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Eliminación de Secuencia , Talasemia alfa , Humanos , Talasemia alfa/genética , Brasil , Masculino , Globinas alfa/genética , Niño , Femenino , Hemoglobina H/genéticaRESUMEN
The complex pathophysiology of sickle cell anemia (SCA) involves intravascular hemolytic processes and recurrent vaso-occlusion, driven by chronic vascular inflammation, which result in the disease's severe clinical complications, including recurrent painful vaso-occlusive episodes. Hydroxyurea, the only drug frequently used for SCA therapy, is a cytostatic agent, although it appears to exert nitric oxide/soluble guanylyl cyclase (sGC) modulating activity. As new drugs that can complement or replace the use of hydroxyurea are sought to further reduce vaso-occlusive episode frequency in SCA, we investigated the effects of the sGC agonists BAY 60-2770 (sGC activator) and BAY 41-2272 (sGC stimulator) in the presence or absence of hydroxyurea on SCA vaso-occlusive mechanisms and cell recruitment both ex vivo and in vivo. These agents significantly reduced stimulated human SCA neutrophil adhesive properties ex vivo in association with the inhibition of surface ß2-integrin activation. A single administration of BAY 60-2770 or BAY 41-2272 decreased tumor necrosis factor cytokine-induced leukocyte recruitment in a mouse model of SCA vaso-occlusion. Importantly, the in vivo actions of both agonists were significantly potentiated by the coadministration of hydroxyurea. Erythroid cell fetal hemoglobin (HbF) elevation is also a major goal for SCA therapy. BAY 41-2272 but not BAY 60-2770 at the concentrations employed significantly induced γ-globin gene transcription in association with HbF production in cultured erythroleukemic cells. In conclusion, sGC agonist drugs could represent a promising approach as therapy for SCA, for use either as stand-alone treatments or in combination with hydroxyurea. SIGNIFICANCE STATEMENT: This preclinical study demonstrates that stimulators and activators of sGC are potent inhibitors of the adhesion and recruitment of leukocytes from humans and in mice with sickle cell anemia (SCA) and may represent a promising approach for diminishing vaso-occlusive episode frequency in SCA. Hydroxyurea, a drug already frequently used for treating SCA, was found to potentiate the beneficial effects of sGC agonists in in vivo studies, implying that these classes of compounds could be used alone or in combination therapy.
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Anemia de Células Falciformes/tratamiento farmacológico , Anemia de Células Falciformes/metabolismo , Hidroxiurea/farmacocinética , Guanilil Ciclasa Soluble/metabolismo , Animales , Benzoatos/farmacología , Compuestos de Bifenilo/farmacología , Línea Celular Tumoral , Modelos Animales de Enfermedad , Células Eritroides/efectos de los fármacos , Células Eritroides/metabolismo , Hemoglobina Fetal/metabolismo , Humanos , Hidrocarburos Fluorados/farmacología , Células K562 , Masculino , Ratones , Ratones Endogámicos C57BL , Pirazoles/farmacología , Piridinas/farmacología , Enfermedades Vasculares/tratamiento farmacológico , Enfermedades Vasculares/metabolismo , Vasodilatadores/farmacologíaRESUMEN
The impaired bioavailability of endogenous nitric oxide (NO) in sickle cell anemia (SCA) may be influenced by polymorphisms in the endothelial nitric oxide synthase gene (eNOS). We compared allelic/genotypic frequencies of the eNOS polymorphisms T-786C, VNTR4a/b and G894T between 89 adult SCA patients and 100 healthy controls, and investigated the relationship between these SNPs and markers of hemolysis [lactate dehydrogenase (LDH), indirect bilirubin (IB) and reticulocyte counts], inflammation [interleukins IL-1ß, IL-6, IL-8, Tumor Necrosis Factor (TNF-α) and C-reactive protein (CRP)] and endothelial dysfunction (ED) [soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble intercellular adhesion molecule-1 (sICAM-1), soluble L-selectin (sL-selectin), von Willebrand Factor (vWF) antigen and D-dimers] in the patients. The frequencies of the mutant -786C allele and -786C/C genotype were significantly higher in patients (p = 0.02 and p = 0.04, respectively) but not significantly correlated with the markers. For VNTR4a/b and G894T, the allelic/genotypic frequencies did not statistically differ between patient and control groups. Patients carrying the 4a allele and those with the 894G/G genotype showed a significant decrease in IB (p = 0.02 and p = 0.04, respectively), and only patients with the 4a allele exhibited reduced IL-1ß (p = 0.01). The correlation profiles between markers of inflammation and ED significantly differed between patients carrying the mutant alleles and those with wild-type genotypes. This appears to be the first report on the relationship between eNOS gene polymorphisms and markers of hemolysis, inflammation and ED in Brazilian SCA patients. Our results indicate that the SNPs analyzed may influence the phenotypic variability of these patients.
