Hemoglobin analysis after overload training in athletes / Análise de hemoglobina após treinamento de sobrecarga em atletas / Análisis de la hemoglobina tras el entrenamiento de sobrecarga en atletas

ABSTRACT Introduction: The cardiovascular system provides athletes with the proper conditions for blood circulation, ensuring the stability and normal metabolism of the body's internal environment during exercise. Objective: Investigate the effect of overload training on the hemoglobin of male taekwondo athletes. Methods: Twenty-one male taekwondo athletes (level 2 or higher) were selected and trained for four weeks, five days per week, with an initial load intensity of 60% of the maximum heart rate and a weekly intensity increase of 10%. Before training and on every weekend during training, hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), reticulocyte count (Ret) were checked, RBC volume distribution width (RDW), hemoglobin content distribution width (HDW), mean reticulocyte volume (MCVr), mean reticulocyte hemoglobin concentration (CHCMr), serum iron (Fe) and ferritin (Fer). Results: After four weeks of increasing load training, athletes showed a progressive and significant decrease in Hb (P<0.01), manifested as exercise-induced hypohemoglobin, and MCV, MCH, MCHC, CHCMr, HDW, and serum ferritin were significantly or extremely decreased(P<0.) 05, P<0.01); RDW increased significantly (P<0.05); Changes in Ret and serum iron showed no statistical significance (P>0.05). Correlation analysis found that hemoglobin concentration had the highest correlation with MCHC and CHCMr. Conclusion: Four-week incremental load training can induce exercise-induced hypohemoglobin in male taekwondo athletes, its changes being most correlated with MCHC and CHCMr, but without significant correlation with serum Fe, RDW, HDW, and MCV. Level of Evidence: Therapeutic Studies -Investigation of Outcomes.

RESUMO Introdução: O sistema cardiovascular fornece aos atletas as condições propícias para a circulação sanguínea, garantindo a estabilidade e o metabolismo normal do ambiente interno corporal durante o exercício. Objetivo: Investigar o efeito do treinamento em sobrecarga sobre a hemoglobina dos atletas masculinos de Taekwondo. Métodos: Vinte e um atletas masculinos de taekwondo (nível 2 ou superior) foram selecionados e treinados durante 4 semanas, 5 dias por semana, com uma intensidade de carga inicial de 60% da frequência cardíaca máxima e um aumento de intensidade semanal de 10%. Antes do treinamento e em todos os fins de semana durante o treinamento, foi verificada a hemoglobina (Hb), volume corpuscular médio (MCV), hemoglobina corpuscular média (MCH), concentração corpuscular média de hemoglobina (MCHC), contagem de reticulócitos (Ret), largura de distribuição do volume de hemácias (RDW), largura de distribuição do conteúdo de hemoglobina (HDW), volume médio de reticulócitos (MCVr), concentração média de hemoglobina reticulócitos (CHCMr), ferro sérico (Fe) e ferritina (Fer). Resultados: Após 4 semanas em treinamento de carga crescente, os atletas mostraram uma diminuição progressiva e significativa em Hb(P<0,01), manifestada como hipohemoglobina induzida por exercício, e MCV, MCH, MCHC, CHCMr, HDW, e ferritina sérica foram significativamente ou extremamente diminuídos(P<0). 05, P<0,01); RDW aumentou significativamente (P<0,05); Alterações de Ret e ferro sérico não apresentaram significância estatística (P>0,05). A análise de correlação constatou que a concentração de hemoglobina teve a maior correlação com MCHC e CHCMr. Conclusão: O treinamento de carga incremental de quatro semanas pode induzir hipohemoglobina induzida por exercício em atletas taekwondo masculinos, sendo suas alterações mais correlacionadas com MCHC e CHCMr, mas sem correlação significativa com Fe sérico, RDW, HDW e MCV. Nível de evidência: Estudos Terapêuticos -Investigação dos Resultados.

RESUMEN Introducción: El sistema cardiovascular proporciona a los deportistas las condiciones propicias para la circulación sanguínea, garantizando la estabilidad y el metabolismo normal del medio corporal interno durante el ejercicio. Objetivo: Investigar el efecto del entrenamiento de sobrecarga en la hemoglobina de los atletas masculinos de taekwondo. Métodos: Se seleccionaron 21 atletas masculinos de taekwondo (de nivel 2 o superior) y se entrenaron durante 4 semanas, 5 días a la semana, con una intensidad de carga inicial del 60% de la frecuencia cardíaca máxima y un aumento semanal de la intensidad del 10%. Antes del entrenamiento y cada fin de semana durante el mismo, se comprobó la hemoglobina (Hb), el volumen corpuscular medio (MCV), la hemoglobina corpuscular media (HCM), la concentración de hemoglobina corpuscular media (MCH) y el recuento de reticulocitos (Ret), Anchura de distribución del volumen de glóbulos rojos (RDW), anchura de distribución del contenido de hemoglobina (HDW), volumen reticulocitario medio (MCVr), concentración media de hemoglobina reticulocitaria (CHCMr), hierro sérico (Fe) y ferritina (Fer). Resultados: Después de 4 semanas de entrenamiento con carga creciente, los atletas mostraron una disminución progresiva y significativa de la Hb (P<0,01), que se manifestó como hipohemoglobina inducida por el ejercicio, y el MCV, el MCH, el MCHC, el CHCMr, el HDW y la ferritina sérica disminuyeron de forma significativa o extrema (P<0,01). 05, P<0,01); el ADE aumentó significativamente (P<0,05); los cambios en el Ret y el hierro sérico no mostraron significación estadística (P>0,05). El análisis de correlación encontró que la concentración de hemoglobina tenía la mayor correlación con MCHC y CHCMr. Conclusión: El entrenamiento de carga incremental de cuatro semanas puede inducir hipohemoglobina inducida por el ejercicio en atletas masculinos de taekwondo, siendo sus cambios los más correlacionados con MCHC y CHCMr, pero sin correlación significativa con el Fe sérico, RDW, HDW y MCV. Nivel de evidencia: Estudios terapéuticos - Investigación de resultados.

