RESUMO
In the present study, a first validated and green spectrofluorimetric approach for its assessment and evaluation in different matrices was investigated. After using an excitation wavelength of 345 nm, Roxadustat (ROX) demonstrates a highly native fluorescence at an emission of 410 nm. The influences of experimental factors such as pH, diluting solvents, and different organized media were tested, and the most appropriate solvent choice was ethanol. It was confirmed that there was a linear relationship between the concentration of ROX and the relative fluorescence intensity in the range 60.0-1000.0 ng ml-1, with the limit of detection and limit of quantitation, respectively, being 17.0 and 53.0 ng ml-1. The mean recoveries % [±standard deviation (SD), n = 5] for pharmaceutical preparations were 100.11% ± 2.24%, whereas for plasma samples, they were 100.08 ± 1.08% (±SD, n = 5). The results obtained after the application of four greenness criteria, Analytical Eco-Scale metric, NEMI, GAPI, and AGREE metric, confirmed its eco-friendliness. In addition, the whiteness meter (RGB12) confirmed its level of sustainability. The International Council for Harmonisation (ICH) criteria were used to verify the developed method through the study in both spiked plasma samples and content uniformity evaluation. An appropriate standard for various applications in industry and quality control laboratories was developed.
Assuntos
Hematínicos , Humanos , Limite de Detecção , Espectrometria de Fluorescência/métodos , Eritropoese , Concentração de Íons de Hidrogênio , Solventes/química , Comprimidos/química , IsoquinolinasRESUMO
OBJECTIVE: We previously reported improved neurodevelopment at 2 and 4 years among preterm infants treated with erythropoietin or darbepoetin, known as erythropoiesis-stimulating agents (ESAs). We now characterize longitudinal outcomes through 6 years. METHODS: Children randomized to ESAs or placebo were evaluated at 6 years. Healthy-term children served as controls. Tests of cognition and executive function (EF) were performed. RESULTS: Cognitive/EF scores remained similar between 4 and 6 years within each group (ESA: 43 children; placebo: 17 children; term: 21 children). ESA recipients scored higher than placebo on Full-Scale IQ (94.2 ± 18.6 vs. 81.6 ± 16.7, p = 0.022), and Performance IQ (97.3 ± 16.2 vs. 81.7 ± 15.2, = 0.005). Aggregate EF trended better for the ESA group. Term controls scored better than placebo on all measures. ESA and term controls scored similarly on cognitive and EF tests. CONCLUSION: ESA recipients had better outcomes than placebo recipients, and were similar to term children. ESAs may improve long-term cognition and executive function in preterm infants.
Assuntos
Hematínicos , Lactente , Criança , Recém-Nascido , Humanos , Hematínicos/uso terapêutico , Recém-Nascido Prematuro , Darbepoetina alfa/uso terapêutico , Cognição , EritropoeseRESUMO
In a standard situation, a quantitative systems pharmacology model describes a "reference patient," and the model parameters are fixed values allowing only the mean values to be described. However, the results of clinical trials include a description of variability in patients' responses to a drug, which is typically expressed in terms of conventional statistical parameters, such as standard deviations (SDs) from mean values. Therefore, in this study, we propose and compare four different approaches: (1) Monte Carlo Markov Chain (MCMC); (2) model fitting to Monte Carlo sample; (3) population of clones; (4) stochastically bounded selection to generate virtual patient populations based on experimentally measured mean data and SDs. We applied these approaches to generate virtual patient populations in the QSP model of erythropoiesis. According to the results of our research, stochastically bounded selection showed slightly better results than the other three methods as it allowed the description of any number of patients from clinical trials and could be applied in the case of complex models with a large number of variable parameters.