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Anemia de Células Falciformes/enzimología , Anemia de Células Falciformes/genética , Productos de Degradación de Fibrina-Fibrinógeno/análisis , Hemólisis , Molécula 1 de Adhesión Intercelular/sangre , Óxido Nítrico Sintasa de Tipo III/genética , Polimorfismo de Nucleótido Simple , Molécula 1 de Adhesión Celular Vascular/sangre , Factor de von Willebrand/análisis , Adulto , Alelos , Anemia de Células Falciformes/sangre , Anemia de Células Falciformes/epidemiología , Bilirrubina/sangre , Biomarcadores/sangre , Brasil/epidemiología , Estudios de Casos y Controles , Citocinas/sangre , Femenino , Frecuencia de los Genes , Haplotipos , Humanos , Inflamación/sangre , L-Lactato Deshidrogenasa/sangre , Masculino , Recuento de Reticulocitos , Adulto JovenRESUMEN
Hereditary anemias are a group of heterogeneous disorders including hemolytic anemias and hyporegenerative anemias, as congenital dyserythropoietic anemia (CDA). Causative mutations occur in a wide range of genes leading to deficiencies in red cell production, structure, or function. The genetic screening of the main genes is important for timely diagnosis, since routine laboratory tests fail in a percentage of the cases, appropriate treatment decisions, and genetic counseling purposes. A conventional gene-by-gene sequencing approach is expensive and highly time-consuming, due to the genetic complexity of these diseases. To overcome this problem, we customized a targeted sequencing panel covering 35 genes previously associated to red cell disorders. We analyzed 36 patients, and potentially pathogenic variants were identified in 26 cases (72%). Twenty variants were novel. Remarkably, mutations in the SPTB gene (ß-spectrin) were found in 34.6% of the patients with hereditary spherocytosis (HS), suggesting that SPTB is a major HS gene in the Southeast of Brazil. We also identified two cases with dominant HS presenting null mutations in trans with α-LELY in SPTA1 gene. This is the first comprehensive genetic analysis for hereditary anemias in the Brazilian population, contributing to a better understanding of the genetic basis and phenotypic consequences of these rare conditions in our population.
