Relación del equivalente de hemoglobina reticulocitaria (Ret-He) con diferentes categorías según la concentración de hemoglobina en candidatos a donar sangre

Objetivo: determinar la relación entre el Ret-He con diferentes categorías según la concentración de hemoglobina en candidatos a donar sangre. Materiales y Métodos: Se realizó un estudio transversal a partir de la revisión de 227 historias clínicas de candidatos a donar sangre durante el mes de diciembre del 2021 en el Servicio de Banco de Sangre y Hemoterapia del Hospital Cayetano Heredia. Se categorizó en 3 grupos según los niveles de hemoglobina: Grupo 1: ≥12,5 g/dl (mujeres) - ≥13,5 g/dl (varones). Grupo 2: 12 g/dl ≥ mujeres <12.5 g/dl y 13 g/dl ≥ varones<13.5 g/dl. Grupo 3: <12 g/dl (mujeres) - <13 g/dl (varones). Se tomaron datos de hematocrito, hemoglobina, VCM, HCM, CHCM y Ret-He. Para el análisis bivariado se utilizó el análisis de varianza (ANOVA) y se consideró un nivel de significancia del 5 %. Resultados: La mediana de Ret-He fue mayor en el grupo 1 que en el grupo 2 (33.5 frente a 32.5; p=0.002) y en el grupo 3 (33.5 frente a 27.8; p<0.001). Se encontró relación estadísticamente significativa entre los grupos estudiados y Ret-He (p<0.001), además de una correlación positiva moderada fuerte entre Ret-He y los índices eritrocitarios. Conclusiones: Se encontró una asociación significativa entre los grupos basados en categorías de hemoglobina y la Ret-He en candidatos a donar sangre.

Objective: to determine the relationship between Ret-He with different categories according to hemoglobin concentration in candidates for blood donation. Methods: A cross-sectional study was carried out based on the review of 227 medical records of candidates to donate blood during the month of December 2021 in the Blood Bank and Hemotherapy Service of the Cayetano Heredia Hospital. They were categorized into 3 groups according to hemoglobin levels: Group 1: ≥12.5 g/dl (females) - ≥13.5 g/dl (males). Group 2: 12 g/dl ≥ females <12.5 g/dl and 13 g/dl ≥ males<13.5 g/dl. Group 3: <12 g/dl (females) - <13 g/dl (males). Hematocrit, hemoglobin, VCM, HCM, CHCM and Ret-He data were collected. For the bivariate analysis, analysis of variance (ANOVA) was used and a significance level of 5% was considered. Results: The median Ret-He was higher in group 1 than in group 2 (33.5 vs. 32.5; p=0.002) and in group 3 (33.5 vs. 27.8; p<0.001). A statistically significant relationship was found between the groups studied and Ret-He (p<0.001), in addition to a moderate-strong positive correlation between Ret-He and erythrocyte indices. Conclusions: An association was found between groups based on hemoglobin categories and Ret-He in blood donation candidates.

Evaluation of RET-He values as an early indicator of iron deficiency anemia in pregnant women

Abstract Introduction During pregnancy, women are at an increased risk of developing iron-deficiency anemia. Objective The objective of this study was to assess the diagnostic performance of the reticulocyte hemoglobin equivalent (RET-He) in the early detection of iron-deficiency anemia in a group of pregnant women and to establish a reference range for this parameter in a group of control individuals. Method: A total of 60 patients and 130 control subjects were included in the study. Blood samples collected from the subjects were submitted to a complete blood count and a serum ferritin test and the data were analyzed by comparing the groups and ROC curves. Results The reference range found for the RET-He was between 29.75pg and 38.24pg, with a median of 35pg. The receiver operating characteristic (ROC) curve analysis for the ferritin parameter showed an area under the curve of 0.732 for the RET-He, 0.586 for hemoglobin, 0.551 for the mean corpuscular hemoglobin concentration and 0.482 for the mean corpuscular volume. Conclusion Early diagnosis of iron deficiency anemia in pregnancy is essential to prevent damage to both maternal and fetal health. The RET-He presents an excellent potential as an auxiliary tool for the diagnosis of iron deficiency in pregnant women.

A cross-sectional study of reticulocyte count and hemoglobin content of umbilical cord blood with relation to maternal factors in newborns of Sikkim

Background: The hematological values of the new born is amenable to certain changes under the influence of fetal, maternal, and environmental factors. Aims and Objectives: The aims of this study was to study the effect of maternal factors on hemoglobin content and reticulocyte count in umbilical cord blood of newborns of Sikkim. Materials and Methods: A total of 150 cord blood samples from newborns delivered at CRH Hospital and STNM Govt. Hospital, Gangtok, Sikkim were analyzed. Two ml of cord blood was taken in an ethylene diamine tetra acetic acid tube under aseptic condition and within 6 h: Hemoglobin estimation and Reticulocyte count was done. Results: The mean reticulocyte count was 2.48 ± 0.87% in normal vaginal delivery (NVD) and in C/S was 3.06 ± 0.97%. The mean hemoglobin content was 13.9 ± 2.6 g/dL in NVD and in C/S was 16.1 ± 3.4 g/dL. P < 0.05 for cord hemoglobin and reticulocytes in relation to mode of delivery was found to be highly significant. The mean reticulocyte count was 2.85 ± 1.0% in normal pregnancy and in complication of pregnancy was 2.9 ± 0.70%. The mean hemoglobin content was 15.36 ± 3.31 g/dL in normal pregnancy and in complication of pregnancy was 16.1 ± 3.4 g/dL. The reticulocyte count was found to be statistically significant in relation to complication of pregnancy. The mean mother’s hemoglobin was 11.16 ± 1.27 when cord hemoglobin content was <14 g/dL and was 11.88 ± 1.24 g/dL when cord hemoglobin content was >14 g/dL. There was statistical significance between the cord hemoglobin content and the hemoglobin content of the mother. Conclusion: Maternal anemia leads to lower circulating hemoglobin in new born. Cord blood screening is a useful means for identification of anemia in neonatal period. High reticulocyte count in neonates denotes active erythropoiesis which might help to defer transfusion unless there were other special reasons for proceeding.

