Reticulocyte Hemoglobin Research Articles

Nearly Half of Extreme Preterm Infants Are Iron Deficient at 4-6 Months of Corrected Age Despite Prophylactic Iron Supplementation

Background: Iron deficiency (ID) is the most prevalent micronutrient deficiency globally and has been associated with negative neurodevelopmental and behavioral outcomes in infants and young children. Susceptibility to ID is heightened in preterm infants due to lower iron reserves at birth, rapid growth during the initial weeks of life, immature erythropoiesis, and frequent blood samplings during hospitalization. To mitigate this problem, international bodies recommend prophylactic iron therapy from 2-6 weeks of age until at least 6-12 months in preterm infants. Currently, the literature regarding iron status in extremely preterm infants (EPI, infants born less than 28 weeks gestational age) who receive prophylactic iron supplementation after birth is limited. Considering the potential long-term detrimental effects of ID on the developing brain of these high-risk preterm infants, examining the prevalence and risk factors for ID are crucial to implement prevention strategies. Objective: To investigate the prevalence and risk factors associated with ID in EPI Methods: A retrospective cohort study was conducted using a population-based Provincial Perinatal Follow-Up database on all live EPIs born in Nova Scotia between 2005 and 2018. Infants with congenital malformations, chromosomal anomalies, blood disorders or those who died before outcome assessment were excluded. Prophylactic iron supplementation (2-3 mg/kg/day) starting at 2-4 weeks of chronological age was a standard of care for all included infants. Further, the dosage of iron was adjusted regularly during the hospital stay based on serum ferritin (SF) levels. Discharge instructions recommended continuing iron prophylaxis until 9-12 months corrected age (CA). At 4-6 months CA, all infants underwent blood tests to evaluate hemoglobin, and SF levels. ID was defined as SF levels below 20g/L or 12g/L at 4 and 6 months CA, respectively. A univariate analysis was performed by using a series of single-variable logistic regression models to identify the factors associated with presence of ID. Factors with a p-value < 0.20 in the univariate analysis were entered into a multivariable risk model using a backward selection procedure. Variables with a p-value < 0.05 were retained. Results: Among 146 infants, 67 (45.9%) had ID. Ferritin (16.4 µg/L vs 50.0 µg/L, p < .001) and reticulocyte hemoglobin equivalent (28.4pg vs 31.6pg, p < .001) were significantly lower in the ID group than non-ID group. The prevalence of ID decreased over time, from 59.7% in 2005-2011 to 40.3% in 2012-2018 (p = 0.52). Table 1 compares the antenatal and neonatal characteristics of the ID and non-ID groups. Table 2 compares sociodemographic variables and clinical variables at 4-6 months CA between two groups. Breastfeeding (exclusive or partial) versus formula alone at a 4-6 months CA was identified as an independent factor protective for ID (Odds: 0.2 (0.1 - 0.6), p = 0.003). Iron therapy at follow-up was also protective for ID but did not remain significant in the final model. Of note, iron therapy was stopped in 40 (27.4%) EPI before 4-6 months CA. Supplemental iron intake at 4-6 months CA was significantly lower in the infants receiving exclusive formula feeding (66.1%) compared with those breastfed (99.1%, p = 0.006). Conclusion(s): Despite prophylactic iron supplementation, nearly half of the EPI had ID at 4-6 months CA, and iron therapy was discontinued in roughly a quarter of all EPI before 4-6 months CA. Breastfeeding at 4-6 months follow-up was found to be protective for ID. High prevalence of ID in EPI highlights the need for further research in this area to avert the negative neurodevelopmental consequences of ID on the developing brain of these high-risk infants. Closer monitoring of iron status and early identification of children at high risk for ID is imperative to maximize the benefits of prophylactic iron supplementation in EPI. Further, there is lack of sufficient data for iron requirements in EPI leading to a variability in iron therapy recommendations by different professional organizations. Future prospective studies to investigate specific iron requirements of EPI as well as risk factors for ID in this high-risk vulnerable population are essential to optimize their neurodevelopmental outcomes.

Iron Deficiency Prior to Discharge in Very Low Birth Weight Infants: Screening with Reticulocyte Hemoglobin Content.

This study aimed to assess the iron status prior to discharge in very low birth weight (VLBW) infants utilizing reticulocyte hemoglobin content (CHr) and evaluate the impact of delayed cord clamping (DCC) on iron status. This is a retrospective analysis of VLBW infants from two tertiary level of care Neonatal Intensive Care Units. The primary outcome was the proportion of VLBW infants with low CHr (<29 pg) prior to discharge. Hematologic parameters were also compared between infants who received or did not receive DCC. Infants with a positive newborn screen for hemoglobin Bart's were excluded. Among the 315 infants included, 99 infants (31.4%) had low CHr prior to discharge. The median (interquartile range) CHr prior to discharge was 30.8 pg (28.4-39 pg). DCC was performed in 46.7% of infants. Hemoglobin at birth, discharge, and CHr prior to discharge were higher and the need for blood transfusion and the number of infants with low CHr prior to discharge were lower in the DCC group. Approximately 31.4% of VLBW infants had low CHr near the time of discharge suggesting they were iron deficient. DCC improved hematological parameters prior to discharge in VLBW infants. CHr content can be used to guide iron supplementation in VLBW infants to potentially improve their iron status and long-term neurocognitive outcomes. · DCC was associated with an improved hemoglobin and iron status at discharge in VLBW infants.. · CHr is an early and reliable marker for iron deficiency.. · Approximately one in three VLBW infants can be iron deficient at the time of discharge..

