38 results on '"Costa FF"'
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2. Use of adipose derived stem cells accelerates the healing process in third-degree burns.
- Author
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Ribeiro M, Santos KC, Macedo MR, de Souza GA, Neto FIA, Araujo GHM, Cavalcante DR, Costa FF, de Sá Ferreira G, Peixoto LA, de Miranda Moraes J, and Vulcani VAS
- Subjects
- Rats, Animals, Rats, Wistar, Skin pathology, Wound Healing radiation effects, Stem Cells metabolism, Stem Cells pathology, Burns pathology, Soft Tissue Injuries, Low-Level Light Therapy methods
- Abstract
Introduction: Burns are defined as a traumatic injury, usually of thermal origin, that affects the epithelial and adjacent tissue and is classified according to the depth reached. Tissue repair involved in this type of injury is often a challenge both due to its severity and the multiplicity of complications. Regenerative medicine has focused on the use of low-level laser photobiomodulation therapy (LLLT) and adipose-derived stem cells (ADSC), especially in the early stages of the process, to promote better healing and shorten repair time. Therefore, aim of this study was to evaluate the action of LLLT (660 nm) and ADSC in the repair process of burned skin tissue and investigate the association of the techniques (LLLT and ADSC)., Materials and Methods: An in vivo study was carried out using 96 rats (Wister) with a scald burn model at a temperature of 95ºC, exposing the animal's back for 14 s. Animals were randomized into seven groups and three periods, five, 14 and 21 days. The groups included GC: Control group, ADSC-: Group treated with CD49d negative cells, ADSC+ : Group treated with positive CD49d cells, CULT: Group treated with conventional isolation cells, LLLT: Group treated only with LLLT Low Power Laser, ADSC-LLLT: Group treated with CD49d negative cells and LLLT. ADSC+LLLT: Group treated with positive CD49d cells and LLLT. The groups treated with LLLT (660 nm; 5 J/cm2) received irradiation three times a week, on alternate days for five, 14 and 21 days, according to the time of biopsy. ADSC-treated groups received one to three applications of the cells in a total volume of 1000 μL starting soon after the surgical debridement of the burn. Photographic monitoring was carried out at 5, 14 and 21 days after the beginning of the experiment to assess the degree of lesion contraction. Macroscopic, morphometric and histopathological analyzes were performed., Results: We showed significant re-epithelialization as well as an improvement in the healing process in the ADSC+, LLLT and ADSC+LLLT groups. We observed effects in the reduction of the inflammatory phase, increase in angiogenesis, decrease in oedema, greater collagen deposition, and better organization of the extracellular matrix compared to the other treatments. Moreover, the immunomagnetic separation of ADSC cells through the expression of the CD49d protein proved to be a useful means to obtain a more homogeneous population of cells with a role in tissue regeneration compared to the ADSC- and CULT groups., Conclusion: In conclusion, the association of ADSC+ with LLLT was effective in accelerating the burn repair process, stimulating cell proliferation and formation of more normal skin tissue., Competing Interests: Declaration of Competing Interest M.R. has received research grants through National Council for Scientific and Technological Development (CNPq) scholarship from Brazil., (Copyright © 2023 Elsevier Ltd and International Society of Burns Injuries. All rights reserved.)
- Published
- 2024
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3. Antitumoral activity of liraglutide, a new DNMT inhibitor in breast cancer cells in vitro and in vivo.
- Author
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Chequin A, Costa LE, de Campos FF, Moncada ADB, de Lima LTF, Sledz LR, Picheth GF, Adami ER, Acco A, Gonçalves MB, Manica GCM, Valdameri G, de Noronha L, Telles JEQ, Jandrey EHF, Costa ET, Costa FF, de Souza EM, Ramos EAS, and Klassen G
- Subjects
- ADAM Proteins genetics, Animals, Antigens, CD genetics, Breast Neoplasms pathology, Cadherins genetics, Cell Line, Tumor, DNA Methylation drug effects, Estrogen Receptor alpha genetics, Female, Humans, Mice, Promoter Regions, Genetic, Antineoplastic Agents therapeutic use, Breast Neoplasms drug therapy, DNA (Cytosine-5-)-Methyltransferase 1 antagonists & inhibitors, Enzyme Inhibitors therapeutic use, Liraglutide therapeutic use
- Abstract
Breast cancer (BC) is the most frequently diagnosed female cancer and second leading cause of death. Despite the discovery of many antineoplastic drugs for BC, the current therapy is not totally efficient. In this study, we investigated the potential of repurposing the well-known diabetes type II drug liraglutide to modulate epigenetic modifications in BC cells lines in vitro and in vivo via Ehrlich mice tumors models. The in vitro results revealed a significant reduction on cell viability, migration, DNMT activity and displayed lower levels of global DNA methylation in BC cell lines after liraglutide treatment. The interaction between liraglutide and the DNMT enzymes resulted in a decrease profile of DNA methylation for the CDH1, ESR1 and ADAM33 gene promoter regions and, consequently, increased their gene and protein expression levels. To elucidate the possible interaction between liraglutide and the DNMT1 protein, we performed an in silico study that indicates liraglutide binding in the catalytic cleft via hydrogen bonds and salt bridges with the interdomain contacts and disturbs the overall enzyme conformation. The in vivo study was also able to reveal that liraglutide and the combined treatment of liraglutide and paclitaxel or methotrexate were effective in reducing tumor growth. Moreover, the modulation of CDH1 and ADAM33 mouse gene expression by DNA demethylation suggests a role for liraglutide in DNMT activity in vivo. Altogether, these results indicate that liraglutide may be further analysed as a new adjuvant treatment for BC., (Copyright © 2021 Elsevier B.V. All rights reserved.)
- Published
- 2021
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4. Synthesis and pharmacological evaluation of pomalidomide derivatives useful for sickle cell disease treatment.
- Author
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de Melo TRF, Dulmovits BM, Fernandes GFDS, de Souza CM, Lanaro C, He M, Al Abed Y, Chung MC, Blanc L, Costa FF, and Dos Santos JL
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- Dose-Response Relationship, Drug, Humans, Molecular Structure, Structure-Activity Relationship, Thalidomide chemical synthesis, Thalidomide chemistry, Thalidomide therapeutic use, Anemia, Sickle Cell drug therapy, Thalidomide analogs & derivatives
- Abstract
Fetal hemoglobin (HbF) induction constitutes a valuable and validated approach to treat the symptoms of sickle cell disease (SCD). Here, we synthesized pomalidomide-nitric oxide (NO) donor derivatives (3a-f) and evaluated their suitability as novel HbF inducers. All compounds demonstrated different capacities of releasing NO, ranging 0.3-30.3%. Compound 3d was the most effective HbF inducer for CD34
+ cells, exhibiting an effect similar to that of hydroxyurea. We investigated the mode of action of compound 3d for HbF induction by studying the in vitro alterations in the levels of transcription factors (BCL11A, IKAROS, and LRF), inhibition of histone deacetylase enzymes (HDAC-1 and HDAC-2), and measurement of cGMP levels. Additionally, compound 3d exhibited a potent anti-inflammatory effect similar to that of pomalidomide by reducing the TNF-α levels in human mononuclear cells treated with lipopolysaccharides up to 58.6%. Chemical hydrolysis studies revealed that compound 3d was stable at pH 7.4 up to 24 h. These results suggest that compound 3d is a novel HbF inducer prototype with the potential to treat SCD symptoms., (Copyright © 2021 Elsevier Inc. All rights reserved.)- Published
- 2021
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5. Neutrophil extracellular trap regulators in sickle cell disease: Modulation of gene expression of PADI4, neutrophil elastase, and myeloperoxidase during vaso-occlusive crisis.
