rHuEPO Administration Research Articles

New Transcriptomic Biomarkers for Detection of the Recombinant Human Erythropoietin (rHuEPO) MirCERA in Horses.

Detection and monitoring of biomarkers related to doping is particularly suitable for the development of analytical strategies dedicated to indirect detection of banned substances. Previous studies in horses have already allowed the investigation of transcriptomic biomarkers in equine blood associated with reGH and rHuEPO administrations. Our most recent developments continue to focus on the discovery and monitoring of transcriptomic biomarkers for the control of ESAs, and a collaborative study with WADA-accredited doping control laboratories has recently been initiated to conduct a pilot study. In humans, three mRNAs (ALAS2, CA1, and SLC4A1) were previously observed to be differentially expressed after blood doping and were associated with immature red blood cells, the so-called circulating reticulocytes. In horses, circulating reticulocytes are rarely observed even after rHuEPO administration. With the improved primers that detect the equine orthologues of the human mRNAs from the ALAS2, CA1, and SLC4A1 genes, we can now report the first evidence of the detection of the three biomarkers in equine blood. In addition, an upregulation of the mRNA levels of the three genes was observed after analysis of blood samples collected from MirCERA-treated animals, with kinetics similar to those previously documented in humans. Our data suggest that ALAS2 and CA1 are promising indirect biomarkers for the detection of recombinant EPO abuse in horses, as observed in humans.

Erythropoietin as a prophylactic measure against anemia in critically ill patients: A combined prospective and retrospective study

Background & Objective: Critically sick patients often develop moderate to severe anemia, due to various factors. We tested the hypothesis that administering prophylactic recombinant human erythropoietin (rHuEPO) together with early identification and correction of iron deficiency anemia in traumatic brain injury patients would have a positive effect on the incidence of RBCs transfusions.
 Methodology: One hundred and seventeen head-trauma patients were enrolled. They received 40,000 units of prophylactic rHuEPO, starting on day three after admission and continuing at weekly intervals for 6 weeks, with follow-up of iron biomarkers. Any patient developing iron deficiency anemia was prescribed oral or intravenous iron supplement. The incidence of RBCs transfusion was recorded.
 Results: The number of RBCs transfusion was significantly less in the prospective group (27.35%) than the retrospective group (42.86%). Hemoglobin and hematocrit levels were significantly less in the first and second weeks after admission and then began to rise. Serum iron, transferrin saturation, and ferritin were at their lowest levels in the fourth week. Meanwhile, total iron binding capacity was higher in the fourth week and lower in the sixth.
 Conclusion: The administration of prophylactic rHuEPO with early discovery and correction of iron deficiency anemia resulted in a significant reduction in the incidence of RBCs transfusion in the head-trauma patients.
 Abbreviations: rHuEPO: recombinant human erythropoietin, RBC: red blood cell, ICU: intensive care unit, TBI: traumatic brain injury, GCS: Glasgow Coma Scale, APACHE: Acute Physiology and Chronic Health Evaluation
 Citation: Amer AF, Elatrozy HI, El Mourad MB. Erythropoietin as a prophylactic measure against anemia in critically ill patients: A combined prospective and retrospective study. Anaesth. pain intensive care 2022;26(5):633−639.
 DOI: 10.35975/apic.v26i5.1481

Metabolomic Profiling of Recombinant Erythropoietin (rHuEpo) in Trained Caucasian Athletes.

Recombinant human erythropoietin (rHuEpo) is prohibited by the World Anti-Doping Agency but remains the drug of choice for many cheating athletes wishing to evade detection using current methods. The aim of this study was to identify a robust metabolomics signature of rHuEpo using an untargeted approach in blood (plasma and serum) and urine. Longitudinal study. University of Glasgow. Eighteen male participants regularly engaged in predominantly endurance-based activities, such as running, cycling, swimming, triathlon, and team sports, were recruited. Each participant received 50 IU·kg -1 body mass of rHuEpo subcutaneously every 2 days for 4 weeks. Samples were collected at baseline, during rHuEpo administration (over 4 weeks) and after rHuEpo administration (week 7-10). The samples were analyzed using hydrophilic interaction liquid chromatography mass spectrometry. Significant metabolic signatures of rHuEpo administration were identified in all biofluids tested in this study. Regarding metabolomics data, 488 plasma metabolites, 694 serum metabolites, and 1628 urinary metabolites were identified. Reproducible signatures of rHuEpo administration across all biofluids included alterations of pyrimidine metabolism (orotate and dihydroorotate) and acyl-carnitines (palmitoyl-carnitine and elaidic carnitine), metabolic pathways that are associated with erythropoiesis or erythrocyte membrane function, respectively. Preliminary metabolic signatures of rHuEpo administration were identified. Future studies will be required to validate these encouraging results in independent cohorts and with orthogonal techniques, such as integration of our data with signatures derived from other "omics" analyses of rHuEpo administration (eg, transcriptomics).