Accuracy of reticulocyte hemoglobin for diagnosing iron deficiency in former very preterm infants: a population-based cohort study.

Serum ferritin (SF) is commonly used to diagnose iron deficiency (ID) but has limitations. Reticulocyte hemoglobin (Ret-He) is being increasingly used for ID diagnosis. This study aimed to assess accuracy of Ret-He for ID diagnosis in former very preterm infants (VPI) at 4-6 months corrected age (CA). A retrospective population-based cohort study was conducted on all live VPI born between 23 and 30 weeks of gestational age (GA) in Nova Scotia from 2012 to 2018. Infants underwent SF and Ret-He testing at 4-6 months CA. ID was defined using two definitions. The first defined ID as SF < 20 mcg/L at both 4- and 6-months CA, and the second as SF < 30 mcg at at both 4- and 6-months CA. The accuracy of Ret-He for identifying ID was assessed using the area under the receiver operating characteristic curve (AUC). ID was present in 39.7% (62) of 156 infants in the first definition and 59.6% (93) in the second at 4-6 months CA. The AUC of Ret-He for ID diagnosis was 0.64 (p = 0.002) in the first definition and 0.59 (p = 0.04) in the second. The optimal cut-off was 29.4pg in the first and 29.7 in the second definition. The sensitivity, specificity, positive predictive value (PPV), and negative predictive values (NPV) at the 29.4 pg cut-off were 50.0%, 78.7%, 60.8%, and 70.5% for definition 1 and 44.1%, 74.6%, 71.9%, and 47.5% at the 29.7pg cut-off for definition 2. Ret-He had low diagnostic accuracy for ID diagnosis in former VPI. Caution is advised when using Ret-He alone for ID diagnosis. Further research is needed to establish optimal approaches for identifying ID in VPI.

Imbalance of iron needs and supply in patients with acute and chronic heart failure

Abstract Background Iron deficiency (ID) is frequent in patients with chronic and acute heart failure (CHF, AHF) associated with morbidity and mortality. ID can be absolute, defined as depleted iron stores (also named true ID), or functional describing hypoferremia despite adequate iron stores as a result of inflammation-related reduced iron mobilization and restriction in mononuclear cells. Empty iron stores can also be found simultaneously to inflammation-related reduced iron mobilization (combined ID). Purpose We aimed to better characterize ID and iron homeostasis in HF patients applying different biomarkers and to evaluate the accuracy of current ID definition by the ESC to indicate actual tissue iron needs and supply. Methods We performed a retrospective cohort study including 277 AHF and 476 CHF patients treated between 02/2021–05/2022 at the Medical University of Innsbruck. Results AHF patients had higher iron needs and lower iron supply than CHF patients, reflected by increased soluble transferrin receptor (sTfR) and sTfR-Ferritin (sTfR-F) index, and lower ferritin, serum iron, transferrin saturation (TfS), hepcidin and reticulocyte hemoglobin (Ret-Hb). Only 28.0% of CHF and 81.4% of AHF patients with absolute ID as well as 10.5% of CHF and 50.0% of AHF patients with combined ID based on the current ESC definition presented with an elevated sTfR-F index (&amp;gt;2.0 without and &amp;gt;3.2 with inflammation) and/or reduced Ret-Hb (&amp;lt;28pg), being indicative for tissue iron needs. Suppressed hepcidin expression (≤8.6ng/mL) as early marker of iron depletion was found in 80% of CHF and 91% of AHF patients with absolute ID and only in 19% of CHF and 32% of AHF patients with combined ID based on the current ESC definition. When breaking down the ID definition into ferritin and TfS, in AHF an elevated sTfR-F index was found in patients with inflammation and associated with lower ferritin (&amp;lt;200 ng/mL), while a reduced Ret-Hb was primarily found in patients with inflammation, TfS &amp;lt;20% and ferritin &amp;lt;100 ng/mL. In CHF an elevated sTfR-F index and a reduced Ret-Hb were primarily present in patients with ferritin &amp;lt;30 ng/mL and TfS &amp;lt;20% as well as in patients with inflammation, TfS &amp;lt;20% and ferritin &amp;lt;100 ng/mL. (Figure 1) Specificity for defining true ID based on iron needs was increased when including inflammation for defining true ID and reducing the upper limit of ferritin for defining true ID in the setting of inflammation (Figure 2). Conclusion A substantial proportion of patients with ID defined by the current ESC definition, had normal tissue iron supply with no increased tissue iron needs. In accordance with previous studies, results from our study suggest that the current ESC definition might overestimate true ID especially in CHF. A more accurate definition of ID may help to identify those patients with AHF and CHF who truly benefit from intravenous iron supplementation.