Assuntos
Eritropoese , Farmacologia em Rede , Humanos , Cadeias de Markov , Método de Monte CarloRESUMO
OBJECTIVES: This study aimed to aid decision makers by analyzing the impact of introducing biosimilar prescription targets on physician prescribing behavior in the prescription of biologic erythropoiesis-stimulating agents in Germany. METHODS: We combined secondary data of regional level biosimilar prescription targets and secondary data of routinely collected claims data of dispensed prescriptions by physicians operating within the statutory health insurance system in ambulatory care across 7 German regions from 2009 to 2015. Two-way fixed-effects regression analysis was used to identify the average treatment effect of introducing biosimilar prescription targets at the physician level. The main outcome of interest was the share of biosimilar prescriptions on all prescriptions within the substance group. We compared 6 regions that introduced biosimilar prescription targets with 1 region without any prescription target policy. RESULTS: Introducing biosimilar prescription targets increased the average share of biosimilars between 6 percentage points (P < .05) in Hamburg and up to 20 percentage points (P < .001) in Saxony-Anhalt. Stratification of specialists by prescription volume and adoption status indicated heterogeneous effects. We identified similar but higher effects for high-volume prescribers. Disentangling of effects with regard to the composition of biosimilar share suggested that the increase in biosimilar share was driven by increased biosimilar use accompanied by a nonsignificant decrease in original biologics prescriptions. CONCLUSIONS: Prescription targets to alter physician prescribing behavior meet their intended goals by increasing biosimilar share. Physicians partly responded to the policy by decreasing overall prescriptions of the target substance. Prescription targets might be a useful tool, but decision makers need to consider all aspects of potential responses.
Assuntos
Medicamentos Biossimilares , Hematínicos , Médicos , Medicamentos Biossimilares/uso terapêutico , Prescrições de Medicamentos , Eritropoese , Alemanha , Hematínicos/uso terapêutico , Humanos , Padrões de Prática MédicaRESUMO
OBJECTIVES: For anaemic elective surgery patients, current clinical practice guidelines weakly recommend the routine use of iron, but not erythrocyte-stimulating agents (ESAs), except for short-acting ESAs in major orthopaedic surgery. This recommendation is, however, not based on any cost-effectiveness studies. The aim of this research was to (1) systematically review the literature regarding cost effectiveness of preoperative iron and/or ESAs in anaemic, elective surgery patients and (2) update existing economic evaluations (EEs) with recent data. METHODS: Eight databases and registries were searched for EEs and randomized controlled trials (RCTs) reporting cost-effectiveness data on November 11, 2020. Data were extracted, narratively synthesized and critically appraised using the Philips reporting checklist. Pre-existing full EEs were updated with effectiveness data from a recent systematic review and current cost data. Incremental cost-effectiveness ratios were expressed as cost per (quality-adjusted) life-year [(QA)LY] gained. RESULTS: Only five studies (4 EEs and 1 RCT) were included, one on intravenous iron and four on ESAs + oral iron. The EE on intravenous iron only had an in-hospital time horizon. Therefore, cost effectiveness of preoperative iron remains uncertain. The three EEs on ESAs had a lifetime time horizon, but reported cost per (QA)LY gained of 20-65 million (GBP or CAD). Updating these analyses with current data confirmed ESAs to have a cost per (QA)LY gained of 3.5-120 million (GBP or CAD). CONCLUSIONS: Cost effectiveness of preoperative iron is unproven, whereas routine preoperative ESA therapy cannot be considered cost effective in elective surgery, based on the limited available data. Future guidelines should reflect these findings.
Assuntos
Hematínicos , Análise Custo-Benefício , Eritropoese , Hematínicos/uso terapêutico , Humanos , Ferro , Anos de Vida Ajustados por Qualidade de VidaRESUMO
PURPOSE: Erythropoiesis-stimulating agents (ESAs), indicated for treating some patients with chemotherapy-induced anemia (CIA), may increase the risk of tumor progression and mortality. FDA required a Risk Evaluation and Mitigation Strategy (REMS) to mitigate these risks. We assessed REMS impact on ESA administration and red blood cell (RBC) transfusion as surrogate metrics for REMS effectiveness. METHODS: Retrospective cohort study including data from January 1, 2006 to December 31, 2018 for beneficiaries ≥65 years enrolled in Centers for Medicare & Medicaid Services (CMS) Medicare Parts A/B with a cancer diagnosis; patients with other indications for ESA use were excluded. Study time was divided into five periods demarcated by issuance of CMS National Coverage Determination (NCD) (Pre-NCD, Pre-REMS) and REMS milestones (Grace Period, REMS, post-REMS). Study outcomes were monthly proportion of chemotherapy episodes (CTEs) with concomitant ESA administration, with post-CTE ESA administration, and with RBC transfusions. RESULTS: Of 1 778 855 beneficiaries treated with CT, 308742 received concomitant ESA for CIA. The proportion of CTEs with concomitant and post-CTE ESA administration decreased Pre-REMS (9.0 percentage points [pp] and 3.5 pp, respectively). There were no significant post-REMS changes in the proportion of CTEs with concomitant (0.0 pp) and post-CTE ESA administration (0.1 pp). Fluctuation in RBC transfusions was <4 pp throughout the study period. CONCLUSIONS: Medicare beneficiaries showed a substantive decrease in ESA administration after NCD, with minimal impact by the REMS and its removal. Small changes in RBC transfusion over the study period were likely due to a national secular trend.