Erythropoietin as a prophylactic measure against anemia in critically ill patients: A combined prospective and retrospective study.

Background: The aim of this study is to test the hypothesis that administering prophylactic recombinant human erythropoietin (rHuEPO) together with early identification and correction of iron deficiency anemia in traumatic brain injury patients would have an effect on the incidence of RBCs transfusion as a primary outcome.
 Methodology: One hundred and seventeen head-trauma patients were enrolled. They received 40,000 units of prophylactic rHuEPO, starting on day three after admission and continuing at weekly intervals for 6 weeks, with follow-up of iron biomarkers. Any patient developing iron deficiency anemia was prescribed oral or intravenous iron. The incidence of RBCs transfusion was recorded.
 Results: The incidence of RBCs transfusion was significantly less in the prospective group (27.35%) than the retrospective group (42.86%). Hemoglobin and hematocrit levels were significantly less in the first and second weeks after admission and then began to elevate. Serum iron, transferrin saturation, and ferritin were at their lowest levels in the fourth week. Meanwhile, total iron binding capacity was higher in the fourth week and lower in the sixth.
 Conclusion: The administration of prophylactic rHuEPO with early discovery and correction of iron deficiency anemia resulted in a significant reduction in the incidence of RBCs transfusion in head-trauma patients.

Efficacy of Roxadustat on anemia and residual renal function in patients new to peritoneal dialysis

Background Both early correction of anemia and preserving residual renal function (RRF) are reported to improve patient survival. The aim of this study was to explore the efficacy and safety of Roxadustat for treatment of renal anemia in patients new to peritoneal dialysis (PD) and to assess its impact on RRF. Methods A retrospective analysis was performed on 60 initial peritoneal dialysis (PD) patients with renal anemia. Twenty-eight cases were treated with Roxadustat (Roxadustat group) and 32 with recombinant human erythropoietin (control group). Clinical characteristics, hemoglobin (Hb), C-reactive protein, blood lipids, iron metabolism, dialysis adequacy and RRF of the two groups were evaluated and adverse events were recorded. All patients were followed up for at least 40 weeks. Results After 40 weeks of treatment, mean Hb levels were significantly higher from baseline values in both groups, the mean Hb change in Roxadustat group was higher than control group (3.46 ± 1.59 g/dL vs. 2.28 ± 2.27 g/dL, p < 0.05). At 40 weeks, 92.9% patients met the target level of Hb in Roxadustat group and 84.4% in control group. Total iron binding was higher and ferritin was lower in Roxadustat group from baseline values and Roxadustat-induced Hb increases were independent of baseline C-reactive protein levels and history of rhuEPO administration. RRF decreased over time in both groups, the mean RRF change was lower in Roxadustat group than control group (1.15 ± 1.66 mL/min/1.73 m2 vs. 2.31 ± 1.46 mL/min/1.73 m2, p < 0.01). Compared with control group, patients in Roxadustat group had higher levels of total iron binding, 24 h urine volume, total weekly Ccr, and lower systolic pressure, ferritin, C-reactive protein, total cholesterol, LDL. No serious adverse reactions occurred in either group. Conclusion In patients new to PD, Roxadustat effectively and safely improved renal anemia and delay the decline of RRF.

Erythropoietin in children with hemolytic uremic syndrome: a pilot randomized controlled trial.