Enfermedades frecuentes durante el período neonatal. Resultados del hemograma, frotis de sangre periférica, conteo de reticulocitos y conteo de plaquetas

RESUMEN El hemograma, el frotis de sangre periférica, el conteo de reticulocitos y el conteo de plaquetas en el período neonatal son exámenes fundamentales en el diagnóstico y seguimiento de algunas enfermedades más frecuentes en esta fase de la vida como la anemia y las infecciones, las cuales, en ocasiones, pueden causar gran morbilidad y mortalidad en el neonato. Se efectúa una revisión bibliográfica de los resultados del hemograma, lámina periférica, conteo de reticulocitos y conteo de plaquetas en el neonato a término y pretérmino describiendo las variaciones de estos parámetros en el síndrome anémico y la sepsis del recién nacido.

ABSTRACT The complete blood count, the peripheral blood smear, the reticulocyte count and the platelet count in the pathological neonatal period are fundamental tests in the diagnosis and follow-up of some of the most frequent diseases in this phase of life, such as anemia and infections, which can sometimes cause great morbidity and mortality in the newborn. A bibliographic review of the results of the hemogram, peripheral lamina, reticulocyte count and platelet count in term and preterm neonates is carried out, describing the variations of these parameters in the anemia syndrome and sepsis of the newborn.

Hemograma, frotis de sangre periférica, conteo de plaquetas y conteo de reticulocitos en el recién nacido normal y sus variaciones fisiológicas

RESUMEN Se define recién nacido normal a aquel niño con peso superior a 2500 gramos e inferior a 4000 gramos, a término, y que no presenta afección ni presente ni probable en el momento del nacimiento. Los valores del hemograma, la morfología del frotis de sangre periférica, el conteo de reticulocitos y de plaquetas en el recién nacido normal varían según su tiempo de vida y son efecto de diferentes causas fisiológicas; aunque también existen numerosas alteraciones patológicas en estos exámenes de laboratorio en caso de determinadas enfermedades que pueden aparecer en esta etapa de la vida. En los recién nacidos pre término estas cambios son más evidentes. Se realizó una revisión bibliográfica sobre el hemograma, lámina periférica, conteo de plaquetas y conteo de reticulocitos en el recién nacido normal donde se exponen los resultados de estos estudios según su tiempo de nacimiento. Se realizan conclusiones.

ABSTRACT A normal newborn is defined as a child weighing more than 2500 grams and less than 4000 grams, at term, and who does not present affection or possible affection at the time of birth. The values of the hemogram, the morphology of the peripheral blood smear, the reticulocyte and platelet count in the normal newborn vary according to their life span and are the effect of different physiological causes; although there are also numerous pathological alterations in these laboratory tests in the case of certain diseases that may appear at this stage of life. In preterm newborns these changes are more evident. A bibliographic review was carried out on the hemogram, peripheral lamina, platelet count and reticulocyte count in the normal newborn where the results of these studies are presented according to their time of birth. Conclusions are made.

Characteristics of bone marrow compensatory erythropoiesis in hereditary spherocytosis / 中华血液学杂志

Objective: To reveal the compensatory features of bone marrow (BM) erythropoiesis in hereditary spherocytosis (HS) and to explore the effect of diferent hemoglobin levels on this compensation. Methods: Clinical and laboratory data of patients with HS were collected, and the peripheral blood absolute reticulocytes counts value was taken as the surrogate parameter to evaluate the ability of erythropoiesis compensation. BM erythropoiesis compensation in HS with diferent degrees of anemia were evaluated. Results: ①Three hundred and two patients were enrolled, including 115 with compensated hemolytic disease, 74 with mild anemia, 90 with moderate anemia, and 23 with severe anemia. ②Hemoglobin (HGB) was negatively correlated with serum erythropoietin in the decompensated hemolytic anemia group (EPO; rs=-0.585, P<0.001) . ③The median absolute reticulocyte count (ARC) of HS patients was 0.34 (0.27, 0.44) ×10(12)/L, up to 4.25 times that of normal people. The maximum ARC was 0.81×10(12)/L, about 10 times that of normal people. The median ARC of patients with compensated hemolytic disease was 0.29 (0.22, 0.38) ×10(12)/L, up to 3.63 times that of normal people. The median ARC of patients with hemolytic anemia was 0.38 (0.30, 0.46) ×10(12)/L, which was significantly higher than the patients with compensated hemolytic disease, up to 4.75 times that of normal people (z=4.999, P=0.003) . ④ ARC was negatively correlated with HGB in the compensated hemolytic disease group (rs=-0.177, P=0.002) and positively correlated with HGB in the decompensated hemolytic anemia group (rs=0.191, P=0.009) . There was no significant difference in the ARC among patients with mild, moderate, and severe anemia (χ(2)=4.588, P=0.101) . ⑤The median immature reticulocyte production index of the mild, moderate, and severe anemia groups was 13.1% (9.1%, 18.4%) , 17.0% (13.4%, 20.8%) , and 17.8% (14.6%, 21.8%) , respectively; the mild anemia group had lower index values than the moderate and severe anemia groups (P(adj) values were both<0.05) , but there was no significant difference between the latter groups (P(adj)=1.000) . The median immature reticulocyte count of patients in the mild, moderate, and severe groups was 5.09 (2.60, 7.74) ×10(10)/L, 6.24 (4.34, 8.83) ×10(10)/L, and 7.00 (3.07, 8.22) ×10(10)/L, respectively; there was no significant difference among the groups (χ(2)=3.081, P=0.214) . Conclusion: HGB can be maintained at a normal level through bone marrow erythropoiesis, while red blood cells are reduced in HS. However, once anemia develops, the bone marrow exerts its maximum erythropoiesis capacity and does not increase, regardless of anemia aggravation or serum EPO increase.

