bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antithymocyte Globulin (ATG) Plus Cyclosporine (CSA) for Aplastic Anemia (AA) and Low-Risk Myelodysplastic Syndrome (MDS).
Authors: Lee Je-Hwan , Choi Seong-Jun , Lee Jung-Hee , Seol Miee , Kim Keun Hee , Park Jae-Hoo , Kim Hawk , Joo Young-Don , Lee Won-Sik , Zang Dae Young , Kim Hyo Jung , Lee Kyoo Hyung 
Published: October 2019

Abstract
We investigated the effectiveness of immunosuppressive therapy using ATG plus CSA in adult patients with AA and low-risk MDS. Between APR 2000 and JAN 2005, 93 patients, 82 AA and 11 MDS, from 4 institutes in Korea were included in this prospective study. Patients received one of 3 types of ATG according to the availability in Korea: horse ATG (Atgam®, Upjohn, Kalamazoo, MI; 40 mg/kg/d × 4 days), rabbit ATG (Thymoglobuline®, IMTIX-SANGSTAT, Lyon, France; 2.5 mg/kg/d × 5 days), or horse ALG (Lymphoglobuline®, IMTIX-SANGSTAT; 10 mg/kg/d × 5 days). Methylprednisolone 2 mg/kg/d was given before each dose of ATG. Oral CSA 3 mg/kg twice a day was administered for 3–6 months. In 38 patients, the HLA phenotyping results were available. The median disease duration before ATG/CSA therapy was 31 days (range, 3–3126). Eight patients were not evaluable for response because of early deaths in 4, early follow-up loss in 3, and clonal evolution to AML in 1 within 60 days after treatment. Of 85 patients who were evaluable for response, 46 (54.1%) showed response to ATG/CSA therapy: partial response in 23 and complete response in 23. Responses were observed in 44 (56.7%) of 75 AA patients and in 3 (30.0%) of 10 low-risk MDS patients (P=0.103). Patients with HLA B40 phenotype showed higher response (66.7% vs. 32.1%, P=0.043), but other HLA phenotypes did not show any significant influence on response. Cumulative incidence of response was 55.5% at 2-year. Multivariate analysis indicated that disease duration before ATG/CSA therapy (≤ 90 days vs. > 90 days; OR 0.154; P=0.001) and type of ATG (horse ATG vs. rabbit ATG; OR 0.235; P=0.006) were independent predictive factors for response. Nine patients, 7 AA and 2 MDS, relapsed and cumulative incidence of relapse was 25.9% at 5-year. Clonal or disease evolution occurred in 3 patients with AA (PNH, MDS, and AML in 1 of each) and in 3 patients with MDS (RAEB-1 in 2 and RAEB-2 in 1). Eighteen patients died and actuarial survival was 69.8% at 5-year: 76.9% in AA and 36.0% in MDS. No patient who attained at least PR died. Our results demonstrated the effectiveness of ATG/CSA therapy in AA and low-risk MDS. The effects of rabbit ATG was significantly inferior in our study probably due to dose problem rather than ATG type itself. The influence of HLA phenotype on response to ATG/CSA should be further investigated.
Endothelial-Dependent Vasodilation Is Impaired in Children with Sickle Cell Disease (SCD).
Authors: De Montalembert Mariane , Aggoun Yacine , Niakate Assa , Szezepanski Isabelle , Bonnet Damien 
Published: October 2019

Abstract
Impairment of endothelium-dependent vasodilation has been reported in steady-state SCD adult patients, and could contribute to the occurrence of vaso-occlusive events. In order to study whether vascular tone abnormalities could be also observed in SCD children, we enrolled 21 SCD steady-state children (18 homozygous SS, 3 S-b0 thalassemics); 12 males, 9 females, mean aged 10.4 ± 3.3 yrs, mean Hb level: 7.6 ± 1.0 g/dL. None had had a transfusion in the 3 previous months, nor were treated with hydroxyurea. These SCD children were compared to a control group of 23 Afro-Caribbean AA and AS controls matched for age and gender. Blood pressure and vascular function parameters were measured after the child had been recumbent for at least 10 minutes. Flow-mediated (FMD) and nitroglycerine-mediated (GTNMD) dilation of the brachial artery were examined in cases and controls using echotracking techniques. Additionally, intima-media thickness (IMT) and mechanical properties of the common carotid artery were measured using previously described methods.