Potential value of differential expression of CDC42 gene in adult and umbilical cord blood reticulocytes / 中国输血杂志

【Objective】 To explore critical regulatory genes in the hemoglobin switch process by analyzing transcriptomic data from the GSE6236, GSE17639 and GSE35102 datasets. 【Methods】 The mRNA expression profiles of the three datasets were downloaded from the GEO database and gene annotation was performed using the AnnoProbe package.The remove-BatchEffect function of the Limma package was used to remove batch effects. Weighted gene co-expression network analysis (WGCNA) was used to explore the most relevant modular genes in reticulocytes. The receiver operating characteristic curve (ROC) was used to assess the value of differential genes in differentiating between cord blood and adult peripheral blood reticulocytes. The GSE35102 dataset was used to validate changes in differential gene expression during hemoglobin transformation. Finally, real-time quantitative PCR was used to verify differential gene expression in cord blood and adult peripheral blood reticulocytes. 【Results】 Twelve genes showed differential expression in reticulocytes from cord blood and adult peripheral blood ( |logFC|≥1.5, P<0.05). WGCNA found that genes in the blue module were most strongly associated with reticulocytes (R2 =0.76,P<0.001). Of the five genes that overlapped between the two, only CDC42 showed differential expression in the combined dataset (t =3.776, P<0.001) and was able to better differentiate between reticulocytes in cord blood and adult peripheral blood. The expression of CDC42 varied significantly during the hemoglobin transformation process (Z = -2.908, P<0.01), and was significantly lower in adult reticulocytes compared to reticulocytes from cord blood (t =7.824, P <0.001). 【Conclusion】 The CDC42 gene is involved in the hemoglobin switching of reticulocytes and could be a potential therapeutic target for sickle cell disease.

Value of immature reticulocyte fraction in anemias and other hematologic conditions: a brief narrative review

Novos parâmetros hematológicos, como a fração de reticulócitos imaturos (IRF), tendem a se tornar ferramentas importantes na prática clínica. O IRF identifica os reticulócitos mais imaturos, que contêm grande quantidade de ácido ribonucleico, sendo um importante parâmetro para avaliar a atividade da medula óssea, em tempo real, para o diagnóstico diferencial das anemias, acompanhamento do seu tratamento, e para o acompanhamento ou recuperação da medula óssea em diversas condições clínicas. No entanto, ainda há um longo caminho a percorrer antes que a IRF possa ser usada na prática clínica. Assim sendo, é urgente estabelecer os valores de referência e padronizar as metodologias utilizadas pelos diferentes analisadores hematológicos e como expressar seus resultados. Esta revisão narrativa fornece uma perspectiva crítica sobre o IRF e seu potencial para o uso clínico, bem como suas limitações.

New hematological parameters, such as immature reticulocyte fraction (IRF), tend to become important tools in clinical practice. IRF identifies the most immature reticulocytes that contain a large amount of ribonucleic acid, being an important parameter to evaluate bone marrow activity in real time for differential diagnosis of anemias, monitoring of its treatment, and for follow-up or bone marrow recovery in various clinical conditions. However, there is still a long way to go before IRF can be used in clinical practice. Thus, it is urgent to establish reference values and to standardize of the methodologies used by different hematological analyzers and how to express the results. This narrative review provides a critical perspective on IRF, its potential of clinical use and limitations.

Reticulocyte parameters for monitoring anemia in patients with chronic kidney disease on peritoneal dialysis / Parâmetros reticulocitários para monitoramento de anemia em pacientes com doença renal crônica em diálise peritoneal

ABSTRACT Introduction: Anemia is a complication with impact on morbidity and mortality in chronic kidney disease (CKD) patients. Current markers for the diagnosis and monitoring of anemia in CKD are limited by the interrelation between erythropoiesis, iron stores, inflammation, and the resistance to treatment with erythropoiesis stimulant agents (ESA). Objective: The aim of this study was to analyze the role of immature reticulocyte fraction (IRF) and hemoglobin concentration in reticulocytes (RET-He) by the hematology analyser Sysmex XN-5000 in the monitoring of CKD anemia in peritoneal dialysis patients. Methods: This was a prospective, observational multicenter study which compared IRF and RET-He with parameters recommended by the guidelines. Inflammatory biomarkers were analyzed by the Luminex® Multiplexing Instruments system. Thirty-five patients (59 ± 13 years old; 51% men) were included in the analysis. Results: Hemoglobin was 12.2 ± 2 g/dl; 87% had resistance to ESA. Patients with erythropoietin resistance index (ERI) in the upper quartile presented a significantly higher of IRF and a lower percentage of iron deficiency (12%) compared to ferritin (82%) and transferrin saturation index (STI) (51%). Interleukin-6 (IL-6) levels correlated with the percentage of medium fluorescence reticulocyte (MFR) (r = 0.45, p < 0.03). Hemoglobin values after 60 and 180 days were consistently higher in the group of patients with a IRF% lower than 10.5. Conclusion: IRF and RET-He may add value in the iron deficiency investigation, as well as in the identification of patients with ERI. Due to the restricted number of patients analyzed in this study, future studies should be encouraged in larger populations and with prospective follow-up, to validate our findings.

