RESUMEN

Increasing fetal hemoglobin (HbF) provides clinical benefit in patients with sickle cell disease (SCD). We recently identified heme-regulated inhibitor (HRI, EIF2AK1), as a novel HbF regulator. Because HRI is an erythroid-specific protein kinase, it presents a potential target for pharmacologic intervention. We found that maximal HbF induction required >80% to 85% HRI depletion. Because it remains unclear whether this degree of HRI inhibition can be achieved pharmacologically, we explored whether HRI knockdown can be combined with pharmacologic HbF inducers to achieve greater HbF production and minimize potential adverse effects associated with treatments. Strongly cooperative HbF induction was observed when HRI depletion was combined with exposure to pomalidomide or the EHMT1/2 inhibitor UNC0638, but not to hydroxyurea. Mechanistically, reduction in the levels of the HbF repressor BCL11A reflected the cooperativity of HRI loss and pomalidomide treatment, whereas UNC0638 did not modulate BCL11A levels. In conjunction with HRI loss, pomalidomide maintained its HbF-inducing activity at 10-fold lower concentrations, in which condition there were minimal observed detrimental effects on erythroid cell maturation and viability, as well as fewer alterations in the erythroid transcriptome. When tested in cells from patients with SCD, combining HRI depletion with pomalidomide or UNC0638 achieved up to 50% to 60% HbF and 45% to 50% HbF, respectively, as measured by high-performance liquid chromatography, and markedly counteracted cell sickling. In summary, this study provides a foundation for the exploration of combining future small-molecule HRI inhibitors with additional pharmacologic HbF inducers to maximize HbF production and preserve erythroid cell functionality for the treatment of SCD and other hemoglobinopathies.

Asunto(s)

Anemia de Células Falciformes , Hemoglobina Fetal , Anemia de Células Falciformes/tratamiento farmacológico , Eritrocitos Anormales , Células Eritroides , Hemoglobina Fetal/genética , Humanos , Hidroxiurea/farmacología

RESUMEN

BACKGROUND AND PURPOSE: Leukoerythroblastic reaction (LER) is characterized by the presence of immature erythroid cells and myeloid precursors (metamyelocytes, myelocytes, promyelocytes, myeloblasts, and blasts) as well as, exclusively in myelofibrotic disorders, teardrop cells in the peripheral blood (J Pathol Bacteriol, 42, 1936, 541; Semaine Med, 22, 1902, 373). Research on how to interpret LER and its meaning in clinical practice is scarce, and there is no consensus on the diagnostic criteria. We summarize the current evidence with the aim of clarifying the knowledge on this subject. METHODS: We conducted a comprehensive search of the PubMed-MEDLINE, EMBASE and ELSEVIER databases, the Cochrane Library, Google Scholar, and medical journals to identify relevant papers. RESULTS: Our search identified 425 papers, of which, 35 (11 trials and 24 case reports) ultimately met the inclusion criteria. These showed two principal groups of diseases associated with leukoerythroblastosis (LEB), corresponding to solid and hematological malignancies. The other etiologies, in order of frequency, were hemolytic diseases, infection, and others, while hemorrhage was only reported in the trials group. CONCLUSION: The literature on LER is scarce and heterogeneous. The etiological factors of LER are diverse, and its presence in malignant disease is an indicator of disease progression and an adverse prognosis suggesting poor survival. In those cases where LER had neither hematological nor solid neoplasms, its manifestation, prognosis and its impact on our daily clinical practice are unknown.

Asunto(s)

Susceptibilidad a Enfermedades , Células Eritroides/patología , Leucocitos/patología , Trastornos Mieloproliferativos/diagnóstico , Trastornos Mieloproliferativos/etiología , Animales , Manejo de la Enfermedad , Humanos , Anamnesis , Trastornos Mieloproliferativos/terapia

RESUMEN

Reversing the developmental switch from fetal hemoglobin (HbF, α2γ2) to adult hemoglobin (HbA, α2ß2) is an important therapeutic approach in sickle cell disease (SCD) and ß-thalassemia. In healthy individuals, SCD patients, and patients treated with pharmacologic HbF inducers, HbF is present only in a subset of red blood cells known as F cells. Despite more than 50 years of observations, the cause for this heterocellular HbF expression pattern, even among genetically identical cells, remains unknown. Adult F cells might represent a reversion of a given cell to a fetal-like epigenetic and transcriptional state. Alternatively, isolated transcriptional or posttranscriptional events at the γ-globin genes might underlie heterocellularity. Here, we set out to understand the heterogeneity of HbF activation by developing techniques to purify and profile differentiation stage-matched late erythroblast F cells and non-F cells (A cells) from the human HUDEP2 erythroid cell line and primary human erythroid cultures. Transcriptional and proteomic profiling of these cells demonstrated very few differences between F and A cells at the RNA level either under baseline conditions or after treatment with HbF inducers hydroxyurea or pomalidomide. Surprisingly, we did not find differences in expression of any known HbF regulators, including BCL11A or LRF, that would account for HbF activation. Our analysis shows that F erythroblasts are not significantly different from non-HbF-expressing cells and that the primary differences likely occur at the transcriptional level at the ß-globin locus.