Blood pressures in SCD and control children were comparable. IMT was not different in SCD children (0.42 ± 0.06 mm) vs controls (0.42 ± 0.03 mm). Systolic and diastolic diameters of the common carotid artery were significantly higher in SCD children than in controls (respectively, 6.8 ± 0.8 vs 5.7 ± 0.4, p<0.001, and 5.7 ±0.7 vs 5.0 ± 0.4 mm, p< 0.005) but the stiffness of the common carotid artery was not different in SCD children and in controls. Finally, FMD was significantly decreased in SCD children vs controls (5.6 ± 0.2 vs 8.0 ± 0.2 %, p: 0.008), while GTNMD was comparable in SCD patients and controls. We hypothesize that the increase of the carotid artery diameters, without modification of the distensibility, is related to the higher cardiac output secondary to anemia. Interestingly, we observed a significant decrease of the endothelial-dependent flow-mediated vasodilation in SCD children, which had never been reported before. This could result of impaired NO bioavailability, correlated with enhanced arginase activity and hemolysis rate. Oral arginine supplementation in SCD children could be an appropriate treatment.
Evaluation of the CapillaryS 2 CE System for Hemoglobinopathy Investigations.
Authors: McFarlane Andrew G. , Halchuk Linda M. , Eng Barry , Waye John , Crowther Mark A. 
Published: October 2019

Abstract
Traditional diagnosis of hemoglobinopathies rely on separation and accurate quantification of hemoglobin (Hb) fractions using alkaline and acid electrophoresis, isoelectric focusing (IEF) and/or High Performance Liquid Chromatography (HPLC). Recent reports have suggested capillary electrophoresis (CE) as an alternate method for screening. We evaluated a new CE system the Sebia CapillaryS 2 (Evry Cedex, France) and compared it to IEF and HPLC. For this evaluation samples referred to our laboratory for routine hemoglobinopathy screening or as cord bloods sent for confirmation of a variant detected as a result of newborn screening were analyzed using all three techniques. IEF was performed with the Resolve IEF kit (Perkin Elmer,Wallac Oy, Finland). HPLC was done on the BioRad Variant II (Munich, Germany) using the Hb A2/Hb A1c Dual program. The CE was performed using the “Capillarys Hemoglobin(E) kit”. Suspected rare variants were further investigated with DNA investigations, including PCR and direct nucleotide sequencing. Variant hemoglobins were detected in 156 of 764 samples (Table 1). The correlation between IEF, HPLC and CE was excellent. One variant (Hb Toulon) was not detected by the CE but ran with Hb F. The CE system did not report the Hb F value on samples with very low (< 1.0%) Hb F by HPLC. Two cases of Hb Constant Spring were identified by CE but not by either IEF or HPLC. The correlation of both Hb F and Hb A2 reported between the CE and HPLC systems were excellent (R > 0.99 and 0.98, Figure 1 and Figure 2 respectively). The positive bias of Hb A2 seen in HPLC when Hb S is present was lower in the CE. However, the CE system demonstrated a negative bias for Hb A2 if Hb C was present. CE was the only system capable of separating Hb A2 from Hb E. We conclude that CE is comparable to both IEF and HPLC for separation and quantification of hemoglobin fractions; this system has the added advantage of reporting Hb A2 in the presence of Hb E and consistently identifies Hb Constant Spring, which is not easily detected on IEF or HPLC. Further this CE system is user friendly and holds promise as a tool for newborn screening and routine laboratory hemoglobinopathy investigations.