RESUMEN Introducción: La anemia es una complicación con impacto en la morbimortalidad en pacientes con enfermedad renal crónica (ERC). Los biomarcadores usados para el diagnóstico y seguimiento de la anemia en la ERC están limitados por la interrelación entre eritropoyesis, depósitos de hierro, inflamación y resistencia al tratamiento con agentes estimulantes de la eritropoyesis (AEE). Objetivo: El objetivo de este estudio fue analizar el papel de la fracción de reticulocitos inmaduros (IRF) y el equivalente de hemoglobina en reticulocitos (RET-He) mediante el analizador hematológico Sysmex XN-5000 en el seguimiento de la anemia por ERC en pacientes en diálisis peritoneal. Métodos: Estudio prospectivo, observacional multicéntrico que comparó IRF y RET-He con los parámetros recomendados por las guías. Los biomarcadores inflamatorios fueron analizados por el sistema Luminex® Multiplexing Instruments. Este estudio incluyó a 35 pacientes (59 ± 13 años; 51% hombres). Resultados: La hemoglobina fue de 12,2 ± 2 g/dl; el 87% tenía resistencia a AEE. Los pacientes con índice de resistencia a la eritropoyetina (IRE) en el cuartil superior tenían un IRF significativamente más alto y un porcentaje más bajo de deficiencia de hierro (12%) en comparación con la ferritina (82%) y las ITS (51%). Los niveles de interleucina-6 (IL-6) se correlacionaron con el porcentaje de reticulocitos de fluorescencia media (MFR) (r = 0,45, p < 0,03). Los valores de hemoglobina después de 60 y 180 días, fueron consistentemente más altos en el grupo de pacientes con IRF% inferior a 10,5. Conclusión: IRF y RET-He pueden agregar valor en la investigación de ferropenia, así como en la identificación de pacientes con ERI. Debido al número limitado de pacientes analizados en este estudio, se deben impulsar estudios futuros en poblaciones más grandes y con seguimiento prospectivo, para validar nuestros hallazgos.

RESUMO Introdução: Anemia é uma complicação com impacto na morbidade e na mortalidade de pacientes com doença renal crônica (DRC). Os biomarcadores utilizados no diagnóstico e no monitoramento de anemia na DRC são limitados devido à inter-relação entre eritropoiese, estoque de ferro, inflamação e resistência à terapêutica com agentes estimuladores da eritropoiese (AEE). Objetivo: O objetivo deste estudo foi analisar o papel dos marcadores fração de reticulócitos imaturos (IRF) e concentração de hemoglobina nos reticulócitos (RET-He) do analisador hematológico Sysmex XN 5000 no monitoramento da anemia em pacientes em diálise peritoneal. Métodos: Estudo prospectivo, observacional e multicêntrico que comparou IRF e RET-He com parâmetros laboratoriais recomendados pelos guidelines. Biomarcadores inflamatórios foram analisados pelo sistema Luminex® Multiplexing Instruments. Este estudo incluiu 35 pacientes (59 ± 13 anos; 51% homens). Resultados: Os valores de hemoglobina foram 12,2 ± 2 g/dl; 87% apresentaram resistência a AEE. Pacientes com índice de resistência à eritropoietina (IRE) no quartil superior apresentaram valores significativamente maiores de IRF e menor porcentagem de deficiência de ferro (12%) em comparação com pacientes com ferritina (82%) e índice de saturação de transferrina (IST) (51%). Os níveis de interleucina 6 (IL-6) correlacionaram-se com a porcentagem de reticulócitos de fluorescência média (MFR) (r = 0,45, p < 0,03). Valores de hemoglobina após 60 e 180 dias foram consistentemente mais altos no grupo de pacientes com IRF% menor que 10,5. Conclusão: IRF e RET-He podem agregar valor na investigação da deficiência de ferro, bem como na identificação do índice de existência à eritropoietina (ERI). Devido ao número restrito de pacientes analisados neste trabalho, estudos futuros devem ser estimulados em populações maiores e com acompanhamento prospectivo, para validação dos nossos achados.

Diagnóstico diferencial de las anemias hemolíticas / Differential diagnosis of hemolytic anemias

Introducción: El término hemólisis hace referencia a la destrucción de los eritrocitos y ocurre en un amplio rango de condiciones clínicas fisiológicas y patológicas. Es empleado para definir situaciones en la que la vida media de los eritrocitos está disminuida por causas mecánicas, tóxicas, autoinmunes o infecciosas. Objetivo: Describir los principales marcadores de hemólisis que se encuentran variablemente alterados en las diferentes formas de anemias hemolíticas. Métodos: Se realizó una revisión de la literatura, en inglés y español, a través del sitio web PubMed y el motor de búsqueda Google Académico de artículos publicados en los últimos 10 años. Se hizo un análisis y resumen de la información. Análisis y síntesis de la información: La hemoglobina es el marcador más directo de la gravedad clínica en las enfermedades hemolíticas. Sus valores pueden estar muy próximos a los valores de referencia en las formas ligeras (Hb > 100 g/L) o significativamente reducidos en las moderadas (Hb entre 80-100 g/L), graves (Hb entre 60-80 g/L) y muy graves (Hb < 60 g/L). Sin embargo, existen otros marcadores esenciales para diferenciar las formas de presentación aguda y crónica, la hemólisis extravascular de la intravascular y la presencia de signos extrahematológicos tales como: los reticulocitos y esquistocitos, la deshidrogenasa láctica, la haptoglobina, la bilirrubina, la ferritina y la hemosiderinuria. Conclusiones: Los parámetros hemolíticos pueden estar diferencialmente alterados en varias condiciones lo cual ayuda en la realización del diagnóstico diferencial de las anemias hemolíticas(AU)