Asunto(s)

Eritroblastos/metabolismo , Hemoglobina Fetal/biosíntesis , Hemoglobina A/metabolismo , Adulto , Anemia de Células Falciformes/sangre , Anemia de Células Falciformes/genética , Línea Celular , Separación Celular/métodos , Células Cultivadas , Eritroblastos/clasificación , Eritroblastos/efectos de los fármacos , Células Eritroides/clasificación , Células Eritroides/metabolismo , Hemoglobina Fetal/genética , Perfilación de la Expresión Génica , Hemoglobina A/genética , Humanos , Hidroxiurea/farmacología , Proteómica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Talidomida/análogos & derivados , Talidomida/farmacología

RESUMEN

OBJECTIVE: To validate multilineage score system correlating results of flow cytometry, cytogenetics, cytomorphology and histology from samples of patients with suspected myelodysplastic syndrome or cytopenia of unknown origin. METHODS: A retrospective study analyzing laboratory data of 49 patients with suspected myelodysplastic syndrome or cytopenia of unknown origin, carried out between May and September 2017. The inclusion criteria were availability of flow cytometry results, and at least one more method, such as morphology, histology or cytogenetics. Thirty-eight patients were classified as diagnosis of myelodysplastic syndromes, whereas 11 were classified as normal. Patients were evaluated based on score systems, Ogata score and flow cytometry multilineage score. RESULTS: Comparing the scores obtained in the Ogata score and the multilineage score, it was observed that in four cases the Ogata score was zero or 1 point, while the multilineage score was higher than 3 points. In addition, in 12 cases with Ogata score of 2, the multilineage score was greater than 3. CONCLUSION: The flow cytometry multilineage score system demonstrated to be more effective in dysplasia analysis, by assessing the erythroid, monocytic, granulocytic and precursor cell lineages, apart from the parameters evaluated by the Ogata score.

Asunto(s)

Citometría de Flujo/normas , Síndromes Mielodisplásicos/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Células de la Médula Ósea/patología , Niño , Preescolar , Análisis Citogenético/métodos , Análisis Citogenético/normas , Células Eritroides/patología , Femenino , Citometría de Flujo/métodos , Granulocitos/patología , Humanos , Lactante , Masculino , Persona de Mediana Edad , Monocitos/patología , Estándares de Referencia , Reproducibilidad de los Resultados , Estudios Retrospectivos , Adulto Joven

RESUMEN

ABSTRACT Objective To validate multilineage score system correlating results of flow cytometry, cytogenetics, cytomorphology and histology from samples of patients with suspected myelodysplastic syndrome or cytopenia of unknown origin. Methods A retrospective study analyzing laboratory data of 49 patients with suspected myelodysplastic syndrome or cytopenia of unknown origin, carried out between May and September 2017. The inclusion criteria were availability of flow cytometry results, and at least one more method, such as morphology, histology or cytogenetics. Thirty-eight patients were classified as diagnosis of myelodysplastic syndromes, whereas 11 were classified as normal. Patients were evaluated based on score systems, Ogata score and flow cytometry multilineage score. Results Comparing the scores obtained in the Ogata score and the multilineage score, it was observed that in four cases the Ogata score was zero or 1 point, while the multilineage score was higher than 3 points. In addition, in 12 cases with Ogata score of 2, the multilineage score was greater than 3. Conclusion The flow cytometry multilineage score system demonstrated to be more effective in dysplasia analysis, by assessing the erythroid, monocytic, granulocytic and precursor cell lineages, apart from the parameters evaluated by the Ogata score.

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RESUMO Objetivo Validar ficha de escore multilinhagem correlacionando resultados obtidos de citometria de fluxo, citogenética, citomorfologia e histologia de amostras de pacientes com suspeita de síndrome mielodisplásica ou citopenias a esclarecer. Métodos Estudo retrospectivo de análise de dados laboratoriais de 49 pacientes com suspeita clínica de síndrome mielodisplásica ou citopenias a esclarecer realizado entre maio e setembro de 2017. Os critérios de inclusão foram a disponibilidade de resultados de citometria de fluxo e de, pelo menos, outra metodologia, entre morfologia, histologia, ou citogenética. Trinta e oito pacientes foram classificados como diagnosticados com síndromes mielodisplásicas enquanto 11 foram classificados como normais. Os pacientes foram avaliados utilizando sistemas de escore, escore de Ogata e ficha multilinhagem. Resultados Comparando as pontuações obtidas no escore de Ogata e na ficha multilinhagem, observou-se que, em quatro casos, o score de Ogata foi zero ou 1 ponto, enquanto, pela ficha multilinhagem, a pontuação foi superior a 3 pontos. Além disso, em 12 casos com escore de Ogata 2, a pontuação pela ficha multilinhagem foi superior a 3. Conclusão A ficha multilinhagem demonstrou ser mais eficaz na análise de displasia por avaliar as linhagens eritroide, monocítica, granulocítica e células precursoras, além dos parâmetros avaliados no escore de Ogata.