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Anemia Diseritropoyética Congénita/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Mutación , Espectrina/genética , Esferocitosis Hereditaria/genética , Brasil , Femenino , Humanos , MasculinoAsunto(s)
Anemia Hemolítica , Hemoglobinas Anormales , Mutación , Adulto , Anemia Hemolítica/sangre , Anemia Hemolítica/genética , Anemia Hemolítica/patología , Anemia Hemolítica/terapia , Brasil , Femenino , Hemoglobinas Anormales/química , Hemoglobinas Anormales/genética , Hemoglobinas Anormales/metabolismo , HumanosRESUMEN
AIM: To evaluate the frequency of post-treatment apical periodontitis associated with root filled teeth with at least one untreated root canal. METHODOLOGY: Eight hundred and seven cone beam computed tomography images containing at least one root filled tooth were selected from a collection of 1543 images from Brazilian individuals. Scans were taken using ICAT Classic devices (Imaging Sciences, Hatfield, PA, USA) in a private oral radiology clinic from January to April 2015. All root filled teeth were analysed for the presence of missed canals and apical periodontitis. The chi-square and odds ratio tests were used to verify if there were an association and risk relationship between the occurrence of untreated canals and apical periodontitis. RESULTS: A total of 2294 teeth with evidence of root fillings were identified. Two hundred and eighty-one teeth had at least one untreated missed canal (12%). The frequency of apical periodontitis in teeth with at least one untreated canal was significantly greater in comparison to teeth with all canals treated (274/281, 98% versus 1736/2013, 86%) (P < 0.01). The odds for apical periodontitis to be present was 6.25 times greater for teeth with an untreated canal. The mesiobuccal roots of maxillary first molars had the greatest frequency of untreated canals (114/154, 74%), with the second mesiobuccal canal being the most frequently missed (n = 106/114, 93%). CONCLUSION: Root filled teeth with at least one missed canal had a high prevalence of post-treatment apical periodontitis.
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Periodontitis Periapical , Brasil , Cavidad Pulpar , Humanos , Obturación del Conducto Radicular , Raíz del DienteRESUMEN
BACKGROUND: Sickle leg ulcer (SLU) is a chronic and debilitating complication of sickle cell disease (SCD) associated with huge physical and psychosocial discomfort. The occurrence of SLU has remained steady despite successful preventive strategies and advances in SCD care. Although multifactorial factors have been implicated in SLU, these are not fully understood, and data on how these relate to young Nigerian SCD patients are scanty. AIMS: This study aims to evaluate the sociodemographic, clinical, and laboratory markers of SLU in a young Nigerian SCD cohort. PATIENTS AND METHODS: This study involved 109 young SCD patients and 67 healthy peers. The sociodemographic and laboratory parameters of the participants were examined in addition to the evaluation of the SCD cohort for SLU. RESULTS: Only the HbSS patients had SLU. This was found in six of them giving a prevalence of 5.9% (6/101). Their median age was 17, range 14-21 years. There was a preceding history of trauma in 4 (66.7%), and this included a case of traditional scarifications for local therapeutic purposes. Two of the three (66.7%) males with SLU also had priapism (P = 0.0132). Patients with SLU were older, had less frequent bone pain crises, and significantly belonged to the low socioeconomic class (P < 0.05). Although patients with SLU had relatively higher lactate dehydrogenase, platelet count, aspartate transaminase, bilirubin, white blood cell, and lower Hb concentration and HbF, these did not attain statistical significance (P > 0.05). CONCLUSION: This study confirms that SLU is common among young SCD patients with HbSS genotype, low socioeconomic background, and older age. It also suggests that SLU could be more related to hemolysis-associated SCD phenotypes among the patients.
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Anemia de Células Falciformes/complicaciones , Bilirrubina/sangre , Población Negra/estadística & datos numéricos , Úlcera de la Pierna/etiología , Adolescente , Adulto , Anciano , Anemia de Células Falciformes/sangre , Anemia de Células Falciformes/epidemiología , Biomarcadores , Estudios de Cohortes , Femenino , Humanos , Úlcera de la Pierna/sangre , Úlcera de la Pierna/epidemiología , Masculino , Persona de Mediana Edad , Nigeria/epidemiología , Recuento de Plaquetas , Prevalencia , Priapismo , Índice de Severidad de la Enfermedad , Factores Socioeconómicos , Adulto JovenRESUMEN
Human hemoglobin (Hb) Coimbra (ßAsp99Glu) is one of the seven ßAsp99 Hb variants described to date. All ßAsp99 substitutions result in increased affinity for O2 and decreased heme-heme cooperativity and their carriers are clinically characterized by erythrocytocis, caused by tissue hypoxia. Since ßAsp99 plays an important role in the allosteric α1ß2 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 α1ß2 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.