- Author
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Hounkpe BW, Chenou F, Domingos IF, Cardoso EC, Costa Sobreira MJV, Araujo AS, Lucena-Araújo AR, da Silva Neto PV, Malheiro A, Fraiji NA, Costa FF, Bezerra MAC, Santos MNN, and De Paula EV
- Abstract
Background: Recent evidence suggests that generation of neutrophil extracellular traps (NETosis), one of the components of immunothrombosis, is associated with the pathogenesis of both venous thromboembolism and sickle cell disease (SCD). NETosis is a complex process regulated by several proteins such as peptidyl arginine deaminase 4 (PADI4), neutrophil elastase (ELANE), and myeloperoxidase (MPO). Among these regulators, PADI4 is responsible of histone citrullination, an essential step for NETosis. Accordingly, its inhibition has been recently cited as a promising therapeutic strategy for diseases such as SCD. Although attractive, this strategy requires supportive evidence of its role in the pathogenesis of SCD., Patients and Methods: Patients from two independent cohorts were enrolled in this study. Samples were obtained at steady state (53 patients) or during acute episodes of vaso-occlusive crisis (VOC; 28 patients) in patients from cohort 1. mRNA was extracted from granulocytes to analyze PADI4 , ELANE , and MPO expression by qPCR. Furthermore, plasma activity of PADI4 was assessed from an independent cohort in 15 patients, within 24 hours from admission for VOC. Race-matched healthy individuals from the same geographic regions were used as controls for each cohort., Results and Conclusions: Higher levels of gene expression of PADI4 and ELANE were observed during VOC. Furthermore, plasma activity of PADI4 was higher in acute VOC when compared to healthy individuals. These results demonstrate that NETosis regulators are modulated during acute VOC, and pave the way for studies of PADI4 inhibition as a therapeutic strategy for acute VOC in SCD., (© 2020 The Authors. Research and Practice in Thrombosis and Haemostasis published by Wiley Periodicals LLC on behalf of International Society on Thrombosis and Haemostasis (ISTH).)
- Published
- 2020
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6. Evaluation of oxidative stress-related genetic variants for predicting stroke in patients with sickle cell anemia.
- Author
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Domingos IF, Pereira-Martins DA, Borges-Medeiros RL, Falcao DA, Hatzlhofer BL, Brewin JN, Gardner K, Mendonca TF, Cavalcanti MS, Cunha AF, Anjos AC, Rodrigues ES, Kashima S, Cruz PR, Melo MB, Menzel S, Araujo AS, Costa FF, Bezerra MA, and Lucena-Araujo AR
- Subjects
- Adult, Aged, Humans, Oxidative Stress genetics, Ultrasonography, Doppler, Transcranial, Anemia, Sickle Cell complications, Anemia, Sickle Cell genetics, Stroke diagnostic imaging, Stroke epidemiology, Stroke genetics, alpha-Thalassemia
- Abstract
Overt stroke in adults with sickle cell anemia (SCA) continues to be a major cause of morbidity and mortality, while no evidence-based strategy for prevention has been reached so far. Although transcranial Doppler ultrasonography represents the most important tool for identifying young patients with SCA at risk of primary stroke, strategies for stroke prediction in adulthood remain challenging. Emerging data suggest that oxidative stress may exert a pivotal role in the pathogenesis of ischemic brain injury. Combining these pieces of evidences with the well-known genetic contribution to the development of stroke in SCA, we hypothesized that genetic variants related to the biology of oxidative stress could be used to identify adult patients at higher risk of stroke. Overall, 499 unrelated patients with SCA aged >18 years were genotyped for SOD2 Val16Ala (rs4880), GPX3 T-568C (rs8177404), GPX3 T-518C (rs8177406), GPX3 T-65C (rs8177412), and CAT01 C-262 T (rs1001179) polymorphisms, along with α-thalassemia status and β-globin gene haplotypes. Of these, only the SOD2 Val16Ala polymorphism was associated with stroke. SOD2 Val16Ala polymorphism was independently associated with risk of stroke (odds ratio: 1.98; 95% confidence interval [CI]: 1.18-3.32; P = .009) and with the long-term cumulative incidence of stroke (hazard ratio: 2.24, 95% CI: 1.3-3.9; P = .004). In summary, we provide evidence that oxidative stress-related genetic variants, in particular, the SOD2 Val16Ala polymorphism, may represent a simple and inexpensive alternative for identifying patients at risk of stroke., Competing Interests: Declaration of Competing Interest The authors have no competing financial interests to declare., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
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7. Synthesis and evaluation of resveratrol derivatives as fetal hemoglobin inducers.
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Bosquesi PL, Melchior ACB, Pavan AR, Lanaro C, de Souza CM, Rusinova R, Chelucci RC, Barbieri KP, Fernandes GFDS, Carlos IZ, Andersen OS, Costa FF, and Dos Santos JL
- Subjects
- Analgesics therapeutic use, Animals, Cells, Cultured, Constriction, Pathologic chemically induced, Constriction, Pathologic drug therapy, Disease Models, Animal, Humans, Interleukin-1beta metabolism, Lipopolysaccharides pharmacology, Macrophages cytology, Macrophages drug effects, Macrophages metabolism, Male, Mice, Nitric Oxide metabolism, Resveratrol pharmacology, Resveratrol therapeutic use, Structure-Activity Relationship, Tumor Necrosis Factor-alpha metabolism, Analgesics chemical synthesis, Resveratrol analogs & derivatives
- Abstract
Resveratrol (RVT) derivatives (10a-i) were designed, synthesized, and evaluated for their potential as gamma-globin inducers in treating Sickle Cell Disease (SCD) symptoms. All compounds were able to release NO at different levels ranging from 0 to 26.3%, while RVT did not demonstrate this effect. In vivo, the antinociceptive effect was characterized using an acetic acid-induced abdominal contortion model. All compounds exhibited different levels of protection, ranging from 5.9 to 37.3%; the compound 10a was the most potent among the series. At concentrations between 3.13 and 12.5 µM, the derivative 10a resulted in a reduction of 41.1-64.3% in the TNF-α levels in the supernatants of macrophages that were previously LPS-stimulated. This inhibitory effect was higher than that of RVT used as the control. In addition, the compound 10a and RVT induced double the production of the gamma-globin chains (γG + γA), compared to the vehicle, using CD34+ cells. Compound 10a also did not induce membrane perturbation and it was not mutagenic in the in vivo assay. Thus, compound 10a emerged as a new prototype of the gamma-globin-inducer group with additional analgesic and anti-inflammatory activities and proving to be a useful alternative to treat SCD symptoms., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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8. Differences in heme and hemopexin content in lipoproteins from patients with sickle cell disease.