Introduction: The term hemolysis refers to the destruction of erythrocytes, a process occurring in a wide range of physiological and pathological clinical conditions. The term is used to define situations in which mean erythrocyte lifespan is reduced due to mechanical, toxic, autoimmune or infectious causes. Objective: Describe the main markers of hemolysis found to be variably altered in the different forms of hemolytic anemias. Methods: A review was conducted of the literature about the topic published in English and Spanish in the website PubMed and the search engine Google Scholar in the last 10 years. Data were analyzed and summarized. Data analysis and synthesis: Hemoglobin is the most direct marker of clinical severity in hemolytic diseases. Its values may be very close to reference levels in mild disease (Hb > 100 g/l), whereas they will be significantly reduced in moderate (Hb 80-100 g/l), severe (Hb 60-80 g/l) and very severe disease (Hb < 60 g/l). However, other markers are also essential to distinguish acute from chronic presentation, extravascular from intravascular hemolysis, and the presence of extrahematological signs such as reticulocytes and schistocytes, lactate dehydrogenase, haptoglobin, bilirubin, ferritin and hemosiderinuria. Conclusions: Differentially altered hemolytic parameters may be found in several conditions, which makes them useful for the differential diagnosis of hemolytic anemias(AU)

The spectrum of Evans syndrome: a literature review

Evans syndrome (ES) is an autoimmune disorder characterized by the simultaneous or sequential development of autoimmune hemolytic anemia (AIHA) and immune thrombocytopenia (ITP) and/or immune neutropenia in the absence of any underlying cause. Evans syndrome is a rare disorder although the exact frequency is unknown. No sex predilection is known and Evans syndrome has been described in all ethnic groups and at all ages. Classification of ES includes primary, with this being an exclusion diagnosis with no underlying condition, and secondary in the presence of an underlying disease. Clinical features are associated with anemia and thrombocytopenia including pallor, weakness, fatigue, jaundice, petechiae, ecchymosis, gingivorrhagia and epistaxis. First, a detailed history must be taken from the patient to determine the risk factors for developing ES then a family history of immune disorders along with a thorough physical examination. The management of Evans syndrome remains a challenge. Steroids with and without IVIG are recommended as front-line therapy. Red blood cell/platelet transfusion is indicated only in severe symptomatic patients due to the risk of exacerbations. Splenectomy may also be considered a second-line treatment.

Avaliação do conteúdo de hemoglobina do reticulócito em pacientes anêmicos sob tratamento dialítico / Evaluation of the reticulocyte hemoglobin content in anemic patients in dialytic treatment

A anemia é uma complicação importante na doença renal crônica (DRC), culminando com o aumento da morbidade e mortalidade, tornando-se fundamental a busca de marcadores hematológicos que permitam seu diagnóstico precoce. Este artigo teve como objetivo revisar na literatura estudos que investigaram a associação entre o conteúdo de hemoglobina contida nos reticulócitos com a anemia em pacientes com DRC em tratamento dialítico e gerar evidências de sua importância na prática clínica. Foi realizada a busca nas bases de dados eletrônicas: Medline e Web of Science. O período de busca definido foi de janeiro de 1997 a dezembro de 2017. O conteúdo de hemoglobina dos reticulócitos avalia a hemoglobina contida nos reticulócitos, sendo que estes, após liberação da medula óssea, permanecem no sangue periférico de um a quatro dias antes de completar sua maturação, conferindo-lhe maior especificidade em refletir a disponibilidade de ferro aos precursores hematopoiéticos, além de fornecer uma avaliação precoce da resposta eritropoética frente ao tratamento de ferro IV. Os artigos selecionados nessa revisão demonstraram que o conteúdo de hemoglobina do reticulócito, além de refletir precocemente a disponibilidade de ferro aos precursores hematopoiéticos, o mesmo não sofre interferência de citocinas inflamatórias, tornando-o um marcador eficaz no monitoramento da cinética do ferro em pacientes em diálise.

Anemia is an important complication in chronic kidney disease (CKD), culminating with the increase in morbidity and mortality, making it fundamental to search for hematological markers that allow its early diagnosis. This review aimed to review in the literature studies that investigated the association between hemoglobin content in reticulocytes and anemia in patients with CKD in dialysis and to generate evidence of its importance in clinical practice. The hemoglobin content of the reticulocytes evaluates the hemoglobin contained in the reticulocytes, which, after release of the bone marrow, remain in the peripheral blood 1 to 4 days before their maturation completes, giving it greater specificity in reflecting the availability of iron to the reticulocytes. Hematopoietic precursors, in addition to providing an early assessment of the erythropoietic response to IV iron treatment. The articles selected in this review have demonstrated that the reticulocyte hemoglobin content in addition to early reflection of the availability of iron to the hematopoietic precursors does not suffer interference from inflammatory cytokines, making it an effective marker in the monitoring of iron kinetics in dialysis patients.

Hemoglobina de reticulocito y su importancia en el diagnóstico temprano de anemia ferropénica / Reticulocyte hemoglobin and its importance in early diagnosis of iron deficiency anemia

Resumen Introducción: El contenido de hemoglobina de reticulocitos (CHr), es un parámetro en la biometría hematológica automatizada que proporciona información sobre el contenido de hierro, por ello se ha utilizado como un marcador de la biodisponibilidad del hierro en la eritropoyesis, permite su detección en una etapa temprana de la anemia ferropénica y otras patologías como inflamación crónica, enfermedad renal crónica; además realizar monitoreo de terapias con eritropoyetina y hierro. Objetivo: Exponer la aplicabilidad de la CHr como un parámetro en el diagnóstico precoz de la anemia por deficiencia de hierro, así como su medición e interpretación. Materiales y métodos: Se realizó la revisión de artículos científicos en inglés y español en las bases de datos PubMed, ScienceDirect, LILACS y Medline, usando descriptores validados en Medical Subject Headings (MeSH), considerando periodo de publicabilidad del 80% inferior a 5 años. Resultados: Se describe la importancia, aplicabilidad, determinación e interpretación de este parámetro como biomarcador específico hemático temprano en el diagnóstico de deficiencia de hierro antes de presentarse cambios morfológicos eritroides. Conclusiones: La CHr es un parámetro de gran utilidad en el diagnóstico temprano de anemia ferropénica y otras patologías como deficiencia funcional de hierro, estados de inflamación crónica y enfermedad renal crónica.

