patients (SS RBC) than in RBC of normal individuals, and were further increased in RBC of SS + HU patients. cGMP levels correlated with fetal haemoglobin (HbF) levels in SS/SS + HU patients, but not with reticulocyte count. Furthermore, NO‐stimulated sGC activity, following incubation of cells with a NO donor, was significantly greater in SS RBC than in normal RBC. These results demonstrate, for the first time, an increased metabolism of NO mediated by sGC in the SS RBC, which is further increased by hydroxyurea. Augmentation of cGMP levels by NO in erythroid cells may constitute a mechanism for induction of HbF and other erythrocyte functions and represent a possible therapeutic target for treatment of sickle cell disease.

Influence of blood pressure profile on frailty phenotype in community-dwelling elders in Brazil - FIBRA study.

Frailty is a clinical condition associated with pathological aging and biological vulnerability. In the spectrum of events related to frailty, aging of the cardiocirculatory system and abnormalities in arterial blood pressure (BP) partly explain the changes in tissue perfusion and, potentially, the decrease in physiological reserves. This study investigated the relationship between BP levels, systemic arterial hypertension (SAH) and the frailty phenotype by analyzing frailty criteria in a cross-sectional model into the FIBRA network, a populational sample of community-dwelling elders in Southeastern Brazil. Study participants with ≥65 years were selected by probabilistic sampling of residents in the urban area of the municipality of Campinas (n=900). Considering frailty as a whole and the difference between genders, there was a greater proportion of frail or pre-frail individuals among women than men. Analysis of individual frailty criteria showed that weight loss and fatigue were more common among women (18.3% vs. 12.5%, p=0.034 and 22.5% vs. 11.9%, p<0.001, respectively). Comparison of individuals with or without SAH failed to reveal any differences related to frailty criteria. Nevertheless, averages of diastolic blood pressure (DBP) and mean arterial blood pressure values were lower among elderly individuals with reduced grip strength, physical activity and the frailty classification as a whole (OR 0.986, IC 0.975-0.997) (for every 1 mmHg reduction in MBP values, the likelihood of being frail increased 1.4%). Our findings corroborate the relationship between BP values and frailty in the elderly and contribute to an understanding of the pathophysiological mechanisms of the syndrome.

Systemic lupus erythematosus in patients with sickle cell disease

Sickle cell disease (SCD) is a prevalent genetic disorder that includes sickle cell anemia (hemoglobin SS), hemoglobin SC, and hemoglobin Sb-thalassemia. Patients with SCD present with a defective activation of the alternate pathway of the complement system that increases the risk of capsulate bacteria infection and failure to eliminate antigens, predisposing these patients to autoimmune diseases. The authors describe three patients with SCD that developed systemic lupus erythematosus (SLE). In all patients, SLE diagnosis was delayed because symptoms were initially attributable to SCD. Physicians should be alerted to the possible development of SLE in patients with SCD to not delay the diagnosis and start appropriate treatment.

Gene expression profiles of erythroid precursors characterise several mechanisms of the action of hydroxycarbamide in sickle cell anaemia

Hydroxycarbamide (HC) (or hydroxyurea) has been reported to increase fetal haemoglobin levels and improve clinical symptoms in sickle cell anaemia (SCA) patients. However, the complete pathway by which HC acts remains unclear. To study the mechanisms involved in the action of HC, global gene expression profiles were obtained from the bone marrow cells of a SCA patient before and after HC treatment using serial analysis of gene expression. In the comparison of both profiles, 147 differentially expressed transcripts were identified. The functional classification of these transcripts revealed a group of gene categories associated with transcriptional and translational regulation, e.g. EGR‐1, CENTB1, ARHGAP4 and RIN3, suggesting a possible role for these pathways in the improvement of clinical symptoms of SCA patients. The genes involved in these mechanisms may represent potential tools for the identification of new targets for SCA therapy.