- Author
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Vendrame F, Olops L, Saad STO, Costa FF, and Fertrin KY
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- Adolescent, Adult, Aged, Case-Control Studies, Female, Humans, Male, Middle Aged, Young Adult, Anemia, Sickle Cell blood, Heme metabolism, Hemopexin metabolism, Lipoproteins blood, Lipoproteins chemistry
- Abstract
Background: High blood cholesterol is associated with atherogenesis and endothelial dysfunction. The latter is present in hemolytic diseases, such as sickle cell anemia, whose carriers have hypocholesterolemia and low incidence of coronary artery disease., Objective: We aimed to characterize cholesterol fractions in patients with sickle cell disease and explore the relationship among lipoproteins, varying degrees of hemolysis, and its biomarkers., Methods: We recruited 37 healthy individuals, 39 with hemoglobin SC disease, and 40 with sickle cell anemia and quantified cholesterol fractions, heme resulting from hemoglobin breakdown, and its main scavenger protein hemopexin., Results: Hypocholesterolemia was most significant in patients with sickle cell anemia, and cholesterol levels correlated positively with hemopexin. Nevertheless, patients still had higher relative low-density lipoprotein (LDL) oxidation than healthy subjects. Analysis of lipoproteins isolated by density ultracentrifugation showed that the LDL fraction contained higher concentrations of heme than the high-density lipoprotein (HDL) fraction, whereas HDL contained more hemopexin than LDL, albeit greatly reduced in patients., Conclusion: Our findings show that the abnormally low lipoprotein levels in sickle cell anemia correlate with hemolysis markers, particularly with hemopexin concentrations, along with significant reduction of this heme scavenger in HDL fractions. This may suggest an important role for HDL in the defense against heme-induced endothelial dysfunction in hemolytic diseases., (Copyright © 2018 National Lipid Association. Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
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9. Understanding the molecular basis of the high oxygen affinity variant human hemoglobin Coimbra.
- Author
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Jorge SE, Bringas M, Petruk AA, Arrar M, Marti MA, Skaf MS, Costa FF, Capece L, Sonati MF, and Estrin D
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- 2,3-Diphosphoglycerate pharmacology, Allosteric Regulation, Heme metabolism, Hemoglobins, Abnormal genetics, Humans, In Vitro Techniques, Kinetics, Models, Molecular, Molecular Dynamics Simulation, Oxygen metabolism, Phytic Acid pharmacology, Protein Interaction Domains and Motifs, Protein Structure, Quaternary, Hemoglobins, Abnormal chemistry, Hemoglobins, Abnormal metabolism
- Abstract
Human hemoglobin (Hb) Coimbra (βAsp99Glu) is one of the seven βAsp99 Hb variants described to date. All βAsp99 substitutions result in increased affinity for O
2 and decreased heme-heme cooperativity and their carriers are clinically characterized by erythrocytocis, caused by tissue hypoxia. Since βAsp99 plays an important role in the allosteric α1β2 interface and the mutation in Hb Coimbra only represents the insertion of a CH2 group in this interface, the present study of Hb Coimbra is important for a better understanding of the global impact of small modifications in this allosteric interface. We carried out functional, kinetic and dynamic characterization of this hemoglobin, focusing on the interpretation of these results in the context of a growth of the position 99 side chain length in the α1β2 interface. Oxygen affinity was evaluated by measuring p50 values in distinct pHs (Bohr effect), and the heme-heme cooperativity was analyzed by determining the Hill coefficient (n), in addition to the effect of the allosteric effectors inositol hexaphosphate (IHP) and 2,3-bisphosphoglyceric acid (2,3-BPG). Computer simulations revealed a stabilization of the R state in the Coimbra variant with respect to the wild type, and consistently, the T-to-R quaternary transition was observed on the nanosecond time scale of classical molecular dynamics simulations., (Copyright © 2017 Elsevier Inc. All rights reserved.)- Published
- 2018
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10. Stathmin 1 expression in plasma cell neoplasms.
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Machado-Neto JA, Pericole FV, Costa FF, Traina F, and Olalla Saad ST
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- 2017
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11. Telomere length correlates with disease severity and inflammation in sickle cell disease.
- Author
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Colella MP, Santana BA, Conran N, Tomazini V, Costa FF, Calado RT, and Saad STO
- Abstract
Background: Telomeres, the ends of linear chromosomes, shorten during mitotic cell division and erosion may be aggravated by inflammation or proliferative and oxidative stress. As the bone marrow is under hyperproliferative pressure in sickle cell disease and several tissues are submitted to chronic inflammation, this study sought to determine the telomere length of patients with sickle cell disease., Methods: The mean telomere length was measured in peripheral blood leukocytes by quantitative polymerase chain reaction. The age-adjusted telomere to single copy gene ratio was compared between 91 adult sickle cell disease patients and 188 controls., Results: Sickle cell disease patients had significantly shorter telomeres than the controls (p-value<0.0001). Moreover, among sickle cell disease genotypes, Hb SS patients had significantly shorter telomeres compared to Hb SC and Hb Sβ patients (p-value<0.0001). Patients on hydroxyurea also had shorter telomeres in comparison to those off the drug (p-value=0.02). A positive correlation was observed between telomere length and hemoglobin level (r=0.3; p-value=0.004), whereas negative correlations were detected between telomere length and lymphocyte count (r=-0.3; p-value=0.005) and interleukin-8 serum levels (r=-0.4; p-value=0.02)., Conclusions: The findings of this study indicate that telomeres are short in sickle cell disease patients and that telomere erosion directly correlates with disease genotype, inflammation markers, and the use of hydroxyurea., (Copyright © 2017 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier Editora Ltda. All rights reserved.)
- Published
- 2017
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12. A randomized trial of amlodipine in addition to standard chelation therapy in patients with thalassemia major.
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Fernandes JL, Loggetto SR, Veríssimo MP, Fertrin KY, Baldanzi GR, Fioravante LA, Tan DM, Higa T, Mashima DA, Piga A, Coelho OR, Costa FF, and Saad ST
- Subjects
- Administration, Oral, Adolescent, Adult, Child, Double-Blind Method, Drug Therapy, Combination, Female, Humans, Iron metabolism, Male, Middle Aged, Myocardium metabolism, Prognosis, Young Adult, Amlodipine therapeutic use, Chelation Therapy, Vasodilator Agents therapeutic use, beta-Thalassemia drug therapy
- Abstract
Cardiovascular disease resulting from iron accumulation is still a major cause of death in patients with thalassemia major (TM). Voltage-gated calcium-channel blockade prevents iron entry into cardiomyocytes and may provide an adjuvant treatment to chelation, reducing myocardial iron uptake. We evaluated whether addition of amlodipine to chelation strategies would reduce myocardial iron overload in TM patients compared with placebo. In a multicenter, double-blind, randomized, placebo-controlled trial, 62 patients were allocated to receive oral amlodipine 5 mg/day or placebo in addition to their current chelation regimen. The main outcome was change in myocardial iron concentration (MIC) determined by magnetic resonance imaging at 12 months, with patients stratified into reduction or prevention groups according to their initial T2* below or above the normal human threshold of 35 ms (MIC, 0.59 mg/g dry weight). At 12 months, patients in the reduction group receiving amlodipine (n = 15) had a significant decrease in MIC compared with patients receiving placebo (n = 15) with a median of -0.26 mg/g (95% confidence interval, -1.02 to -0.01) vs 0.01 mg/g (95% confidence interval, -0.13 to 0.23), P = .02. No significant changes were observed in the prevention group (treatment-effect interaction with P = .005). The same findings were observed in the subgroup of patients with T2* <20 ms. Amlodipine treatment did not cause any serious adverse events. Thus, in TM patients with cardiac siderosis, amlodipine combined with chelation therapy reduced cardiac iron more effectively than chelation therapy alone. Because this conclusion is based on subgroup analyses, it needs to be confirmed in ad hoc clinical trials. This trial was registered at www.clinicaltrials.gov identifier as #NCT01395199., (© 2016 by The American Society of Hematology.)
- Published
- 2016
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13. Familial forms of multiple sclerosis and neuromyelitis optica at an MS center in Rio de Janeiro State, Brazil.