Assuntos
Anemia , Antineoplásicos , Hematínicos , Idoso , Anemia/induzido quimicamente , Anemia/tratamento farmacológico , Anemia/epidemiologia , Antineoplásicos/efeitos adversos , Transfusão de Sangue , Eritropoese , Hematínicos/efeitos adversos , Humanos , Medicare , Estudos Retrospectivos , Avaliação de Risco e Mitigação , Estados Unidos/epidemiologiaRESUMO
One of the urgent problems of nursing premature babies is the timely prediction, diagnosis and treatment of anemia, the frequency of which reaches 90%. Of particular importance is the study of reticulocytic blood parameters in deep-premature newborns, since the correct assessment of hemograms is crucial in the management of this category of children. To determine the characteristics of red blood cells and reticulocyte parameters of venous blood hemogram in premature newborns 24-32 weeks of gestation in the dynamics of the neonatal period. 111 newborns were examined at 24-32 weeks of gestation in the early neonatal period (on day 3-7) and at the age of 1 month of life. Along with standard diagnostic procedures, in accordance with current clinical recommendations and standards, 28 parameters of erythrocyte and reticulocyte hemogram parameters were determined for children. Venous blood was examined using an automatic hematological analyzer ADVIA 2120i, Siemens, USA. In deep-premature newborns in the early neonatal period, there is a high activity of erythropoiesis, respectively, the severity of respiratory and metabolic disorders with rejuvenation of reticulocytic subpopulations. Negative values of Delta hemoglobin were found against the background of a decrease in the average amount of hemoglobin in reticulocytes in children 24-27 weeks of gestation, which characterizes the lowest values of iron availability for erythropoiesis in this category of newborns. It is shown that a decrease in reticulocyte counts and normochromia in all examined newborns by the age of one month are accompanied by high levels of immature reticulocyte fraction, while a third of children still have limited iron availability for erythropoiesis.
Assuntos
Eritropoese , Reticulócitos , Criança , Contagem de Eritrócitos , Hemoglobinas/análise , Humanos , Recém-Nascido , Contagem de Reticulócitos , Reticulócitos/químicaRESUMO
PURPOSE: Erythropoiesis-stimulating agents (ESAs), are used for treating chronic kidney disease (CKD)-related anemia, contributing to CKD costs. The study was aimed at investigating direct healthcare costs of CKD patients treated with ESAs and the potential savings achievable by increasing the use of biosimilars and preventing inappropriate ESA use. METHODS: A multi-center, cohort study was conducted using claims databases of five large Italian geographic areas. Yearly mean direct healthcare costs per patient were estimated, stratifying by CKD stage. The total yearly cost and potential savings related to ESA use were estimated: (a) considering 25/50/75% of originator ESA substitution with biosimilars; (b) eliminating inappropriate ESA dispensing. RESULTS: During the study period, the ESA-related yearly mean cost represented 17% of total yearly costs in stage I-III, decreasing to 13% in stage IV-V and 6% in dialysis. Among originator users, assuming a 25% of biosimilar uptake, the annual cost-savings of ESA treatment would represent 10.5% of total ESA costs in CKD stage I-V and 7.7% in dialysis. Among incident ESA users for which hemoglobin levels were available, 9% started inappropriately ESA treatment, increasing to 62.0% during the first year of maintenance therapy. Hypothesizing prevention of the first inappropriate ESA dispensing, the total yearly cost-savings would amount to 35 772, increasing to 167 641 eliminating the inappropriate dispensing during maintenance therapy. CONCLUSIONS: Higher use of lowest cost ESA, prevention of inappropriate ESA use as well as other strategies aimed at slowing down the progressive renal impairment are essential for minimizing clinical and economic burden of CKD.