A Phase 1b Double-Blind, Placebo-Controlled Study of DISC-0974, an Anti-Hemojuvelin Antibody, in Patients with Non-Dialysis Dependent Chronic Kidney Disease and Anemia

Hepcidin, a central regulator of iron homeostasis, is pathologically elevated in patients with non-dialysis-dependent chronic kidney disease (NDD-CKD) and anemia. Elevations in hepcidin contribute to the onset, maintenance, and severity of anemia. DISC-0974 is an investigational, first-in-class, monoclonal antibody that blocks hemojuvelin (HJV), a co-receptor in the bone morphogenetic protein-signaling pathway regulating hepcidin expression. In a healthy volunteer study, DISC-0974 demonstrated dose-dependent reductions in serum hepcidin, increases in serum iron with doses up to 56 mg administered subcutaneously (SC), and increasing trends in reticulocyte count, reticulocyte hemoglobin, mean corpuscular hemoglobin, total hemoglobin (Hgb), and red blood cell count. In a rat model of anemia in CKD, DISC-0974 decreased hepcidin, increased serum iron, and normalized hemoglobin (Wu et al., ASH 2022). DISC-0974-103 is a Phase 1b, double-blind, placebo-controlled, single-ascending dose study (NCT05745883) to assess the safety, tolerability, pharmacokinetics (PK), and pharmacodynamics (PD) of DISC-0974 in patients with NDD-CKD and anemia. Eligible participants include patients over 18 years of age with NDD-CKD, stages 2-5, hemoglobin (Hgb) &amp;lt;10.5 g/dL in women and &amp;lt;11.0 g/dL in men, serum ferritin ≥100 μg/L, and transferrin saturation (TSAT) ≤30%. Major exclusionary criteria include: concomitant treatment with therapeutic oral iron, intravenous iron, erythropoietin-stimulating agents, or blood transfusions; splenectomy; and nutritional, genetic, infectious, or autoimmune causes of anemia. Approximately 32 participants will be enrolled. On Day 1, participants will receive a single dose of DISC-0974 SC with evaluations for safety, PK, and PD through 57 days of follow-up. Dose escalation is planned across 4 cohorts consisting of 8 participants each, randomized 3:1 active treatment to placebo, treated at single doses of 28, 40, 60, and 90 mg SC DISC-0974. Safety Review Committee assessments are planned to gate escalation decisions. Primary endpoints include adverse events (AEs), clinical laboratory assessments, vital signs, physical examinations, and electrocardiograms. Secondary endpoints include changes in serum hepcidin-25, iron, TSAT, ferritin, Hgb, and other hematology biomarkers. Enrollment is currently ongoing, and available study results will be presented at the meeting.

A Phase 1b Trial of DISC-0974, an Anti-Hemojuvelin Antibody, in Patients with Myelofibrosis and Anemia

Hepcidin, a central regulator of iron homeostasis, is pathologically elevated in patients with myelofibrosis (MF) and anemia. Chronic elevations in hepcidin contribute to the onset and severity of anemia. DISC-0974 is an investigational, first-in-class, monoclonal antibody that blocks hemojuvelin, a co-receptor in the bone morphogenetic protein-signaling pathway driving hepcidin expression. Preclinical studies have shown that DISC-0974 suppresses hepcidin and increases serum iron. A healthy volunteer study has demonstrated dose-dependent reductions in serum hepcidin, increases in serum iron with doses up to 56 mg administered subcutaneously, and increasing trends in reticulocyte count, reticulocyte hemoglobin, mean corpuscular hemoglobin, total hemoglobin (Hgb), and red blood cell count. In this Phase 1b/2a, open-label, multiple-ascending dose study (NCT05320198), we are assessing the safety, tolerability, pharmacokinetics (PK), and pharmacodynamics (PD) of DISC-0974 in patients with MF and anemia. Eligible participants include patients over 18 years of age with intermediate-2 or higher-risk MF and anemia. MF is required to be confirmed using the World Health Organization 2016 criteria. Anemia is defined per protocol as Hgb &amp;lt; 10 g/dL or transfusion dependence, as defined by the International Working Group for Myelofibrosis Research and Treatment (IWG-MRT). A stable dose of hydroxyurea and/or Janus kinase (JAK) inhibitor is allowed on trial. Major exclusion criteria include: liver iron concentration ≥ 7 mg/g dry weight; splenectomy; nutritional, genetic, infectious, or autoimmune causes of anemia. Dose escalation is based on a Bayesian optimal interval design with accelerated titration. Safety Review Committee assessments gate escalation decisions. Dosing occurs monthly for a total of 6 doses. Primary endpoints include safety and tolerability assessments of adverse events (AEs), clinical laboratory assessments, vital signs, physical examinations, and electrocardiograms. Secondary endpoints include anemia response as defined by the IWG-MRT, transfusion response as defined by a decrease of 50% in transfusion requirement over any 8-week period, standard PK parameters and PD metrics including serum hepcidin-25, iron, transferrin saturation, ferritin, and hematology biomarkers. All data are summarized using descriptive statistics. Initial data show a dose-related increase in pharmacokinetic parameters. DISC-0974 treatment led to dose-dependent reductions in serum hepcidin-25 and increases in serum iron levels. There have been no serious AEs or AEs leading to study withdrawal. Most AEs were mild, transient, and unrelated to DISC-0974. As of August 1, 2023, enrollment and dose escalation are ongoing. Safety, PK, and PD data from at least the first two cohorts will be presented.