The efficacy of recombinant human erythropoietin (rHuEPO) in sparing red blood cell (RBC) transfusions in children with hemolytic uremic syndrome related to Shiga toxin-producing Escherichia coli (STEC-HUS) is uncertain. We conducted a pilot randomized controlled open trial between December 2018 and January 2021. Children were randomized to the intervention (subcutaneous rHuEPO 50 U/kg three times weekly until discharge + RBC transfusion if hemoglobin ≤ 7g/dL and/or hemodynamic instability) or to the control arm (RBC transfusion if hemoglobin ≤ 7g/dL and/or hemodynamic instability). Primary outcome was the number of RBC transfusions received during hospitalization. Secondary outcomes were to explore whether baseline EPO levels were adequate to the degree of anemia, to correlate selected acute phase parameters with the number of RBC transfusions, and to assess possible adverse events. Twelve patients per arm were included; they were comparable at recruitment and throughout the disease course. Median number of RBC transfusions was similar between groups (1.5, p = 0.76). Most patients had baseline EPO levels adequate to the degree of anemia, which did not correlate with the number of transfusions (r = 0.19, p = 0.44). Conversely, baseline (r = 0.73, p = 0.032) and maximum lactic dehydrogenase levels (r = 0.78, p = 0.003), creatinine peak (r = 0.71, p = 0.03) and dialysis duration (r = 0.7, p = 0.04) correlated significantly with RBC requirements. No side effects were recorded. In children with STEC-HUS, the administration of rHuEPO did not reduce the number of RBC transfusions. Larger studies addressing higher doses and similar severity of kidney failure at rHuEPO initiation (e.g. at start of dialysis) are warranted. ClinicalTrials.gov identifier: NCT03776851. A higher resolution version of the Graphical abstract is available as Supplementary information.

Integrating Whole Blood Transcriptomic Collection Procedures Into the Current Anti-Doping Testing System, Including Long-Term Storage and Re-Testing of Anti-Doping Samples.

Introduction: Recombinant human erythropoietin (rHuEPO) administration studies involving transcriptomic approaches have demonstrated a gene expression signature that could aid blood doping detection. However, current anti-doping testing does not involve collecting whole blood into tubes with RNA preservative. This study investigated if whole blood in long-term storage and whole blood left over from standard hematological testing in short-term storage could be used for transcriptomic analysis despite lacking RNA preservation. Methods: Whole blood samples were collected from twelve and fourteen healthy nonathletic males, for long-term and short-term storage experiments. Long-term storage involved whole blood collected into Tempus™ tubes and K2EDTA tubes and subjected to long-term (i.e., ‒80°C) storage and RNA extracted. Short-term storage involved whole blood collected into K2EDTA tubes and stored at 4°C for 6‒48 h and then incubated at room temperature for 1 and 2 h prior to addition of RNA preservative. RNA quantity, purity, and integrity were analyzed in addition to RNA-Seq using the MGI DNBSEQ-G400 on RNA from both the short- and long-term storage studies. Genes presenting a fold change (FC) of >1.1 or < ‒1.1 with p ≤ 0.05 for each comparison were considered differentially expressed. Microarray analysis using the Affymetrix GeneChip® Human Transcriptome 2.0 Array was additionally conducted on RNA from the short-term study with a false discovery ratio (FDR) of ≤0.05 and an FC of >1.1 or < ‒1.1 applied to identify differentially expressed genes. Results: RNA quantity, purity, and integrity from whole blood subjected to short- and long-term storage were sufficient for gene expression analysis. Long-term storage: when comparing blood tubes with and without RNA preservation 4,058 transcripts (6% of coding and non-coding transcripts) were differentially expressed using microarray and 658 genes (3.4% of mapped genes) were differentially expressed using RNA-Seq. Short-term storage: mean RNA integrity and yield were not significantly different at any of the time points. RNA-Seq analysis revealed a very small number of differentially expressed genes (70 or 1.37% of mapped genes) when comparing samples stored between 6 and 48 h without RNA preservative. None of the genes previously identified in rHuEPO administration studies were differently expressed in either long- or short-term storage experiments. Conclusion: RNA quantity, purity, and integrity were not significantly compromised from short- or long-term storage in blood storage tubes lacking RNA stabilization, indicating that transcriptomic analysis could be conducted using anti-doping samples collected or biobanked without RNA preservation.