Abstract Introduction: The reticulocyte hemoglobin content (CHr) is a parameter in automated hematological biometrics, which can provide information on the iron content. So it has been used as a marker of the bioavailability of iron in the erythropoiesis, it allows its detection at an early stage of iron deficiency anemia and other pathologies such as chronic inflammation, chronic kidney disease; in addition to monitoring therapy with erythropoietin and iron. Objective: To expose the applicability of CHr as a parameter in the early diagnosis of iron deficiency anemia, as well as its measurement and interpretation. Materials and methods: The review of scientific articles in English and Spanish was carried out in the PubMed, ScienceDirect, LILACS and Medline databases, using descriptors validated in Medical Subject Headings (MeSH), considering the publication period of 80% less than 5 years. Results: The importance, applicability, determination and interpretation of this parameter is described as an early specific biomarker in the blood in the diagnosis of iron deficiency before presenting morphological changes occurring during terminal erythroid differentiation. Conclusions: CHr is a very useful parameter in the early diagnosis of iron deficiency anemia and other pathologies such as functional deficiency, chronic inflammation states and chronic renal disease.

Citometría de flujo en reticulocitos de sangre periférica como indicador de inestabilidad cromosómica en pacientes con gliomas de alto grado / Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients

Resumen Introducción. La cuantificación de la inestabilidad cromosómica es un parámetro importante para evaluar la genotoxicidad y la radiosensibilidad. Las técnicas convencionales requieren cultivos celulares o laboriosos análisis microscópicos de cromosomas o núcleos. La citometría de flujo en reticulocitos ha surgido como una alternativa para los estudios in vivo, ya que reduce los tiempos de análisis e incrementa hasta en 20 veces el número de células analizables. Objetivos. Estandarizar los parámetros de citometría de flujo requeridos para seleccionar y cuantificar reticulocitos micronucleados (RET-MN) a partir de muestras de sangre periférica, y cuantificar la frecuencia de esta subpoblación anormal como medida de inestabilidad citogenética en sendas poblaciones de voluntarios sanos (n=25) y pacientes (n=25) recién diagnosticados con gliomas de alto grado antes de iniciar el tratamiento. Materiales y métodos. Las células sanguíneas se marcaron con anti-CD71-PE para reticulocitos, anti- CD61-FITC para la exclusión de plaquetas y yoduro de propidio para detectar el ADN en reticulocitos. La fracción celular MN-RETCD71+ se seleccionó y se cuantificó con un citómetro de flujo automático. Resultados. Se describió detalladamente la estandarización de los parámetros citométricos, con énfasis en la selección y la cuantificación de la subpoblación celular MN-RETCD71+. Se establecieron los niveles basales de MN-RETCD71+ en la población de control y en los pacientes se encontró un incremento de 5,2 veces antes de iniciar el tratamiento (p<0,05). Conclusión. Los resultados evidenciaron la utilidad de la citometría de flujo acoplada a la marcación de las células RETCD71+ como método eficiente para cuantificar la inestabilidad cromosómica in vivo. Se sugieren posibles razones del incremento de micronúcleos en células RETCD71+ de pacientes con gliomas.

Abstract Introduction: The quantification of chromosomal instability is an important parameter to assess genotoxicity and radiosensitivity. Most conventional techniques require cell cultures or laborious microscopic analyses of chromosomes or nuclei. However, a flow cytometry that selects the reticulocytes has been developed as an alternative for in vivo studies, which expedites the analytical procedures and increases up to 20 times the number of target cells to be analyzed. Objectives: To standardize the flow cytometry parameters for selecting and quantifying the micronucleated reticulocytesCD71+ (MN-RET) from freshly drawn peripheral blood and to quantify the frequency of this abnormal cell subpopulation as a measure of cytogenetic instability in populations of healthy volunteers (n =25), and patients (n=25), recently diagnosed with high-grade gliomas before the onset of treatment. Materials and methods: Blood cells were methanol-fixed and labeled with anti-CD-71-PE for reticulocytes, antiCD-61-FITC for platelet exclusion, and propidium iodide for DNA detection in reticulocytes. The MN-RETCD71+ cell fraction was selected and quantified with an automatic flow cytometer. Results: The standardization of cytometry parameters was described in detail, emphasizing the selection and quantification of the MN-RETCD71+ cellular fraction. The micronuclei basal level was established in healthy controls. In patients, a 5.2-fold increase before the onset of treatment was observed (p <0.05). Conclusion: The data showed the usefulness of flow cytometry coupled with anti-CD-71-PE and anti- CD-61-FITC labeling in circulating reticulocytes as an efficient and high resolution method to quantify chromosome instability in vivo. Finally, possible reasons for the higher average of micronuclei in RETCD71+ cells from untreated high-grade glioma patients were discussed.

Niveles de eritropoyetina y reticulocitos en residentes de bajas alturas migrantes a medianas alturas / Erythropoietin and reticulocyte levels in lowland residents migrating to middle altitudes

RESUMEN Introducción: La eritropoyetina es una alfa globulina glicosilada con producción renal en más del 90% en la vida adulta. Es la principal hormona en el mantenimiento constante de la masa eritrocitaria, aunque existen modificaciones en sus niveles asociados con el tabaquismo, anemias, EPOC y la migración de bajas a medianas o altas alturas. Esto último desencadena un proceso hipóxico que puede llegar a producir mal agudo de montaña. Objetivo: Describir el comportamiento de la eritropoyetina, el recuento de reticulocitos y su influencia, en procesos de adaptación a la altura. Metodología: Estudio descriptivo de corte transversal que incluyó 11 participantes provenientes de bajas alturas a quienes se les determinó la concentración sérica de eritropoyetina y el recuento de reticulocitos en un periodo de 28 días. Resultados: Ocho de los participantes presentaron un ascenso progresivo en los niveles séricos de eritropoyetina, uno mantuvo una curva plana y dos presentaron comportamiento atípico respecto a lo reportado en la literatura. Conclusión: La eritropoyetina es un factor fundamental que marca el comienzo de la eritropoyesis, cuya finalidad es mejorar el aporte de oxígeno en procesos de adaptación a la altura. Además, la hipoxia es un factor determinante en el inicio y desarrollo del mal agudo de montaña. El recuento de reticulocitos depende del estímulo proliferativo y anti-apoptótico de la eritropoyetina, así como de las concentraciones séricas de vitamina B12, hierro y ácido fólico.