Hemoglobinopathy Detected Hemoglobinopathy Number β Thalassemia 99 Hb S trait 55 Cord with Hb Barts 28 Hb C trait 13 Hb E trait 12 Hb D trait 8 α variant (not yet identified) 5 Hb Barts + Hb S trait 4 Hb Barts + Hb D trait 3 Hb S disease 3 Hb Barts + Hb E trait 2 Hb E +β Thalassemia 2 Hb H disease 2 Hb Constant Spring 2 Hb J Baltimore 2 Hb Toulon 2 δ variant (not identified) 2 Hb S + Hb C 1 Hb Barts + Hb C trait 1 Hb Q Thailand + --SEA/αα 1 Homozygous Hb E + Hb Constant Spring 1 Hb S + Hb Kenya 1 Hb J Roviga 1 Hb Beograd 1 Hb G Coushatta 1 Hb Sirian 1 Hb Titusville 1 B variant (not identified yet) 1 Total Abnormal 255 No hemoglobinopathy detected 509 Total 764 Hb F: HPLC vs CE Hb F: HPLC vs CE Hb A2: HPLC vs CE Hb A2: HPLC vs CE
High Calcium Requirement for Phosphatidylserine Exposure in Sickle Cell Disease.
Authors: DeJong Kitty , Miller Annabeth M. , Emerson Renee K. , Kuypers Frans A. 
Published: October 2019

Abstract
While the adverse effects of red blood cells (RBC) that expose phosphatidylserine (PS) in the sickle cell circulation are well recognized, the mechanism that underlies the formation of these cells is not well understood. Diminished flippase activity is found in a subset of sickle cells, and while this explains how PS persists in the outer RBC monolayer, it does not explain how the phospholipids are randomized across the membrane bilayer. Rapid scrambling of phospholipids, resulting in PS exposure, requires elevation of intracellular free Ca++ levels by Ca++ influx into the RBC. In this study we assessed how much Ca++ is required for PS exposure, and how this relates to previous findings of elevated Ca++ influx during deoxygenation of sickle cells. We loaded RBC for 30 minutes at 37°C with accurately established levels of intracellular Ca++ by balancing CaCl2 solutions with the chelator BAPTA, followed by Annexin V labeling under conditions that scrambling did not continue. In both normal and sickle RBC from either human or mouse, PS exposure was only found at or above 10 microM Ca++. Flippase inhibition with vanadate did not change this, indicating that phospholipid scrambling had not been initiated under these conditions. Scramblase sulfhydryl modification with NEM reduced the Ca++ requirement to lower than 1 microM, as did treatment with the scramblase stimulator oligomycin. Further experiments showed that Ca++ needs to be present during the entire duration of the scrambling process, and removal of Ca++ arrests the scrambling process. With 1 microM Ca++ in oligomycin-treated (scramblase stimulated) RBC and in absence of flippase activity (using vanadate), it took 5 minutes at 37°C to obtain a significant (>2%) population of PS-exposing cells. These conditions are unlikely met in all sickle cells. In addition, these experiments showed that the flippase is not inhibited by 1 or 100 microM Ca++, despite previous reports suggesting that Ca++ loading inhibits the flippase. Taken together, our data indicate that PS scrambling requires more Ca++ influx than has been reported for deoxygenated sickle cells (50–100 nM). We suggest that a small percentage of individual sickle cells contain Ca++ levels of 1 microM or above, which would lead to PS exposure in those RBC that have sustained flippase inhibition and scramblase modifications similar to those achieved with NEM or oligomycin. We have previously reported evidence for such scramblase modifications, as well as reduced flippase activity in a subset of sickle cells. Our data indicate that the degree of Ca++ influx may be the determining parameter for the extent of PS exposure in sickle cells.
Plasma Concentrations of Asymmetric Dimethylarginine, an Endogenous Nitric Oxide Synthetase Inhibitor, Are Elevated in Sickle Cell Patients but Do Not Increase during Painful Crises.
Authors: Landburg Pearl , Teerlink Tom , de Jong Sigrid , Muskiet Frits A. , Duits Ashley J. , Schnog John-John B. 