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Papais-Alvarenga RM, Pereira FF, Bernardes MS, Papais-Alvarenga M, Batista E, Paiva CA, Santos CM, and Vasconcelos CC
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- Adolescent, Adult, Brazil epidemiology, Cross-Sectional Studies, Disability Evaluation, Female, Humans, Magnetic Resonance Imaging, Male, Middle Aged, Neurologic Examination, Young Adult, Family Health, Multiple Sclerosis epidemiology, Neuromyelitis Optica epidemiology
- Abstract
Objective: To describe familial forms of demyelinating diseases from an MS referral center in Río de Janeiro State, Brazil., Methods: A descriptive, cross-sectional study was done to identify familial IIDD cases in Hospital da Lagoa, a public hospital where 75% of patients with IIDD who live in Rio de Janeiro state, located in the Southeast region of Brazil, are referred. The diagnoses of all consecutive patients followed in 2011 were reviewed to apply new diagnostic criteria (Wingerchuk et al., 2008). The diagnosis of IIDD was confirmed based on clinical history, neurological examination, MRI of the skull and spinal cord, CSF analysis and investigation of IgG NMO antibodies. The cases that had at least one other relative with IIDD were selected for the study., Results: Familial forms were found only in the multiple sclerosis (MS) and neuromyelitis optica syndrome (NMOSD) categories. 23 MS families were identified, 60.86% with first degree kinship. It has a Caucasian preponderance, 90% of whom were white. The frequency of early onset was 15% and 20% of the MSf cases have progressive primary course., Conclusion: The frequency of familial cases of IIDD was 6.12% among MS patients and 2.8% in NMO spectrum syndromes., (Copyright © 2015. Published by Elsevier B.V.)
- Published
- 2015
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14. Acute hemolytic vascular inflammatory processes are prevented by nitric oxide replacement or a single dose of hydroxyurea.
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Almeida CB, Souza LE, Leonardo FC, Costa FT, Werneck CC, Covas DT, Costa FF, and Conran N
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- Anemia, Sickle Cell blood, Anemia, Sickle Cell drug therapy, Anemia, Sickle Cell pathology, Animals, Cell Movement drug effects, Disease Models, Animal, Hemolysis drug effects, Humans, Hydrazines antagonists & inhibitors, Hydrazines pharmacology, Inflammation blood, Inflammation drug therapy, Inflammation pathology, Leukocytes metabolism, Leukocytes pathology, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Nitric Oxide Donors antagonists & inhibitors, Nitric Oxide Donors pharmacology, Primary Cell Culture, Tumor Necrosis Factor-alpha pharmacology, Viscosity, Water pharmacology, Cyclic N-Oxides pharmacology, Free Radical Scavengers pharmacology, Hemoglobins metabolism, Hydroxyurea pharmacology, Imidazoles pharmacology, Leukocytes drug effects, Nitric Oxide metabolism
- Abstract
Hemolysis and consequent release of cell-free hemoglobin (CFHb) impair vascular nitric oxide (NO) bioavailability and cause oxidative and inflammatory processes. Hydroxyurea (HU), a common therapy for sickle cell disease (SCD), induces fetal Hb production and can act as an NO donor. We evaluated the acute inflammatory effects of intravenous water-induced hemolysis in C57BL/6 mice and determined the abilities of an NO donor, diethylamine NONOate (DEANO), and a single dose of HU to modulate this inflammation. Intravenous water induced acute hemolysis in C57BL/6 mice, attaining plasma Hb levels comparable to those observed in chimeric SCD mice. This hemolysis resulted in significant and rapid systemic inflammation and vascular leukocyte recruitment within 15 minutes, accompanied by NO metabolite generation. Administration of another potent NO scavenger (2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) to C57BL/6 mice induced similar alterations in leukocyte recruitment, whereas hemin-induced inflammation occurred over a longer time frame. Importantly, the acute inflammatory effects of water-induced hemolysis were abolished by the simultaneous administration of DEANO or HU, without altering CFHb, in an NO pathway-mediated manner. In vitro, HU partially reversed the Hb-mediated induction of endothelial proinflammatory cytokine secretion and adhesion molecule expression. In summary, pathophysiological levels of hemolysis trigger an immediate inflammatory response, possibly mediated by vascular NO consumption. HU presents beneficial anti-inflammatory effects by inhibiting rapid-onset hemolytic inflammation via an NO-dependent mechanism, independently of fetal Hb elevation. Data provide novel insights into mechanisms of hemolytic inflammation and further support perspectives for the use of HU as an acute treatment for SCD and other hemolytic disorders., (© 2015 by The American Society of Hematology.)
- Published
- 2015
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15. Somatic mutations of calreticulin in a Brazilian cohort of patients with myeloproliferative neoplasms.
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Machado-Neto JA, de Melo Campos P, de Albuquerque DM, Costa FF, Lorand-Metze I, Olalla Saad ST, and Traina F
- Published
- 2015
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16. Investigating alpha-globin structural variants: a retrospective review of 135,000 Brazilian individuals.
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Kimura EM, Oliveira DM, Jorge SE, Ribeiro DM, Zaccariotto TR, Santos MN, Almeida V, Albuquerque DM, Costa FF, and Sonati Mde F
- Abstract
Background: Brazil has a multiethnic population with a high diversity of hemoglobinopathies. While screenings for beta-globin mutations are far more common, alterations affecting alpha-globin genes are usually more silent and less well known. The aim of this study was to describe the results of a screening program for alpha-globin gene mutations in a representative sample of the Southeastern Brazilian population., Methods: A total of 135,000 individuals, including patients with clinical suspicion of hemoglobinopathies and their family members, randomly chosen individuals submitted to blood tests and blood donors who were abnormal hemoglobin carriers were analyzed. The variants were screened by alkaline and acid electrophoreses, isoelectric focusing and cation-exchange high performance liquid chromatography (HPLC) and the abnormal chains were investigated by reverse-phase high performance liquid chromatography (RP-HPLC). Mutations were identified by molecular analyses, and the oxygen affinity, heme-heme cooperativity and Bohr effect of the variants were evaluated by functional tests., Results: Four new and 22 rare variants were detected in 98 families. Some of these variants were found in co-inheritance with other hemoglobinopathies. Of the rare hemoglobins, Hasharon, Stanleyville II and J-Rovigo were the most common, the first two being S-like and associated with alpha-thalassemia., Conclusion: The variability of alpha-globin alterations reflects the high degree of racial miscegenation and an intense internal migratory flow between different Brazilian regions. This diversity highlights the importance of programs for diagnosing hemoglobinopathies and preventing combinations that may lead to important clinical manifestations in multiethnic populations., (Copyright © 2015 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier Editora Ltda. All rights reserved.)
- Published
- 2015
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17. Reduced plasma angiotensin II levels are reversed by hydroxyurea treatment in mice with sickle cell disease.