Roxyscan: A Novel Method to Assess Red Blood Cell Resilience to Oxidative Stress in Sickle Cell Disease

Background: Red blood cells (RBCs) are biconcave-shaped cells with viscoelastic membranes that are optimally adapted for oxygen delivery and gas exchange during their normal lifespan of 120 days. Reactive oxygen species (ROS) continuously challenge the viability of RBCs. Despite an arsenal of compounds to protect the RBC, including Glutathione (GSH), Vitamin E, superoxide dismutase (SOD) and Catalase, damage can easily be inflicted that will disrupt cellular deformability, structure, and membrane properties. The susceptibility to oxidative stress in individuals with Sickle Cell Disease (SCD) is believed to be linked to multiple factors. These include the release of heme and iron during hemolysis, inflammation processes, and a reduction in antioxidant defenses. Methods:Susceptibility of RBCs to oxidative stress was assessed with the RoxyScan. This novel application of the Lorrca MaxSis (RR Mechatronics) measures rheological behavior of RBCs in response to cumene hydroperoxide (CuHP, 100 µM) exposure during continuous shear (30 Pa). For this we used the Lorrca's peristaltic pump and suction needle, directing the blood and CuHP to the instrument's cup and mixing them for 30 seconds (s) before starting the run. Deformability of RBCs (EI, elongation index) was measured over time. PoX is defined as the time where a 10% decrease in EI is reached. EI start: EI t=0. EI final: EI t=1600s, ΔEI (EIstart-EIfinal). RoxyScan measurements were performed on blood samples from 16 adult SCD patients (HbSS, HbS/β0 or HbS/β+) and 19 healthy controls (HC). Outcome measurement PoX (calculated from curve fitted data) was correlated to laboratory parameters from complete blood count (Cell Dyn Sapphire, Abbott), RBC sickling tendency (Point of Sickling, PoS, as assessed by oxygen gradient ektacytometry on the Lorrca MaxSis), and levels of HbF/HbS (Tosoh G8). Results:RBCs from untreated HC show a stable EI over time (Fig 1A). Upon exposure to CuHP a similar decrease in EI is seen in 19 HC (Fig 1A), mean decrease 0.055, range (0.017 to 0.10). The mean and median calculated PoX in HC is 1723s and 1702s respectively. In contrast, SCD RBC showed an earlier and more pronounced decrease in EI in response to CuHP exposure (representative example, Fig 1A). Comparing the PoX between HC and SCD revealed significant differences: mean PoX HC: 1723s (SD 329s), SCD: 725s (SD 472s), ***p&amp;lt;0.0001, indicating that SCD RBCs are more susceptible to oxidative stress. Correlating PoX to routine laboratory parameters identified a correlation with Hb levels (***p=0.002), inverse correlation with reticulocyte count (***p=0.0014) and hemoglobin distribution width (HDW, ***p&amp;lt;0.001) (Fig 1B). Similarly strong inverse correlations were found for EI start and EI end (***p&amp;lt;0.001) (Fig 1B). PoX also showed an inverse correlation with red cell distribution width (RDW, ns p=0.07) and mean corpuscular volume (MCV, ns p=0.77). Interestingly, PoX showed a significant inverse correlation with PoS (***p=0.004), indicating that SCD RBCs which are more susceptible to oxidative stress tend to sickle at a higher oxygen tension. PoX showed no significant correlations with HbS/HbF levels (Fig 1B). Conclusion: In this study we demonstrate the applicability of the RoxyScan as a novel method to assess RBC resilience to oxidative stress. When applied to SCD, RBCs show an earlier and more pronounced response to oxidative stress than RBCs from HC. The inverse correlation of PoX, the major outcome parameter of this technique, with reticulocyte count could indicate a generally increased susceptibility to oxidative stress due to a higher level of hemolysis. The correlation with Hb levels affirms this. PoX showed other strong and significant correlations with routine laboratory parameters, such as HDW, implying that the greater variability in hemoglobin content is associated with higher susceptibility to oxidative stress. On a functional level, it is interesting to find strong correlations of PoX with PoS, showing a direct link between RBC sickling tendency and ability to withstand oxidative stress. On the other hand, HbS and HbF levels did not correlate significantly to PoX. Overall, our study highlights the potential use of the RoxyScan and its associated biomarkers in the analysis of the RBC's response to oxidative stress. Further research is in progress to investigate the clinical applicability of this new technique.

The effect of postoperative intravenous iron in anaemic, older cardiac surgery patients on disability-free survival (AGE ANEMIA study): study protocol for a multi-centre, double-blind, randomized, placebo-controlled trial

