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Abstract 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.

Background In a simulation based on a pharmacokinetic model we demonstrated that increasing the erythropoiesis stimulating agents (ESAs) half-life or shortening their administration interval decreases hemoglobin variability. The benefit of reducing the administration interval was however lessened by the variability induced by more frequent dosage adjustments. The purpose of this study was to analyze the reticulocyte and hemoglobin kinetics and variability under different ESAs and administration intervals in a collective of chronic hemodialysis patients. Methods The study was designed as an open-label, randomized, four-period cross-over investigation, including 30 patients under chronic hemodialysis at the regional hospital of Locarno (Switzerland) in February 2010 and lasting 2 years. Four subcutaneous treatment strategies (C.E.R.A. every 4 weeks Q4W and every 2 weeks Q2W, Darbepoetin alfa Q4W and Q2W) were compared with each other. The mean square successive difference of hemoglobin, reticulocyte count and ESAs dose was used to quantify variability. We distinguished a short- and a long-term variability based respectively on the weekly and monthly successive difference. Results No difference was found in the mean values of biological parameters (hemoglobin, reticulocytes, and ferritin) between the 4 strategies. ESAs type did not affect hemoglobin and reticulocyte variability, but C.E.R.A induced a more sustained reticulocytes response over time and increased the risk of hemoglobin overshooting (OR 2.7, p = 0.01). Shortening the administration interval lessened the amplitude of reticulocyte count fluctuations but resulted in more frequent ESAs dose adjustments and in amplified reticulocyte and hemoglobin variability. Q2W administration interval was however more favorable in terms of ESAs dose, allowing a 38% C.E.R.A. dose reduction, and no increase of Darbepoetin alfa. Conclusions The reticulocyte dynamic was a more sensitive marker of time instability of the hemoglobin response under ESAs therapy. The ESAs administration interval had a greater impact on hemoglobin variability than the ESAs type. The more protracted reticulocyte response induced by C.E.R.A. could explain both, the observed higher risk of overshoot and the significant increase in efficacy when shortening its administration interval. Trial registration ClinicalTrials.gov: NCT01666301

Aims To derive reference values for red cell variables and platelet counts from a cohort of infants sampled at precise ages during the first 13 months of life. Methods Blood counts, reticulocyte counts and zinc protoporphyrin concentrations were obtained from healthy term infants of North European ancestry at 2, 5 and 13 months of age. Results Mean cell volume (MCV) and mean cell haemoglobin (MCH) values did not differ significantly between 5 and 13 months and MCH concentration was unaffected by age. Values of all other variables at any one age differed significantly from those at the other two. Haemoglobin, mean cell haemoglobin, zinc protoporphyrin and platelet values (95% ranges) at 2 (n=119), 5 (n=97) and 13 months (n=42) were, respectively, 91–125, 101–129 and 105–133 g/L; 28.6–33.1, 24.5–28.7 and 24.3–28.7 pg; 36–116, 25–91 and 27–57 micromol/mol haem; and 216–658, 241–591 and 209–455×109/L. At 2 and 5 months, respectively, 26.9% and 10.8% of subjects had platelet counts >500×109/L. Reticulocyte counts at 2 months and MCV and MCH values at 5 months were significantly higher in girls. In boys, red cell distribution width values were significantly higher at 5 months, and zinc protoporphyrin values at both 2 and 5 months. Conclusions These findings indicate the value of obtaining reference data at precise ages during infancy and confirm and extend earlier reports indicating a gender difference in laboratory measures used to assess iron status in early infancy.

Reticulocytes are immature erythrocytes that have been released into the peripheral circulation after being extruded from the bone marrow. However, it has not been investigated whether these reticulocyte indicators are helpful in treating conditions linked to abnormal haemoglobin variants. In the current investigation, participants with abnormal haemoglobin variants were evaluated for the utility of reticulocyte parameters using a high-end haematology analyser. We conducted a prospective cross-sectional study among 217 participants with abnormal haemoglobin variants at the Eastern Regional Hospital, Koforidua. Statistical analyses were performed with IBM SPSS V.26.0 (Chicago, IL, USA) and R programming language version 4.2.3. A p-value of < 0.05 was considered statistically significant for all analyses. We found a significant association between age groups and anaemia severity (p = 0.016). Haemoglobin levels varied significantly among abnormal haemoglobin variants (p < 0.05), with the “SF” phenotype showing the lowest levels. Production defects accounted for 1.4% of cases, while maturation defects were present in 91.7%. An increase in low-fluorescence reticulocytes was linked to a 0.03 g/dL rise in haemoglobin (β = 0.03, p < 0.05), whereas increases in high- and medium-fluorescence reticulocytes led to declines of 0.069 g/dL and 0.052 g/dL, respectively (p < 0.05). Haemoglobin levels were significantly lower in participants with maturation abnormalities (p < 0.01). The study found relationships between the severity of anaemia, age, and haemoglobin phenotypes in individuals with abnormal haemoglobin variants. Given the predominance of maturation defects, management aimed at addressing ineffective erythropoiesis is necessary. The differences in how reticulocyte subpopulations affect haemoglobin levels imply that reticulocyte indices may be helpful biomarkers for monitoring and guiding therapeutic approaches.