- Author
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dos Santos AF, Almeida CB, Brugnerotto AF, Roversi FM, Pallis FR, Franco-Penteado CF, Lanaro C, Albuquerque DM, Leonardo FC, Costa FF, and Conran N
- Subjects
- Anemia, Sickle Cell physiopathology, Angiotensin-Converting Enzyme 2, Animals, Disease Models, Animal, Dose-Response Relationship, Drug, Enzyme-Linked Immunosorbent Assay, Hydroxyurea administration & dosage, Kidney metabolism, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Peptidyl-Dipeptidase A genetics, Real-Time Polymerase Chain Reaction, Receptor, Angiotensin, Type 1 genetics, Receptor, Angiotensin, Type 2 genetics, Anemia, Sickle Cell drug therapy, Angiotensin II blood, Gene Expression Regulation drug effects, Hydroxyurea pharmacology, Renin-Angiotensin System drug effects
- Abstract
Aims: Sickle cell disease (SCD) pathogenesis leads to recurrent vaso-occlusive and hemolytic processes, causing numerous clinical complications including renal damage. As vasoconstrictive mechanisms may be enhanced in SCD, due to endothelial dysfunction and vasoactive protein production, we aimed to determine whether the expression of proteins of the renin-angiotensin system (RAS) may be altered in an animal model of SCD., Main Methods: Plasma angiotensin II (Ang II) was measured in C57BL/6 (WT) mice and mice with SCD by ELISA, while quantitative PCR was used to compare the expressions of the genes encoding the angiotensin-II-receptors 1 and 2 (AT1R and AT2R) and the angiotensin-converting enzymes (ACE1 and ACE2) in the kidneys, hearts, livers and brains of mice. The effects of hydroxyurea (HU; 50-75mg/kg/day, 4weeks) treatment on these parameters were also determined., Key Findings: Plasma Ang II was significantly diminished in SCD mice, compared with WT mice, in association with decreased AT1R and ACE1 expressions in SCD mice kidneys. Treatment of SCD mice with HU reduced leukocyte and platelet counts and increased plasma Ang II to levels similar to those of WT mice. HU also increased AT1R and ACE2 gene expression in the kidney and heart., Significance: Results indicate an imbalanced RAS in an SCD mouse model; HU therapy may be able to restore some RAS parameters in these mice. Further investigations regarding Ang II production and the RAS in human SCD may be warranted, as such changes may reflect or contribute to renal damage and alterations in blood pressure., (Copyright © 2014 Elsevier Inc. All rights reserved.)
- Published
- 2014
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18. Guidelines on the treatment of anemia of chronic renal failure using recombinant human erythropoietin: Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular Guidelines Project: Associação Médica Brasileira - 2014.
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da Silva Araújo A, de Castro Lobo CL, Covas DT, Costa FF, Medeiros L, Cançado RD, Gualandro SF, and Saad ST
- Published
- 2014
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19. Influence of cone-beam computed tomographic scan mode for detection of horizontal root fracture.
- Author
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Costa FF, Pinheiro LR, Umetsubo OS, dos Santos O Jr, Gaia BF, and Cavalcanti MG
- Subjects
- Cone-Beam Computed Tomography methods, Dental Alloys chemistry, Humans, Image Processing, Computer-Assisted statistics & numerical data, Imaging, Three-Dimensional statistics & numerical data, Mandible diagnostic imaging, Observer Variation, Phantoms, Imaging, Post and Core Technique instrumentation, Radiation Dosage, Sensitivity and Specificity, Tooth Socket diagnostic imaging, Cone-Beam Computed Tomography statistics & numerical data, Tooth Fractures diagnostic imaging, Tooth Root injuries, Tooth, Nonvital diagnostic imaging
- Abstract
Introduction: The purpose of the present study was (1) to test the accuracy of a small-volume cone-beam computed tomographic (CBCT) device in detecting horizontal root fractures (HRFs) in teeth with and without an intracanal metallic post (IMP) and (2) to investigate the use of 2 different acquisition protocols of a CBCT device for HRF diagnosis., Methods: Forty endodontically treated teeth with and without an IMP were examined using PreXion 3D CBCT scanner (Terarecon, San Mateo, CA) with a 5-cm high and 5-cm diameter cylinder at 0.10-mm voxel reconstruction. Two observers analyzed the samples to determine the presence and location of HRFs., Results: Sensitivity values ranged from 0.40-0.80. The most favorable results were found for the samples with no IMP observed using the protocol of a higher number of x-ray projections (0.70-0.80). Accuracy in the groups with an IMP ranged from 75%-90% in the 1024 x-ray projection protocol (HI-HI group) versus 70%-85% for the same samples examined in the 512 x-ray projection protocol (HI-STD group). Intraobserver agreement ranged from relevant to perfect concordance for both protocols (HI-HI = Kappa: 0.60-1.00 and HI-STD = Kappa: 0.55-0.89). Interobserver agreement ranged from moderate to perfect concordance for both protocols (HI-HI = Kappa: 0.79-0.89 and HI-STD = Kappa: 0.42-0.76)., Conclusions: Even though there are statistically significant differences for the protocol with the higher number of x-ray projections, we found high accuracy, sensitivity, sensibility, and intra- and interobserver agreement in detecting HRFs for both Prexion 3D protocols., (Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
20. Hb Southampton [B106(G8)Leu→PRO, CTG→CCG] in a Uruguayan woman.
- Author
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Pereira JA, López P, Costa FF, Sans M, and Sonati Mde F
- Abstract
Hemoglobin Southampton (also known as hemoglobin Casper) is a rare hemoglobin structural variant resulting from a substitution of a leucine residue for proline at codon beta106 [beta106(G8)Leu→Pro, CTG→CCG]. It is very unstable and associated with severe hemolytic anemia. We detected this mutation in a 37-year-old Uruguayan woman with a history of severe chronic hemolytic anemia since her childhood. According to our knowledge this is the first time that this variant has been found in the Uruguayan population.
- Published
- 2013
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- View/download PDF
21. Hydroxyurea and a cGMP-amplifying agent have immediate benefits on acute vaso-occlusive events in sickle cell disease mice.
- Author
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Almeida CB, Scheiermann C, Jang JE, Prophete C, Costa FF, Conran N, and Frenette PS
- Subjects
- 3',5'-Cyclic-AMP Phosphodiesterases antagonists & inhibitors, 3',5'-Cyclic-AMP Phosphodiesterases metabolism, Acute Disease, Anemia, Sickle Cell chemically induced, Anemia, Sickle Cell metabolism, Animals, Cell Adhesion drug effects, Cell Communication, Disease Models, Animal, Endothelium, Vascular cytology, Endothelium, Vascular drug effects, Endothelium, Vascular metabolism, Erythrocytes cytology, Erythrocytes drug effects, Female, Humans, Leukocyte Rolling, Leukocytes cytology, Leukocytes drug effects, Male, Mice, Mice, Inbred C57BL, Tumor Necrosis Factor-alpha toxicity, Vascular Diseases chemically induced, Vascular Diseases metabolism, Anemia, Sickle Cell drug therapy, Antisickling Agents therapeutic use, Cyclic GMP metabolism, Hydroxyurea therapeutic use, Pyrazoles pharmacology, Pyrimidines pharmacology, Vascular Diseases drug therapy
- Abstract
Inhibition of leukocyte adhesion to the vascular endothelium represents a novel and important approach for decreasing sickle cell disease (SCD) vaso-occlusion. Using a humanized SCD-mouse-model of tumor necrosis factor-α-induced acute vaso-occlusion, we herein present data demonstrating that short-term administration of either hydroxyurea or the phosphodiesterase 9 (PDE9) inhibitor, BAY73-6691, significantly altered leukocyte recruitment to the microvasculature. Notably, the administration of both agents led to marked improvements in leukocyte rolling and adhesion and decreased heterotypic red blood cell-leukocyte interactions, coupled with prolonged animal survival. Mechanistically, these rheologic benefits were associated with decreased endothelial adhesion molecule expression, as well as diminished leukocyte Mac-1-integrin activation and cyclic guanosine monophosphate (cGMP)-signaling, leading to reduced leukocyte recruitment. Our findings indicate that hydroxyurea has immediate beneficial effects on the microvasculature in acute sickle-cell crises that are independent of the drug's fetal hemoglobin-elevating properties and probably involve the formation of intravascular nitric oxide. In addition, inhibition of PDE9, an enzyme highly expressed in hematopoietic cells, amplified the cGMP-elevating effects of hydroxyurea and may represent a promising and more tissue-specific adjuvant therapy for this disease.
- Published
- 2012
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22. Hydroxyurea is associated with reductions in hypercoagulability markers in sickle cell anemia.