BackgroundPostoperative anaemia is common in older cardiac surgery patients and often caused by iron deficiency. Anaemia may negatively affect recovery after cardiac surgery. This study aims to determine the efficacy of treatment of postoperative iron deficiency anaemia (IDA) with intravenous iron (IVI) on disability 90 days after cardiac surgery in older patients.MethodsThis is a randomized placebo-controlled double-blind multi-centre trial. In total, 310 patients aged ≥ 70 years with moderate IDA on postoperative day 1 (haemoglobin 85–110 g/L and ferritin concentration < 100 μg/L or iron saturation < 20%) after uncomplicated elective cardiac surgery (aortic valve repair or coronary artery bypass graft surgery) will be included. Patients will be randomly allocated to receive either IVI (ferric derisomaltose) or placebo (sodium chloride 0.9%) on postoperative day 1 in a 1:1 ratio, stratified by centre and type of cardiac surgery. The primary outcome is disability measured by the 12-item World Health Organization Disability Assessment score 2.0 after 90 days. Secondary outcome measures are the number of postoperative red blood cell (RBC) transfusions, change in reticulocyte haemoglobin content (pg) from randomization to hospital discharge, Hb levels at discharge, hospital complications, dyspnoea (assessed with the Rose Dyspnoea Scale) and health-related quality of life (HRQL) (assessed with The Older Persons and Informal Caregivers-Short Form (TOPICS-SF) questionnaire) after 90 days and days alive and out of hospital after 90 days. Lastly, the functional outcomes (e.g. steep ramp or 6-min walk test) and Hb level after 90 days will be assessed as an exploratory endpoint.DiscussionThe results of this study will demonstrate whether early treatment of postoperative IDA with IVI improves disability at 90 days in older cardiac surgery patients.Trial registrationClinicalTrials.gov NCT04913649. Registered on June 4, 2021.

Reticulocyte hemoglobin equivalent as a marker to assess iron deficiency: A large pediatric tertiary care hospital study.

Detection of iron deficiency (ID) remains challenging. We aimed to evaluate the performance of reticulocyte hemoglobin equivalent (Ret-He) as a potential diagnostic marker to assess ID and iron deficiency anemia (IDA) in a large pediatric cohort. A total of 3158 patients (aged 15 days to 19 years with a median age of 8.5 years; 60.2% female) were retrospectively studied. Statistical analysis was performed (a) to evaluate relationship of Ret-He with other relevant complete blood count and iron panel parameters; (b) to compare the levels of Ret-He in ID and IDA groups to a control group; and (c) to assess sensitivity and specificity of Ret-He in ID, IDA, and anemia without ID groups. Ret-He values were significantly positively correlated to ferritin and transferrin saturation (TSAT). The median Ret-He was significantly lower in ID. A Ret-He cutoff of ≤30.0 pg distinguished cases of ID from the control group with a sensitivity of 90.2%, specificity of 59.5%, and area under curve (AUC) of 0.88. Ret-He showed better diagnostic performance in the IDA group and acceptable performance for ID without anemia. The sensitivity, specificity, and AUC were 90.1%, 80.9%, and 0.93 for IDA at cutoff value of ≤27.4 pg, and 80.8%, 51.1%, and 0.70 for ID without anemia at cutoff value of ≤30.8 pg, respectively. Our large pediatric tertiary care hospital study demonstrates that Ret-He is a reliable marker to help confirm IDA in pediatric population. However, further studies are needed for its use to capture the early stages of ID.

Diagnostic performance of automated red cell parameters in predicting bone marrow iron stores.

The aim of the study was to determine the diagnostic performance of novel automated red cell parameters for estimating bone marrow iron stores. The study was a retrospective single-centre study based on data from an automated haematology analyser and results of bone marrow iron staining. Red cell parameters were measured on a Sysmex XN-series haematology analyser. Bone marrow iron stores were assessed semiquantitatively bycytochemical reaction according to Perls. The analysis included 429 bone marrow aspirate smears from 393 patients. Median age of patients was 67 years, 52 % of them were female. The most common indication for bone marrow examination was a plasma cell dyscrasia (n=104; 24 %). Median values of percentage of hypochromic and hyperchromic red blood cells (%HYPO-He, %HYPER-He), reticulocyte haemoglobin equivalent (RET-He) and microcytic red blood cells (MicroR) were statistically significantly different between cases with iron deplete and iron replete bone marrow. In a logistic regression model, ferritin was the best predictor of bone marrow iron stores (AUC=0.891), outperforming RET-He and %HYPER-He (AUC=0.736 and AUC=0.722, respectively). In a combined model, ferritin/MicroR index achieved the highest diagnostic accuracy (AUC=0.915), outperforming sTfR/log ferritin index (AUC=0.855). While single automated red cell parameters did not show improved diagnostic accuracy when compared to traditional iron biomarkers, a novel index ferritin/MicroR has the potential to outperform ferritin and sTfR/log ferritin index for predicting bone marrow iron stores. Further research is needed for interpretation and implementation of novel parameters and indices, especially in the context of unexplained anaemia and myelodysplastic syndromes.

Relationship between Iron Metabolic Parameters and Platelet Counts in Blood Donors

To investigate the correlation of iron metabolic parameters with platelet counts in blood donors. A total of 400 blood donors who met requirements of apheresis platelet donation were collected, and their hematological parameters were analyzed. The donors were divided into low ferritin group and normal group, the differences of hematological parameters between the two groups were compared, and the correlation of iron metabolic parameters and routine hematology parameters with platelet counts were analyzed. Whether male or female, low ferritin group had higher platelet counts than normal group (P < 0.01). Among the iron metabolic parameters, the platelet counts was negatively correlated with serum ferritin (SF), serum iron (SI), and transferrin saturation (TSAT) (r =-0.162, r =-0.153, r =-0.256), and positively correlated with total iron binding capacity (TIBC) and unsaturated iron binding capacity (UIBC) (r =0.219, r =0.294) in female blood donors. Platelet counts was also negatively correlated with SF, SI and TSAT (r =-0.188, r =-0.148, r =-0.224) and positively correlated with UIBC (r =0.220) in male blood donors. Among the routine hematology parameters, platelet counts was negatively correlated with mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and reticulocyte hemoglobin equivalent (Ret-He) in female blood donors (r =-0.236, r =-0.267, r =-0.213, r =-0.284). Platelet counts was also negatively correlated with MCH, MCHC and Ret-He in male blood donors (r =-0.184, r =-0.221, r =-0.209). In blood donors with low C-reactive protein level, the lower the iron store capacity, the lower the iron utilization, and the platelet counts tends to rise.