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Hemoglobinas Anormales/química , Hemoglobinas Anormales/metabolismo , 2,3-Difosfoglicerato/farmacología , Regulación Alostérica , Hemo/metabolismo , Hemoglobinas Anormales/genética , Humanos , Técnicas In Vitro , Cinética , Modelos Moleculares , Simulación de Dinámica Molecular , Oxígeno/metabolismo , Ácido Fítico/farmacología , Dominios y Motivos de Interacción de Proteínas , Estructura Cuaternaria de ProteínaRESUMEN
BACKGROUND: The Consumo Alimentar e Atividade Fisica de Escolares (CAAFE) questionnaire is an online research tool that has been developed to enable the self-report of physical activity and diet by Brazilian schoolchildren aged 7-10 years. Formative research was conducted with nutritionists during the development of the web-based questionnaire. The suggestions and insights obtained were used to design a tool to monitor schoolchildren's food consumption based on the concept of healthy and unhealthy food indicators. The present study aimed to report the focus group discussions conducted with nutritionists concerning the CAAFE questionnaire. METHODS: Focus group discussions were conducted using a semi-structured questionnaire, and these were then analysed thematically. RESULTS: Twenty-four nutritionists participated (four focus groups; average per group: six people); the majority (n = 22) had experience with 7-10-year-old children. Four themes emerged: (i) healthy and unhealthy food indicators; (ii) suggestions for the online instrument; (iii) potential applications; and (iv) challenges for its construction. CONCLUSIONS: Comments made by nutritionists enabled the construction of an instrument that is able to answer questions related to food consumption in schools and at home.
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Dieta , Conducta Alimentaria , Evaluación Nutricional , Nutricionistas , Encuestas y Cuestionarios , Adulto , Brasil , Niño , Grupos Focales , Humanos , Internet , Masculino , Persona de Mediana Edad , Competencia Profesional , Investigación CualitativaRESUMEN
Sickle cell retinopathy (SCR) develops in up to 30% of sickle cell disease patients (SCD) during the second decade of life. Treatment for this affection remains palliative, so studies on its pathophysiology may contribute to the future development of novel therapies. SCR is more frequently observed in hemoglobin SC disease and derives from vaso-occlusion in the microvasculature of the retina leading to neovascularization and, eventually, to blindness. Circulating inflammatory cytokines, angiogenic factors, and their interaction may contribute to the pathophysiology of this complication. Angiopoietin (Ang)-1, Ang-2, soluble vascular cell adhesion molecule-1, intercellular adhesion molecule (ICAM)-1, E-selectin, P-selectin, IL1-ß, TNF-α, pigment epithelium derived factor (PEDF) and vascular endothelial growth factor plasmatic levels were determined in 37 SCD patients with retinopathy, 34 without retinopathy, and healthy controls. We observed that sICAM-1 is significantly decreased, whereas PEDF is elevated in HbSC patients with retinopathy (P=0.012 and P=0.031, respectively). Ang-1, Ang-2 and IL1-ß levels were elevated in SCD patients (P=0.001, P<0.001 and P=0.001, respectively), compared to controls, and HbSS patients presented higher levels of Ang-2 compared to HbSC (P<0.001). Our study supports the possible influence of sICAM-1 and PEDF on the pathophysiology of retinal neovascularization in SCD patients.