Asunto(s)

Humanos , Masculino , Femenino , Lactante , Preescolar , Niño , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Adulto Joven , Síndromes Mielodisplásicos/patología , Citometría de Flujo/normas , Estándares de Referencia , Biopsia , Células de la Médula Ósea/patología , Monocitos/patología , Reproducibilidad de los Resultados , Estudios Retrospectivos , Análisis Citogenético/métodos , Análisis Citogenético/normas , Células Eritroides/patología , Citometría de Flujo/métodos , Granulocitos/patología , Persona de Mediana Edad

RESUMEN

Beta-thalassaemia (BT) is classified according to blood transfusion requirement as minor (BTMi), intermedia (BTI) and major (BTM). BTM is the most severe form, requiring regular transfusions while transfusion need is only occasional in BTI. Differential gene expression between patients has not been assessed so far. Here, we evaluated the global gene expression profiles during differentiation of human erythroid cells of two patients carrying the same mutation [CD39, (C → T)], though displaying different phenotypes (BTI and BTM). Considering the role of reactive oxygen species (ROS) in the pathophysiology of thalassaemia, we focused on differentially expressed genes involved in metabolic pathways triggered by ROS, such as inflammation and apoptosis, and, from these, we selected the Apurinic/Apyrimidinic Endodeoxyribonuclease 1 (APEX1) and High Mobility Group Box1 (HMGB1) genes, whose role in BT is not well established. An in-depth expression analysis of transcriptional and protein levels in patients carrying a range of mutations associated with BT phenotypes indicated that APEX1 was increased in both BTI and BTM. Furthermore, higher amounts of HMGB1 was found in the plasma of BTI patients. Our findings suggest that these proteins have important roles in BT and could represent new targets for further studies aiming to improve the management of the disease.

Asunto(s)

ADN-(Sitio Apurínico o Apirimidínico) Liasa/genética , Proteína HMGB1/genética , Estrés Oxidativo , Transducción de Señal , Transcriptoma , Talasemia beta/genética , Talasemia beta/metabolismo , Adulto , Apoptosis , Apirasa/metabolismo , Biomarcadores , Estudios de Casos y Controles , Diferenciación Celular/genética , Biología Computacional/métodos , ADN-(Sitio Apurínico o Apirimidínico) Liasa/metabolismo , Células Eritroides/citología , Células Eritroides/metabolismo , Femenino , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Redes Reguladoras de Genes , Proteína HMGB1/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Fenotipo , Especies Reactivas de Oxígeno/metabolismo , Talasemia beta/diagnóstico

RESUMEN

Reactivation of fetal hemoglobin (HbF) production benefits patients with sickle cell disease and ß-thalassemia. To identify new HbF regulators that might be amenable to pharmacologic control, we screened a protein domain-focused CRISPR-Cas9 library targeting chromatin regulators, including BTB domain-containing proteins. Speckle-type POZ protein (SPOP), a substrate adaptor of the CUL3 ubiquitin ligase complex, emerged as a novel HbF repressor. Depletion of SPOP or overexpression of a dominant negative version significantly raised fetal globin messenger RNA and protein levels with minimal detrimental effects on normal erythroid maturation, as determined by transcriptome and proteome analyses. SPOP controls HbF expression independently of the major transcriptional HbF repressors BCL11A and LRF. Finally, pharmacologic HbF inducers cooperate with SPOP depletion during HbF upregulation. Our study implicates SPOP and the CUL3 ubiquitin ligase system in controlling HbF production in human erythroid cells and may offer new therapeutic strategies for the treatment of ß-hemoglobinopathies.

Asunto(s)

Células Eritroides/metabolismo , Hemoglobina Fetal/genética , Proteínas Nucleares/metabolismo , Proteínas Represoras/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Adulto , Femenino , Humanos , Masculino , Proteínas Nucleares/genética , Proteínas Represoras/genética , Adulto Joven