Comparison of automated flowcytometric reticulocyte analysis with manual reticulocyte count

Background: Reticulocytes are young or immature red blood cells released from bone marrow and that contain remanants of ribonucleic acid (RNA) and ribosomes. Reticulocyte count (RC) is the index of erythropoietic activity within bone marrow. The reticulocyte counting methods at clinical laboratories are currently divided into manual and automated.Methods: A total of 500 samples of study cases were processed by manual method using New Methylene Blue (NMB) and automated method based on flowcytometry by PENTRA XLR HORIBA hematology analyzer. All quality control parameters were evaluated and values obtained by both methods were compared using various statistical methods.Results: Automated hematology analyzer provides excellent precision and linearity with no significant carryover. On comparing manual and automated RC method good method correlation was found (correlation coefficient r-0.865), however individual case wise percent deviation between manual and automated RC and CRC varied significantly. In addition within run precision calculated for automated RC differed significantly from manual count. The mean of difference between duplicate readings (150 samples) of manual and automated RC (<5%) were 0.3 and 0.01 respectively while 6.3 and 0.15 respectively for >5% RC. Thus, automated method was found to be more precise than the manual RC.Conclusions: The manual count method for RC associated with significant imprecision compared to flowcytometric method mostly based on interobserver variation and the smaller number of cell being counted. In contrast, the automated method is rapid, easy to operate, count higher number of cells with precise measurement.

Role of IV administered Iron sucrose with recombinant erythropoietin in the treatment of moderate and severe iron deficiency anemia in the third trimester of pregnancy

Background: Iron deficiency Anemia in pregnancy is one of the most common and intractable nutritional problems in the world today. The objective of this study was to investigate the therapeutic efficacy and safety of rHuEPO combined with IV iron sucrose, in the treatment of pregnant women in third trimester with moderate and severe iron deficiency anemia and whether addition of erythropoietin will increase the rate of rise of Hb without compromising on the safety of the therapy.Methods: 60 pregnant women in the third trimester, diagnosed as cases of moderate and severe iron deficiency anemia were enrolled in this study with 30 subjects in each of the 2 groups. Recombinant Erythropoietin 2000 IU s/c and Inj Iron sucrose 100 mg slow intravenously in 100 ml 0.9% NS over 1 hr on alternate days was administered to the case group and the control group was administered only iron sucrose slow IV in the same dose on alternate days till target Hb (11gm%) was reached. Efficacy measures were reticulocyte count, increase in Hb/week, time to target Hb level and need for continued therapy after 4 weeks.Results: In the case group, the increases in Hb were greater after 1 week of treatment and this was found to be significant (P < .01), the median duration of therapy was shorter in the case group (22 versus 34 days), with more patients reaching the target hemoglobin level by 4 weeks as opposed to 7 weeks in the control group. Average rise in Hb/week was much more in the case group. The groups did not differ with respect to maternal and fetal safety parameters.Conclusions: Iron sucrose plus rhEPO is an effective treatment for iron deficiency anemia in pregnancy probably because of a synergistic action, with rhEPO stimulating erythropoiesis and iron sucrose delivering iron for hemoglobin synthesis.

Subclinical Malaria and Reticulocyte Count in Apparently Healthy Female Undergraduate Students in Rivers State University, Port Harcourt, Nigeria

Aims: The study was aimed at determining subclinical malaria and estimating reticulocyte count in apparently healthy female undergraduate students of Rivers State University, Port Harcourt.Study Design:This is a non-randomized, comparative case-control study.Place and Duration of Study:The study was conducted using female students residing at the hostels of Rivers State University, Port Harcourt. Analysis was carried out at the Haematology Laboratory, Department of Medical Laboratory Science, Rivers State University, Port Harcourt, Nigeria, between July and August, 2018. Methodology:For the subjects used in this study, a total of 32 students (32%) that were diagnosed of having Plasmodium falciparum malariainfection were used as test subjects, while a total of 68 students (68%) that were diagnosed to be Plasmodium falciparumnegative, and withoutOriginal Research Article malaria, were used as control.Thick and thin blood films examination using Giemsa staining technique was used to detect and calculate the malaria parasite density while a thinbloodfilm examination using new methylene blue staining technique was used to evaluate the reticulocyte count in the blood.Results:The reticulocyte count of test subjects (subjects with Plasmodium falciparummalaria) was 0.15 ± 0.04%and that of control subjects (subjects without any malaria parasite) was 0.31 ± 0.08%. The test subjects had significantly lower reticulocyte count (p ˂ 0.0001) than the control subjects. The age range “15-19” years had the highest malaria parasite density of 0.52 ± 0.18%, while “25-29” years had the least parasite density of 0.33 ± 0.24. There was no statistical variation in malaria parasite density according to age ranges (p = 0.13; p ˃ 0.05). However, the age range of “15-19” years had the lowest reticulocyte count as most of the female students within this age group were diagnosed to have been infected with malaria parasite.Conclusion:This study revealed that reticulocyte counts of malaria (Plasmodium falciparum) infected individuals decreased when compared to those without malaria parasite and this decreasewas statistically significant. There was no statistical significant variation in malaria parasite density irrespective of age ranges. Prophylaxis for malaria in such settings would be an efficient means of preventing infectious reservoirs and higher rates of subclinical malaria infection.