Published: October 2019

Abstract
Plasma concentrations of asymmetric dimethylarginine (ADMA), an endogenously produced inhibitor of nitric oxide (NO) synthase, are elevated and related to endothelial activation in clinically asymptomatic patients with sickle cell disease (SCD). However, ADMA concentrations have not been defined during acute vaso-occlusive complications. We determined plasma ADMA and arginine concentrations in 96 samples (40 collected in the clinically asymptomatic state and 56 collected at presentation with a painful crisis) of 44 HbSS patients (mean age 32±13 years, m:f=23:21) and in 39 samples (22 collected in the clinically asymptomatic state and 17 collected at presentation with a painful crisis) of 22 HbSC patients (mean age 30±14 years, m:f=10:12). Thirty-five race, sex and age matched HbAA blood donors served as healthy controls. Plasma ADMA concentrations were elevated in asymptomatic sickle cell patients as compared to healthy controls (HbSS: mean 0.7±0.1 umol/L, HbSC mean 0.5±0.1 umol/L, HbAA mean 0.4±0.1 umol/L; p<0.001 one way ANOVA). Plasma arginine concentrations were lower in asymptomatic HbSS patients as compared to asymptomatic HbSC patients and healthy controls (HbSS: mean 59.6±25.8 umol/L, HbSC mean 75.5±25.8 umol/L, HbAA mean 74.0±27.1 umol/L; p=0.03 one way ANOVA). Ratio’s of ADMA to arginine (ADMA/Arginine × 1000) were highest in HbSS patients (HbSS: mean 16.2±11.6, HbSC mean 7.8±2.9, HbAA mean 6.3±3.2; p=0.001 one way ANOVA). In both HbSS and HbSC patients, neither ADMA (p=0.17/p=0.57), arginine (p=0.13/p=0.18) nor the ADMA/arginine ratio (p=0.88/p=0.08) differed between the asymptomatic phase and at presentation with a painful crisis. In conclusion, even though ADMA is elevated in SCD and therefore may well be involved in the development of chronic organ complications by limiting NO availability, ADMA concentrations are not affected during acute painful sickle cell crises.
Prediction of Adverse Outcomes in Children with Sickle Cell Anemia: Testing the CSSCD Early Prediction Model in the Dallas Newborn Cohort.
Authors: Quinn Charles T. , Lee Nancy J. , Shull Elizabeth P. , Buchanan George R. 
Published: October 2019

Abstract
The Cooperative Study of Sickle Cell Disease (CSSCD) reported that dactylitis, severe anemia, and leukocytosis in very young children with SCD increased the risk of later adverse outcomes, including death, stroke, recurrent acute chest syndrome (ACS), and frequent pain (NEJM2000;342:83). Specifically, in the CSSCD infant cohort, the occurrence of dactylitis in the first year of life (“early dactylitis”) and a steady-state hemoglobin (Hgb) concentration <7 g/dL in the second year of life (“severe anemia”) together predicted a high risk (0.36 probability) of a later adverse outcome. A steady-state leukocyte count of approximately 20,000/mm3 or higher in the second year of life (“leukocytosis”) in combination with either early dactylitis or severe anemia also predicted a high risk of an adverse outcome. The testing of this model in other cohorts has not been reported, so we evaluated its performance characteristics in the Dallas Newborn Cohort (Blood2004;103:4023). We studied all cohort members with sickle cell anemia (SS) or sickle-β0-thalassemia (Sβ0) who had 5 or more years of follow-up and complete laboratory data and medical records. We searched our electronic database and patients’ medical records to ascertain the occurrence of the three predictors (early dactylitis, severe anemia, and leukocytosis) and the four adverse events [death; clinically overt first stroke; frequent pain (average of 2 or more painful events/year for the entire follow-up period, with ≥2 events/year for 3 consecutive years); and recurrent ACS (average of 1 or more episode/year with ≥1 episode/year for 3 consecutive years)]. We only considered hospitalizations for pain in this analysis, unlike the CSSCD, because we do not systematically track episodes of pain treated only at home or in outpatient facilities. We studied a total of 119 children (68% male, 97% SS) who had a mean follow-up of 9.7 years (range 5 – 15.4). Twenty-four (20%) had early dactylitis, 5 (4%) had severe anemia, and 15 (12.6%) had leukocytosis. Eighteen subjects (15.1%) experienced adverse events: 2 (1.7%) died, 13 (10.9%) had a stroke, 3 (2.5%) had frequent pain, and 1 (0.8%) had recurrent ACS. Sensitivity and specificity of the model were 0 and 97%. Positive and negative predictive values were 0 and 84%. We used binary logistic regression to further evaluate the model. The whole model test was not significant (P=0.21), and R2 was 0.044. The area under the ROC curve was 0.64 (Figure), and better-performing cut-points for Hgb and leukocyte count could not be found. In summary, the CSSCD early prediction model had very low sensitivity and positive predictive value in the Dallas Newborn Cohort. Specificity and negative predictive value were high, however. This retrospective analysis has several limitations and biases, but it suggests that the CSSCD model does not well predict adverse outcomes in the Dallas Newborn Cohort.