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Anemia de Células Falciformes/sangre , Proteínas del Ojo/sangre , Molécula 1 de Adhesión Intercelular/sangre , Factores de Crecimiento Nervioso/sangre , Neovascularización Retiniana/sangre , Serpinas/sangre , Adulto , Anemia de Células Falciformes/complicaciones , Angiopoyetina 1/sangre , Angiopoyetina 2/sangre , Estudios de Casos y Controles , Femenino , Humanos , Interleucina-1beta/sangre , Masculino , Persona de Mediana Edad , Neovascularización Retiniana/etiología , Factor de Necrosis Tumoral alfa/sangreRESUMEN
O objetivo deste trabalho foi avaliar a variação do perfil proteico e do cálcio solúvel na coagulação do leite pelo etanol nas temperaturas de 4ºC, 10ºC, 15ºC e 20ºC. Amostras de leite de 61 animais foram avaliadas quanto à estabilidade ao etanol nas concentrações de 66 a 92 por cento (v/v) nas temperaturas de 4ºC, 10ºC, 15ºC e 20ºC. Três amostras, após 24 horas de armazenamento a 4ºC, foram ultracentrifugadas em quadruplicata (40.000 x g) a 4ºC e a 20ºC, respectivamente, por 60 minutos. Em seguida, o sobrenadante foi retirado e submetido à análise do cálcio solúvel pela técnica via úmida (digestão nitroperclórica) e leitura em espectrofotômetro de absorção atômica. O perfil proteico foi analisado pela técnica de eletroforese capilar empregando kit específico para determinação proteica. Os resultados mostraram uma correlação positiva entre o aumento da temperatura das amostras e a estabilidade do leite frente às diferentes concentrações de etanol. A porcentagem de cálcio solúvel no sobrenadante após ultracentrifugação foi maior nas amostras tratadas a 4ºC (P<0,05). As amostras ultracentrifugadas na temperatura de 4ºC apresentaram quantidades superiores de β-caseína no sobrenadante em comparação com as amostras tratadas a 20ºC. O abaixamento da temperatura favoreceu a migração da β-caseína e do cálcio coloidal para a fase solúvel do leite, o que possivelmente favoreceu o aumento da instabilidade das amostras no teste do etanol. Os resultados sugerem que a temperatura ideal para a realização de teste de estabilidade do leite frente ao etanol deveria ser de 21ºC...
The aim of this study was to evaluate the variation in protein profile and soluble calcium in milk coagulation by ethanol at 4ºC, 10ºC, 15ºC and 20ºC. Milk samples from 61 dairy cows were evaluated for stability of ethanol concentrations from 66 to 92 percent (v/v) at temperatures of 4°C, 10°C, 15°C and 20°C. Three samples were ultracentrifuged (40,000 x g) after 24 hours of storage at 4°C and 20°C, respectively, for 60 minutes. Their supernatants were removed and subjected to analyses of soluble calcium through nitro-perchloric digestion and atomic absorption spectrophotometry. The protein profiles were determined by capillary electrophoresis using a specific kit for protein determination. The results showed a positive correlation between the increase in temperature of the samples and the stability of milk against various concentrations of ethanol. The percentage of soluble calcium in the supernatant after centrifugation was higher in samples treated at 4°C (P<0.05). The samples ultracentrifuged at 4°C showed higher amounts of β-casein in the supernatant compared with samples stored at 20°C. The lowering of the temperature favored the migration of β-casein and colloidal calcium to the soluble phase of milk, which may also have favored the instability of milk in the ethanol test. According to the results, the milk sample temperature for the ethanol stability test should be 21ºC...
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Animales , Calcio/química , Etanol/efectos adversos , Proteínas de la Leche/química , Ultracentrifugación , Leche/metabolismo , Temperatura de TransiciónRESUMEN
The purpose of this study was to test the precision and accuracy of three-dimensional (3D) linear measurements for Le Fort I osteotomy, obtained from multi-slice computed tomography (MSCT) and cone beam computed tomography (CBCT) scans. The study population consisted of 11 dried skulls submitted to 64-row MSCT and CBCT scans. Three-dimensional reconstructed images (3D-CT) were generated, and linear measurements (n=11) based on anatomical structures and landmarks of interest for Le Fort I osteotomy were performed independently by two oral and maxillofacial radiologists, twice each, using Vitrea software; this allows true 3D measurement on 3D-CT images. The results demonstrated no statistically significant differences between the inter-examiner and intra-examiner analyses, and physical and true 3D linear measurements using MSCT and CBCT images. Regarding examiner accuracy, no statistically significant differences were found for the comparisons among the physical and the MSCT and the CBCT linear measurements by either examiner. For examiners 1 and 2, the analysis intra-examiner correlation coefficient ranged from 0.87 to 0.96 and 0.82 to 0.98, respectively, using MSCT, and from 0.84 to 0.98 and 0.80 to 0.98, respectively, using CBCT, indicating almost perfect agreement for all analyses performed. 3D linear measurements obtained from MSCT and CBCT images were considered precise and accurate for Le Fort I osteotomy and thus accurate and helpful for Le Fort I osteotomy planning.