RESUMEN

ß-thalassemia is a worldwide distributed monogenic red cell disorder, characterized by an absent or reduced beta globin chain synthesis. The unbalance of alpha-gamma chain and the presence of pathological free iron promote severe oxidative damage, playing crucial a role in erythrocyte hemolysis, exacerbating ineffective erythropoiesis and decreasing the lifespan of red blood cells (RBC). Catalase, glutathione peroxidase and peroxiredoxins act together to protect RBCs from hydrogen peroxide insult. Among them, peroxiredoxins stand out for their overall abundance and reactivity. In RBCs, Prdx2 is the third most abundant protein, although Prdxs 1 and 6 isoforms are also found in lower amounts. Despite the importance of these enzymes, Prdx1 and Prdx2 may have their peroxidase activity inactivated by hyperoxidation at high hydroperoxide concentrations, which also promotes the molecular chaperone activity of these proteins. Some studies have demonstrated the importance of Prdx1 and Prdx2 for the development and maintenance of erythrocytes in hemolytic anemia. Now, we performed a global analysis comparatively evaluating the expression profile of several antioxidant enzymes and their physiological reducing agents in patients with beta thalassemia intermedia (BTI) and healthy individuals. Furthermore, increased levels of ROS were observed not only in RBC, but also in neutrophils and mononuclear cells of BTI patients. The level of transcripts and the protein content of Prx1 were increased in reticulocyte and RBCs of BTI patients and the protein content was also found to be higher when compared to beta thalassemia major (BTM), suggesting that this peroxidase could cooperate with Prx2 in the removal of H2O2. Furthermore, Prdx2 production is highly increased in RBCs of BTM patients that present high amounts of hyperoxidized species. A significant increase in the content of Trx1, Srx1 and Sod1 in RBCs of BTI patients suggested protective roles for these enzymes in BTI patients. Finally, the upregulation of Nrf2 and Keap1 transcription factors found in BTI patients may be involved in the regulation of the antioxidant enzymes analyzed in this work.

Asunto(s)

Células Eritroides/metabolismo , Peroxirredoxinas/metabolismo , Talasemia beta/metabolismo , Talasemia beta/patología , Adolescente , Adulto , Western Blotting , Niño , Preescolar , Eritrocitos/citología , Eritrocitos/metabolismo , Femenino , Humanos , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/metabolismo , Masculino , Persona de Mediana Edad , Neutrófilos/citología , Neutrófilos/metabolismo , Oxidación-Reducción , Peroxirredoxinas/genética , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Adulto Joven

RESUMEN

CONTEXT AND OBJECTIVE: Microcytic anaemia results from defective synthesis of haemoglobin in the erythroid precursors, causing a reduction in its mean corpuscular volume (MCV). The most common causes of microcytosis, without the increase in HbA2 levels, are iron deficiency anaemia (IDA) and α-thalassemia. The aim of this study was to identify the causes of microcytic anaemia and evaluate the haematological parameters from blood donors deemed ineligible (due to the low haematocrit level) that would differentiate the IDA and α-thal, whether isolated or in association. METHODS: Genomic DNA was submitted to the polymerase chain reaction multiplex for the diagnosis of the most common allele deletions of α-thal and erythrogram and in order to verify haematological parameters. Iron deficiency (ID) was determined through the measurement of serum ferritin. RESULTS: Of the 204 samples, 82 (40.2%) were identified with ID, 24 (17.8%) with α-thal and 10 (4.9%) with ID associated with α-thal. In the α-thal with ID group haemoglobin (Hb), MCV, mean corpuscular Hb concentration (MCHC) and mean corpuscular Hb (MCH) values were significantly lower compared to the isolated α-thal. In the group with ID Hb, MCV, MCHC and MCH values were significantly lower compared to those with isolated α-thal. The α-thal with ID group, showed Hb, MCV, MCHC and MCH significantly reduced when compared to those with IDA. CONCLUSIONS: This study showed that the values of haematological parameters, especially haematocrit, Hb, MCV, MCH, MCHC and red blood cell distribution width (RDW), are lower in patients with IDA, especially when associated with α-thal and therefore it may be useful to discriminate between the different types of microcytic anaemia.

Asunto(s)

Anemia/sangre , Donantes de Sangre , Índices de Eritrocitos , Células Eritroides/metabolismo , Hemoglobina A2/metabolismo , Adulto , Células Eritroides/patología , Femenino , Hematócrito , Humanos , Masculino

RESUMEN

If cultured in appropriate conditions, such as supplementing culture media with costly cytokines and growth factors, hematopoietic stem/progenitor cells (HSPCs) from different origins have shown to be an adequate source of erythroid cells. This requirement turns erythroid cells production into a complicated process to be scaled-up for future applications. The aim of our work was to genetically modify HSPCs with human erythropoietin (hEPO) sequence by lentiviral transgenesis in order for cells to secrete the hormone into the culture medium. Initially, we evaluated erythroid differentiation in colony forming units (CFU) assays and further analyzed cell expansion and erythroid differentiation throughout time in suspension cultures by flow cytometry and May-Grünwald-Giemsa staining. Additionally, we studied hEPO production and its isoforms profile. The different assessment approaches demonstrated erythroid differentiation, which was attributed to the hEPO secreted by the HSPCs. Our data demonstrate that it is possible to develop culture systems in which recombinant HSPCs are self-suppliers of hEPO. This feature makes our strategy attractive to be applied in biotechnological production processes of erythroid cells that are currently under development.