Haematological Indicators of Response to Erythropoietin Therapy in Chronic Renal Failure Patients on Haemodialysis: Impact of Angiotensin-Converting Enzyme rs4343 Gene Polymorphism.

PurposeThis is the first cross-sectional study studying the changes in haematological indicators of the response to recombinant human erythropoietin (rHuEPO) therapy in chronic renal failure (CRF) patients on haemodialysis (HD) stratified according to ACE G2350A (rs4343) gene polymorphism.DesignAn observational cross-sectional study.SettingNephrology department and Biochemistry and molecular biology department, faculty of medicine, Cairo University.PatientsA total of 256 CRF patients on HD for at least six months (162 male and 103 female) and 160 healthy subjects (122 male and 38 female) were recruited in the current study after signing a consent form. ACE G2350A (rs4343) Insertion/Deletion (I/D) was tested, the association between ACE G2350A (RS4343) gene polymorphisms and patients response to rHuEpo was evaluated.ResultsACE G2350A (rs4343) I/D was the most prevalent genotype, while I/I genotype was the lowest prevalent among patient or control subjects included in the study. D allele is the most prevalent allele, either among patients or the control group. Hemoglobin (Hb) level in patients with I/I and Deletion/Deletion (D/D) genotype was significantly higher compared to those with I/D genotype (P = 0.012 and P = 0.005, respectively). Serum iron in the I/D genotype was significantly higher than those with either I/I or D/D genotype (P = 0.045 and P = 0.018, respectively). Angiotensin-converting enzyme (ACE) content, total leukocytic count (TLC), and soluble erythropoietin receptor (sEpoR) were independent predictors of Hb level. The ACE gene, TLC, and serum iron were the independent factors that may affect the Haematocrit (Hct) level. ACE G2350A (rs4343) gene polymorphisms may affect the HD patient’s responses to rHuEPOs.ConclusionIn HD patients, screening for ACE G2350A (rs4343) gene polymorphisms before rHuEpo administration may help predict patient response.

Micro‐doses of Recombinant Human Erythropoietin Enhance Endurance Performance While Indirect Detection by The Athlete Biological Passport Is Improved by Including The Immature Reticulocyte Fraction

Recombinant human erythropoietin (rHuEPO) is ergogenic but micro-doses remain a challenge for anti-doping authorities. Here, we investigated the hypothesis that frequent micro-doses of rHuEPO enhance maximal oxygen consumption (VO2max) and time trial performance and that the combination of immature reticulocyte fraction (IRF) and the Athlete's Biological Passport (ABP) would enhance detection compared to the ABP alone. In a randomized, double-blinded, placebo-controlled design, 34 trained (VO2max: 52 ± 7 ml×min-1×kg-1, mean ± SD) adults received either rHuEPO (rHuEPO: epoetin-β, 9 IU/kg, n = 16) or placebo (PLA: 0.9% NaCl, n = 18) three times a week for four weeks. Before and after the intervention, VO2max, mean power output during a 400-kcal time trial and total hemoglobin mass (tHb) were measured. Venous blood samples were collected before, each week during rHuEPO administration, and 3, 5 and 10 days after the last injection. Hematological values were evaluated using dedicated ABP software. VO2max increased by 4.7% (p<0.01) and tHb increased by 6.8% (p<0.001) in the rHuEPO group when compared to the PLA group where no changes were evident. Mean power output during the time trial increased by 4.0% (p<0.01) in the rHuEPO group with no changes observed in the PLA group. Inclusion of IRF thresholds improved ABP sensitivity from 47% to 73%, yielding a peak sensitivity during the first two weeks of the rHuEPO treatment and 10 days after cessation of treatment. In conclusion, these findings show that four weeks of rHuEPO micro-doses are sufficient to induce considerable ergogenic effects on both VO2max and time trial performance in trained adults. The inclusion of IRF thresholds into the ABP improved sensitivity during treatment and the viability of including IRF into the ABP should be considered although more studies are needed.