Soluble transferrin receptor concentration compared with reticulocyte hemoglobin content as a reliable marker of iron deficiency anemia in hemodialysis patients

Abstract Introduction Adequate iron stores are necessary for successful erythropoiesis-stimulating agents (ESAs) therapy in hemodialysis (HD) patients. However, iron status estimation in HD patients remains challenging, as most parameters are affected by inflammation. We aimed to compare the performance of soluble transferrin receptor concentration (sTfR) and reticulocyte hemoglobin content (CHr) as potential markers for iron status assessment. Methods This was a cross-sectional study that included 60 anemic patients with kidney failure on regular HD for a minimum of 3 months. Patients were in the maintenance phase of the ESAs with stable doses for at least 4 weeks and at least 3 weeks free of iron therapy. Patients were classified into two groups; group I: 18 patients with noniron deficiency anemia (NIDA); transferrin saturation (TSAT) greater than 20% and ferritin greater than 200 ng/ml and group II: 42 patients with iron deficiency anemia (IDA); transferrin saturation less than 20% and/or ferritin less than 200 ng/ml. CHr and sTfR were evaluated as reliable markers for diagnosing iron deficiency. Results CHr was significantly lower while sTfR was significantly higher in patients with IDA. Based on the receiver operating characteristic (ROC) curve, the area under the curve (AUC) of sTfR was significantly larger than the area under the curve of CHr; however, this had a borderline significance. Conclusion sTfR has a significantly higher diagnostic power for diagnosis of IDA than CHr.

Role of reticulocyte hemoglobin equivalent in early detection of iron deficiency anemia in regular hemodialysis patients

Role of reticulocyte hemoglobin equivalent in early detection of iron deficiency anemia in regular hemodialysis patients

Reticulocyte hemoglobin content as a marker of iron deficiency in premature newborns with very low birth weight. A simple tool for diagnosing iron deficiency

Reticulocyte hemoglobin content (RET-He) is a promising marker of iron deficiency (ID) in newborns. Objective: to determine the diagnostic value of RET-He as a marker of ID in premature newborns with very low birth weight (VLBW). We conducted a single-center retrospective cohort study, which included 66 premature infants admitted to the National Medical Research Center for Obstetrics, Gynecology and Perinatology named the Academician V.I. Kulakov of the Ministry of Healthcare of the Russian Federation. Data were obtained from January 2016 to December 2018. The gestational age ranged from 29 to 32 weeks. Laboratory examination included blood tests on the 1st and 3rd day of life, then every 10–14 days until the day of life, then every 10–14 days the Institute of Neonatology and Pediatrics; discharge from hospital, and the measurements of serum iron, ferritin, transferrin on the 7th until the discharge from hospital. This clinical study was approved by the Biomedical Research Ethics Committee (Minutes No.12 of 17 November 2016) and the Scientific Council (Minutes No.19 of 29 November 2016) of the National Medical Research Center for Obstetrics, Gynecology and Perinatology named after the Academician V.I. Kulakov of Ministry of Healthcare of the Russian Federation. RET-He was the highest at birth and declined gradually thereafter in premature newborns reaching the lowest values after 3 weeks of life (median (interquartile range) 28.4 (25.8–34.8) pg (on the 1st day of life – 40.0 (35.7–41.9) pg and 33.5 (29.2–36.6) pg at the time of discharge). A low RET-He level was associated with low reticulocytes, with no changes in hemoglobin. There was a positive correlation between RET-He and MCH. D-He decreased from 1 to 42 days of life as a marker of increasing anemia. There was a negative correlation between RET-He and Hypo-He (p &lt; 0.005). Starting from 42 days of life, or by the time of discharge, 32% of premature infants (n = 21) had a low ferritin level and 77% (n = 51) of premature infants had a low RET-He level, of which 21 infants developed ID (a positive correlation between RET-He and ferritin after 42 days of life (r = 0.34, p = 0.046)). There was no correlation between RET-He and ferritin in newborns without ID. Also, there were no correlations between RET-He and iron and RET-He and transferrin. After 42 days of life, RET-He less than 28.4 pg was a marker of ID (sensitivity 83.3% and specificity 93.7%). Low RET-He, D-He, RBC-Hе and high microR, Hypo-He were the earliest markers of ID in premature infants which predicted a decrease in serum iron and ferritin levels. RET-He, D-Не and Hypo-He are biomarkers with accurate diagnostic value of ID in premature infants with VLBW.

