Causes of Malfunction of Bioprostheses Inserted Percutaneously in the Aortic Valve Position in Patients Whose Native Aortic Valve Was Congenitally Bicuspid and Stenotic.

Transcatheter aortic valve implantation (TAVI) has brought in recent years relief of cardiac-induced symptoms to a large number of patients with aortic stenosis. Whether it is better to use TAVI for the treatment of aortic valve stenosis superimposed on a congenitally bicuspid valve has been debated in contrast to its proved usefulness in aortic valve stenosis involving a tricuspid aortic valve. From January 2020 to March 2023, surgical aortic valve replacement of TAVI valve and native aortic valve was done in 6 patients. The clinical findings of the patients and morphologic findings from the surgical specimens submitted to the cardiac pathology department were subsequently examined. All the 6 native aortic valves had bicuspid configuration. The TAVI valve in each patient was excised from 9 to 88 months (mean 36 months) after it had been implanted because of paravalvular leak in 4, severe stenosis of the prosthetic valve in 1, and bioprosthetic cuspal degeneration in 1. Prosthetic valve endocarditis was clinically suspected in 2 patients, but the specimen culture was negative. Before surgical aortic valve replacement, 3 patients experienced stroke after TAVI. All 6 patients had low hemoglobin levels (mean 9.5 mg/100 ml) and low hematocrit levels (mean 29.5%). Reticulocyte count was available in 4 patients and was increased in all (mean 3.5%). When the stenotic native aortic valve configuration is bicuspid, the raphe tends to be calcified first and located perpendicular to the flow of the blood and may prevent the ring of the caged bioprosthesis from being transferred to the aortic wall, which is a requirement for full opening of the lumen of the bioprosthesis. Thus, thorough consideration needs to be made before performing TAVI in patients whose native aortic valve is stenotic and bicuspid.

Regenerative response in dogs naturally and experimentally infected with Babesia rossi.

BACKGROUND: The regenerative response following Babesia rossi infection in dogs is mild, despite severe hemolytic anemia. OBJECTIVE: We aimed to compare the admission absolute reticulocyte count (ARC) and reticulocyte indices in 103 dogs naturally infected with B. rossi with 10 dogs suffering from immune-mediated hemolytic anemia (IMHA) and 14 healthy control dogs. The regenerative response was also evaluated in five dogs experimentally infected with B. rossi. METHODS: This is a retrospective observational study of records generated on the ADVIA 2120 hematology analyzer. RESULTS: The median hematocrits (HCT) of the B. rossi and IMHA groups were significantly lower than the control group (p < .001 for both); however, no differences were seen between the B. rossi and IMHA groups. Compared with the control group, the median ARC was significantly higher in the B. rossi (p = .006) and IMHA (p = .019) groups but significantly lower in the B. rossi group than the IMHA group (p = .041). In the experimentally infected dogs, there was a sudden decrease in the ARC approximately 48 h after the detection of peripheral parasitemia, which was followed by an increase after treatment. Reticulocytes of naturally infected B. rossi dogs were larger, with more variation in cellular volume. The reticulocytes of the experimentally infected dogs decreased in size with decreasing hemoglobin concentrations as the study progressed. CONCLUSIONS: The regenerative response in dogs naturally infected with B. rossi is inadequate, given the severity of the anemia observed, and it might be a result of direct suppressive action by the parasite or host response on the bone marrow.

Umbilical Cord Blood Bilirubin, Albumin, Reticulocyte Count, and Nucleated Red Blood Cells to Predict Subsequent Hyperbilirubinemia in Term Neonates: A Prospective Observational Study.