ABSTRACT Introduction: EPO is a glycosylated alpha globulin produced in more than 90% by kidneys through adult life, being a key hormone that regulated the erythrocytic mass. However, there are some modifications in the levels of this hormone that may be related to smoking, anemia, EPOC and migration to from low to higher altitudes, inducing hypoxic processes. Depending on the individual, it may produced the disease named as acute mountain sickness. Objective: To describe erythropoietin level modifications, reticulocytes count and its influence, on the adaptive process to altitude. Methodology: This is a transversal descriptive study including 11 participants from low altitudes places, whose EPO serum concentration and reticulocytes count was determined during 28 days. Results: Eight participants presented a progressive increase in EPO serum levels, one participant exhibited a constant level and two more showed atypical results according to previous literature. Conclusion: EPO is a key factor for determining the erythropoiesis beginning, as its objective is to improve the oxygen provision during altitude adaptation processes by increasing its concentration in blood due to hypoxic stimulus. Besides, hypoxia is a determinant factor in the beginning and development of acute mountain sickness. The reticulocytes count depends also on the EPO proliferative and anti-apoptotic stimulus, and on the serum concentrations of B12 vitamin, iron and folic acid.

The performance evaluation of reticulocytes detected by XN-3000 automatic blood analyzer / 中国医学装备

Objective:To evaluate the performance of reticulocyte (Ret) detected by Sysmex XN-3000 automatic blood analyzer.Methods: 96 blood samples were collected randomly in clinical detection of reticulocytes, and respectively were detected by XN-3000 automatic blood analyzer and microscopy. The precision, stability, linearity and carryover of Ret in blood samples detected by XN-3000 automatic blood analyzer were evaluated by the correlation analysis of the results from the two methods and the correlation analysis would be completed by microscopy.Results: precision, linear range and sensitivity of Ret measured by XN-3000 were within the allowable range. The stability did not change significantly below 4℃ within 48h (CV<1%). The carryover rate was 0.14%. The results measured by XN-3000 were well compared with microscopy (r=0.979).Conclusion: Sysmex XN-3000 automatic blood analyzer is applied to the clinical detection of large quantities of specimens and it is ideal for measuring Ret.

A 2-month-old boy with hemolytic anemia and reticulocytopenia following intravenous immunoglobulin therapy for Kawasaki disease: a case report and literature review / 소아과

Herein, we report a rare case of hemolytic anemia with reticulocytopenia following intravenous immunoglobulin therapy in a young infant treated for Kawasaki disease. A 2-month-old boy presented with fever lasting 3 days, conjunctival injection, strawberry tongue, erythematous edema of the hands, and macular rash, symptoms and signs suggestive of incomplete Kawasaki disease. His fever resolved 8 days after treatment with aspirin and high dose infusion of intravenous immunoglobulin. The hemoglobin and hematocrit decreased from 9.7 g/dL and 27.1% to 7.4 g/dL and 21.3%, respectively. The patient had normocytic hypochromic anemia with anisocytosis, poikilocytosis, immature neutrophils, and nucleated red blood cells. The direct antiglobulin test result was positive, and the reticulocyte count was 1.39%. The patient had an uneventful recovery. However, reticulocytopenia persisted 1 month after discharge.

Reticulocyte Count Of Nigerian Children With Malaria Infection In The University Of Calabar Teaching Hospital.

Abstracts: Background: This work was sets out to assess the reticulocyte count of Nigerian Children infected with malaria in the University of Calabar Teaching Hospital, comprising of fifty malaria infected children and forty six apparently healthy children. The malaria infected children comprised of 25 males and 25 females aged 1-10 years. Blood samples were collected and analysed for reticulocyte count using the 1% new Methylene Blue supra vital staining method and malaria parasites using, 2% Giemsa stain. The mean reticulocyte count in the malaria infected children (2. 17 ± 0.3%) was significantly higher (P<0.01) than that in the apparently healthy children (1.03 ± 0.3 %). The mean parasites density in infected children was 30,428.76 ± 11,876.85 ul. A comparism of reticulocyte level of infected male (2.204 ± 0.32%) and female children (2.133 ± 0.3 6%) showed no significant difference (p>O.O5). Equally, the reticulocyte levels of infected children between 1-5 years of age (2.1 5 ± 0.34%) was not significantly different (P> 0.05) from that of children aged between 6-10 years (2.18 ± 0.32%). The level of reticulocytosis is a reflection of parasite density in the malaria infected children.