Assuntos
Medicamentos Biossimilares , Hematínicos , Insuficiência Renal Crônica , Estudos de Coortes , Eritropoese , Custos de Cuidados de Saúde , Humanos , Itália , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/tratamento farmacológicoRESUMO
BACKGROUND: Erythroferrone (ERFE) is an erythroid hormone putatively involved in stress erythropoiesis. Its regional clearance and circulating form in humans, as well as levels in normal health and coronary disease remain unclear. METHODS: To establish a reference interval, ERFE was measured in 155 healthy volunteers using the Intrinsic LifeSciences ELISA. To identify trans-organ gradients in ERFE, regional blood sampling was undertaken in patients (n = 13) undergoing clinically indicated cardiac catheterisation. The Intrinsic ELISA was assessed for reproducibility, stability, linearity and possible cross-reactivity, interference and anticoagulant effects. Circulating forms of ERFE were evaluated by HPLC. RESULTS: In healthy individuals, the median concentration of ERFE was 0.51 ng/mL (IQR: 0.12-1.25), with men (n = 78) having higher levels than women (n = 77) (0.67 vs 0.32 ng/mL, p = 0.0001). ERFE concentrations in trans-organ sampling revealed no clear organ of clearance or production. Samples with high endogenous ERFE levels were suppressed by haemoglobin (≥2 g/L), bilirubin (≥200 µmol/L), lipaemia (>1 g/L), and freeze thawing (≥2 cycles), but this was not observed with low ERFE concentrations. Endogenous ERFE immunoreactivity was 46% higher in EDTA plasma compared with serum and lithium heparin plasma. On SE-HPLC, ERFE eluted as intact and cleaved forms. CONCLUSION: We provide a useful reference range for ERFE in EDTA plasma. We found no specific site of secretion or clearance. The Intrinsic ELISA performed adequately but is limited by interference and stability when endogenous levels are high. Circulating forms are multiple and complex.
Assuntos
Doença da Artéria Coronariana/sangue , Hormônios Peptídicos/análise , Hormônios Peptídicos/sangue , Adulto , Idoso , Biomarcadores/sangue , Cateterismo Cardíaco , Cromatografia Líquida de Alta Pressão , Ensaio de Imunoadsorção Enzimática , Eritropoese/fisiologia , Eritropoetina/sangue , Feminino , Ferritinas/sangue , Voluntários Saudáveis , Hepcidinas/sangue , Humanos , Ferro/sangue , Masculino , Pessoa de Meia-Idade , Valores de Referência , Adulto JovemRESUMO
The study was aimed at designing a health exercise program appealing to inactive young men, and then testing the men's metabolic responses to the program using common diagnostic markers of general health. Six men, aged 22-29 years, took a part in training program to increase their motor performance and improve general health conditions. Body composition parameters, clinical chemistry variables (metabolites, albumin, total protein, ferritin, C reactive protein, lipid profile, ions, and selected enzymes activities) and blood morphology parameters were determined. Motor performance measured before and after a 4-month-long macrocycle indicated an increase in endurance, pace, and agility of the participants. Significant differences were found in analyzed enzymes activities. There was a significant increase in C-reactive protein levels from pre- to post-training. Additionally, changes in hematological biomarkers were seen that suggest erythropoiesis might significantly increase, specifically during the last 2-month-long mesocycles. The proposed training program induced small improvements in endurance, pace, and agility. It was also confirmed that changes in aspartate (AST) and alanine (ALT) activities emerge before any increase in creatine kinase (CK) activity that is important in monitoring of the training loads. Observed changes in red blood cell-related parameters suggest increase in erythropoiesis in the second half of the training cycle.
Assuntos
Exercício Físico/fisiologia , Desempenho Psicomotor/fisiologia , Adulto , Biomarcadores/sangue , Composição Corporal/fisiologia , Eritropoese , Promoção da Saúde , Humanos , Masculino , Resistência Física/fisiologia , Adulto JovemRESUMO
In the clinical setting, reticulocytes are used as an index for the hematopoietic function of the bone marrow. Different maturation stages of reticulocytes are early markers for bone marrow hematopoietic stem cell transplantation and bone marrow regeneration after chemotherapy. Therefore, we aimed to establish a method for detecting the different reticulocyte maturation stages. Based on the decreases in mitochondrial membrane potential during reticulocyte maturation, we used MitoTracker Green (MTG)/tetramethylrhodamine, ethylester (TMRE) to identify the different reticulocyte maturation stages and used Hoechst33342 to exclude nucleated cells. The results show that this method was universal and could be applied to detect the proportions of reticulocytes in different samples. Their proportion in normal peripheral blood, a blood deficiency model, bone marrow, and spleen were (6 ± 2)%, (38 ± 4)%, (14 ± 4)%, and (3 ± 1)%, respectively. The results obtained using this method were similar to those obtained using the manual counting method (methylene blue); the correlation was good (R = 0.817; p < .01) and the coefficient of variation was lower for the method established. Moreover, reticulocytes in peripheral blood could be further divided into three distinct maturation stages: R1 (MTGneg/TMREhigh), R2 (MTGhigh/TMREhigh), and R3 (MTGhigh/TMREneg). Reticulocytes in the bone marrow and spleen could be further divided into four distinct maturation stages: R1 (MTGneg/TMREhigh), R2-1 (MTGhigh/TMREhigh/FSbig), R2-2 (MTGhigh/TMREhigh/FSsmall), and R3 (MTGhigh/TMREneg). Based on changes in mitochondrial membrane potential, MTG/TMRE/Hoechst33342 staining could be used to detect reticulocytes in different samples and at different maturation stages with low cost and high accuracy.