INTRODUCTION: Hyperbilirubinaemia is one of the most important causes of re-admission in the early neonatal period. The socioeconomic factors are one of the most common reasons for early discharge in a developing country like India. OBJECTIVES: This study aims to evaluate and analyze the statistical correlation of umbilical cord blood bilirubin, albumin, nucleated red blood cells (nRBC), and reticulocyte count as early predictors of neonatal hyperbilirubinemia. METHOD: A prospective observational study was conducted from November 2015 to April 2017 in a tertiary care hospital in North Karnataka, India. Umbilical cord blood was collected at birth for analysis of bilirubin, albumin, reticulocyte count, and nRBC in term neonates. Total serum bilirubin (TSB) levels were estimated using the VITROS BuBc Slide method at 72 hours of life. Data were analyzed using SPSS version 23 (IBM Corp., Armonk, NY). RESULTS: A total of 200 term neonates were enrolled in the study, out of which 123 completed follow-ups. Of the 66 newborns who had cord bilirubin levels ≥1.75 mg/dl, 23 (34.8%) developed hyperbilirubinemia after 72 hours of life, whereas 10 of the 57 newborns (17.5%) whose cord bilirubin levels <1.75 mg/dl developed hyperbilirubinemia after 72 hours of life. Cord blood albumin of ≥3.75 g/dl was seen in 93 neonates, of which 18 (19.4%) developed hyperbilirubinemia after 72 hours of life and 15 (50%) with <3.75 g/dl developed hyperbilirubinemia after 72 hours of life. Cord reticulocyte count ≥4.95% was seen in 54 neonates; 20 (37.03%) developed hyperbilirubinemia, whereas in 69 neonates with <4.95%, 13 (18.84%) developed hyperbilirubinemia after 72 hours of life. Of the 62 neonates who had cord nRBC ≥3.5%, 28 (45.2%) neonates developed hyperbilirubinemia after 72 hours of life, whereas 5 of the 61 neonates (8.19%) with cord nRBC <3.5% developed hyperbilirubinemia after 72 hours of life. CONCLUSIONS: Cord blood bilirubin, albumin, reticulocyte counts, and nucleated RBC can be used as predictors of subsequent neonatal hyperbilirubinemia.

Reticulocyte haemoglobin equivalent (RET-He) as an early marker of responsiveness to oral iron supplementation.

AIM: We investigated the potential of reticulocyte haemoglobin equivalent (RET-He) as an early marker of responsiveness to iron supplementation. METHODS: Data were obtained from a randomised controlled trial of daily iron supplementation in 356 Cambodian women (18-45 y) who received 60 mg elemental iron for 12 weeks. A fasted venous blood specimen was collected at baseline, 1-week and 12-week timepoints. Whole blood haemoglobin (g/L) and RET-He (pg) were measured using a Sysmex haematology analyser. RET-He measures were evaluated for their predictive ability on haemoglobin response to iron supplementation (defined as ≥10 g/L at 12 weeks). Receiver operating characteristic (ROC) curves were used to assess discrimination performance, and the area under the ROC curve (AUCROC) served as a measure of the ability of each predictor to discriminate between women likely or unlikely to elicit a haemoglobin response. RESULTS: Predictive ability (AUCROC (95% CI)) of baseline, 1-week, and change from baseline to 1-week RET-He on haemoglobin response was 0.70 (0.63 to 0.76), 0.48 (0.41 to 0.56) and 0.81 (0.75 to 0.87), respectively. Based on the Youden index, an absolute increase in RET-He of ~1.1 pg or a percentage increase of ~4.4% over 1 week were optimal thresholds to predict responsiveness to iron supplementation. CONCLUSION: Single timepoint RET-He measures have poor predictive ability; however, change in RET-He after 1 week was a strong predictor of haemoglobin response among Cambodian women receiving 60 mg elemental iron and can be measured easily and quickly after only 1 week of iron therapy.

Development and Pilot Testing of a Comprehensive Mobile Application to Assist Cell Count Determination During Peripheral Smear and Bone Marrow Examination.

BACKGROUND: In the modern era of complete blood count analysis, manual differential count is performed whenever 'flags' are generated by an automated hematology analyzer. Traditionally, tally counters with five or eight keys are used for manual differential count. A few mobile applications are available to perform this task; however, the application features and cell representation are limited. OBJECTIVES: The primary objective of our study was to develop an indigenous, comprehensive mobile application to assist with manual blood cell differential count. The secondary objective was to measure the usability of a newly developed application among undergraduate medical students. MATERIALS AND METHODS: A new mobile application was developed using a Java development kit, Version 11.0.13 (Oracle Corporation, Austin, USA) in Android Studio Dolphin (2021.3.1) (Google, California, USA). The application content was validated by three pathologists with more than five years of experience. The app's usability was tested among 60 participants using a validated mHealth App Usability Questionnaire (MAUQ). The questionnaire had 18 items covering three domains: ease of use, interface & satisfaction, and usefulness. RESULTS: The newly developed application supports peripheral smear WBC differential count, platelet count, reticulocyte count, malaria parasite quantification, and bone marrow differential count. During usability testing, the app was easy to use in 95% (57/60) of participants, time-efficient in 91.7% (55/60), and helpful for healthcare practice learning in 96.7% (58/60). The total mean score was 6.11, indicating high usability. CONCLUSION: A comprehensive mobile application to assist manual differential count with adequate cell representation was developed. The mobile application was easy to use, time-efficient, and valuable among the study participants.