Asunto(s)

Técnicas de Cultivo de Célula/métodos , Diferenciación Celular/genética , Células Eritroides/citología , Eritropoyetina/genética , Eritropoyetina/metabolismo , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/metabolismo , Lentivirus/genética , Biotecnología/métodos , Células Cultivadas , Ensayo de Unidades Formadoras de Colonias , Células Eritroides/metabolismo , Eritropoyetina/biosíntesis , Eritropoyetina/química , Humanos , Lentivirus/metabolismo

Asunto(s)

Médula Ósea/parasitología , Células Eritroides/metabolismo , Malaria Vivax/patología , MicroARNs/metabolismo , Plasmodium vivax/aislamiento & purificación , Humanos , Malaria Vivax/parasitología , Masculino , MicroARNs/genética , Persona de Mediana Edad , Transcriptoma

RESUMEN

Porcine reproductive and respiratory syndrome viruses (PRRSV) present a wide phenotypic and genetic diversity. Experimental infections have demonstrated viral replication, including highly pathogenic strains (HP-PRRSV), in primary lymphoid organs such as the thymus. However, studies of the bone marrow are scarce but necessary to help elucidate the immunobiology of PRRSV strains of differing virulence. In this study, whereas viral RNA was detected within the bone marrow of animals experimentally infected with both low virulent Lelystad (LV) and 215-06 PRRSV-1 strains and with the highly virulent SU1-bel strain, PRRSV positive cells were only occasionally detected in one SU1-bel infected animal. PRRSV RNA levels were associated to circulating virus with the highest levels detected in LV-infected pigs. At 3 dpi, a decrease in the proportion of haematopoietic tissue and number of erythroid cells in all infected groups was associated with an increase in TUNEL or cleaved caspase 3 labelling and higher counts of myeloid cells compared to control. The expression of IL-1α and IL-6 was elevated at the beginning of the infection in all infected animals. The expression of TNF-α was increased at the end of the study in all infected groups with respect to control. Different PRRSV-1 strains induced, presummably by indirect mechanisms and independently of viral load and strain virulence, moderate and sustained hypoplasia of erythroid cells and myeloid cell hyperplasia at early stages of infection. These changes were paralleled by a peak in the local expression of IL-1α, IL-6 and TNF-α in all infected groups.

Asunto(s)

Síndrome Respiratorio y de la Reproducción Porcina/patología , Virus del Síndrome Respiratorio y Reproductivo Porcino/patogenicidad , Animales , Médula Ósea/patología , Médula Ósea/virología , Citocinas/inmunología , Células Eritroides/patología , Células Eritroides/virología , Hiperplasia/patología , Hiperplasia/veterinaria , Hiperplasia/virología , Masculino , Células Mieloides/patología , Células Mieloides/virología , Síndrome Respiratorio y de la Reproducción Porcina/virología , Virus del Síndrome Respiratorio y Reproductivo Porcino/fisiología , Porcinos , Carga Viral , Virulencia , Replicación Viral

RESUMEN

Acute erythroid leukemia (AML M6) is a hematopoietic neoplasm frequently described in cats and mice, rarely in other animal species. This report describes a case of AML M6 in a yearling Thoroughbred filly. Clinically the horse presented marked pale mucous membranes and exercise intolerance. In addition, the owner and referring veterinarian reported a 30-day history of progressive weight loss. The CBC revealed severe anemia and leukopenia by neutropenia. Cytology evaluation obtained from bone marrow fine needle aspirates evidenced inversion of the myeloid: erythroid ratio (0.2), with 48% of the nucleated cells corresponding to rubriblasts. In addition to the gross evidence of anemia, necropsy findings consisted of splenomegaly and lymphadenomegaly. The diagnosis of AML M6B was confirmed histologically due to splenomegaly and lymphadenomegaly, secondary to neoplastic metastasization.(AU)

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Leucemia eritroide aguda (LMA M6) é um neoplasia hematopoiética descrita com frequência em gatos e camundongos, mas raramente nas demais espécies de animais. Esse relato descreve um caso dessa doença em um equino, fêmea, Puro Sangue Inglês de um ano de idade. Clinicamente, o equino apresentou acentuada palidez das mucosas, intolerância ao exercício e o proprietário relatava emagrecimento progressivo. No hemograma, foi observada anemia grave e leucopenia por neutropenia. No mielograma, havia inversão da relação mieloide:eritroide (0,2) e 48% das células nucleadas da medula óssea correspondiam a rubriblastos. Na necropsia, além de evidências de anemia, havia esplenomegalia e linfadenomegalia. O diagnóstico de LMA M6B foi confirmado histologicamente e o aumento do baço e dos linfonodos foi decorrente de metastatização.(AU)

Asunto(s)

Animales , Femenino , Enfermedades de los Caballos , Leucemia Eritroblástica Aguda/diagnóstico , Leucemia Eritroblástica Aguda/veterinaria , Células Eritroides/patología