Assuntos
Contagem de Células/métodos , Citometria de Fluxo/métodos , Potencial da Membrana Mitocondrial/fisiologia , Reticulócitos/citologia , Reticulócitos/fisiologia , Animais , Células Sanguíneas/citologia , Células da Medula Óssea/citologia , Contagem de Eritrócitos/métodos , Eritropoese , Camundongos , Coloração e RotulagemRESUMO
Fibroblast growth factor 23 (FGF23) was initially characterized as an important regulator of phosphate and calcium homeostasis. New research advances demonstrate that FGF23 is also linked to iron economy, inflammation and erythropoiesis. These advances have been fuelled, in part, by the serendipitous development of two distinct FGF23 assays that can substitute for invasive bone biopsies to infer the activity of the three main steps of FGF23 regulation in bone: transcription, post-translational modification and peptide cleavage. This 'liquid bone biopsy for FGF23 dynamics' enables large-scale longitudinal studies of FGF23 regulation that would otherwise be impossible in humans. The balance between FGF23 production, post-translational modification and cleavage is maintained or perturbed in different hereditary monogenic conditions and in acquired conditions that mimic these genetic disorders, including iron deficiency, inflammation, treatment with ferric carboxymaltose and chronic kidney disease. Looking ahead, a deeper understanding of the relationships between FGF23 regulation, iron homeostasis and erythropoiesis can be leveraged to devise novel therapeutic targets for treatment of anaemia and states of FGF23 excess, including chronic kidney disease.
Assuntos
Anemia Ferropriva/genética , Eritropoese/genética , Raquitismo Hipofosfatêmico Familiar/genética , Fatores de Crescimento de Fibroblastos/genética , Inflamação/genética , Ferro/metabolismo , Fosfatos/metabolismo , Insuficiência Renal Crônica/genética , Anemia Ferropriva/tratamento farmacológico , Anemia Ferropriva/metabolismo , Osso e Ossos/metabolismo , Cálcio , Eritropoese/fisiologia , Raquitismo Hipofosfatêmico Familiar/metabolismo , Compostos Férricos/efeitos adversos , Fator de Crescimento de Fibroblastos 23 , Fatores de Crescimento de Fibroblastos/metabolismo , Fatores de Crescimento de Fibroblastos/fisiologia , Humanos , Hipofosfatemia/induzido quimicamente , Inflamação/metabolismo , Maltose/efeitos adversos , Maltose/análogos & derivados , Processamento de Proteína Pós-Traducional , RNA Mensageiro/metabolismo , Insuficiência Renal Crônica/metabolismo , Transcrição GênicaRESUMO
Living high-training low (LHTL) is performed by competitive athletes expecting to improve their performance in competitions at sea level. However, the beneficial effects of LHTL remain controversial. We sought to investigate whether 21 days of LHTL performed at a 3,000 m simulated altitude (fraction of inspired oxygen [FIO2]=14.5%) and at sea level can improve hematological parameters, exercise economy and metabolism, hemodynamic function, and exercise performance compared with living low-training low (LLTL) among competitive athletes. All participants (age = 23.5 ± 2.1 years, maximal oxygen consumption [VO2max] = 55.6 ± 2.5 mL·kg-1·min-1, 3,000 m time trial performance=583.7 ± 22.9 seconds) were randomly assigned to undergo LHTL (n = 12) or LLTL (n = 12) and evaluated before and after the 21 days of intervention. During the 21-day intervention period, the weekly routine for all athletes included 6-day training and 1-day rest. The daily training programs consisted of >4 hours of various exercise programs (i.e., jogging, high-speed running, interval running, and 3,000 m or 5,000-m time trial). The LHTL group resided in a simulated environmental chamber (FIO2 = 14.5%) for >12 hours per day and the LLTL group at sea level under comfortable conditions. The hematological parameters showed no significant interaction. However, LHTL yielded more improved exercise economy, metabolic parameters (oxygen consumption=-152.7 vs 32.4 mL·kg-1·30min-1, η2 = 0.457, p = 0.000; tissue oxygenation index=6.18 vs .66%, η2 = 0.250, p = 0.013), and hemodynamic function (heart rate = -234.5 vs -49.7 beats·30min-1, η2 = 0.172, p = 0.044; stroke volume = 136.4 vs -120.5 mL/30 min, η2 = 0.191, p = 0.033) during 30 minutes of submaximal cycle ergometer exercise corresponding to 80% maximal heart rate before training than did LLTL. Regarding exercise performance, LHTL also yielded more improved VO2max (5.40 vs 2.35 mL·kg-1·min-1, η2 = 0.527, p = 0.000) and 3,000 m time trial performance (-34.0 vs -19.5 seconds, η2 = 0.527, p = 0.000) than did LLTL. These results indicate that compared with LLTL, LHTL can have favorable effects on exercise performance by improving exercise economy and hemodynamic function in competitive runners.