A-139 Investigating the Utility of Reticulocyte Hemoglobin Equivalent for Diagnosis of Iron Deficiency Anemia

Abstract Background Iron deficiency anemia (IDA) is one of the most common leading global health problems, which could cause extreme fatigue, chest pain, headache, and even neurodevelopment deterioration in children. Traditionally, assessing serum ferritin, transferrin, and iron are the tools to diagnose IDA. However, a high false-positive rate of these conventional biomarkers might be observed due to inflammation, infection, or malignancy. Reticulocyte hemoglobin equivalent (RET-He) is a new parameter that reflects the recent functional availability of iron for erythropoiesis, with the advantage of real-time results as part of automated reticulocyte analysis. The present study, therefore, aimed to investigate the utility of RET-He for the diagnosis of patients with IDA. Methods This retrospective study collected laboratory data from patients with both serum ferritin and reticulocyte results from January 2022 through December 2022, and excluded the high white blood cell count (WBC &amp;gt; 9380 cells/μL) and high C-reaction protein (CRP &amp;gt; 0.5 mg/dL). RET-He was measured by Sysmex XN-10 or XN-20 automated hematology analyzer (Sysmex, Kobe, Japan). Serum ferritin was measured by Roche Cobas e801 (Roche Diagnostics, Mannheim, Germany). Iron and total iron binding capacity (TIBC) were measured by Roche Cobas c702 (Roche Diagnostics, Mannheim, Germany). Anemia was defined as hemoglobin concentration &amp;lt;13 g/dL for men and &amp;lt; 12 g/dL for women. Iron deficiency was defined as serum ferritin &amp;lt;15 μg/L. Student T-tests and Pearson correlation were performed, and P-values less than 0.05 were considered statistically significant. Statistical analyses were performed with SPSS software (Version 22.0). Results A total of 2208 patient data were enrolled in this study and were classified into the IDA, iron deficiency, anemia, and control groups according to ferritin level and hemoglobin concentration. The RET-He were 20.6, 27.7, 30.4, and 32.3 pg in the IDA, iron deficiency, anemia, and control groups, respectively. The IDA group showed significantly lower RET-He levels than the control group (P &amp;lt; 0.001). By Pearson correlation analysis, the RET-He showed a weak positive correlation with ferritin and hemoglobin (r = 0.236, 0.274), moderate positive correlation with iron (r = 0.354), and moderate negative correlation with TIBC (r = −0.369). In addition, the average turn-around time (received-reported) of RET-He was 16.4 min, which was shorter than other biochemical markers such as ferritin, iron, and TIBC (57.7, 34.2, 34.4 min). Conclusion The RET-He could be an assessment of the iron actually used for the biosynthesis of hemoglobin and as a useful and rapid tool for diagnosing and monitoring iron deficiency anemia.

Sclerosing angiomatoid nodular transformation of the spleen in a child with anemia: a case report and review of the literature

BackgroundSclerosing angiomatoid nodular transformation of the spleen is a relatively rare benign vascular lesion in both adult and pediatric age groups with unclear etiopathogenesis and variable clinical presentations. Many benign and also malignant splenic masses could mimic sclerosing angiomatoid nodular transformation, both clinically and radiologically. Herein, we report our experience with a case of sclerosing angiomatoid nodular transformation in a 3-year-old girl.Case reportA 3-year-old Iranian girl presented with abdominal pain, back pain, and constipation for 2 weeks. She was being followed up by a pediatrician due to her short stature and persistent anemia. Physical examination showed stable vital signs, short stature, pallor, and a puffy face. Laboratory evaluation showed normochromic normocytic anemia with a normal reticulocyte count, ferritin, and hemoglobin electrophoresis. Radiologic assessments revealed a hypoechoic lesion in the spleen with high vascularity, clinically suspected to be lymphoma. She was operated on, and after partial splenectomy, pathologic evaluation of the spleen showed a solitary, well-demarcated, and unencapsulated dark mass. Microscopic examination revealed micronodular appearance composed of irregular-shaped vascular spaces lined by plump endothelial cells and surrounded by concentric collagen fibers, features in keeping with sclerosing angiomatoid nodular transformation. The patient’s anemia was resolved after surgery, and no clinical or radiologic deficits were noted during the 10-month follow-up visits.ConclusionAlthough sclerosing angiomatoid nodular transformation is exceedingly rare in children, it should be considered a differential diagnosis in pediatric splenic neoplasms with concurrent hematologic manifestations, such as anemia.

Hematinic Potential of Olive Leaf Extract: Evidence from an In Vivo Study in Mice and a Pilot Study in Healthy Human Volunteers.

Natural resources have recently received considerable attention as complementary or alternative hematinic agents. In this regard, olive leaf extract, which is rich in bioactive phenolic compounds, has been reported to induce erythroid differentiation in human hematopoietic stem cells. Therefore, in the present study, we aimed to explore the potential hematinic properties of aqueous olive leaf extract (WOL) in vivo. After 24 days of administering WOL to healthy mice orally, red blood cell (RBC), hematocrit, reticulocyte, and reticulocyte hemoglobin content (CHr) showed a significant increase. Additionally, WOL promoted plasma iron levels and the expression of splenic ferroportin (Fpn), an iron transporter. Additionally, a single-arm pilot study involving a limited number of healthy volunteers was conducted to assess WOL's feasibility, compliance, and potential benefits. Following an 8-week intervention with WOL, RBC count and hemoglobin level were significantly increased. Notably, there were no significant changes in the safety measures related to liver and kidney functions. Furthermore, we identified oleuropein and oleuroside as the active components in WOL to induce erythroid differentiation in the K562 cell line. Altogether, our study presents evidence of the hematinic potential of WOL in the in vivo studies, opening up exciting possibilities for future applications in preventing or treating anemia.