Evaluation of erythrocyte and reticulocyte parameters as indicative of iron deficiency in patients with anemia of chronic disease

Objective: The aim of this study was to evaluate the effectiveness of mature red cell and reticulocyte parameters to identify three conditions: iron deficiency anemia, anemia of chronic disease, and anemia of chronic disease associated with absolute iron deficiency. Methods: Peripheral blood cells from 117 adult patients with anemia were classified accord- ing to iron status, inflammation, and hemoglobinopathies as: iron deficiency anemia (n = 42), anemia of chronic disease (n = 28), anemia of chronic disease associated with iron deficiency anemia (n = 22), and heterozygous β-thalassemia (n = 25). The percentage of microcytic eryth-rocytes, hypochromic erythrocytes, and the levels of hemoglobin in both reticulocytes and mature red cells were determined. Receiver operating characteristic analysis was used to evaluate the accuracy of the parameters in differentiating anemia. Results: There was no difference between the groups of iron deficiency and anemia of chronic disease associated with absolute iron deficiency for any of the parameters. The percentage of hypochromic erythrocytes was the best parameter to identify absolute iron deficiency in patients with anemia of chronic disease (area under curve = 0.785; 95% confidence interval: 0.661-0.909 with sensitivity of 72.7%, and specificity of 70.4%; cut-off value 1.8%). The formula microcytic erythrocyte count minus hypochromic erythrocyte count was very accurate to differentiate iron deficiency anemia from heterozygous β-thalassemia (area under curve = 0.977; 95% confidence interval: 0.950-1.005 with a sensitivity of 96.2%, and specificity of 92.7%; cut-off value 13.8). Conclusion: The erythrocyte and reticulocyte indices are moderately good to identify absolute iron deficiency in patients with anemia of chronic disease.

New red blood cell and reticulocyte parameters and reference values for healthy individuals and in chronic kidney disease / Novos parâmetros e valores de referência eritrocitários e reticulocitários normais e na doença renal crônica

Introduction: The importance of local references values has been well described in the literature; this is because the characteristics of the population may influence the laboratory tests. Objective: To establish the reference range for traditional and extended red blood cell parameters and reticulocyte indices in order to investigate its application in patients with chronic kidney disease (CKD). Materials and methods: 249 blood donors (125 males and 124 females) were selected to establish the reference values. The hemodialysis sample consisted of 62 patients with terminal CKD (48 male and 14 female). All analyzes were performed using the Sysmex XE-5000 automated hematology analyzer. Results: Differences between reference values was observed in relation to gender: red blood cells (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular hemoglobin concentration (MCHC), percentage of hyperchromic red blood cells (%HYPER), percentage of microcytosis (%MICRO), percentage of macrocytosis (%MACRO), absolute reticulocyte count (RET), reticulocyte hemoglobin content (RET-He), immature reticulocyte fraction (IFR), low fluorescence reticulocytes (LFR), medium fluorescence reticulocytes (MFR), and high fluorescence reticulocytes (HFR). Individuals with CKD presented RBC, HGB, HCT, MCHC, red cell distribution width expressed as coefficient of variation (RDW-CV), percentage of hypochromic red blood cells (%HYPO), percentage of reticulocytes (RET%), RET (female group), IFR, LFR, MFR, and HFR results compatible with the anemic state, which can be observed in 91.8% of patients. All studied parameters were in the area under the curve (AUC) > 0.4. In male group, %HYPO (AUC: 0.806) and IFR (AUC: 0.762) presented higher AUC values, while female group presented %HYPO (AUC: 0.806), %HYPER (AUC: 0.815), and IFR (AUC: 0.660). Conclusion: The medical advancement, the development of new techniques and hematological parameters have revealed important information ...

Introdução: A importância dos valores de referências locais tem sido bastante descrita na literatura, isso porque características da população podem influenciar os testes laboratoriais. Objetivos: Estabelecer o intervalo de referência para parâmetros eritrocitários tradicionais e estendidos e índices reticulocitários a fim de investigar sua aplicação em pacientes com doença renal crônica (DRC). Materiais e métodos: Dos doadores de sangue, 249 pacientes foram selecionados para estabelecimento dos valores de referência (125 homens e 124 mulheres); dos pacientes em hemodiálise, a amostra foi composta por 62 indivíduos com DRC terminal (48 homens e 14 mulheres). Todas as análises foram realizadas no avaliador hematológico Sysmex XE-5000. Resultados: Foi observada uma distinção entre os valores de referência em relação ao gênero: células vermelhas do sangue (RBC), hemoglobina (HGB), hematócrito (HCT), concentração de hemoglobina corpuscular média (CHCM), porcentagem de eritrócitos hiper-hemoglobinizados (%HIPER), porcentagem de microcitose (%MICRO), porcentagem de macrocitose (%MACRO), contagem absoluta de reticulócitos (RET), contagem relativa de reticulócitos (RET-He), fração de reticulócitos imaturos (IFR), reticulócitos de baixa fluorescência (LFR), reticulócitos de média fluorescência (MFR) e reticulócitos de alta fluorescência (HFR). Os indivíduos com DRC apresentaram resultados de RBC, HGB, HCT, CHCM, coeficiente de variação do tamanho dos eritrócitos (RDW-CV), %HIPO, RET%, RET (grupo das mulheres), IFR, LFR, MFR e HFR compatíveis com o estado anêmico, que pode ser observado em 91,8%. Todos os parâmetros estudados apresentaram área sob a curva (AUC) > 0,4. Para o grupo dos homens, a %HIPO (AUC: 0,806) e a IFR (AUC: 0,762) apresentaram melhores valores de AUC; já para o grupo das mulheres foram a %HIPO (AUC: 0,806), a %HIPER (AUC: 0,815) e a IFR (AUC: 0,660). Conclusão: Avanço da medicina e surgimento de novas técnicas e parâmetros ...