JTZ-951 (enarodustat), a hypoxia-inducible factor prolyl hydroxylase inhibitor, improves iron utilization and anemia of inflammation: Comparative study against recombinant erythropoietin in rat

Anemia with inflammation-induced defective iron utilization is a pathological condition observed in patients suffering from chronic kidney disease (CKD) or chronic inflammatory disease. There is no reasonable treatment for these conditions, because the effects of erythropoiesis stimulating agents (ESAs) or iron supplementation in the treatment of anemia are insufficient. JTZ-951 (enarodustat) has been characterized as a novel, orally bioavailable inhibitor of hypoxia-inducible factor prolyl hydroxylase (HIF-PH), and has been developed as a novel therapeutic agent for anemia with CKD. In this study, the effects of JTZ-951 on iron utilization during erythropoiesis and on anemia of inflammation were compared with those of recombinant human erythropoietin (rHuEPO) using normal rat and rat model of anemia of inflammation.In normal rats, under conditions in which JTZ-951 and rHuEPO showed similar erythropoietic effect, repeated doses of JTZ-951 induced erythropoiesis while retaining the hemoglobin content in red blood cells, while administration of rHuEPO resulted in decrease in some erythrocyte-related parameters. As for iron-related parameters during erythropoiesis, JTZ-951 exhibited more efficient iron utilization compared to rHuEPO. A single dose of JTZ-951 resulted in decrease in hepcidin expression observed within 24 h after administration, but a single dose of rHuEPO did not.In a rat model of anemia of inflammation (also known as a model with functional iron-deficiency), JTZ-951 showed erythropoietic effect, in contrast with rHuEPO.These results suggest that, unlike rHuEPO, JTZ-951 stimulates erythropoiesis by increasing iron utilization, and improves anemia of inflammation.

Erythropoietin plus iron versus control treatment including placebo or iron for preoperative anaemic adults undergoing non-cardiac surgery.

Erythropoietin plus iron versus control treatment including placebo or iron for preoperative anaemic adults undergoing non-cardiac surgery.

Proof-of-concept study on improved efficacy of rHuEPO administered as a long-term infusion in rats

BackgroundHuman recombinant erythropoietin (rHuEPO) is often used in the treatment of diseases associated with a decreased production of red blood cells (RBC), such as chronic renal failure. rHuEPO is typically administered as an intravenous or subcutaneous (SC) injection every few days. The low minimum effective concentration (MEC) of rHuEPO, compared to the concentrations observed after standard doses, suggests that a low dose of the drug administered as a long-term infusion should be efficacious. This study aimed to compare the efficacy observed after a single subcutaneous administration of rHuEPO with that observed after a long-term infusion of rHuEPO via implanted osmotic pumps at a similar or lower dose.Materials and methodsIn this study three rats received rHuEPO as a single SC injection at a dose of 1350 IU/kg, nine via osmotic pumps at a rate of 0.25, 0.5 and 1 IU/kg and at a total dose of 333 IU/kg, 667 IU/kg, 1333 IU/kg. Three rats served as a control group. The erythropoietin concentrations, RBC count and hemoglobin were measured.ResultsAn increase in RBC count and hemoglobin was observed after SC infusion of rHuEPO. The baseline corrected area under the effect curve for hemoglobin and RBC count was more than 10-times higher for the SC infusion than for a single SC administration with a comparable dose.ConclusionsThis study demonstrates that administration of rHuEPO as a long-term infusion at a rate ensuring MEC allows to achieve a high efficacy of therapy using relatively small doses of the drug.

Screening and confirmatory analysis of recombinant human erythropoietin for racing camels' doping control.

Recombinant human erythropoietin (rHuEPO) belongs to the therapeutic class of erythropoiesis stimulating agents (ESAs) due to its implication in the creation pathway of red blood cells and thus enhancement of oxygenation. Because of this bioactivity, rHuEPO has been considered as a major doping agent in sports competitions for decades. Over the years, doping control laboratories designed several analytical strategies applied to human and animal samples to highlight any misuse. Even though multiple analytical approaches have been reported, none has yet been dedicated to racing camels. Here, we describe an analytical strategy to test camel plasma samples at screening using an ELISA assay and a targeted nano-liquid chromatography-high-resolution tandem mass spectrometry for confirmatory analysis. The method was validated and has been successfully applied to post-race samples, allowing the detection of a positive case of rHuEPO administration.