Correlação entre as contagens de reticulócitos manual e automática em amostras de felinos anêmicos / Correlation between manual and automatized values of reticulocyte count in anemic feline samples

O objetivo do presente estudo foi correlacionar os valores de reticulócitos pontilhados e agregados obtidos por metodologia manual com a metodologia automática de contagem de reticulócitos totais em amostras de sangue de gatos anêmicos, analisados em um contador hematológico com citometria de fluxo. Para isso, 40 amostras de sangue de pacientes felinos anêmicos, independentemente de idade e sexo, foram utilizadas para a determinação das contagens absolutas de reticulócitos totais pela metodologia automatizada por citometria de fluxo fluorescente e pela técnica manual com corante supravital, em duplicata. Na contagem manual, houve a discriminação entre reticulócitos pontilhados e agregados. Para a correlação entre os métodos, foi realizada a análise de regressão de Passing-Bablok. A média do hematócrito dos gatos foi de 15,25%, tendo a maioria dos gatos (32,5%) apresentado anemia moderada (hematócrito = 17,81%). Como resultados, a análise de regressão demonstrou que a correlação entre a contagem absoluta total automática foi superior à contagem manual de reticulócitos agregados (rho= 0,71; P<0,001) do que a contagem absoluta de reticulócitos pontilhados (rho= 0,68; P<0,001). Os resultados apresentados sugerem que a contagem de reticulócitos total absoluta realizada pelo analisador hematológico ProCyte Dx em gatos anêmicos se refere à contagem absoluta de reticulócitos. Dessa maneira, recomenda-se que os valores possam ser utilizados para a avaliação imediata da condição hematológica de gatos anêmicos.(AU)

The aim of this study was to correlate the punctate and aggregated reticulocytes values obtained by manual methodology and the automatic reticulocyte count in 40 blood samples from anemic cats. Total reticulocyte absolute counts were determined by automated fluorescence flow cytometry and manual methods in 40 blood samples obtained from anemic cats. The manual count was obtained by supravital stain in duplicate to each sample and the reticulocyte morphology were discriminated between punctate and aggregates reticulocytes. Passing-Bablok regression analysis was utilized to compare the methods. Most samples were from anemic cat (15,25%) and the hematocrit mean was 17,81%. Regression analysis showed that the correlation between the absolute total automatic counts is higher with aggregated reticulocytes (rho= 0,71; P< 0,001) than with absolute punctate reticulocytes counts (rho= 0, 68, P< 0.001). Results suggest that the ProCyte Dx reticulocytes count in anemic cats is correlated with aggregate reticulocyte count. Thus, the greater amount of RNA and organelles in aggregate reticulocytes generates a cellular complexity and, therefore, greater impregnation of the dye in an automatic count. Thus, the values obtained by the hematologic instrument can be used for the immediate evaluation of the hematological condition in anemic cats.(AU)

Contagem de Reticulócitos na Prática Clínica: um Exame Pouco Utilizado / Reticulocytes Counting in Clinical Practice: a Little Examination Used

Objetivo: Estimar o percentual de contagem de reticulócitos em relação aos hemogramas realizados nos laboratórios de análises clínicas de Divinópolis/MG. Métodos: Trata-se de um estudo descritivo realizado nos laboratórios de análises clínicas do município de Divinópolis/MG inscritos no Cadastro Nacional de Estabelecimento de Saúde (CNES). Foi realizada uma entrevista através de questionário a fim de obter as informações dos números de hemogramas e contagens de reticulócitos realizados durante os últimos 12 meses, do atendimento (ambulatorial ou hospitalar), da instituição (pública ou privada), do porte (pequeno, médio ou grande) e de quais os métodos eram utilizados para realização dos exames. Resultados: Dos 15 laboratórios inscritos no CNES, 11 foram entrevistados (73%). Foram realizados 276.666 hemogramas e 4.248 contagens de reticulócitos nos últimos 12 meses em todos os laboratórios, o que significa o percentual de 1,5% de contagem de reticulócitos em relação aos hemogramas realizados. A maioria dos laboratórios é privado (81%), de médio porte (54%) e realizam atendimento ambulatorial (54%). Para realizar o hemograma, a maioria (81%) utiliza a técnica citometria de fluxo e para a contagem de reticulócitos todos realizam a técnica manual com o reagente azul de cresil brilhante. Conclusão: O percentual de contagem de reticulócitos em relação aos hemogramas realizados em laboratórios do município de Divinópolis/MG é baixo, mesmo sendo o primeiro exame complementar para o diagnóstico diferencial da anemia. Essa baixa solicitação pode estar relacionada com a desvalorização do exame no meio clínico ou pela falta de acurácia e pobre reprodutibilidade dos resultados encontrados. (AU)

Objective: To estimate the percentage of reticulocyte counts in relation to blood count performed in the clinical analysis laboratories in Divinópolis / MG. Methods: This is a descriptive study carried out in the clinical analysis laboratories of Divinópolis/MG registered in the National Register of Health Establishment (CNES). A questionnaire interview was conducted in order to obtain information on the number of blood counts and reticulocyte counts performed during the last twelve months. The questionnaire also aimed to identify whether the institution was public or private, whether it was outpatient or hospital care and which methods were used to perform the tests. Results: Of the 15 laboratories registered in the CNES, 11 were interviewed (73%). 276,666 blood counts and 4,248 reticulocyte counts were performed in the last 12 months in all laboratories, which means the percentage of reticulocyte count of 1.5% in relation to the blood counts performed. Most of the laboratories are private (81%), medium-sized (54%) and carry out outpatient care (54%). To perform the blood count, most (81%) use the flow cytometry technique and for reticulocyte counting all perform the manual technique with the bright blue cresyl reagent. Conclusions: The percentage of reticulocyte counts in relation to blood counts performed in laboratories in the city of Divinópolis/MG is low, even though it is the first complementary exam for the differential diagnosis of anemia. This low request may be related to the devaluation of the reticulocyte in the clinical environment or the lack of accuracy and poor reproducibility of the results found. (AU)