Evaluation of red cell and reticulocyte parameters as indicative of iron deficiency in patients with anemia of chronic disease

Objective: The purpose of this study was to evaluate the effectiveness of mature red cell and reticulocyte parameters under three conditions: iron deficiency anemia, anemia of chronic disease, and anemia of chronic disease associated with absolute iron deficiency. Methods: Peripheral blood cells from 117 adult patients with anemia were classified according to iron status, and inflammatory activity, and the results of a hemoglobinopathy investigation as: iron deficiency anemia (n = 42), anemia of chronic disease (n = 28), anemia of chronic disease associated with iron deficiency anemia (n = 22), and heterozygous β thalassemia (n = 25). The percentage of microcytic red cells, hypochromic red cells, and levels of hemoglobin content in both reticulocytes and mature red cells were determined. Receiver operating characteristic analysis was used to evaluate the accuracy of the parameters in differentiating between the different types of anemia. Results: There was no significant difference between the iron deficient group and anemia of chronic disease associated with absolute iron deficiency in respect to any parameter. The percentage of hypochromic red cells was the best parameter to discriminate anemia of chronic disease with and without absolute iron deficiency (area under curve = 0.785; 95% confidence interval: 0.661–0.909, with sensitivity of 72.7%, and specificity of 70.4%; cut-off value 1.8%). The formula microcytic red cells minus hypochromic red cells was very accurate in differentiating iron deficiency anemia and heterozygous β thalassemia (area under curve = 0.977; 95% confidence interval: 0.950–1.005; with sensitivity of 96.2%, and specificity of 92.7%; cut-off value 13.8). Conclusion: The indices related to red cells and reticulocytes have a moderate performance in identifying absolute iron deficiency in patients with anemia of chronic disease...

Hierro corporal en donantes habituales de un banco de sangre de Medellín-Colombia / Body iron in repetitive donors of a blood bank from Medellìn-Colombia

Introducción: en donantes de sangre es importante conocer el hierro circulante, almacenado y funcional; tradicionalmente solo se incluye la medición de hemoglobina. Objetivo: describir el comportamiento del estado del hierro en donantes habituales según las características demográficas e ingesta de hierro y su relación con la ferritina, la hemoglobina y la hemoglobina reticulocitaria. Métodos: estudio descriptivo transversal que incluyó 70 donantes habituales del Banco de Sangre de la Universidad de Antioquia, seleccionados al azar. Se evaluaron la ferritina, el eritrograma, la hemoglobina reticulocitaria, el estudio coprológico, la actividad física y el consumo de hierro. Se calcularon medidas de resumen, frecuencias y se utilizaron las siguientes pruebas estadísticas t Student, U de Mann Withney, H de Kruskal Wallis, Anova y correlaciones, con un nivel de significación de p<0.05. Resultados: el 60 % de los donantes fueron mujeres y la edad promedio fue 33 años. El promedio de los parámetros eritrocitarios evaluados estuvo dentro de los valores de referencia; 14,3 % (n=10) presentaron ferropenia y 76,9 % (n=50) bajo consumo de hierro/día. Los hombres evidenciaron valores más altos para recuento de eritrocitos, hemoglobina, hematocrito, constantes corpusculares y ferritina. Los adolescentes (18-20 años) tuvieron valores más bajos de ingesta, reticulocitos y ancho de distribución eritrocitaria. La ferritina se correlacionó con la hemoglobina, hemoglobina corpuscular media, concentración de hemoglobina corpuscular media, ancho de distribución eritrocitaria y hemoglobina reticulocitaria. Conclusiones : se evidenciaron adecuados mecanismos reguladores del metabolismo del hierro con eritropoyesis eficiente y se sugieren estudios para evaluar aspectos nutricionales en los donantes y factibilidad de la determinación de ferritina en donantes habituales para verificar reservas adecuadas que compensen las pérdidas de sangre por la donación.

Introduction: in blood donors it is important to determine circulated, stored and functional iron; traditionally hemoglobin is the unique measurement included. Objective: we intended to describe the behavior of iron in repetitive donors according to demographic characteristics as well as iron intake, and its relation to ferritin, hemoglobin and reticulocyte hemoglobin. Methods: cross-sectional study of 70 repetitive donors at the University of Antioquía Blood Bank, selected at random. Ferritin, eritrogram, reticulocyte hemoglobin, stool, physical activity and iron dietary were evaluated. Summary measures and frequencies were calculated and the following tests were used: T Student, U of Mann Whitney, H of Kruskal Wallis, Anova and correlations in SPSS. Results: 60% of donors were women and the mean age was 33 years old. The median of the erythrocyte parameters evaluated was within reference values; 14,3 % (n = 10) showed iron deficiency and 76,9 % (n = 50) had low iron intake/day. Men got higher values for erythrocyte count, hemoglobin, hematocrit, corpuscular constants and ferritin. Teenagers (18 - 20 years old) had lower values in intake, reticulocytes and red blood cell distribution width. Ferritin correlated to hemoglobin, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, red blood cell distribution width and reticulocyte hemoglobin. Conclusion: adequate regulatory mechanisms of iron metabolism with efficient erythropoiesis were evident and we suggest further studies to evaluate nutritional aspects of the blood donors and the feasibility of ferritin determination in regular donors in order to verify adequate reserves to compensate blood caused by donation.

Reticulocyte maturity indices in iron deficiency anemia

Objective: The aim of this study was to analyze the reticulocyte maturity indices (low, medium, and high fluorescence ratios) in iron deficient 1- to 6-year-old children, and identify the prevalence of iron deficiency anemia in this population. Methods: The present study included 39 subjects, divided into two groups: control subjects (n = 33), and subjects with iron deficiency anemia (n = 6). The results were analyzed by Student's t-test for comparison of means. Differences were considered significant when two-tailed p-value < 0.05. Results: Subjects with iron deficiency anemia presented increases in the proportion of mean (10.3 ± 4.7% vs. 6.0 ± 3.4%; p-value = 0.003), and high fluorescence reticulocytes (2.3 ± 0.87% vs. 0.9 ± 0.9%; p-value = 0.03) compared to the control group. The prevalence of anemia in this population was 15% (n = 6). Conclusion: The indices related to immaturity of reticulocytes are higher in the presence of iron deficiency, thus demonstrating a deficiency in the raw material to form hemoglobin and are, therefore, possible early markers of iron deficiency and anemia. We emphasize the need to standardize these indices for use in clinical practice and lab test results. .