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.

Impact of Staining Methods and Human Factors on Accuracy of Manual Reticulocyte Enumeration.

Although peripheral blood reticulocyte enumeration reflects bone marrow functional integrity, which is important for differential diagnosis of hematological diseases, the factors affecting its accuracy have not been adequately addressed. Using 100 consecutive venous blood samples being processed with four supravital staining techniques [i.e., brilliant cresyl blue (BCB), new methylene blue (NMB), and BCB/NMB with Liu's stain] for reticulocyte enumeration, two technologists (senior vs. junior) conducted microscopic counting. The results were compared with those obtained with an automated system (Sysmex XE-5000) that served as the standard. The aims of this study were to identify (1) the technique that gave the most reliable outcome, and (2) possible human factors (i.e., seniority, repeated counting) that may affect the counting results. Analysis showed least bias (i.e., deviation from automated counting) associated with BCB staining, followed by NMB. In addition, the senior observer exhibited a higher bias in counting compared with their junior counterpart. Repeated counting also correlated with a higher rate of bias. Nevertheless, inter-observer consistency was high (intraclass correlation coefficient >0.95) and inter-/intra-observer variations were non-significant (both p > 0.05). Our results supported the use of BCB stain for reticulocyte enumeration and the reliability of manual counting despite the involvement of human factors, which had negligible impacts on the final outcomes.

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.

Assessment of Reticulocyte and Erythrocyte Parameters From Automated Blood Counts in Vaso-Occlusive Crisis on Sickle Cell Disease.

Sickle cell disease is a complex genetic disease involving cell adhesion between red blood cells, white blood cells, platelets and endothelial cells, inducing painful vaso-occlusive crisis (VOC). We assessed reticulocyte and erythrocyte parameters in a cohort of confirmed SCD patients, and investigated whether a combination of these routine laboratory biomarkers of haemolysis could be used to predict VOC development. Reticulocyte and erythrocyte parameters were evaluated using the Sysmex XN-9000 analyser. A total of 98 patients with SCD were included, 72 in steady state and 26 in VOC. Among the 72 patients in steady state, 22 developed a VOC in the following year (median: 3 months [2-6]). The following parameters were increased in SCD patients with VOC development compared to SCD patients without VOC development in the following year: reticulocyte count (94.6 109/L [67.8-128] vs. 48.4 109/L [24.9-87.5]), immature reticulocyte count (259 109/L [181-334] vs. 152 109/L [129-208]) reticulocyte/immature reticulocyte fraction (IRF) ratio (6.63 109/(L*%) [4.67-9.56] vs. 4.94 109/(L*%) [3.96-6.61]), and medium fluorescence reticulocytes (MFR) (19.9% [17.4-20.7] vs. 17.1% [15.95-19.75]). The association of a reticulocyte count of >189.4 109/L and an MFR of >19.75% showed a sensitivity of 81.8% and a specificity of 88% to predict VOC development in the following year. Based on our findings, a combination of routine laboratory biomarkers, as reticulocyte count, immature reticulocyte count and fluorescent reticulocyte fraction at steady state, could be used to predict VOC development in SCD.

[Characteristics of bone marrow compensatory erythropoiesis in hereditary spherocytosis].

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 Between Manual and Automated Methods of Counting Reticulocytes and the Effect of Sample Storage on Reticulocyte Count: A Cross-Sectional Study from Southern India.