RESUMEN

La implementación de nuevas tecnologías y la ampliación de la cobertura a nivel de imágenes, han facilitado encontrar casos incidentales, siendo el más común el adenoma suprarrenal no funcionante. El mielolipoma suprarrenal es una neoplasia benigna, rara, en la mayoría de casos asintomática compuesta por tejido adiposo maduro y elementos hematopoyéticos. Se presenta un caso clínico de una paciente remitida al servicio de radiología por presentar en ecografía previa, una masa en región suprarrenal derecha incidentalmente. La paciente se encontraba asintomática, por lo cual se realiza tomografía axial computarizada abdominal. A la paciente por su ausencia de síntomas se le continúo con seguimiento periódico sin detectar cambios evolutivos (MÉD.UIS. 2014;27(2):105-107) .

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The implementation of new technologies and expanding coverage to level of images, have provided incidental finding cases, the most common nonfunctioning adrenal adenoma. Myelolipoma adrenal benign tumors, rarely, in most cases consisting of asymptomatic mature adipose tissue and hematopoietic elements (myeloid and erythroid cells). We present a case of a patient referred to the radiology department for filing in previous ultrasound a right adrenal mass incidentally region. The patient was asymptomatic, which is done by abdominal CT. A patient by the absence of symptoms is continued with regular monitoring undetected evolutionary changes (MÉD.UIS. 2014;27(2):105-107).

Asunto(s)

Humanos , Femenino , Persona de Mediana Edad , Radiología , Células Eritroides , Tomografía , Ultrasonografía , Hallazgos Incidentales , Neoplasias

RESUMEN

Copper is essential in cell physiology, participating in numerous enzyme reactions. In mitochondria, copper is a cofactor for respiratory complex IV, the cytochrome c oxidase. Low copper content is associated with anemia and the appearance of enlarged mitochondria in erythropoietic cells. These findings suggest a connection between copper metabolism and bioenergetics, mitochondrial dynamics and erythropoiesis, which has not been explored so far. Here, we describe that bathocuproine disulfonate-induced copper deficiency does not alter erythropoietic cell proliferation nor induce apoptosis. However it does impair erythroid differentiation, which is associated with a metabolic switch between the two main energy-generating pathways. That is, from mitochondrial function to glycolysis. Switching off mitochondria implies a reduction in oxygen consumption and ROS generation along with an increase in mitochondrial membrane potential. Mitochondrial fusion proteins MFN2 and OPA1 were up-regulated along with the ability of mitochondria to fuse. Morphometric analysis of mitochondria did not show changes in total mitochondrial biomass but rather bigger mitochondria because of increased fusion. Similar results were also obtained with human CD34+, which were induced to differentiate into red blood cells. In all, we have shown that adequate copper levels are important for maintaining proper mitochondrial function and for erythroid differentiation where the energy metabolic switch plus the up-regulation of fusion proteins define an adaptive response to copper deprivation to keep cells alive.

Asunto(s)

Cobre/deficiencia , Metabolismo Energético , Células Eritroides/metabolismo , Eritropoyesis/fisiología , GTP Fosfohidrolasas/biosíntesis , Dinámicas Mitocondriales/genética , Proteínas Mitocondriales/biosíntesis , Regulación hacia Arriba , Apoptosis/genética , Muerte Celular/genética , Diferenciación Celular/genética , Proliferación Celular , Células Cultivadas , Cobre/metabolismo , Metabolismo Energético/genética , Células Eritroides/citología , Células Eritroides/patología , Eritropoyesis/genética , Humanos , Células K562 , Mitocondrias/genética , Mitocondrias/patología , Simulación de Dinámica Molecular , Regulación hacia Arriba/genética

RESUMEN

It is now recognized that in addition to its activity upon erythroid progenitor cells, erythropoietin (Epo) is capable of stimulating survival of different non-erythroid cells. Since stimulation of erythropoiesis is unwanted for neuroprotection, Epo-like compounds with a more selective action are under investigation. Although the carbamylated derivative of erythropoietin (cEpo) has demonstrated non-hematopoietic tissue protection without erythropoietic effect, little is known about differential mechanisms between Epo and cEpo. Therefore, we investigated signaling pathways which play a key role in Epo-induced proliferation. Here we show that cEpo blocked FOXO3a phosphorylation, allowing expression of downstream target p27(kip1) in UT-7 and TF-1 cells capable of erythroid differentiation. This is consistent with the involvement of cEpo in slowing down G1-to-S-phase progression compared with the effect of Epo upon cell cycle. In contrast, similar antiapoptotic actions of cEpo and Epo were observed in neuronal SH-SY5Y cells. Inhibition and competition assays suggest that Epo may act through both, the homodimeric (EpoR/EpoR) and the heterodimeric (EpoR/ßcR) receptors in neuronal SH-SY5Y cells and probably in the TF-1 cell type as well. Results also indicate that cEpo needs both the EpoR and ßcR subunits to prevent apoptosis of neuronal cells. Based on evidence suggesting that cell proliferation pathways were involved in the differential effect of Epo and cEpo, we went forward to studying downstream signals. Here we provide the first evidence that unlike Epo, cEpo failed to induce FOXO3a inactivation and subsequent p27(kip1) downregulation, which is clearly shown in the incapacity of cEpo to induce erythroid cell growth.