Figure Figure
Anti-Inflammatory Effect of Hydroxyurea Therapy in Sickle Cell Disease.
Authors: Lanaro Carolina , Franco-Penteado Carla F. , Conran Nicola , Saad Sara T.O. , Costa Fernando F. 
Published: October 2019

Abstract
There is growing evidence that inflammatory stresses within the microvasculature may play a significant role in the vaso-occlusion that is characteristic of sickle cell disease. Inflammation, leukocyte adhesion to vascular endothelium, and subsequent endothelial injury appear to contribute to the pathogenesis of SCD. Whilst alterations in a number of inflammatory, anti-inflammatory cytokines and inflammatory biomarkers have been previously related, reports have been conflicting and a conclusive role for cytokines in SCD remains to be established. Furthermore, the effect of hydroxyurea therapy (HU) on the release of inflammatory mediators is not well understood. This study was undertaken to determine plasma levels of IL-8, IL-10, TNF-a and PGE2 in healthy controls, SCD patients in steady-state and in SCD patients on HU therapy (SCDHU; 20–30 mg/kg/day; 3-month minimum treatment duration) using specific ELISA assays (R&D Research). TNF-a levels were significantly higher in SCD individuals when compared to controls (2.95 ± 0.44 pg/ml, n=17; vs 1.43 ± 0.20 pg/ml, n=8, p=0.006). HU therapy, however, was observed to significantly reverse augmented TNF-a levels (SCDHU; 1.61 ± 0.18 pg/ml, n=15, p=0.007). Plasma IL-8 levels were significantly elevated in SCD individuals (16.58 ± 2.52 pg/ml, n=29, p<0.001) compared to healthy controls (5.67 ± 0.62 pg/ml, n=16); however HU therapy did not significantly alter augmented IL-8 in SCD (14.59 ± 1.57 pg/ml, n=22). Interestingly, detectable levels of the anti-inflammatory cytokine, IL-10 (>3.8 pg/ml), were found in 3 (15%) of 20 SCD patients, and 1 (11%) of 9 controls, whilst 7 (50%) of 14 HU-treated patients demonstrated detectable IL-10 levels. Levels of plasma PGE2, an inflammatory biomarker, were significantly increased in SCD patients compared to control individuals (128.35 ± 12.25 pg/ml, n=26; vs 88.45 ± 5.95 pg/ml, n=14, p=0.01) and similar levels were detected in patients treated with HU (143.6 ± 16.75 pg/ml, n=17, p=0.004, compared to control). Data demonstrate that the inflammatory cytokines, TNFa and IL-8, and the inflammatory biomarker, PGE2, are augmented in a population of SCD patients studied during steady-state, contributing to clarify conflicting data. Furthermore, HU therapy reversed augmented TNFa levels, and appears to have an augmenting effect on the anti-inflammatory cytokine, IL-10, suggesting an anti-inflammatory action of HU therapy that may be important for the prevention of vaso-occlusion.
Identification of a New Mutation in the 5′-UTR of Hepcidin Gene in beta-Thalassemia Major (TM) Patients.
Authors: Delbini Paola , Duca Lorena , Nava Isabella , Meo Anna , La Rosa Marina , Cappellini Maria D. 