- Author
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Colella MP, De Paula EV, Conran N, Machado-Neto JA, Annicchino-Bizzacchi JM, Costa FF, Saad ST, and Traina F
- Subjects
- Anemia, Sickle Cell blood, Enzyme-Linked Immunosorbent Assay, Humans, Real-Time Polymerase Chain Reaction, Anemia, Sickle Cell drug therapy, Biomarkers blood, Hydroxyurea therapeutic use
- Published
- 2012
- Full Text
- View/download PDF
23. Use of large-volume cone-beam computed tomography in identification and localization of horizontal root fracture in the presence and absence of intracanal metallic post.
- Author
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Costa FF, Gaia BF, Umetsubo OS, Pinheiro LR, Tortamano IP, and Cavalcanti MG
- Subjects
- Bicuspid diagnostic imaging, Bicuspid injuries, Cadaver, Humans, Metals, Observer Variation, Phantoms, Imaging, Signal-To-Noise Ratio, Tooth, Nonvital diagnostic imaging, Cone-Beam Computed Tomography methods, Post and Core Technique, Tooth Fractures diagnostic imaging, Tooth Root injuries
- Abstract
Introduction: The objective of the study was to evaluate the ability of large-volume cone-beam computed tomography (CBCT) to detect horizontal root fracture and to test the influence of a metallic post., Methods: Through the examination of 40 teeth by large-volume CBCT (20-cm height and 15-cm diameter cylinder) at 0.2-mm voxel resolution, 2 observers analyzed the samples for the presence and localization of horizontal root fracture., Results: The values of accuracy in the groups that had no metallic post ranged from 33%-68%, whereas for the samples with the metallic post, values showed a wide variation (38%-83%). Intraobserver agreement showed no statistically significant difference between the groups with/without metallic post; both ranged from very weak to weak (kappa, 0.09-0.369)., Conclusions: The low accuracy and low intraobserver and interobserver agreement reflect the difficulty in performing an adequate diagnosis of horizontal root fractures through a large-volume CBCT by using a small voxel reconstruction., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
24. Hb S-São Paulo: a new sickling hemoglobin with stable polymers and decreased oxygen affinity.
- Author
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Jorge SE, Petruk AA, Kimura EM, Oliveira DM, Caire L, Suemasu CN, Silveira PA, Albuquerque DM, Costa FF, Skaf MS, Martínez L, and Sonati Mde F
- Subjects
- Amino Acid Substitution, Anemia, Sickle Cell metabolism, Base Sequence, Chromatography, High Pressure Liquid, Electrophoresis, Hemoglobin, Sickle chemistry, Hemoglobin, Sickle genetics, Heterozygote, Humans, Infant, Isoelectric Focusing, Male, Molecular Dynamics Simulation, Molecular Sequence Data, Polymers, Protein Stability, Solubility, Static Electricity, beta-Globins chemistry, beta-Globins genetics, Anemia, Sickle Cell genetics, Hemoglobin, Sickle metabolism, Oxygen metabolism, beta-Globins metabolism
- Abstract
Hb S-São Paulo (SP) [HBB:c.20A>T p.Glu6Val; c.196A>G p.Lys65Glu] is a new double-mutant hemoglobin that was found in heterozygosis in an 18-month-old Brazilian male with moderate anemia. It behaves like Hb S in acid electrophoresis, isoelectric focusing and solubility testing but shows different behavior in alkaline electrophoresis, cation-exchange HPLC and RP-HPLC. The variant is slightly unstable, showed reduced oxygen affinity and also appeared to form polymers more stable than the Hb S. Molecular dynamics simulation suggests that the polymerization is favored by interfacial electrostatic interactions. This provides a plausible explanation for some of the reported experimental observations., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
25. Detection of horizontal root fracture with small-volume cone-beam computed tomography in the presence and absence of intracanal metallic post.
- Author
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Costa FF, Gaia BF, Umetsubo OS, and Cavalcanti MG
- Subjects
- Bicuspid, Cone-Beam Computed Tomography instrumentation, Humans, Metals, Observer Variation, Radiation Dosage, Radiography, Dental methods, Radiology Information Systems, Sensitivity and Specificity, Tooth Root diagnostic imaging, Tooth, Nonvital diagnostic imaging, Cone-Beam Computed Tomography methods, Post and Core Technique, Tooth Fractures diagnostic imaging, Tooth Root injuries
- Abstract
Introduction: The aim of the present study was to test the accuracy of small-volume cone-beam computed tomography (CBCT) scanning in the detection of horizontal root fractures and to assess the influence of a metallic post., Methods: Forty teeth were divided into four groups based on the presence of metallic posts and horizontal root fracture. The teeth were examined by small-volume CBCT scanning at 0.2-mm voxel resolution. Three observers analyzed the samples for the presence of a horizontal root fracture. Sensitivity and specificity were calculated., Results: High values for accuracy (73%-88%) were obtained in the groups without a metallic post, and statistically significant differences were found when the group with a metallic post has been observed (55%-70%). Intraobserver agreement also showed statistically significant differences in the groups with a metallic post., Conclusions: Small-volume CBCT scanning showed high accuracy in detecting horizontal root fracture without a metallic post. However, the presence of a metallic post significantly reduced the specificity and sensitivity of this examination., (Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
26. Screening for hotspot mutations in PI3K, JAK2, FLT3 and NPM1 in patients with myelodysplastic syndromes.
- Author
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Machado-Neto JA, Traina F, Lazarini M, Campos Pde M, Pagnano KB, Lorand-Metze I, Costa FF, and Saad ST
- Subjects
- Adult, Aged, Aged, 80 and over, Female, Genetic Testing, Humans, Male, Middle Aged, Nucleophosmin, Janus Kinase 2 genetics, Mutation genetics, Myelodysplastic Syndromes genetics, Nuclear Proteins genetics, Phosphatidylinositol 3-Kinase genetics, fms-Like Tyrosine Kinase 3 genetics
- Abstract
Introduction: Myelodysplastic syndromes encompass a heterogeneous group of clonal hematopoietic stem cell disorders characterized by ineffective hematopoiesis, refractory cytopenia and a tendency to progress toward acute myeloid leukemia. The accumulation of genetic alterations is closely associated with the progression of myelodysplastic syndromes toward acute myeloid leukemia., Objective: To investigate the presence of mutations in the points most frequent for mutations (hotspot mutations) in phosphatidylinositol-3-kinase (PI3K), Janus kinase 2 (JAK2), FMS-like tyrosine kinase 3 (FLT3) and nucleophosmin (NPM1), which are involved in leukemia and other cancers, in a population of Brazilian MDS patients., Methods: Fifty-one myelodysplastic syndromes patients were included in the study. According to French-American-British classification, the patients were distributed as follows: 31 with refractory anemia, 8 with refractory anemia with ringed sideroblasts, 7 with refractory anemia with excess blasts, 3 with refractory anemia with excess blasts in transformation and 2 with chronic myelomonocytic leukemia. Bone marrow samples were obtained and screened for the presence of hotspot mutations using analysis based on amplification with the polymerase chain reaction, sequencing, fragment size polymorphisms or restriction enzyme digestion. All patients were screened for mutations at the time of diagnosis, and 5 patients were also screened at the time of disease progression., Results: These results show that hotspot mutations in the PI3K, JAK2, FLT3 and NPM1 genes are not common in MDS patients; nevertheless, JAK2 mutations may be present in myelodysplasia during disease progression., Conclusions: These results show that hotspot mutations in the PI3K, JAK2, FLT3 and NPM1 genes are not common in MDS patients; nevertheless, JAK2 mutations may be present in myelodysplasia during disease progression.
- Published
- 2011
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27. ARHGAP21 associates with FAK and PKCzeta and is redistributed after cardiac pressure overload.