Evaluation of the interference of platelets in reticulocyte counting by flow cytometry using thiazole orange / Avaliação da interferência de plaquetas na contagem de reticulócitos por citometria de fluxo, utilizando laranja de tiazol

ABSTRACT Introduction: The reticulocyte count by flow cytometry (FC) - an automated counting method - can present errors due to the presence of interfering factors, contributing to a slight increase in results. However, automated methods have large advantages over the manual method, taken as reference, what justifies efforts to improve their quality. Objective: Evaluate platelet interference with the reticulocyte count by FC, using thiazole orange (TO) (FC/TO). Materials and methods: The method of reticulocyte count by FC/TO and a modified automated equivalent method, which excluded CD61-positive cells (platelets) from analysis (FC/TO/MOD), were compared to the manual method. Conclusion: Results were analyzed according to the recommendations of the Clinical and Laboratory Standards Institute (CLSI) to assess interchangeability between the methods, by linear regression analysis and paired t-test. The exclusion of interfering fragments from result analysis by the modified method produced results in closer proximity to those of the reference method.

RESUMO Introdução: A contagem de reticulócitos por citometria de fluxo (CF) - um método de contagem automatizada - pode apresentar erros devido à presença de interferentes, contribuindo para uma ligeira elevação dos resultados. No entanto, os métodos automatizados possuem grandes vantagens em relação ao manual, tido como referência, o que justifica esforços para a melhoria de sua qualidade. Objetivo: Avaliar a interferência de plaquetas na contagem de reticulócitos por CF, utilizando laranja de tiazol (thiazole orange [TO]) (CF/TO). Materiais e métodos: O método de contagem de reticulócitos por CF/TO e um método equivalente automatizado modificado, no qual se excluíram células CD61-positivas (plaquetas) da análise (CF/TO/MOD), foram comparados com o método manual. Conclusão: Os resultados foram analisados de acordo com as recomendações do Clinical and Laboratory Standards Institute (CLSI) para avaliar a intercambialidade entre os métodos, por meio da análise de regressão linear e do teste t pareado. A exclusão de interferentes da análise dos resultados pelo método modificado demonstrou maior proximidade dos resultados com aqueles do método de referência.

Pattern Of Reticulocyte Count And Hemoglobin Concentration In Pregnant And Non- Pregnant Women In Rural Haryana.

Background & Objective: Anemia is a common nutritional health problem in women’s child bearing age. Pregnant women are at higher risk particularly during 3rd trimester of pregnancy. By definition, it is decrease in number of RBCs below 4.5 million or Hb level below 11.0 gm % and hematocrit < 0.33 in pregnant women. Anemia contributes significantly to maternal mortality and morbidity and also carries a risk factor for infant iron deficiency anemia. Main causes of maternal anemia are blood loss, increased menstrual bleeding and nutritional iron & folate deficiency. Anemia can be estimated by Hb estimation and Reticulocyte count (RC), which is a quantitative measure of Bone marrow production of new RBCs, with a range of 0.5-2.5%(adults) and 2.6% in infants. Reticulocyte Index (RI) is a calculation of ratio b/w level of anemia in response to which, RC has risen. Objective is to study pattern of reticulocyte count in pregnant and non-pregnant women, 2) To compare Hb level in pregnant women in 3 trimesters and non-pregnant women.3) To analyze pattern of reticulocyte count and compare levels of Hb concentration in pregnant women in 3 trimesters and non- pregnant women. Methodology: A total of 100 women with age range of 18-40 yrs were examined (50 pregnant and 50 healthy non pregnant women) presenting to Obstetrical O.P.D. Blood samples from subjects were collected, tested and hematologically analyzed. Both Inclusion and Exclusion criteria strictly followed. Results & Conclusion: Hb level in pregnant women was significantly low as compared to non pregnant women, whereas Reticulocyte count was higher in pregnant women than non pregnant women. Variation of Hb concentration was seen as, high in 3rd Trimester. R C was raised in 2nd Trimester and reached peak in 3rd trimester, while it remained normal or decreased in 1st trimester. Non-pregnant women showed normal range of R C. So values of all red cell parameters in pregnant women was significantly low, as compared to age matched controls except Reticulocyte count, which was raised during pregnancy.

Investigation and Analysis of Reference Intervals and Decision Limits of Reticulocyte Count in China / 现代检验医学杂志

Objective To investigate and analyze the current situation of reference intervals and decision limits of reticulocyte count in China.Methods The information related to upper/lower limits,decision limits,data resources,method principles, instruments and reagents of reticulocyte count in 2014 was collected via the external quality assessment software system based on website.The participants were divided according to their methods and instruments.The SPSS 1 9.0 was used for conducting the data analysis and statistics of results for each group.Results There were 276 laboratories submitting the val-id results.The major resources for reference intervals came from national guide to clinical laboratory procedures (53.99%), instructions of instrument manufacturers (16.30%),textbook (9.78%),instructions of reagents manufacturers (9.06%) and others.The major resources for decision limits were national guide to clinical laboratory procedures (56.06%),instruc-tions of instrument manufacturers (16.67%),textbook (7.58),instructions of reagents manufacturers (7.58%)and others. The medians and averages of upper/lower limits and decision limits among subgroups were close,whereas the P2.5 and P97.5 were significant discrepancies.Only 45.65% and 48.48% of participant laboratories had verified reference intervals and deci-sion limits before clinic use.The lower limits and decision limits among subgroups had nostatistical difference (P>0.05), where as the upper limits of different instrument groups had significant differences (P<0.001).Conclusion The current situation of reference intervals and decision limits of reticulocyte count is unsatisfactory.It is certain to establish a uniformed reference interval and decision limits for reticulocyte count,which provides a reference for the standardization of reticulocyte count testing.