Assuntos
Altitude , Desempenho Atlético/fisiologia , Comportamento Competitivo/fisiologia , Hemodinâmica/fisiologia , Condicionamento Físico Humano/métodos , Condicionamento Físico Humano/fisiologia , Corrida/fisiologia , Metabolismo Energético/fisiologia , Eritropoese/fisiologia , Frequência Cardíaca/fisiologia , Humanos , Masculino , Consumo de Oxigênio/fisiologia , Fatores de Tempo , Adulto JovemRESUMO
We studied the performance of mean reticulocyte hemoglobin content (MCHr), measured on Abbott CELL-DYN Sapphire analyzer for the detection of functional iron deficiency. Patients with anemia and with renal disease were prospectively selected from the outpatient population of our hospital, 512 subjects were studied. Diagnoses and other medical data were retrieved from the hospital information system. MCHr was measured using a CELL-DYN Sapphire hematology analyzer (Abbott Diagnostics). Standard laboratory and statistical tests were used. Receiver operating characteristic (ROC) analysis was used to establish the diagnostic performance of MCHr for detecting iron-restricted erythropoiesis, with transferrin saturation <20% as the gold standard. Patients in the iron deficiency anemia group all had a state of iron depletion and iron-restricted erythropoiesis (median MCHr 25.4 pg). Patients with anemia of chronic disease showed also low MCHr, median 27.6 pg, but not statistically different from the iron deficient group (p = .0585). Renal patients with iron restriction (n = 66) had significantly lower MCHr (p < .0001) than those receiving adequate iron supply (n = 100): median MCHr were 27.9 and 32.5 pg, respectively. ROC analysis gave sensitivity 84.4% and specificity 80.1% with area under curve 0.863 (95% CI 0.823-0.902) at an MCHr cut-off 30.0 pg. MCHr on CELL-DYN Sapphire has equivalent clinical performance for detecting absolute or functional iron deficiency in patients with chronic kidney disease as previously published for another type of analyzer.
Assuntos
Anemia Ferropriva/fisiopatologia , Anemia/fisiopatologia , Eritropoese/fisiologia , Insuficiência Renal Crônica/fisiopatologia , Anemia/complicações , Anemia Ferropriva/complicações , Feminino , Ferritinas/sangue , Hemoglobinas/metabolismo , Humanos , Ferro/metabolismo , Masculino , Curva ROC , Insuficiência Renal Crônica/complicaçõesRESUMO
Single-cell RNA sequencing studies of differentiating systems have raised fundamental questions regarding the discrete versus continuous nature of both differentiation and cell fate. Here we present Palantir, an algorithm that models trajectories of differentiating cells by treating cell fate as a probabilistic process and leverages entropy to measure cell plasticity along the trajectory. Palantir generates a high-resolution pseudo-time ordering of cells and, for each cell state, assigns a probability of differentiating into each terminal state. We apply our algorithm to human bone marrow single-cell RNA sequencing data and detect important landmarks of hematopoietic differentiation. Palantir's resolution enables the identification of key transcription factors that drive lineage fate choice and closely track when cells lose plasticity. We show that Palantir outperforms existing algorithms in identifying cell lineages and recapitulating gene expression trends during differentiation, is generalizable to diverse tissue types, and is well-suited to resolving less-studied differentiating systems.