Background and Objectives: Women are more prone to iron deficiency (ID) anemia when pregnant. The diagnostic use of mean reticulocyte volume (MRV) in identifying ID anemia during pregnancy has not been thoroughly investigated. The objective of this study is to evaluate the effectiveness of MRV in diagnosing ID in pregnant women. Methods and Study Design: Firstly, MRV of 20 healthy female volunteers (healthy group) was measured on specific days for one month. Subsequently, clinical data from 724 pregnant women were thoroughly examined. These women were divided into two groups: 282 with ID (research group) and 442 without ID (control group). Parameters such as MRV, reticulocyte hemoglobin equivalent (RHE), red blood cell volume distribution width- standard deviation (RDW-SD), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), hematocrit (HCT), reticulocyte count (RET), MRV/MCV ratio, and serum ferritin (SF) were analyzed and compared. Results: MRV remained consistent over a period of one month for 20 healthy individuals. In addition, there were significant differences in MRV, RHE, RDW-SD, MCV, MCH, MCHC, HCT, RET, and MRV/MCV between the research group and control group. The receiver operating characteristic (ROC) analysis showed that the areas under the curve (AUCs) for these measures were as follow: 0.840, 0.837, 0.676, 0.654, 0.639, 0.602, 0.571, 0.550, and 0.816, respectively. Ultimately, there was a substantial disparity in MRV prior to and following therapy with oral iron treatments. Conclusions: In healthy women, MRV remains stable and is a reliable ID marker, which can be used to assess oral iron treatment effectiveness during pregnancy.

Rajasekhar Suragani et al . show that a modified activin receptor IIB ligand trap increases red blood cell numbers in mice, rats and monkeys and ameliorates anemia in mice and rats, including in a mouse model of myelodysplastic syndromes. The ligand trap binds to the cytokine GDF11 and acts by inhibiting Smad2/3 signaling, thereby reversing ineffective erythropoiesis. Also in this issue, Michael Dussiot et al . show related findings using a wild-type activin receptor IIA ligand trap. Erythropoietin (EPO) stimulates proliferation of early-stage erythrocyte precursors and is widely used for the treatment of chronic anemia. However, several types of EPO-resistant anemia are characterized by defects in late-stage erythropoiesis 1 , 2 , which is EPO independent 2 , 3 . Here we investigated regulation of erythropoiesis using a ligand-trapping fusion protein (ACE-536) containing the extracellular domain of human activin receptor type IIB (ActRIIB) modified to reduce activin binding. ACE-536, or its mouse version RAP-536, produced rapid and robust increases in erythrocyte numbers in multiple species under basal conditions and reduced or prevented anemia in murine models. Unlike EPO, RAP-536 promoted maturation of late-stage erythroid precursors in vivo . Cotreatment with ACE-536 and EPO produced a synergistic erythropoietic response. ACE-536 bound growth differentiation factor-11 (GDF11) and potently inhibited GDF11-mediated Smad2/3 signaling. GDF11 inhibited erythroid maturation in mice in vivo and ex vivo . Expression of GDF11 and ActRIIB in erythroid precursors decreased progressively with maturation, suggesting an inhibitory role for GDF11 in late-stage erythroid differentiation. RAP-536 treatment also reduced Smad2/3 activation, anemia, erythroid hyperplasia and ineffective erythropoiesis in a mouse model of myelodysplastic syndromes (MDS). These findings implicate transforming growth factor-β (TGF-β) superfamily signaling in erythroid maturation and identify ACE-536 as a new potential treatment for anemia, including that caused by ineffective erythropoiesis.