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Tomografía Computarizada de Haz Cónico , Imagenología Tridimensional , Osteotomía Le Fort , Cráneo/diagnóstico por imagen , Adulto , Cadáver , Femenino , Humanos , Masculino , Persona de Mediana Edad , Interpretación de Imagen Radiográfica Asistida por Computador , Tomografía Computarizada por Rayos XRESUMEN
OBJECTIVE: The purpose of this study was to compare the precision and accuracy of linear measurements for Le Fort I osteotomy performed by two different imaging software programs and obtained from three-dimensional cone beam CT (3D-CBCT) images. METHODS: The study population consisted of 11 dried skulls submitted to CBCT, which generated 3D images. Linear measurements were based on craniometric anatomical landmarks pre-defined by the authors as specifically used for Le Fort I osteotomy and were identified by two radiologists twice each, independently, using Vitrea 3.8.1 (Vital Images Inc., Plymouth, MN) and open-source digital imaging communication in medicine viewer OsiriX 1.2 64-bit (Pixmeo, Geneva, Switzerland). Subsequently, a third examiner made physical measurements using a digital caliper (167 series; Mitutoyo Sul Americana Ltd, Suzano, SP, Brazil). RESULTS: The results demonstrated a statistically significant difference between OsiriX and the gold standard, especially in the pterygoid process (TPtg L = 0.019, LLpPtg R = 0.016 and LLpPtg L = 0.012). Vitrea showed no statistical difference in comparison with the gold standard, and showed a high level of accuracy in all the measurements performed. The major difference found was 0.42 mm (LLpPtg R). Interexaminer analysis ranged from 0.90 to 0.97 using Vitrea and from 0.8 to 0.97 using OsiriX. Intraexaminer correlation coefficient ranged from 0.90 to 0.98 and from 0.84 to 0.98 for Examiners 1 and 2, respectively, using Vitrea and from 0.93 to 0.99 for Examiner 1 and from 0.64 to 0.96 for Examiner 2 using OsiriX. CONCLUSION: Vitrea may be considered as precise and accurate, insofar as it was able to perform all the 3D linear measurements. On the other hand, linear measurements performed using OsiriX were not successful in producing accurate linear measurements for Le Fort I osteotomy.
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Tomografía Computarizada de Haz Cónico , Precisión de la Medición Dimensional , Procesamiento de Imagen Asistido por Computador , Maxilar/diagnóstico por imagen , Osteotomía Le Fort , Adulto , Cadáver , Tomografía Computarizada de Haz Cónico/normas , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador/normas , Imagenología Tridimensional/métodos , Imagenología Tridimensional/normas , Masculino , Persona de Mediana Edad , Planificación de Atención al Paciente , Cráneo/diagnóstico por imagen , Programas Informáticos , Hueso Esfenoides/diagnóstico por imagen , Adulto JovenRESUMEN
We describe here a new frameshift mutation of ß-thalassemia in a Uruguayan family with Italian ancestry [ß48 (-T); HBB:c.146delT]. This frameshift results in formation of premature stop codon (TGA) 40 bp downstream and in a short unstable product that is degraded in the cell.