Assuntos
Algoritmos , Diferenciação Celular/genética , Linhagem da Célula/genética , Análise de Sequência de RNA/estatística & dados numéricos , Análise de Célula Única/estatística & dados numéricos , Animais , Biotecnologia , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Eritropoese/genética , Regulação da Expressão Gênica no Desenvolvimento , Hematopoese/genética , Humanos , Cadeias de Markov , Camundongos , Modelos Biológicos , Modelos EstatísticosRESUMO
Hematopoiesis is arguably one of the best understood stem cell systems; however, significant challenges remain to reach a consensus understanding of the lineage potential, heterogeneity, and relationships of hematopoietic stem and progenitor cell populations. To gain new insights, we performed quantitative analyses of mature cell production from hematopoietic stem cells (HSCs) and multiple hematopoietic progenitor populations. Assessment of the absolute numbers of mature cell types produced by each progenitor cell revealed a striking erythroid dominance of all myeloid-competent progenitors assessed, accompanied by strong platelet reconstitution. All populations with myeloid potential also produced robust numbers of red blood cells and platelets in vivo. Clonal analysis by single-cell transplantation and by spleen colony assays revealed that a significant fraction of HSCs and multipotent progenitors have multilineage potential at the single-cell level. These new insights prompt an erythroid-focused model of hematopoietic differentiation.
Assuntos
Diferenciação Celular , Eritropoese , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/metabolismo , Células-Tronco Multipotentes/citologia , Células-Tronco Multipotentes/metabolismo , Animais , Biomarcadores , Linhagem da Célula , Ensaio de Unidades Formadoras de Colônias , Hematopoese , Transplante de Células-Tronco Hematopoéticas , Imunofenotipagem , Camundongos , Modelos BiológicosRESUMO
BACKGROUND: In conditions associated with inflammation, biochemical parameters alone could be inadequate for assessing iron status. We investigated the potential utility of mean reticulocyte hemoglobin content (MCHr) in the assessment of the erythropoiesis status in inflammatory bowel disease (IBD). METHODS: We recruited 124 anemic outpatients with IBD. Serum iron, transferrin and ferritin were tested. Complete blood counts were performed on a CELL-DYN Sapphire analyzer (Abbott Diagnostics). Differences among groups were assessed using analysis of variance, considering Pâ¯<â¯0.05 to be significant. Receiver operating characteristic analysis was used to assess the diagnostic performance of MCHr for detecting iron deficient erythropoiesis. The reference used as an indicator of insufficient iron availability was transferrin saturation <20%. RESULTS: Overall, 47.6% of the patients had iron deficiency anemia (IDA) and 31.5% anemia of chronic disease (ACD), while the others (20.9%) had mixed anemia. Patients with ACD or mixed anemia showed functional iron deficiency: normal or high ferritin and low MCHr. The area under curve was 0.858 (95% CI 0.742-0.942), considering a cut off 30.3â¯pg, the sensitivity was 82.2%, specificity 83.3%. CONCLUSIONS: MCHr provides information on iron availability in IBD patients. It is a reliable test to assess iron supply for erythropoiesis.
Assuntos
Anemia Ferropriva/diagnóstico , Eritropoese , Hemoglobinas/análise , Doenças Inflamatórias Intestinais/complicações , Reticulócitos/química , Adulto , Idoso , Anemia Ferropriva/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROCRESUMO
AIM: Erythropoiesis-stimulating agents (ESAs) are particularly used to treat dialysis patients suffering from anemia due to renal failure. Since March 2014, ESAs have no longer been funded on top-of-diagnosis-related groups (DRGs) in French hospitals and are funded via DRGs. There are two ways to fund dialysis in French hospitals: the 'DRG for dialysis in session' and the 'off-dialysis DRG', which is not a DRG and consists in a supplement tariff specific to dialysis for patients hospitalized for another main reason than dialysis. The aim of this study is to assess the impact of this funding change on the dialysis activity and on the budget of the 37 University Public University Hospitals in Paris (10% of the hospitalizations in France). MATERIALS & METHODS: A before-after study (March-September 2013 vs March-September 2014) was conducted. Medical activity data (and ESAs consumption data) were used to assess the number and costs of DRGs associated with ESAs use. As we do not have access to the whole dialysis activity over the period studied, two hypotheses were considered: the proportion of the dialysis activity was constant between the two periods (Hypothesis A); the dialysis activity was correlated to ESA consumptions delivered by hospital pharmacies to healthcare units (Hypothesis B). A budget impact analysis was also conducted taking into account the evolution of DRG costs and ESA prices. RESULTS: The number of dialysis 'DRG for dialysis in session' with ESA consumption have increased by 5% (Hypothesis A) and decreased by 9% (Hypothesis B) between the two periods while the volume of 'off-dialysis DRG' with ESA consumption increased by 2% (Hypothesis A) and by 9% (Hypothesis B). The budget impact was -1.02 million (Hypothesis A) and -0.7 million (Hypothesis B) leading to a loss for our hospitals. CONCLUSION: There is no significant impact of the change of funding of ESAs on the hospital activity. The DRG-based payment is negative for the budget of our hospitals but is positive for the French National Health Insurance. Indeed, with the price decrease, the ESAs are more costly for hospitals (not funded on top-of-DRG), but less costly for the society.