Reticulocyte count: comparison among methods / Contagem de reticulócitos: comparação entre métodos

Introduction: The automated counting of reticulocytes has some advantages over the manual method routinely used in clinical laboratories. Technological innovations provide more statistically reliable results, while optimizing the time to perform this test. However, the cost for implementing the automated procedure in laboratory routines still constitutes a barrier to its use in small- and medium-size Brazilian laboratories. Objective: This study evaluated the performance of a new laboratory protocol for reticulocyte counting by flow cytometry using acridine orange (FC/AO), compared with the manual method and with another automated one by flow cytometry using the commercial kit BD Retic-Count (FC/RC) Conclusion: The results showed that, besides being comparable to the manual method, still considered standard, the evaluated new protocol is economically more advantageous than the automated methods currently available, and its cost is comparable to that of the manual method for laboratories that already have appropriate equipment and infrastructure. .

Introdução: A contagem automatizada de reticulócitos apresenta vantagens em relação ao método manual, rotineiramente utilizado em laboratórios clínicos. Inovações tecnológicas permitem resultados estatisticamente mais confiáveis, além de otimizarem o tempo para realização desse exame. No entanto, o custo para aplicação do procedimento automatizado em rotinas laboratoriais ainda constitui uma barreira para sua implementação em laboratórios brasileiros de pequeno e médio porte. Objetivo: O presente estudo avaliou o desempenho de um protocolo laboratorial para contagem de reticulócitos por citometria de fluxo utilizando acridine orange (CF/AO), comparando-o com o método manual e o automatizado por citometria de fluxo utilizando o kit comercial BD Retic-Count (CF/RC). Conclusão: Os resultados mostraram que, além de ser comparável com o método manual, ainda considerado padrão, o protocolo avaliado é economicamente mais vantajoso do que os métodos automatizados atualmente disponíveis, sendo o seu custo comparável com o do método manual para laboratórios que já apresentam aparelhagem e infraestrutura adequadas. .

Contagem de reticulócitos por citometria de fluxo utilizando acridine orange: validação de um novo protocolo / Flow cytometry reticulocyte counting using acridine orange: validation of a new protocol

Introduction: Currently, the reticulocyte counting is a challenge for clinical laboratories in Brazil, mainly for the ordinary ones, which still use the manual method. This method has some limitations, since it consists of a laborious method, time consuming, with low accuracy. Objectives: This study has developed and evaluated the performance of a New Laboratory Protocol for flow cytometry (FC) reticulocytes counting using acridine orange (AO) as dye, aiming to standardize a more precise, easy, fast implementation, and low cost protocol. After standardization of the New Protocol (FC/AO), it was compared with the manual method. The results were analyzed according to the recommendations of the National Committee for Clinical Laboratory Standards (NCCLS), now known as Clinical and Laboratory Standards Institute (CLSI), to evaluate the interchangeability of methods in linear regression analysis and paired t test, besides other quality control tests. Conclusion: Based on these results concerning to the correlation between the methods and the tests related to quality control, we can admit that FC/AO for reticulocyte counting shows undeniable advantages when compared to the preexisting manual method...

Introdução: Atualmente, a contagem de reticulócitos representa um desafio para os laboratórios clínicos no Brasil, principalmente os de pequeno e médio porte, nos quais ainda se utiliza o método manual. Este método apresenta algumas limitações, classificando-se como tedioso, demorado e de baixa precisão. Objetivos: O presente estudo desenvolveu e avaliou o desempenho de um novo protocolo laboratorial para contagem de reticulócitos por citometria de fluxo (CF) utilizando acridine orange (AO) como corante, visando padronizar um protocolo preciso, de fácil e rápida execução e custo acessível. Após a padronização do novo protocolo desenvolvido (CF/AO), fez-se a comparação com o método manual. Os resultados foram analisados de acordo com recomendações do National Committee for Clinical Laboratory Standards (NCCLS), atualmente Clinical and Laboratory Standards Institute (CLSI), para avaliar a intercambialidade entre os métodos, por meio da análise de regressão linear e teste t pareado, além de outros testes de controle de qualidade. Conclusão: Diante dos resultados obtidos referentes à correlação entre os métodos e os testes voltados ao controle de qualidade, pode-se admitir que o CF/AO estabelecido para contagem de reticulócitos possui vantagens inegáveis quando comparado com o método manual...

Interpretación del hemograma automatizado: claves para una mejor utilización de la prueba / Interpretation of automated blood count: the key to a better use of the test

Resumen: El hemograma es una de las pruebas que más se solicita al laboratorio clínico y, sin duda, una de las que más aporta al clínico en la evaluación del paciente. Desde el punto de vista técnico, se reconocen seis tipos de hemograma y el tipo VI es el que ofrece mayor cantidad de parámetros de utilidad clínica, los cuales se obtienen a partir de una amplia combinación de tecnologías, como el enfoque hidrodinámico y la citometría de flujo. En este módulo se describen los parámetros que componen el hemograma tipo VI, la utilidad clínica, los valores de referencia y algunos aspectos que se deben tener en cuenta al interpretar los resultados y tomar decisiones médicas; además, se hace énfasis en los nuevos parámetros disponibles en el hemograma tipo VI, como el recuento de reticulocitos y los índices derivados, el recuento de granulocitos inmaduros y de plaquetas reticuladas, entre otros, los cuales son parámetros de gran utilidad en diferentes escenarios clínicos y hacen que este hemograma sea el de elección para la práctica médica actual.