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Salud de la Familia , Mutación del Sistema de Lectura , Globinas beta/genética , Talasemia beta/genética , Adulto , Codón sin Sentido , Exones , Femenino , Eliminación de Gen , Heterocigoto , Humanos , Italia , Linaje , Estabilidad Proteica , Uruguay , Población Blanca , Globinas beta/análisis , Globinas beta/metabolismo , Talasemia beta/sangre , Talasemia beta/metabolismoRESUMEN
BCR-ABL-negative myeloproliferative neoplasms (MPNs) are most frequently characterized by the JAK2V617F gain-of-function mutation, but several studies showed that JAK2V617F may not be the initiating event in MPN development, and recent publications indicate that additional alterations such as chromatin modification and microRNA (miRNA) deregulation may have an important role in MPN pathogenesis. Here we report that 61 miRNAs were significantly deregulated in CD34+ cells from MPN patients compared with controls (P<0.01). Global miRNA analysis also revealed that polycythemia vera (JAKV617F) and essential thrombocythemia (JAK2 wild type) patients have significantly different miRNA expression profiles from each other. Among the deregulated miRNAs, expression of miR-134, -214 and -433 was not affected by changes in JAK2 activity, suggesting that additional signaling pathways are responsible for the deregulation of these miRNAs in MPN. Despite its upregulation in MPN CD34+ and during normal erythropoiesis, both overexpression and knockdown studies suggest that miR-433 negatively regulates CD34+ proliferation and differentiation ex vivo. Its novel target GBP2 is downregulated during normal erythropoiesis and regulates proliferation and erythroid differentiation in TF-1 cells, indicating that miR-433 negatively regulates hematopoietic cell proliferation and erythropoiesis by directly targeting GBP2.
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Biomarcadores de Tumor/genética , Diferenciación Celular , Proliferación Celular , Células Eritroides/citología , MicroARNs/genética , Trastornos Mieloproliferativos/genética , Antígenos CD34/metabolismo , Células Cultivadas , Células Eritroides/metabolismo , Eritropoyesis/fisiología , Proteínas de Unión al GTP/antagonistas & inhibidores , Proteínas de Unión al GTP/genética , Proteínas de Unión al GTP/metabolismo , Perfilación de la Expresión Génica , Humanos , Janus Quinasa 2/genética , Luciferasas/metabolismo , Mutación/genética , Trastornos Mieloproliferativos/metabolismo , Trastornos Mieloproliferativos/patología , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Mensajero/genética , ARN Interferente Pequeño/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Phosphatidylinositol phosphate kinases (PIPKs) are enzymes that participate in diverse intracellular signaling pathways. They are classified into 3 functionally distinct subfamilies - PIPKI (α, ß, γ), PIPKII (α, ß, γ), and PIPKIII - located in various subcellular compartments. Recently, the PIPKIIα and ß-globin genes were found to be overexpressed in reticulocytes from 2 siblings with hemoglobin H disease, suggesting a possible relationship between PIPKIIa and the production of globins. The main aim of this study was to determine the expression profiles of PIPK genes in healthy individuals during in vitro erythropoiesis using quantitative real-time polymerase chain reaction and to compare these profiles with profiles of globin genes. Our results showed that expression of all PIPKs increases as the cells differentiate, coinciding with the expression profiles of globins. Analysis of the effects of globins on PIPK genes revealed that they varied significantly between the globins, the most noticeable being the effect of α-globin on PIPKIIα (P < 0.0001) and γ-globin on PIPKIIγ (P < 0.0001). The relationship between the expression of PIPKs and globin genes was statistically significant, particularly between PIPKIIα and α-globin (P = 0.0002) and PIPKIIγ and ß-globin (P < 0.0001). Linear correlation analysis revealed a strong relationship between PIPKIIα and α-globin genes. This study is the first to establish the expression profiles of PIPK genes during in vitro erythropoiesis in healthy individuals and suggests a parallel between the expression of PIPK and globin genes, reinforcing the hypothesis that they may be related.