- Author
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Borges L, Bigarella CL, Baratti MO, Crosara-Alberto DP, Joazeiro PP, Franchini KG, Costa FF, and Saad ST
- Subjects
- Active Transport, Cell Nucleus, Animals, Cell Line, Cell Nucleus, Disease Models, Animal, Hypertrophy, Left Ventricular pathology, Male, Mice, Myocytes, Cardiac metabolism, Myocytes, Cardiac ultrastructure, Phosphorylation, Pressure, Rats, Rats, Inbred SHR, Rats, Wistar, Adaptor Proteins, Signal Transducing metabolism, Focal Adhesion Kinase 1 metabolism, Hypertrophy, Left Ventricular etiology, Hypertrophy, Left Ventricular metabolism, Molecular Chaperones metabolism
- Abstract
ARHGAP21 is highly expressed in the heart, which demonstrates activity over Cdc42 and interacts with proteins of the cytoskeleton and adherent junctions. The main cause of cardiac hypertrophy is mechanical stimulus; therefore we analyzed ARHGAP21 expression after acute mechanical stress in the myocardium and its association with FAK and PKCzeta. We demonstrated that ARHGAP21 is relocated to Z-lines and costameres after pressure overload, and interacts with PKCzeta and FAK in control rats (sham), rats submitted to aortic clamping and spontaneously hypertensive rats (SHR). Co-transfection using ARHGAP21 and PKCzeta constructions demonstrated that ARHGAP21 associates with PKCzeta-GST and endogenous FAK. Pulldown assay showed that ARHGAP21 binds to the C-terminal region of FAK. Moreover, ARHGAP21 binds to PKCzeta phosphorylated on Thr410 in sham and SHR. However, ARHGAP21 only binds to FAK phosphorylated on Tyr925 of SHR. Additionally, PKCzeta is phosphorylated by mechanical stimuli. These results suggest that ARHGAP21 may act as a signaling or scaffold protein of FAK and PKCzeta signaling pathways, developing an important function during cardiac stress.
- Published
- 2008
- Full Text
- View/download PDF
28. Human leukocyte formin: a novel protein expressed in lymphoid malignancies and associated with Akt.
- Author
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Favaro PM, de Souza Medina S, Traina F, Bassères DS, Costa FF, and Saad ST
- Subjects
- Amino Acid Sequence, Cell Line, Tumor, Cytoskeletal Proteins genetics, Formins, HL-60 Cells, Humans, Jurkat Cells, Molecular Sequence Data, Proto-Oncogene Proteins c-akt, Sequence Analysis, Protein, Cytoskeletal Proteins chemistry, Cytoskeletal Proteins metabolism, Gene Expression Regulation, Neoplastic, Leukemia, Lymphocytic, Chronic, B-Cell metabolism, Leukocytes, Mononuclear metabolism, Protein Serine-Threonine Kinases, Proto-Oncogene Proteins metabolism
- Abstract
The very large family of Formin proteins is involved in processes such as morphogenesis, embryonic differentiation, cell polarity, and cytokinesis. A novel human gene from the Formin family, denominated human leukocyte formin gene, was cloned. The cDNA of the gene was determined to be 3959bp long with an open reading frame of 3302bp and computational analysis located this gene on chromosome 17, suggesting that it is composed of 27 exons. Northern blot analysis revealed a restricted expression of mRNA in the thymus, spleen, and peripheral blood leukocytes in normal human tissues. Western blot analysis demonstrated that the protein encoded by this gene is overexpressed in lymphoid malignancies; cancer cell lines and peripheral blood leukocyte from chronic lymphocytic leukemia (CLL) patients. Furthermore, the human leukocyte formin protein was observed to associate with Akt, a critical survival regulator in many different cell types.
- Published
- 2003
- Full Text
- View/download PDF
29. ARHGAP10, a novel human gene coding for a potentially cytoskeletal Rho-GTPase activating protein.
- Author
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Bassères DS, Tizzei EV, Duarte AA, Costa FF, and Saad ST
- Subjects
- Base Sequence, Brain metabolism, Cell Differentiation genetics, Cloning, Molecular, Cytoskeletal Proteins metabolism, DNA, Complementary analysis, DNA, Complementary genetics, GTPase-Activating Proteins biosynthesis, HL-60 Cells, Humans, Molecular Sequence Data, Muscles metabolism, Organ Specificity, rhoA GTP-Binding Protein, Cytoskeletal Proteins genetics, GTPase-Activating Proteins genetics, Genome, Human
- Abstract
Rho-GTPase activating proteins (Rho-GAPs) are negative regulators of Rho-GTPase signaling pathways related to actin cytoskeleton dynamics, cell proliferation, and differentiation. We have identified a novel human gene, termed ARHGAP10, that codes for a 1957-aminoacid Rho-GAP, containing a PDZ, a PH, and a Rho-GAP domain. The cDNA is 7118 bp long and has an open reading frame of 5874 bp. A computational analysis located this gene on chromosome 10 band 10p12.32 suggesting that it is composed of 25 exons. Northern analysis revealed that it is widely expressed, with high levels in brain and muscle. Real-time quantitative PCR analysis confirmed an increase in ARHGAP10 expression during differentiation of HL-60 cells with all-trans-retinoic acid and hematopoietic stem cells with erythropoietin, suggesting that this gene could play a role in normal hematopoiesis. The fact that this gene is highly expressed in muscle and brain, which are highly differentiated tissues, further supports the hypothesis that ARHGAP10 is important for cell differentiation.
- Published
- 2002
- Full Text
- View/download PDF
30. Mechanical properties of stored red blood cells using optical tweezers.
- Author
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Huruta RR, Barjas-Castro ML, Saad ST, Costa FF, Fontes A, Barbosa LC, and Cesar CL
- Subjects
- Elasticity, Humans, Image Processing, Computer-Assisted, Models, Biological, Viscosity, Blood Preservation, Erythrocyte Deformability, Erythrocyte Membrane physiology, Lasers, Micromanipulation instrumentation
- Published
- 1998
31. Beta-spectrin Promiss-ao: a translation initiation codon mutation of the beta-spectrin gene (ATG --> GTG) associated with hereditary spherocytosis and spectrin deficiency in a Brazilian family.
- Author
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Bassères DS, Vicentim DL, Costa FF, Saad ST, and Hassoun H
- Subjects
- Adult, Brazil, DNA Mutational Analysis, Female, Genes, Dominant, Humans, Male, Pedigree, Spectrin deficiency, Codon genetics, Peptide Chain Initiation, Translational genetics, Point Mutation, Spectrin genetics, Spherocytosis, Hereditary genetics
- Published
- 1998
32. Band 3 Campinas: a novel splicing mutation in the band 3 gene (AE1) associated with hereditary spherocytosis, hyperactivity of Na+/Li+ countertransport and an abnormal renal bicarbonate handling.