Role of the erythropoietin receptor in Lung Cancer cells: erythropoietin exhibits angiogenic potential.

Background: Recombinant human erythropoietin (rHuEPO), a hormone regulating the proliferation and differentiation of erythroid cells, is one of the prescription drugs used to treat cancer-associated anemia. However, administration of rHuEPO to cancer patients has been reported to be associated with decreased survival, and the mechanism by which it acts remains controversial. The present study aimed to investigate the expression of the EPO-receptor in lung cancer cell lines and whether rHuEPO treatment affected its growth and migration. Moreover, the angiogenic effects of rHuEPO were also explored in vivo.Methods: Expression of the EPO-receptor in lung cancer cell lines was measured by Western blotting and enzyme linked immunosorbent assays (ELISAs). Proliferation of the lung cancer cells was monitored in the presence of rHuEPO. Human umbilical vein endothelial cells (HUVECs) were used for tube formation assays in vitro, and transwell migration assays were performed to detect migration under rHuEPO treatment. Matrigel plug technology was employed to observe the angiogenic effects in both nude mice and Matrigel-containing lung cancer cell lines H838 or H1975. Microvessel density (MVD) was measured using CD31 Immunohistochemistry (IHC) staining.Results: EPO-receptor (EPO-R) was only detected in the cell lines H838 and H1339 by ELISA. However, the EPO-R protein was detected in all cell lines by Western blotting, which is in contradiction to the ELISA results. Proliferation and migration were not affected by rHuEPO treatment. However, rHuEPO promoted HUVEC tube formation in vitro and significantly induced the formation of new blood vessels in vivo. Furthermore, rHuEPO did not antagonize the inhibitory effects of Afatinib (epidermal growth factor receptor-tyrosine kinase inhibitor; EGFR-TKI) in simultaneous treatment with rHuEPO. In a 3D cell co-culture model, rHuEPO did not enhance the secretion of vascular endothelial growth factor (VEGF) in lung cancer cells or human lung fibroblast cell line MRC-5.Conclusions: We have shown that the role of EPO goes beyond erythropoiesis, also playing a strong role in angiogenesis by participating in new blood vessel formation in lung cancer models. Thus, rHuEPO may raise the risk of thrombosis and metastasis in vivo. Additionally, our results suggest that studies using commercially available EPO-R antibodies should be reexamined; some of these antibodies may not in fact recognize EPO-R.

Effect of preoperative recombinant human erythropoietin administration on inflammatory lung injury due to cardiopulmonary bypass

Objective To discuss the effects of preoperative use of recombinant human erythropoietin (rHuEPO) on inflammatory lung injury patients after cardiopulmonary bypass (CPB) under extracorporeal circulation. Methods A total of 54 patients who were scheduled to receive elective open cardiac operations under CPB were enrolled. They were divided into two groups(n=27). rHuEPO group: patients were titrated intravenously with rHuEPO (100 IU/kg, diluted into 50 ml normal saline) for consecutive three days at the same time period before operation. Control group: equivalent normal saline in place of rHuEPO was given to patients via venous injection. Peak airway pressure (Ppeak), plateau pressure (Pplat), and pulmonary dynamic compliance (Cdyn) were well monitored and recorded before CPB (T1), immediately after chest closure (T2), and 2, 4 h and 6 h after surgery (T3, T4, and T5). At T1, T2, T5, and 12, 24, 48 h, and 72 h after surgery (T6, T7, T8, and T9), arterial oxygenation indices (OI) were also recorded. Furthermore, at pre-rHuEPO treatment (T0), T1, T2, T5, T6, T7, T8, and T9, the plasma levels of the tumor necrosis factor (TNF)-α and interleukin (IL)-1β and IL-10 were measured. The time of mechanical ventilation and stay in intense care unit (ICU), discharge time and hospitalization stay were also recorded. Results Compared with the control group, Cdyn in the rHuEPO group at T2-T5 were significantly increased (P<0.05), whereas Ppeak and Pplat were decreased (P<0.05). OI values of rHuEPO group at T2 and T5-T9 were significantly higher than that of control group(P<0.05). It was demonstrated that at any time point after closing chest, the plasma concentrations of TNF-α, IL-1β, and IL-10 in both groups were relatively elevated compared with those before, while the concentrations of TNF-α and IL-1β in the rHuEPO group were lower than those in control group (P<0.05). Compared with the control group, patients receiving rHuEPO treatment underwent an apparent reduction in both mechanical ventilation and staying time in ICU (P<0.05). Conclusions Preoperative rHuEPO administration can attenuate the inflammatory lung injury after CPB via the anti-inflammation pathway and ameliorate pulmonary functions. Key words: Recombinant human erythropoietin; Cardiopulmonary bypass; Lung injury; Inflammatory response