Asunto(s)

Eritropoyetina/análogos & derivados , Eritropoyetina/farmacología , Transducción de Señal/efectos de los fármacos , Apoptosis/efectos de los fármacos , Apoptosis/genética , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Procesos de Crecimiento Celular/efectos de los fármacos , Procesos de Crecimiento Celular/genética , Línea Celular Tumoral , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/genética , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Células Eritroides/efectos de los fármacos , Células Eritroides/metabolismo , Eritropoyesis/efectos de los fármacos , Eritropoyesis/genética , Proteína Forkhead Box O3 , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/metabolismo , Fase G1/efectos de los fármacos , Fase G1/genética , Humanos , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Fosforilación , Receptores de Eritropoyetina/genética , Receptores de Eritropoyetina/metabolismo , Fase S/efectos de los fármacos , Fase S/genética , Transducción de Señal/genética

RESUMEN

BACKGROUND: Rangelia vitalii is a tick-transmitted piroplasm that causes both hemolytic and hemorrhagic disease in dogs in Brazil. OBJECTIVE: The aim of this study was to evaluate the response of the bone marrow in dogs experimentally infected with R vitalii during the acute stage of the disease. METHODS: For this study, 2 groups of a total of 12 young dogs were used. Group A was composed of healthy dogs (n = 5), and group B consisted of animals infected with R vitalii (n = 7). Blood samples were collected on days 0, 10, 20, and 30 post-inoculation and stored in EDTA tubes for a full hematology profile, including a reticulocyte count. On days 10 and 20, bone marrow samples were collected, stained, and examined. RESULTS: In infected dogs anemia was identified on days 10 and 20 post-inoculation (P < .01), and on day 20 reticulocytosis was present. Infected dogs had leukopenia due to neutropenia and eosinopenia, along with lymphocytosis and monocytosis, when compared with control animals. In bone marrow, the myeloid:erythroid ratio was significantly decreased (P < .05) in infected dogs due to increased erythroid precursors. CONCLUSIONS: Dogs experimentally infected with R vitalii develop regenerative extravascular hemolytic anemia accompanied by erythroid hyperplasia in the bone marrow. During the acute phase of the disease, leukopenia due to neutropenia and eosinopenia suggests intense tissue recruitment of these cells in response to the endothelial damage caused by this parasite.

Asunto(s)

Anemia Hemolítica/veterinaria , Babesia/clasificación , Babesiosis/veterinaria , Médula Ósea/patología , Enfermedades de los Perros/parasitología , Anemia Hemolítica/parasitología , Animales , Antiprotozoarios/uso terapéutico , Babesiosis/sangre , Babesiosis/tratamiento farmacológico , Babesiosis/parasitología , Babesiosis/patología , Diminazeno/análogos & derivados , Diminazeno/uso terapéutico , Enfermedades de los Perros/sangre , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/patología , Perros , Células Eritroides/patología , Femenino , Hiperplasia/veterinaria

RESUMEN

Sca1 is a surface marker of haematopoietic stem cell but its role in erythropoiesis is still largely unknown. In this work we evaluated the ability of Sca1⁺ cells to differentiate into cells of the erythrocytic lineage. We performed FACS analysis of complete and purified Sca1⁺ bone marrow cells from C3H/HeNHsd mice and measured the expression of CD71 and Terr119 to evaluate the stages in erythroid development. Definitive erythropoiesis was evident within the complete bone marrow, while only proerythroblasts were found in Sca1⁺ cells, suggesting that Sca1 is a negative regulator of erythropoiesis. We also used FDCP-mix cells and their PU.1 and SCL transfectants. The PU.1 transfectant showed significantly increased expression of Sca1 and was not induced to differentiate into red blood cells, while the SCL transfectant showed significantly lower expression of Sca1 and produced red blood cells. The results of this study suggest that increased Sca1 expression on erythropoietic precursors inhibits erythroid differentiation.