- Author
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Lima PR, Gontijo JA, Lopes de Faria JB, Costa FF, and Saad ST
- Subjects
- Anion Exchange Protein 1, Erythrocyte deficiency, Anion Exchange Protein 1, Erythrocyte metabolism, Codon genetics, DNA Mutational Analysis, DNA, Complementary genetics, Diuretics pharmacology, Erythrocyte Membrane chemistry, Female, Furosemide pharmacology, Humans, Introns genetics, Ion Transport drug effects, Kidney Tubules, Collecting drug effects, Kidney Tubules, Collecting metabolism, Lithium blood, Male, Pedigree, Polymorphism, Single-Stranded Conformational, RNA Splicing, Sodium blood, Anion Exchange Protein 1, Erythrocyte genetics, Antiporters metabolism, Bicarbonates metabolism, Frameshift Mutation, Kidney metabolism, Spherocytosis, Hereditary metabolism
- Abstract
We have studied the molecular defect underlying band 3 deficiency in one family with hereditary spherocytosis using nonradioactive single strand conformation polymorphism of polymerase chain reaction (PCR) amplified genomic DNA of the AE1 gene. By direct sequencing, a single base substitution in the splicing donor site of intron 8 (position + 1G --> T) was identified. The study of the cDNA showed a skipping of exon 8. This exon skipping event is responsible for a frameshift leading to a premature stop codon 13 amino acids downstream. The distal urinary acidification test by furosemide was performed to verify the consequences of the band 3 deficiency in alpha intercalated cortical collecting duct cells (alphaICCDC). We found an increased basal urinary bicarbonate excretion, associated with an increased basal urinary pH and an efficient distal urinary acidification. We also tested the consequences of band 3 deficiency on the Na+/H+ exchanger, by the measurement of Na+/Li+ countertransport activity in red blood cells. The Na+/Li+ countertransport activity was increased threefold to sixfold in the patients compared with the controls. It is possible that band 3 deficiency in the kidney leads to a decrease in the reabsorption of HCO3- in alphaICCDC and anion loss, which might be associated with an increased sodium-lithium countertransport activity.
- Published
- 1997
33. Eleven novel mutations in the factor VIII gene from Brazilian hemophilia A patients.
- Author
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Arruda VR, Pieneman WC, Reitsma PH, Deutz-Terlouw PP, Annichino-Bizzacchi JM, Briët E, and Costa FF
- Subjects
- Base Sequence, Brazil, Exons genetics, Hemophilia A ethnology, Humans, Molecular Sequence Data, Point Mutation, Polymerase Chain Reaction, Polymorphism, Single-Stranded Conformational, Sequence Deletion, Silver Staining, Factor VIII genetics, Hemophilia A genetics, Mutation
- Abstract
The molecular characterization of the mutations in hemophilia A patients is hampered by the large size of the factor VIII gene and the great heterogeneity of mutations. In this study, we have performed a protocol involving multiplex polymerase chain reaction in which 19 exons were amplified in four different combinations followed by nonradioactive single-strand conformational polymorphism (SSCP) to screen for mutations. Southern blotting was used to detect inversion of the factor VIII gene resulting from recombination between copies of the gene A (F8A) located in intron 22 of the factor VIII gene and two copies close telomeric region of X chromosome. Forty-two hemophilia A patients (21 with severe and 21 with mild-to-moderate disease) were studied. The inversion of factor VIII occurred in 13 of 21 patients affected by severe hemophilia A. One patient showed a large extra band in addition to the three bands observed after Southern blotting with the F8A probe. An abnormal electrophoretic pattern of SSCP was detected in 85% and 50% of the patients affected by mild-to-moderate and severe disease, respectively. Sixteen different mutations were identified. Eleven mutations were novel and comprised 9 point mutations and 2 small deletions. This study shows that the methodology used is safe and rapid and has potential for detecting almost all of the genetic defects of the studied hemophilia A patients.
- Published
- 1995
34. Glucose-6-phosphate dehydrogenase deficiency in sickle cell disease by DNA analysis.
- Author
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Saad ST, Costa FF, Salles TS, Sonatti MF, and Figueiredo MS
- Subjects
- Adolescent, Adult, Alleles, Anemia, Sickle Cell complications, Anemia, Sickle Cell genetics, Base Sequence, Brazil epidemiology, Child, Child, Preschool, Comorbidity, DNA Mutational Analysis, Female, Gene Frequency, Globins genetics, Glucosephosphate Dehydrogenase genetics, Glucosephosphate Dehydrogenase Deficiency complications, Glucosephosphate Dehydrogenase Deficiency genetics, Humans, Male, Middle Aged, Molecular Sequence Data, Prevalence, Anemia, Sickle Cell epidemiology, Glucosephosphate Dehydrogenase Deficiency epidemiology
- Published
- 1995
35. An insertional frameshift mutation of the beta-spectrin gene associated with elliptocytosis in spectrin nice (beta 220/216).
- Author
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Tse WT, Gallagher PG, Pothier B, Costa FF, Scarpa A, Delaunay J, and Forget BG
- Subjects
- Adolescent, Amino Acid Sequence, Base Sequence, Elliptocytosis, Hereditary blood, Exons, Female, Humans, Male, Molecular Sequence Data, Oligonucleotide Probes, Pedigree, Protein Conformation, Reference Values, Elliptocytosis, Hereditary genetics, Frameshift Mutation, Mutagenesis, Insertional, Spectrin genetics
- Abstract
Spectrin Nice (beta 220/216) is a spectrin variant associated with a shortened beta chain found in a patient with elliptocytosis. The shortened beta chain (beta' chain) appeared as an additional band of approximately 216 Kd on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was defective in its ability to be phosphorylated. There were increased amounts of spectrin dimers in crude spectrin extracts from the propositus and the association constant of spectrin dimer self-association was decreased. There was an associated increase of the alpha I 74-Kd fragment from the alpha chain after partial trypic digestion of spectrin. To identify the underlying molecular defect, we analyzed cDNA for beta spectrin obtained by polymerase chain reaction amplification of reverse-transcribed reticulocyte messenger RNA from peripheral blood of the propositus. DNA sequencing of individual as well as pooled subclones showed that two extra bases (GA) are inserted in codon no. 2046 in one allele of the beta-spectrin gene. The insertion results in a frameshift mutation and generates an aberrant C-terminus truncated by about 4 Kd, consistent with the estimated size of the beta' chain observed. By allele-specific oligonucleotide hybridization, the insertion was shown to be present in the propositus and absent in his parents, confirming a previous proposal that it is a de novo mutation. The determination of the location of the mutation in spectrin Nice points to specific regions of the beta-spectrin chain where phosphorylation may occur. A model is proposed to describe the interaction between the alpha- and beta-spectrin chains and to explain the effects of the mutation found in spectrin Nice on the trypsin digestion pattern of its associated alpha chain.
- Published
- 1991
36. The Brazilian type of nondeletional A gamma-fetal hemoglobin has a C----G substitution at nucleotide -195 of the A gamma-globin gene.
- Author
-
Costa FF, Zago MA, Cheng G, Nechtman JF, Stoming TA, and Huisman TH
- Subjects
- Base Sequence, Brazil, Chromosome Deletion, DNA genetics, Humans, Molecular Sequence Data, Mutation, Globins genetics
- Published
- 1990
37. Haematological toxicity associated with agricultural chemicals in Brazil.
- Author
-
Lorand IC, Souza CA, and Costa FF
- Subjects
- Adolescent, Adult, Anemia, Aplastic chemically induced, Brazil, Child, Female, Humans, Male, Agricultural Workers' Diseases chemically induced, Hematologic Diseases chemically induced, Hydrocarbons, Chlorinated, Insecticides poisoning
- Published
- 1984
- Full Text
- View/download PDF
38. Effects of piracetam and iodoacetamide on erythrocyte sickling.
- Author
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Costa FF, Zago MA, and Bottura C
- Subjects
- Anemia, Sickle Cell drug therapy, Animals, Antisickling Agents, Erythrocyte Aging drug effects, Humans, In Vitro Techniques, Rats, Thalassemia drug therapy, Anemia, Sickle Cell blood, Erythrocytes, Abnormal drug effects, Iodoacetamide therapeutic use, Iodoacetates therapeutic use, Piracetam therapeutic use, Pyrrolidinones therapeutic use, Thalassemia blood
- Published
- 1979
- Full Text
- View/download PDF
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