Understanding red blood cell parameters in the context of the frailty phenotype: interpretations of the FIBRA (Frailty in Brazilian Seniors) study

Frailty and anemia in the elderly appear to share a common pathophysiology associated with chronic inflammatory processes. This study uses an analytical, cross-sectional, population-based methodology to investigate the probable relationships between frailty, red blood cell parameters and inflammatory markers in 255 community-dwelling elders aged 65 years or older. The frailty phenotype was assessed by non-intentional weight loss, fatigue, low grip strength, low energy expenditure and reduced gait speed. Blood sample analyses were performed to determine hemoglobin level, hematocrit and reticulocyte count, as well as the inflammatory variables IL-6, IL-1ra and hsCRP. In the first multivariate analysis (model I), considering only the erythroid parameters, Hb concentration was a significant variable for both general frailty status and weight loss: a 1.0g/dL drop in serum Hb concentration represented a 2.02-fold increase (CI 1.12-3.63) in an individuals chance of being frail. In the second analysis (model II), which also included inflammatory cytokine levels, hsCRP was independently selected as a significant variable. Each additional year of age represented a 1.21-fold increase in the chance of being frail, and each 1-unit increase in serum hsCRP represented a 3.64-fold increase in the chance of having the frailty phenotype. In model II reticulocyte counts were associated with weight loss and reduced metabolic expenditure criteria. Our findings suggest that reduced Hb concentration, reduced RetAbs count and elevated serum hsCRP levels should be considered components of frailty, which in turn is correlated with sarcopenia, as evidenced by weight loss.

Determining the C-Reactive Protein Level in Patients With Different Clinical Forms of Chagas Disease

Chagas disease is caused by Trypanosoma cruzi and remains a health problem in the developing countries of South America. The condition leads to cardiac conduction disturbances and chronic heart failure. In this study, 136 individuals were evaluated by the Chagas Disease Study Group of the Hospital de la Universidad Estatal de Campinas in Brazil to determine the relationship between chronic heart failure and the serum C-reactive protein (CRP) level. When patients were stratified according to the different clinical presentations of Chagas disease, it was found that the CRP levels in those with severe heart disease and non-Chagasic cardiopathy were significantly higher than in controls or those with mild heart disease (P< .05), even when participants were stratified by age (i.e. <40 and > or =40 years). There was a direct linear correlation between age and CRP level, such that the older the individual, the higher the CRP level. These data provide further evidence for an association between chronic inflammation and the development of heart failure. Although CRP elevations are not exclusively related to Chagas disease, the CRP level may be a useful marker for the progression of Chagas disease to a more advanced phase.

IDENTIFICATION OF DIFFERENTIALLY EXPRESSED GENES INDUCED BY HYDROXYUREA IN RETICULOCYTES FROM SICKLE CELL ANAEMIA PATIENTS

1 The major effect associated with hydroxyurea (HU) treatment of sickle cell anaemia (SCA) patients is an increase in fetal haemoglobin (HbF) synthesis, which inhibits the polymerization of haemoglobin S. 2 Hydroxyurea improves clinical symptoms by reducing the frequency of pain and vaso‐occlusive crises, acute chest syndrome, transfusion requirements and hospitalization. 3 The molecular mechanisms responsible for HU‐mediated induction of fetal globin transcription are not completely understood. Therefore, the aim of the present study was to identify differentially expressed genes participating in these mechanisms. 4 We established two suppression subtractive hybridization (SSH) libraries from reticulocytes obtained from SCA patients either not on or on HU treatment. The gene expression of some of the genes identified was subsequently evaluated by real‐time polymerase chain reaction (PCR). 5 Genes identified with altered expression included SUDS3, FZD5 and PHC3, which may be associated with the regulation of globin expression. 6 This is the first demonstration of an association between HU treatment and the expression of genes identified in erythroid cells.

Molecular identification of Sicilian (deltaß)º-thalassemia associated with ß-thalassemia and hemoglobin S in Brazil

We describe the clinical and molecular characteristics of two unrelated Brazilian families with an association of the Sicilian form of (ds)o-thalassemia with hemoglobin S and s-thalassemia. Direct sequencing of the s-globin gene showed only the hemoglobin S mutation in patient 1 and the s-thalassemia IVS1-110 in patient 2. The other allele was deleted in both patients and PCR of DNA samples of the breakpoint region of both patients showed a band of approximately 1,150 bp, expected to be observed in the DNA of carriers of Sicilian (ds)o-thalassemia. The nucleotide sequence of this fragment confirmed the Sicilian deletion. There are few reports concerning the Hb S/(ds)o-thalassemia association and patient 2 is the first reported case of Sicilian type of (ds)o-thalassemia in association with s-thalassemia documented at the molecular level.