Abstract: The complete blood count (CBC) is one of the most requested tests in the clinical laboratory, and without a doubt, it is one of the tests that provides some of the most useful information for physicians taking care of patients. From a technical perspective, there are six types of complete blood counts, and the type-VI CBC offers the greatest amount of useful parameters, which are obtained using different technologies integrated into a single platform, including hydrodynamic focusing and flow cytometry. In this review article, the parameters included in the complete blood count are described, including their clinical usefulness, reference ranges and other issues that should be taken into account during the time of analyzing the results and during the entire medical decision- making process. In addition, the new parameters available with the type-VI CBC are emphasized, for example the reticulocyte count and derived parameters, immature granulocytes count, and immature platelet count, among others, which are extremely useful in different clinical scenarios and make the type-VI CBC of choice for current medical practice standards.

Serum Hepcidin Level as an Early Marker of Hematopoietic Activity after Allogeneic Peripheral Blood Stem Cell Transplantation / 임상검사와정도관리

BACKGROUND: Hepcidin plays a central role in the regulation of iron metabolism, and hepatic iron production is stimulated by iron load and inflammation. Recent animal studies have shown that hepcidin levels increase when hematopoiesis is blocked. We aimed to monitor pre- and post-stem cell transplantation hepcidin levels and evaluate its association with hematologic recovery. METHODS: The study group comprised 12 patients with hematologic malignancies (7 with AML, 4 with ALL, and 1 with refractory anemia with excess blasts-2) undergoing allogeneic peripheral blood stem cell transplantation (PBSCT). One day before and 3 days, 1 week, 2 weeks, 4 weeks, and 8 weeks after PBSCT, reticulocyte count and levels of Hb, ferritin, and C-reactive protein were monitored; serum hepcidin-25 was measured by ELISA. RESULTS: The median serum hepcidin-25 levels (ng/mL) were significantly higher until 1 week after PBSCT (103.6, 103.3, and 96.5) than those at 2, 4, and 8 weeks after PBSCT (63.9, 53.9, and 56.6, respectively). The reticulocyte count also significantly increased from 2 weeks after PBSCT. The hepcidin level showed an inverse correlation with reticulocyte count (r=-0.56, P or =63.9) tended to demonstrate lower Hb recovery at 8 weeks than patients with low hepcidin levels did (P=0.15), but without any differences in the incidence of complications. CONCLUSIONS: These findings indicate that hepcidin production is associated with erythropoietic activity and that hepcidin level may be used as an early marker of hematopoietic recovery in PBSCT.

Correction of Pseudoreticulocytosis in Leukocytosis Samples Using the Sysmex XE-2100 Analyzer Depends on the Type and Number of White Blood Cells

BACKGROUND: The reticulocyte count is a good marker of erythropoietic activity of the bone marrow. In the mid-1990s, automated flow cytometric analysis replaced microscopy for the quantification of reticulocytes. Leukocytosis cases with an erroneously high reticulocyte count and a high immature reticulocyte fraction (IRF) have been reported. In this study, we analyzed reticulocyte counts in leukocytosis samples, in an effort to identify a correction method. METHODS: The study comprised of 21 samples from 16 leukocytosis patients. Results of reticulocyte analyses obtained using a XE-2100 hematology analyzer (Sysmex, Japan) were compared with those obtained using the supravital staining technique, which is a reference method. If the samples showed erroneously high reticulocyte counts and IRF, they were reanalyzed after serial dilution with isotonic solution. RESULTS: Five samples from 4 patients showed erroneously elevated reticulocyte counts and/or IRF on the XE-2100 analyzer. They displayed abnormal reticulocyte scattergrams, with 4 of 5 cases indicated by a flag. The white blood cell (WBC) fractions overlapped with the reticulocyte regions, especially with the IRF. Diagnoses and blast counts were variable when such errors occurred; WBC counts varied from 218.19x10(9)/L to 725.14x10(9)/L. The errors were corrected by simple dilution with isotonic solution. However, the corrective WBC counts differed according to individual cases. CONCLUSIONS: When leukocytosis samples exhibit an abnormal reticulocyte scattergram with a flag, or an abnormally high IRF, we recommend the dilution of the sample with isotonic solution to a WBC count of about 100.00x10(9)/L, followed by reanalysis of the reticulocyte count and reticulocyte scattergram.

Is erythropoietin an acceptable treatment modality for routine use in preventing anaemia of prematurity.

Objectives: To find out whether routine use of erythropoietin (EPO) would reduce transfusion requirements in infants <1.5 kg and/or <32 weeks at birth and compare haemoglobin and reticulocyte counts of EPO treated and untreated groups. Methods: Seventy eight infants were randomized. Cases received subcutaneous EPO 300 units/kg/day,thrice weekly starting before 72 hours of birth for 14 days. All infants were given oral iron and folic acid with initiation of enteral feedings and this was continued during the entire treatment period. Results: EPO treated group acquired high reticulocyte counts earlier than control group with maximal difference in 2nd week but there was no statistically significant difference between groups. There was a gradual decline in haemoglobin values in both cases and controls; though this was less marked in the EPO treated group, the difference was not statistically significant. There was no statistically significant difference in total transfusion requirements between infants who received EPO and the control group. Conclusion: There is no benefit from routine use of EPO during the first two weeks of life in preventing anaemia of prematurity.