Serum ferritin values in neonates

To compare serum ferritin and RET-He values among extremely low gestational age neonates ELGANs with other markers of iron-deficient erythropoiesis. This is a secondary analysis of the NICHD Darbepoetin Trial. Study data from placebo recipients who had a serum ferritin, a RET-He, and a mean corpuscular volume (MCV) measurement within a 24-hour period were analyzed for correlation. Mixed linear regression models showed no association between ferritin and RET-He at both early (β = 0.0016, p = 0.40) and late (β = -0.0001, p = 0.96) time points. Positive associations were observed between RET-He and MCV at baseline, early, and late time points (p < 0.01, =0.01, <0.001, respectively), while ferritin was not associated with MCV at any time point. Our study shows that RET-He is better correlated with MCV as a marker of iron-limited erythropoiesis than ferritin. The results suggest that ferritin is limited as a marker of iron sufficiency in premature infants. FDA IND Number 100138; ClinicalTrials.gov number NCT03169881; NRN ID number NICHD-NRN-0058 (Darbe).

Risk of Hemolytic Anemia in IBD Patients with Glucose-6-Phosphate Dehydrogenase Deficiency Treated with Mesalamine: Results of a Retrospective-Prospective and Ex Vivo Study.

Mesalamine is one of the most-used drugs in inflammatory bowel disease (IBD), especially ulcerative colitis. Regulatory agencies have listed mesalamine as an unsafe drug in subjects with glucose-6-phosphate dehydrogenase (G6PD) deficiency based on the risk of hemolysis, although scientific evidence is lacking. The occurrence of acute and/or chronic hemolytic anemia in IBD patients with G6PD deficiency exposed to mesalamine was evaluated. In this multicenter study, IBD patients with G6PD deficiency (cases) receiving mesalamine were retrospectively evaluated for acute, and prospectively for chronic, hemolysis. The presence of hemolytic anemia was based on red blood cell and reticulocyte count, hemoglobin, lactate dehydrogenase, unconjugated bilirubin, and haptoglobin levels. Cases were compared with controls (IBD patients with normal G6PD). A total of 453 IBD patients (mean age 52.1 ± 16.0 years; 58.5% female) were enrolled. Ulcerative colitis was present in 75% of patients. G6PD deficiency was detected in 17% of patients. Oral mesalamine was used in 67.9% of ulcerative colitis and in 32.4% of Crohn's disease cases. None of the 78 IBD patients with G6PD deficiency receiving mesalamine underwent hospitalization or specific treatment for acute hemolytic anemia. Relevant differences in chronic hemolysis markers were not observed in 30 cases compared with 112 controls receiving mesalamine (≤4500 mg/day). Marker modifications were also observed in mesalamine-free cases, consistent with the basal rate of erythrophagocytosis in G6PD deficiency. Ex vivo experiments showed the release of methemoglobin by G6PD deficient RBCs upon mesalamine challenge, only above 2.5 mg/mL, a concentration never reached in the clinical setting. This study provides, for the first time, evidence that mesalamine is safe in G6PD deficiency at a dosage of up to 4500 mg/day.

Reference intervals for reticulocyte indices, immature reticulocyte fraction, and the percentage of hypochromic red blood cells in adult large breed dogs using the ADVIA 2120 hematology analyzer.

Reticulocyte indices are used to characterize anemia, including the identification of regeneration. In people, the immature reticulocyte fraction (IRF), percentage of hypochromic red blood cells (%HYPO-RBC), and other reticulocyte indices have been used as earlier indicators of erythropoiesis and as valuable monitoring tools in the assessment of various therapies. The reference intervals (RI) of the IRF and %HYPO-RBC have not been reported in dogs. The objective of this study was to establish RIs for novel variables (IRF, %HYPO-RBC, and CH-delta) and assess RIs for more commonly reported reticulocyte indices in healthy dogs. RIs were calculated from blood results retrospectively collected from 106 client-owned healthy dogs at the time of induction into a blood donor program using the ADVIA 2120 hematology analyzer (Siemens Healthcare Diagnostics). For the calculation of RIs, appropriate tests were applied for outlier detection and normality assessment. For variables normally distributed, RIs and their respective 90% confidence intervals (CIs) were calculated using parametric methods, while for variables not normally distributed, robust methods were used and bootstrapping for calculating the 90% CIs. The following RIs were established: reticulocyte hemoglobin content (CHr) 24.5-28 pg, mean reticulocyte volume (MCVr) 85.9-99.3 fL, mean corpuscular hemoglobin concentration of reticulocytes (CHCMr) 271.0-306.3 g/L, IRF 10.4%-43.5%, CH-delta 0.5-4.3 pg, and percentage of hypochromic red blood cells (%HYPO-RBC) 0.10%-0.80%. The results of this study provide RIs for novel reticulocyte variables. Further studies are required to determine the clinical utility of IRF, %HYPO-RBC, and CH delta as early indicators of erythropoietic activity in canine patients.