Effects of EPO on Blood Parameters and Running Performance in Kenyan Athletes

Recombinant human erythropoietin (rHuEpo) administration enhances oxygen carrying capacity and performance at sea level. It remains unknown whether similar effects would be observed in chronic altitude-adapted endurance runners. The aim of this study was to assess the effects of rHuEpo on hematological and performance parameters in chronic altitude-adapted endurance runners as compared to sea level athletes. Twenty well-trained Kenyan endurance runners (KEN) living and training at approximately 2150 m received rHuEpo injections of 50 IU·kg body mass every 2 d for 4 wk and responses compared with another cohort (SCO) that underwent an identical protocol at sea level. Blood samples were obtained at baseline, during rHuEpo administration and 4 wk after the final injection. A maximal oxygen uptake (V˙O2max) test and 3000-m time trial was performed before, immediately after and 4 wk after the final rHuEpo injection. Hematocrit (HCT) and hemoglobin concentration (HGB) were higher in KEN compared to SCO before rHuEpo but similar at the end of administration. Before rHuEpo administration, KEN had higher V˙O2max and faster time trial performance compared to SCO. After rHuEpo administration, there was a similar increase in V˙O2max and time trial performance in both cohorts; most effects of rHuEpo were maintained 4 wk after the final rHuEpo injection in both cohorts. Four weeks of rHuEpo increased the HGB and HCT of Kenyan endurance runners to a lesser extent than in SCO (~17% vs ~10%, respectively) and these alterations were associated with similar improvements in running performance immediately after the rHuEpo administration (~5%) and 4 wk after rHuEpo (~3%).

Role of Human Recombinant Erythropoietin (rHuEPO) in Perinatal Asphyxia-a randomized controlled trial

Background : Perinatal asphyxia is an insult to the fetus or newborn infant due to lack of oxygen (hypoxia) and/or a lack of perfusion (ischemia) to various organs, which will manifest as difficulty in establishing spontaneous respiration evident by delayed cry after birth, at least after one minute. World-wide, perinatal asphyxia accounts for about 900,000 deaths each year. In Bangladesh it is a major cause of neonatal death. A substantial proportion of the children that survive suffer late effects such as cerebral palsy and epilepsy.&#x0D; Objective : To determine the efficacy of erythropoietin in improving the neurological outcome of term neonates with perinatal asphyxia (HIE stage II and III).&#x0D; Materials and methods : A Randomized Controll Trial was carried out in the Neonatal ward and NICU of Dhaka Shishu Hospital from 1st April 2014 to 30th Sep 2015. A total 68 neonates with perinatal asphyxia (both HIE stage II and III) who fulfill the inclusion criteria were enrolled and randomly assigned to intervention group (n=35) and control group (n=33). Intervention group received rHuEPO 300- 500 U/kg/dose daily subcutaneously for 5 days within first 48 hours of birth along with the standard treatment protocol and control group received standard treatment protocol only.&#x0D; Results : Baseline clinical characteristics, USG of brain during hospital stay were almost similar in both groups. Statistically significant effect was noted in seizure control, tolerance of oral feeding, hospital stay and neurological outcome at 3 months of age (p=008). USG of brain at 3 months of age also improved significantly (p=0.027).&#x0D; Conclusion : This study demonstrates the effectiveness of early administration of rHuEPO to term neonates with moderate to severe asphyxia, beneficial effect on short term outcomes like seizure control, tolerance of oral feeding and neurological outcome at 3 months of age. A large multicenter study would be done for further evaluation of these findings.&#x0D; Northern International Medical College Journal Vol.10(1) Jul 2018: 330-334

The Performance Effects of Microdose Recombinant Human Erythropoietin Administration and Carbon Monoxide Rebreathing.