Published: October 2019

Abstract
Hepcidin is a peptide hormone produced in the liver that acts as negative regulator of iron absorption from the enterocytes and of iron release from macrophages. Iron overload and inflammation up-regulate hepcidin synthesis, while anaemia and hypoxia suppress hepcidin expression. Thalassaemia Major (TM) is a hereditary haemolytic anaemia requiring long-life blood transfusions treatment with consequent iron overload. In β-thalassemias is a disorder in which hepcidin is regulated by opposing influences of ineffective erythropoiesis and concomitant iron overload. In order to get further insights on iron regulation in thalassemias, we screened hepcidin and HFE genes in fourty-three TM regularly transfused patients and sixty control subjects. Blood from TM was taken at least 48 hours after chelation therapy and just before blood transfusion. DNA was prepared from peripheral blood, according to standard protocols. Hepcidin and HFE sequences were amplified with PCR using specific primers and PCR products were sequenced, after purification, in a automatic sequencer. Moreover in all patients serum pro-hepcidin was evaluated by ELISA competitive binding assay (DRG,Germany); iron status was evaluated by serum ferritin (SF), percentage of transferrin saturation (TS) by standard procedures and non transferrin bound iron (NTBI) in serum by HPLC after nitrilotriacetic acid (NTA) chelation. Serum IL-6 as inflammation marker was measured by lateral flow immunoassay (Milenia QuickLine, Germany). Molecular analysis detected an undescribed G→T mutation at position +23 of the 5′-untranslated region in two unrelated TM patients; no mutations were found in control subjects. The probands have been regularly transfused since the age of 1 year, receiving 2–3 units of packed red cells and treated with Deferoxamine 40 mg/Kg/day 6 days/week. The first patient, wild type for HFE mutation, was a 29-years-old compound heterozygous IVS II-745/IVS I-110 man. The SF was 1052 ng/ml, TS 94% and NTBI 1.77 μM. Serum pro-hepcidin value was in normal range (213 ng/ml). The proband’s father was heterozygous for the same hepcidin mutation and showed signs of mild iron overload (SF 491 ng/ml and NTBI 0.50 μM). The second patient was a 26-years-old homozygous β039 man with high levels of SF (4346 ng/ml), TS (169%) and NTBI (2.10 μM), while serum pro-hepcidin was 269 ng/ml. HFE analysis revealed a homozygous genotype for H63D mutation. The patient’s mother was heterozygous for hepcidin and H63D mutation and showed mild iron overload (SF 500 ng/ml; NTBI 0.22 μM) whereas the father, heterozygous only for H63D, had normal iron status. According to recent findings (Bridle et al, 2003) our results indicate that hepcidin mutation in association with H63D synergizes the effect on iron homeostasis and it could be responsible for the development of marked iron overload poorly responsive to chelation therapy in β-thalassemia patients.
Plasma Hemoglobin: Potential Sources.
Authors: Westerman Maxwell , Pizzey Arnold , Hirschman Jocelyn , Cerino Mario , Well-Weiner Yonit , Ramotar Prya , Eze Ada , Lawrie Andrew , Purdy Gordon , Mackie Ian , Porter John 
Published: October 2019

Abstract
Plasma hemoglobin(Hb) is a measure of circulating red blood cell(RBC) destruction and is considered to be the basic indicator of intravascular hemolysis. We have examined the effects of splenectomy on levels of plasma Hb and circulating RBC-derived vesicles in patients with thalassemia intermedia (TI) and compared the results to patients with sickle cell anemia (SCA). Plasma Hb levels in splenectomized patients with TI were 48.5 ± 3.7 mg/ml (5)(Mn ± SEM)(No.of patients) and vesicle levels were 11.29 ± 1.12 x 10 3 /ul blood (9). In contrast, plasma Hb levels in patients with SCA were (14.52 ± 3.29)(21) and vesicle levels were 13.2 ± 2.57)(34). Plasma Hb levels and vesicle levels are closely associated in TI and SCA (r=0.79, p=0.01[9]; r=0.58, p=0.006[21] respectively). The finding that plasma Hb levels in patients with TI and SCA, both asplenic, differ in their relationships to corresponding and similar vesicle levels, suggests that other hemolytic factors may contribute to plasma Hb levels. Of importance would be intramedullary hemolysis which is considerable in TI. Vesiculation, which may occur with intramedullary hemolysis does not appear to contribute to circulating vesicle levels. The ratio of plasma Hb levels to vesicle counts would be a marker to distinguish intramedullary hemolysis from intravascular hemolysis. Similar considerations may apply to measures of lactic dehydrogenase (LDH) which is also an indicator of RBC destruction and intravascular hemolysis. The findings suggest that the contribution of intramedullary hemolysis as well as the contribution of intravascular hemolysis should be considered in measurements of plasma Hb.