Assuntos
Hematínicos/economia , Diálise Renal/economia , Idoso , Anemia/tratamento farmacológico , Anemia/economia , Custos e Análise de Custo , Economia Hospitalar , Eritropoese/efeitos dos fármacos , Feminino , França , Financiamento da Assistência à Saúde , Hospitalização/economia , Hospitalização/estatística & dados numéricos , Humanos , Falência Renal Crônica/complicações , Falência Renal Crônica/economia , Masculino , Pessoa de Meia-IdadeRESUMO
The toxic effects of combined cisplatin/docetaxel therapy cycles on erythroid and granulocytic hematopoietic lineages as well as their intercycle recovery were examined in patients with stage III-IV non-small-cell lung carcinoma. Responsiveness of the blood system to this therapy remained at a high level. Combined therapy pronouncedly activated the key elements of the erythroid and granulocytic hematopoietic lineages leading to accumulation of immature and mature myelokaryocytes in the bone marrow, enlargement of the medullary pool of mature neutrophils, and increase in the count of medullary erythroid and granulocytic precursor cells under conditions of their accelerated maturation.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/metabolismo , Granulócitos/metabolismo , Neoplasias Pulmonares/metabolismo , Antraciclinas/farmacologia , Antineoplásicos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Cisplatino/farmacologia , Dissacarídeos/farmacologia , Docetaxel , Doxorrubicina/farmacologia , Eritropoese/efeitos dos fármacos , Granulócitos/citologia , Granulócitos/efeitos dos fármacos , Células-Tronco Hematopoéticas/efeitos dos fármacos , Células-Tronco Hematopoéticas/metabolismo , Humanos , Macrolídeos/farmacologia , Compostos de Nitrosoureia/farmacologia , Compostos Organoplatínicos/farmacologia , Taxoides/farmacologiaRESUMO
PURPOSE: Evaluate changes in use of erythropoiesis-stimulating agents (ESAs) and red blood cell transfusion in cancer patients receiving myelosuppressive chemotherapy following regulatory and reimbursement actions. METHODS: Calendar year patient cohorts (2005-2013) with breast, colorectal, lung, multiple myeloma, non-Hodgkin lymphoma, ovarian, or prostate cancer and receiving myelosuppressive chemotherapy were identified within the Marketscan database. Incidence of ESA treatment and transfusion were estimated in each year, as was median number of ESA administrations. Clinical characteristics associated with ESA administration and transfusions were evaluated by using multivariable logistic regression. Additionally, annual new ESA user cohorts within the Oncology Services Comprehensive Electronic Records database (2011-2014) were examined to assess hemoglobin levels at ESA initiation. RESULTS: Across all tumor types, ESA use decreased substantially (breast cancer: 53.7 to 3.2%; lung cancer: 66.0 to 13.3%, non-Hodgkin lymphoma: 39.8 to 3.8%), transfusion use increased (2 to 5.5%, 5.5 to 18.2%, and 4.5 to 9.1%, respectively), and median number of ESA administrations declined. Across all tumor types, proportion of patients initiating an ESA with hemoglobin >10 g/dL was <10% from 2011 onward. In recent years, cancer patients who are older, female, and have chronic kidney disease or moderate or severe liver disease were most likely to receive ESAs. CONCLUSION: Subsequent to important regulatory and reimbursement ESA-related actions, total ESA exposure among cancer patients receiving myelosuppressive chemotherapy declined substantially. Today, fewer patients receive ESA therapy, and among those treated, more are initiated at hemoglobin levels <10 g/dL and are exposed for a shorter duration, consistent with current product labeling.