Frequent, low doses of recombinant human erythropoietin (rHuEpo) have been shown to increase the oxygen carrying capacity of an athlete and enhance endurance performance, although its effect on repeated sprint ability (RSA) remains unknown. If the mechanisms behind improved RSA performance reside within the augmented O2 carrying capacity, then carbon monoxide (CO) inhalation should inhibit RSA. Purpose: The aim of this study was to assess the effects on maximal oxygen uptake (V˙O2max) and RSA of two interventions known to differentially influence blood oxygen carrying capacity. Methods: Fourteen endurance-trained individuals were administered microdoses of rHuEpo (20-40 IUkg) or placebo twice per week for 7 wk using a randomized, crossover design. V˙O2max and RSA were measured at baseline and after rHuEpo administration. Total hemoglobin mass (tHb-mass) was measured twice at baseline (14 and 7 d before the first injection), three times during rHuEpo administration (10, 24, and 38 d after the first rHuEpo injection) and twice after the cessation of rHuEpo administration (7 and 21 d after the final injection) using the optimized CO rebreathing method. V˙O2max and RSA also were assessed in a separate cohort of 11.

Biomolecular changes on mice caput femur after human recombinant erythropoietin administration

Introduction: Osteonecrosis is a process of death of bone tissue which also causes disturbance in the bone’s blood vessel supply. Administration of recombinant human erythropoietin (rHuEPO) may affect cell proliferation by stimulating angiogenesis and it has been proven that cell proliferation during osteogenesis depends heavily on the formation of new blood vessels. Therefore, EPO has a role in bone regeneration. Methods: This research is an experimental study using Randomized post-test control only group design. Rats were divided into 2 groups, P0 which received dexamethasone injection for 5 weeks and group P1 which received dexamethasone injection combined with rHuEpo for 5 weeks. On the last day of the fifth week, the femoral head of the rats were taken for assessment of VEGF (Vascular Endothelial Growth Factor), BMP-2 (Bone Morphogenic Protein-2), osteocytes, osteoblasts and adipocytes levels. The examination results were then analyzed by using SPSS. Results: From the immunohistochemical and histopathologic examination and further SPSS analysis, it is shown that the number of osteocyte in the femoral heads of rats injected with dexamethasone and rHuEpo were significantly higher than those injected with dexamethasone alone (p <0.05). This was also the case with the number of osteoblasts (p <0.05). The expression of BMP-2 and VEGF were also significantly higher in rats injected with dexamethasone and rHuEPO (p <0.05). However, the number of adipocytes in the femoral heads of the rats injected with dexamethasone and rHuEpo were significantly lower than those injected with dexamethasone alone (p <0.05). Conclusion: Administration of rHuEPO in rats induced with dexamethasone was shown to increase the number of osteoblast, osteocyte, BMP-2 expression, and VEGF, but the number of adipocyte was found to be lower than in rats injected with dexamethasone alone.

Short-term administration of recombinant human erythropoietin decreases B cell number in human peripheral blood

ObjectivesThere are conflicting results on the influence of recombinant human erythropoietin (rHuEPO) administration to lymphocytes, especially to B cells. MethodsWe analyzed peripheral white blood cell (WBC) subsets in patients who received one bolus administration of rHuEPO. 119 autologous blood donors were enrolled in this study. Fourty-nine out of them were treated with rHuEPO. Blood samples were obtained before the first phlebotomy and one week later before the second one. By flow cytometry, we measured the numbers of WBC, lymphocytes, dendritic cells, CD4+ T cells, CD8+ T cells, natural killer (NK) cells, B cells, monocytes, and neutrophils, further details of B cell subsets. ResultsIn the EPO-treatment group, absolute numbers of lymphocytes, especially CD8+ T cells, NK cells, and B cells, significantly decreased after rHuEPO administration. In B cell subsets, absolute numbers of naïve B cells and IgD−CD27− B cells significantly decreased. Other B cell subsets, such as transitional B cells, memory B cells, and marginal zone B cells, also showed a decreasing trend. ConclusionThese findings suggest that a single administration of rHuEPO can influence human immune system via reduction of B cell number in peripheral blood.