Normal ranges of peripreral RET-He and reticulocyte grouping parameters in healthy adults in Wenzhou / 中华检验医学杂志

Objective To investigate the normal ranges of RET-He and reticulocyte grouping parameters of healthy adults in Wenzhou. Methods A total of 1 000 samples from healthy adults were collected in Wenzhou from August 2008 to February 2010. There were 506 males and 494 females. The individuals were stratified based on age into three groups:20 to 40-year-old group (n = 341 ), ＞40 to 60-year-old group (n =316), and ＞60-year-old group (n =343). All blood samples were drawn in EDTA-K2 anticoagulated blood. Peripheral RET-He and reticulocyte grouping parameters ( IRF, LFR, MFR, HFR)were determined by Sysmex XE-2100 automated hematology analyzer. The instrument was calibrated and validated before testing All specimens were analyzed within 2 hours. RET-He, RET absolute value and RET percentage were normal distribution, the normal range was established by using ((x)-1.96s,(x)+ 1.96 s)method. IRF, LFR, MFR and HFR data were skewed distribution,the normal range was established by using (P2.5 ,P97.5 )percentile method. Results The mean and normal range of RET-He were (33.91 ± 1.77) pg and (30.44 - 37.38) pg in male, (33.97 ± 1.85) pg and (30.34 - 37.60) pg in female,respectively. There was no significant difference between the two groups ( t =-0.533, P＞0.05 ). The mean and normal range of RET absolute value were (0.051±0.023) × 1012/L and (0.006 -0.096)×1012/L in male,which were (0. 037 ±0. 026) × 1012/L and (0 -0. 088) × 1012/L in female, there was significant difference between the two groups ( t = 8. 409, P ＜ 0. 05 ) . The mean and normal range of RET percentage in male were(1.041 ±0.459)% and (0. 141 -1.941)% , and in female were (0.869±0.603)% and (0-2. 051 )%, which showed significant difference (t =5.074,P ＜0. 05). The normal ranges of LFR,MFR,HFR,IRF in male were 0. 951 (0.866 -0.988) ,0.046(0.010 -0.114) ,0. 003(0 -0.023) ,0. 050(0.012-0.134), and in female were 0.096(0.880 -0.991) ,0.039(0.008 -0.113) ,0.002(0-0.016) ,0.040(0.009-0.121), respectively. There were significant differences between the two groups (Z values were -5.044, -4.793, -3.559, -5.075, respectively,all P＜0.05). The means of RET-He in 20 to 40-year-old group, ＞40 to 60-year-old group, ＞ 60-year-old group and all age group were (33.38±1.49),(34.36 ±1.46), (34.12±2.21) and (33.94 +1.81) pg, respectively. The normal ranges were(30.46-36.30), (31.50 -37.22), (29.79 - 38.45) and (30.39 - 37.49) pg, which showed statistical difference( F =28. 072,P ＜0. 05). In addition, the normal range of RET-He in 20 to 40-year-old group was lower than that of ＞ 40 to 60-year-old group and ＞ 60-year-old group ( q values were 9. 970, 7. 791,respectively, P＜0. 05). The normal ranges of LFR, MFR, HFR, IRF in ＞ 60-year-old group were 0.961(0. 878 -0. 992) ,0. 037(0. 007 -0. 105 ) ,0. 002(0 -0. 020) and 0. 039(0. 008 -0. 117) ,while they were 0.953(0. 867 -0.990), 0.045(0.009 -0. 116) ,0.003(0 -0.019) ,0. 050(0. 010 -0. 133) in 20 to 40 -year-old group , and 0. 951 (0.855 -0.989) ,0.047 (0.010 - 0.133 ) ,0.003 ( 0-0.020), 0.049 (0.011-0.149)in ＞40 to 60-year-old group, which showed statistical difference (Z values were -3. 949, -4.236,-4. 373, -4. 973, - 2. 747, - 3. 275, - 3.901, - 4. 185, respectively, all P ＜ 0. 05). There was nodifference between male and female for RET-He, the same normal ranges could be used, but there was significant difference between different age groups. LFR, MFR, HFR and IRF were statistically different between different age groups and different sex groups. The normal ranges should be established by gender and age. Conclusion The normal ranges of RET-He and reticulocyte grouping parameters about healthy adults in Wenzhou are established, which plays an important role in clinical value of RET parameters.

Reference Intervals of Reticulocyte Parameters using ADVIA 120 / 대한진단검사의학회지

BACKGROUND: Reticulocytes are circulating immature erythrocytes at the stage just before maturity of the erythrocyte. The maturation time for reticulocyte is 3.5 to 4 days including one day of circulation; therefore, circulating reticulocytes represent marrow erythropoietic activity that is faster than any other hematologic item. To take advantage of reticulocyte parameters, we should first identify the reference ranges and understand the basic information of the parameters. METHODS: We selected healthy subjects with normal Hb, WBC and platelet count in an annual health surveillance of employees of Boramae Hospital. In total, 285 subjects (74 males and 211 females) were analyzed using ADVIA 120. The analyzed parameters were reticulocyte count, reticulocyte indices and reticulocyte maturity indices. The reference intervals were obtained with the central 95% of the distribution. RESULTS: The reference intervals: reticulocyte count 41.8-121.3 (106/ L), 0.9-2.5 (%); MFR (middle fluorescence reticulocyte) 1.3-9.8 (%); HFR (high fluorescence reticulocyte) 0-1.5%; MCVr (MCV for reticulocyte) 100.6-112.7 (fL); CHCMr (cellular hemoglobin concentration mean for reticulocyte) 29.5-33 (g/dL); RDWr 8-10.5 (%); CHr (cellular hemoglobin content for reticulocyte) 30.8-35.5 (pg). According to the maturation of reticulocytes, the volume decreased 20%, CHCM increased 11%, and CH decreased 6%. CONCLUSIONS: The reticulocyte parameters are available for realtime assessment of marrow erythropoietic activity and clinically they will help in the diagnosis and treatment of various anemias.