AimWe investigated the potential of reticulocyte haemoglobin equivalent (RET-He) as an early marker of responsiveness to iron supplementation.MethodsData 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.ResultsPredictive 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.ConclusionSingle 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.

Background Measurement of reticulocyte hemoglobin equivalent (RET‐He) is rapid, convenient, and cost‐effective. Yet, researches on its performance in diagnosing iron deficiency with concurrent inflammation are limited. Hence, this study investigated RET‐He value in various states, including inflammation, and evaluated its diagnostic performance in iron status assessment. Methods Retrospectively, 953 clinical data and laboratory results—complete blood count, reticulocyte count, RET‐He, and serum ferritin—were reviewed. Patients on iron therapy were excluded. Iron status was defined by serum ferritin as the reference method. RET‐He among populations was investigated. Its diagnostic performance and optimal cutoff were determined by ROC analysis. Results Three population groups were classified: healthy control, iron deficiency anemia (IDA), and non‐ID anemia. Significantly, RET‐He value in IDA was lower than that of healthy control, anemia of inflammation, and chronic kidney disease (P < .0001). Low RET‐He was also observed in IDA with concomitant inflammation despite normal‐to‐high serum ferritin levels. No significant difference was observed between RET‐He values in pure IDA and thalassemia (P = .57). ROC curve analysis revealed AUC of 0.876 (P < .0001) at cutoff 30 pg, by which IDA was discriminated with 74.2% sensitivity and 97.4% specificity. Applying cutoff ≤30 pg, IDA can be diagnosed with 96% sensitivity, 97.4% specificity, 80% PPV, and 99.6% NPV. Hence, RET‐He >30 pg signifies a non‐IDA state. Conclusion In addition to convenience and cost‐effectiveness, RET‐He cutoff >30 pg can be potentially used to exclude IDA due to its excellent diagnostic sensitivity and specificity.

AimsTo assess changes in reticulocyte impedance volume, conductivity and light scatter (reticulocyte population data or RPD) obtained with the volume, conductivity and laser light scatter (VCS) technology in healthy subjects and in different erythropoietic states.MethodsBlood samples were analysed with the Beckman–Coulter LH750 system, using the VCS method, from a group of 40 healthy subjects and three groups of patients with different types of untreated or treated anaemia: 24 cases of iron deficiency at the time of diagnosis, 16 patients with iron deficiency anaemia during intravenous iron administration, and 57 patients with chronic kidney disease undergoing dialysis and administration of rHu-erythropoietin and intravenous iron.ResultsRPD data were reproducible. Average mean channels for volume were 50.9 for controls and 49.2, 55.7 and 64.0 for the three patient groups, respectively. Average mean channels for conductivity were 52.0 for controls and 59.8, 55.7 and 56.1 for the three patient groups. Average mean channels for light scatter were 108.4 for controls and 113.3, 117.3 and 128.2 for the three patient groups. SD data indicated increased dispersion in the patient groups.ConclusionWhen values in the patient groups are compared with reference values obtained in healthy controls, the main differences are found in impedance volume and light scatter, which were both increased in patients with stimulated erythropoiesis. These data indicate the opportunity to further evaluate the clinical usefulness of RPD in haematological diseases.

Objective G6PD deficiency is a potentially life-threatening condition in neonates presenting with hyperbilirubinemia. This study aims to identify clinical and laboratory predictors of G6PD deficiency in neonates presenting with hyperbilirubinemia. Methods This was a retrospective study of 227 term neonates admitted to Heyuan People's Hospital from January 2019 to October 2023. Hematological parameters and bilirubin were compared between those with G6PD deficiency and those with normal G6PD. Results Term neonates with G6PD deficiency had higher levels of total bilirubin, indirect bilirubin, mean corpuscular volume, mean corpuscular hemoglobin, immature reticulocyte fraction, high-fluorescence reticulocyte ratio, medium-fluorescence reticulocyte ratio, and content of reticulocytes than those with normal G6PD, but lower levels of red blood cells, hemoglobin, hematocrit, and low-fluorescence reticulocyte ratio. Medium-fluorescence ratios (OR = 1.291, P  = 0.028) independently predicted G6PD deficiency in neonates. The optimal cut-off value for medium-fluorescence ratios was > 18.55%. The area under the curve for diagnosing G6PD deficiency was 0.924 (95% confidence interval: 0.886–0.962, P  < 0.0001), with a sensitivity of 82.6% and specificity of 86.2%. Conclusion MFR emerged as a potentially valuable predictor for G6PD deficiency in neonates.