Reticulocyte count is a basic test in hematology. This study was done to compare manual and automated methods and to study the effect of sample storage on reticulocyte count. Analyses of samples (n = 86) were done at 2, 6, 24 and 48 h after blood collection. Manual counting was done from both freshly prepared slide and stored slide by microscopy on new methylene blue stained smears. Automated enumeration was on Sysmex XT-2000i analyser (Ret search II). The values of immature reticulocyte fraction (IRF) and low fluorescence reticulocytes (LFR) were also recorded. Comparison between two methods was done by Spearman's correlation and Mann-Whitney test. Effect of storage was analysed by repeated measures ANOVA. There was strong positive correlation between both manual and automated methods at 2, 6, 24 and 48 h. The differences between the manual and automated methods were not significant at 2, 6 and 24 h (p 0.975, 0.967 and 0.227). The difference between the freshly prepared slide and stored slide were significant at 6, 24 and 48 h (p 0.015, 0.004 and 0.001). The change in reticulocyte count with time, decrease in IRF and increase in LFR were not significant up to 6 h but were significant at 24 and 48 h after blood collection. Both the methods were accurate and correlated well with each other. Freshly prepared smears for manual counting were better than counting on stored slide. Up to 6 h after blood collection results obtained by both methods are acceptable.

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.

Reference intervals of automated reticulocyte count and immature reticulocyte fraction in a pediatric population.

INTRODUCTION: Reticulocytes are erythroid precursors that develop into mature erythrocytes, and they are an important tool to assess erythropoietic activity, as their count indicates the balance between the cells released from the bone marrow, their stage of maturity, and their rate of development into mature erythrocytes. Considering the described biological variability of the absolute reticulocyte count (ARC) and the immature reticulocyte fraction (IRF) and the limited information available on these hematological parameters in children, this study determined the reference intervals (RIs) of these parameters in a healthy pediatric population. METHODS: A retrospective, observational, and analytical study was designed to establish RIs for the ARC and the IRF according to age and sex. An indirect sampling method was applied to a mixed database of complete blood counts from children aged 2 months to 18 years, using the truncated maximum likelihood indirect method for reference interval estimation. Percentiles were calculated to obtain bimodal RIs. RESULTS: From a total of 190,812 samples, 6,814 were selected. Gender stratification was not necessary for the ARC and the IRF but they required partitioning into six and two age groups, respectively. CONCLUSION: This study determined, by an indirect sampling method, RIs for the ARC and the IRF in a pediatric population according to age and sex.

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.

Pancytopenia Resulting From Low-Dose Methotrexate Use: A Diagnostic Challenge.

Rheumatoid arthritis (RA) is a common autoimmune disease primarily affecting small joints which leads to crippling erosion of the articular cartilage and bone. It is associated with complications related to both its disease course and treatment. Methotrexate (MTX) is a folate antagonist responsible for modulating cell-specific signaling pathways and inhibiting the proinflammatory properties of major cell lineages involved in the pathogenesis of RA. It is considered to be the first-line agent in RA because of its disease-modifying ability and safety profile at low doses. This case report discusses how a middle-aged female presented with severe bone marrow suppression secondary to MTX toxicity, an unusual presentation at the usual low-dose regimen. Her presentation overlapped with several other conditions, especially with Felty's syndrome, a rare complication of RA, characterized by the triad of splenomegaly, neutropenia, and RA. Other differentials included hemophagocytic lymphohistiocytosis, hematologic neoplasms, drug reaction, and infection. Therefore, it was essential to exclude all possible differentials before initiating therapy. We found the corrected reticulocyte count coupled with a good response to leucovorin to be an effective way to differentiate MTX-induced pancytopenia from other possible hematologic diagnoses without the use of a bone marrow biopsy. Additionally, our case incidentally demonstrated a potential interaction between piperacillin/tazobactam and MTX.

Interindividual Biological Variability of Reticulocytes and Their Maturation Fractions in the Pediatric Population.

OBJECTIVES: Because published data about the variability of reticulocyte counts in children are scarce, the interindividual biological variability of the automated reticulocyte count and its maturation fractions according to age and sex were analyzed. METHODS: A retrospective, observational, analytical study was designed to establish and compare normal values of the automated reticulocyte count and its maturation fractions in different age and sex groups. The sample was drawn from results of CBC counts performed in children aged between 2 months and 18 years using an indirect sampling methodology. RESULTS: A total of 9,362 CBC counts were analyzed. Automated reticulocyte count decreased between 2 months and 3 years of age and slowly increased thereafter, showing higher values in girls up to the age of 9 years, and equalized by sex thereafter. Immature reticulocyte fraction increased until 7 months of age; decreased progressively until 4 years of age; and then showed a discreet but constant rise, with significantly higher values in boys older than 1 year. The low-fluorescence fraction was relatively steady, with significantly higher values in girls aged 8 months and older. CONCLUSIONS: The automated reticulocyte count and its maturation fractions show significant variations related to age and sex in pediatric patients.