Asunto(s)

Antígenos Ly/metabolismo , Células Eritroides/metabolismo , Células Precursoras Eritroides/metabolismo , Eritropoyesis , Células Madre Hematopoyéticas/metabolismo , Proteínas de la Membrana/metabolismo , Animales , Línea Celular , Eritrocitos/metabolismo , Glicoforinas/biosíntesis , Ratones , Ratones Endogámicos C3H , Receptores de Transferrina

RESUMEN

In mammalian cells, heme can be degraded by heme-oxygenases (HO). Heme-oxygenase 1 (HO-1) is known to be the heme inducible isoform, whereas heme-oxygenase 2 (HO-2) is the constitutive enzyme. Here we investigated the presence of HO during erythroid differentiation in human bone marrow erythroid precursors and K562 cells. HO-1 mRNA and protein expression levels were below limits of detection in K562 cells. Moreover, heme was unable to induce HO-1, at the protein and mRNA profiles. Surprisingly, HO-2 expression was inhibited upon incubation with heme. To evaluate the physiological relevance of these findings, we analyzed HO expression during normal erythropoiesis in human bone marrow. Erythroid precursors were characterized by lack of significant expression of HO-1 and by progressive reduction of HO-2 during differentiation. FLVCR expression, a recently described heme exporter found in erythroid precursors, was also analyzed. Interestingly, the disruption in the HO detoxification system was accompanied by a transient induction of FLVCR. It will be interesting to verify if the inhibition of HO expression, that we found, is preventing a futile cycle of concomitant heme synthesis and catabolism. We believe that a significant feature of erythropoiesis could be the replacement of heme breakdown by heme exportation, as a mechanism to prevent heme toxicity.

Asunto(s)

Células de la Médula Ósea/enzimología , Eritropoyesis , Hemo Oxigenasa (Desciclizante)/metabolismo , Hemo-Oxigenasa 1/metabolismo , Adulto , Animales , Células de la Médula Ósea/citología , Células de la Médula Ósea/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Células Eritroides/citología , Células Eritroides/efectos de los fármacos , Células Eritroides/enzimología , Eritropoyesis/efectos de los fármacos , Glicoforinas/metabolismo , Hemo/farmacología , Hemo Oxigenasa (Desciclizante)/antagonistas & inhibidores , Hemoglobinas/biosíntesis , Humanos , Células K562 , Proteínas de Transporte de Membrana/metabolismo , Ratones , Monocitos/citología , Monocitos/efectos de los fármacos , Monocitos/enzimología , Receptores Virales/metabolismo

RESUMEN

Os aspectos epidemiológicos, clínicos, hematológicos e anatomopatológicos da leucemia eritroide aguda (LMA M6) foram estudados em 10 gatos que morreram em consequência dessa condição. Os resultados obtidos diferem daqueles previamente descritos na literatura nos seguintes aspectos: 1) a doença ocorreu na forma de um modelo bimodal relacionado à idade dos gatos afetados, em que 50 por cento tinham 1-3 anos de idade e 50 por cento tinham 10 anos de idade ou mais; 2) quase todos os gatos afetados (87,5 por cento) demonstravam policromasia, possivelmente decorrente de eritropoese extramedular; 3) em todos os casos havia múltiplos focos de células hematopoéticas, principalmente eritropoeticas, em múltiplos órgãos, que incluíam baço (85,7 por cento), linfonodos (71,4 por cento), fígado (57,1 por cento) e rim (28,6 por cento); 4) em alguns casos (28,6 por cento) esses focos podiam ser vistos macroscopicamente, na forma de metástases, mas sempre diferiam histologicamente da medula óssea quanto à proporção dos precursores eritroides envolvidos; 5) em pelo menos um caso ocorreu um continuum patologicum até outra forma de LMA (LMA M4), um fenômeno denominado "infidelidade de linhagem". Esse artigo discute essas diferenças e reforça os critérios fundamentais para se estabelecer o diagnóstico definitivo dessa que é a forma mais importante de leucemia em gatos na nossa região.(AU)

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The epidemiological, clinical, hematological, and pathological aspects of acute erythroid leukemia (LMA M6) were studied in 10 cats that died from this condition. The results obtained in the present study differ from those previously described in the literature regarding the following aspects: 1) The disease occurred as a bimodal model related to the age of affected cats, in which 50 percent were1-3-years-old and 50 percent were 10 years-old or above; 2) almost all affected cats (87.5 percent) displayed polychromasia, possible resulting from extramedullar erythropoiesis; 3) in all cases there were multiple foci of hematopoietic cells, mainly erythropoietic, in multiple organs including spleen (85.7 percent), lymph nodes (71.4 percent), liver (57.1 percent), and kidney (28.6 percent); 4) in some cases (28.6 percent) these foci could be observed on gross examination as metastasis, but they always histologically differ from bone marrow in regard to the proportion of erythroid precursors involved; 5) at least in one case a continuum patologicum occurred with the condition evolving to another form of LMA (LMA M4), a phenomenon coined as "lineage infidelity". This paper discuss these differences and stresses the fundamental criteria to establish a definitive diagnosis of this condition, which is the most import form of leukemia in cats from our region.(AU)

Asunto(s)

Animales , Gatos/metabolismo , Leucemia/diagnóstico , Leucemia/veterinaria , Células Eritroides/citología