The rare t(6;8) (q27;p11) translocation in a case of chronic myeloid neoplasm mimicking polycythemia vera

A distinct entity associated with 8p11 rearrangement, characterised by chronic myeloid hyperplasia and fast progression to high grade lymphoma (B or T) or acute myeloid leukaemia (AML), was described by MacDonald et al. [1] and Inhorn et al. [2]. The 8p11 region was the site of a recurrent breakpoint in disease, which was combined with nine different partners, 6q27, 7q34, 9q33, 12p11, 12q15, 13q12, 17q23, 19q13, 22q22 [1–15]. The FGFR1 gene, located on 8p11 region, was described as fused to FGFR1OP, TIF1, CEP1, FGFROP2, CPSF6, ZNF198, MYO18A, HERV-K or BCR genes in each distinct translocation [3–9,12– 15]. All resulting fusion proteins had constitutive tyrosine kinase activity and activated multiple signal transduction pathways [3,9]. The entity was recently recognised as 8p11 myeloproliferative syndrome (MPS) in the 2008 WHO classification system for chronic myeloid neoplasm (CMN) [16]. Among the specific rearrangements of the 8p11 MPS, the t(6;8) was a recurrent translocation in only eight cases [10–15], including three cases previously diagnosed as polycythemia vera (PV) based on clinical features, blood count and analysis of bone marrow aspirate [12,14,15]. We present, herein, a case of CMN with a t(6;8)(q27;p11), which was initially diagnosed as having PV in July 2003. The patient was a 12-yearold child. She was referred to our service because of polycythemia and leukocytosis. Plethora was the unique abnormality found at physical examination. Hematological data were as follows: hemoglobin, 18.7 g/dL; hematocrit, 58.6%; VCM, 70 fL; HCM, 22.3 pg; CHCM, 31.9 g/dL; reticulocyte, 1.4%; WBC, 14.5 6 10/L (metamyelocyte: 2.0%, band: 10.0%, segmented: 52.0%, eosinophil: 2.0%, basophil: 7.0%, lymphocyte: 23.0%; monocyte: 4.0%); and platelets, 240.0 6 10/L. Serum measurements were as follows: iron: 43.0 mg/dL (range: 49.0–151.0), total iron binding capacity: 443.0 mg/dL (range: 242.0–450.0), ferritin: 22.4 ng/mL (range: 10.0–200.0), vitamin B12: 1,521.0 pg/mL (range: 271.0–976.0), and creatinine: 0.7 mg/dL (range: 0.3–1.0). The Cr-EDTA-glomerular filtration rate was 114.0 mL/min/1.73 m (range: 100.0+20.0) and arterial oxygen saturation was 97.0%. Normal hemoglobin electrophoresis and negative tests for unstable hemoglobin were also obtained. Computed tomography scan revealed mild splenomegaly. Bone marrow (BM) smear and biopsy revealed hyperplasia of the erythroid, megakaryocytic and granulocytic lineages without fibrosis or eosinophilia; myeloid/erythroid ratio was 1.9 (range: 2.0– 4.0). No abnormalities of the BCR and ABL genes were seen by reverse transcriptase polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH) technique performed on interphase nuclei, using the LSI BCR/ABL ES 32-191022 probe (Vysis, Downers Grove, IL). The diagnosis of PV was

Resiliência psicológica: fator de proteção para idosos no contexto ambulatorial

INTRODUCTION: Among elderly, psychological resilience is associated with depression, physical health, daily living activities and self-perceived health. OBJECTIVE: To describe the relationships between functionality, depressive symptomatology and resilience between resilient and non resilient groups. METHOD: Descriptive study, 59 patients from the Geriatric Ambulatory/HC Unicamp, age 69-91 years. Instruments: scales of activities of daily living ADL, instrumental activities of daily living IADL, depressive symptoms GDS, Mini-Mental Exam MEEM and resilience. Measurements used were: frequency, position and dispersion, comparisons among categorical variables Chi-square and Fisher Exact, numerical variables Mann-Whitney, Krushal-Wallis, co-relations among variables and linear regression multivariate analysis. RESULTS: Mostly women 80.4%, 70-89 years, income ≤ 2 minimum wage, 43.1% illiterate, 57% showed preserved IADL independence in six or seven items, 43% not preserved partial or total help in at least 2 items, 31.4% showed depressive symptomatology suggestive of depression. The most resilient >66 within 75 scores showed average of 5.2±2.1 IADL, in relation to the least resilient 3.6±2.4; p=0.017. The least resilient showed average of 6.4±4.2 depressive symptoms, the most resilient showed average of 2.6±2.6;p=0.001. Negative co-relation between resilience and depressive symptomatology r=-0.688; p 5 depressive symptons tended to show low scores in resilience linear regression multivariate analysis. CONCLUSION: Resilience is an important protector factor for elders assisted in ambulatory, with a relative degree of dependency and evidence of depressive symptoms.