Hyperhemolysis Syndrome in a Patient With Sickle Cell Disease and Acute Chest Syndrome.

Sickle cell anemia patients often present to the hospital with acute vaso-occlusive pain crisis. Symptoms can include, but are not limited to, chest pain, abdominal pain, and musculoskeletal pain. These symptoms are brought about due to the pathology of the disease. Abnormal hemoglobin S causes red blood cells to band together, otherwise known as "sickling." These patients also often present with very low hemoglobin levels on initial evaluation. In most cases, packed red blood cell transfusions are needed in order to replenish these patient's functional hemoglobin supply. Unfortunately, transfusing sickle cell patients can lead to an unwanted consequence, that of hyperhemolysis syndrome, in which blood transfusions prompt further hemolysis of the already sickled red blood cells. When this complication arises, caution must be exercised in deciding the next steps of treatment.

Novel toxicity of tris(1,3-dichloro-2-propyl) phosphate in adult male rats.

The widespread use of tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) as a flame retardant has led to its release to the environment. Thus, the toxicological effects of TDCIPP on humans and animals are of importance. For better understanding of its potential toxicities, TDCIPP (250, 500, or 650 mg/kg/day) or vehicle control was administrated orally to adult male Wistar-Imamichi rats for 7 days. After the final administration of compounds, organ weights, histopathology, blood biochemistry, and hematology were examined. Hepatic toxicity was observed at doses ≥ 500 mg/kg/day of TDCIPP, and renal toxicity was observed at 650 mg/kg/day. The anti-androgenic activity of TDCIPP was previously confirmed in vitro and in vivo, but weights of epididymis, an androgen-dependent organ, were not affected by TDCIPP treatment in adults. Serum alkaline phosphatase activity was significantly decreased in all TDCIPP-treated rats independent of dose. Hemoglobin concentration, hematocrit, red blood cell count, and reticulocyte count were decreased in all TDCIPP-treated rats, but mean corpuscular volume, total iron-binding capacity, and serum iron were normal, suggesting that renal anemia was caused by TDCIPP. Together with previous reports on effects of anti-androgenic substances on red blood cell indices, anemia caused by TDCIPP could be due to its anti-androgenic activity. These considerations will contribute to further assessment of the toxicity of the compound.

Study of Cord Blood Levels of Erythropoietin, Bilirubin and Reticulocyte Count as Early Predictors of Neonatal Hyperbilirubinemia.

BACKGROUND: Neonatal hyperbilirubinemia is a serious neonatal problem which has hazardous effects on the neonates when the level of indirect bilirubin is increased to the levels that could cause kernicterus. AIMS: The aim of this research is to study the cord blood levels of erythropoietin (EPO), bilirubin and reticulocyte count (RC) as early predictors of neonatal hyperbilirubinemia. METHODS: This is a case-control study, which was conducted at Tanta University Hospital (TUH) from July 2016 to March 2018 on 90 neonates. The studied neonates were divided into 2 groups: Group 1 (45 neonates) who developed pathological hyperbilirubinemia and required treatment and group 2(45 neonates) who did not develop pathological hyperbilirubinemia and did not require treatment. Cord blood levels of EPO, bilirubin and RC were measured in all the studied neonates in both groups. RESULTS: There was a significant difference between both groups with regard to cord blood bilirubin (CBB), hemoglobin, EPO and RC levels where the P. value is 0.001*,0.027, *0.001*&0.001*respectively. There was a significant positive correlation between cord blood EPO levels and both CBB and cord blood RC with r=0.610 and 0.579, respectively and P. value is 0.001* & 0.001* respectively. With regard to ROC curve, there were high cord blood EPO levels where the cut off value was 22.5 mIU/ml while the sensitivity and specificity were 96 and89, respectively. In the cord blood RC, the cut off value was 5.7% while the sensitivity and specificity were 93 and 85, respectively, and lastly, CBB where the cut off value was 1.8 mg/dl while the sensitivity and specificity were 89 and 78 respectively. CONCLUSION: Cord blood levels of EPO, bilirubin and RC were increased in cases of pathological neonatal hyperbilirubinemia. RECOMMENDATION: Cord blood levels of EPO, bilirubin and RC could be used for early prediction of pathological neonatal hyperbilirubinemia.