bluuebunny/crossref_metadata_2025_split huggingface dataset:
Intravascular Hemolytic Anemia with Acanthocytosis Following Alternative Treatment with Ozone, UV Light, and Perflurocarbon
Authors: Hymes Kenneth B. , Huynh Chan 
Published: April 2020

Abstract
Alternative medicine has become more common as patients seek approaches to diseases where traditional medicine has failed. Treatment with ozone has been purported to have benefits for a variety of infectious, inflammatory and neoplastic conditions. Treatment can be administered by the intra arterial, intravenous, intra rectal and subcutaneous routes as well as ozonated autohaemotherapy. We describe a 44 year old woman who received treatment at an alternative medical center for recurrent breast cancer including laetrile, perflurocarbon emulsion, high dose ascorbic acid, vitamin K and extracorporeal treatment of her blood with ozone and ultraviolet light. After receiving her second treatment, she presented to our hospital with a syncopal episode and was found to be anemic (Hb 4 gm/dl). The LDH was 8X&gt; ULN, the serum haptoglobin was undetectable, and the reticulocyte count was increased. The peripheral blood smear showed aniscocytosis, poikilocytosis and polychromasia. There was also severe acanthocytosis. Heinz bodies were not detected. The patient refused blood products and after 48h of hospitalization, her Hb rose to 7gm and she was discharged. She did not return for follow-up evaluation.
Acanthocytosis has been associated with impairment of cholesterol membrane fluidity seen with acquired hepatic disease as well as some congenital diseases. Oxidative stress can lead to peroxidation of membrane phospholipids, and if the intrinsic repair mechanism of the RBC is overwhelmed, the RBCs transform into acanthocytes. To date, there has been no literature describing hemolysis associated with any of the individual treatments which this patient received; however the combination of treatments exposed the RBC’s to multiple oxidative stresses which might lead to lipid peroxidation and formation of acanthocytes. This case illustrates the potential problems that face clinicians when encountering patients who seek alternative therapy.
High Resolution Tertiary and Quaternary Structural Changes in Deoxy and Liganded Hemoglobin E
Authors: Hirsch Rhoda Elison , Malashkevich Vladimir , Balazs Tatiana C. , Almo Steven C. 
Published: April 2020

Abstract
Hemoglobin (Hb) E (β26 Glu→Lys), known to be more unstable than HbA in vitro, remains an enigma in terms of its contributions to red blood cell (RBC) pathophysiological mechanisms. EE individuals exhibit a mild, chronic anemia while HbE-β thalassemia individuals show a range of clinical manifestations, including high morbidity and death, often resulting from cardiac disease. To determine pathophysiological consequences originating from HbE, we created a transgenic mouse expressing exclusively human HbE with equivalent α- and β-chains [Chen et al., Blood 106(11):892a, 2005]. This mice model exhibited hematological characteristics similar to human EE individuals: microcytic RBC with low MCV, low MCH but normal MCHC; target RBC; mild anemia with low Hb, low HCT, elevated reticulocytes; and decreased osmotic fragility. These RBC parameters correlated with indications of mild RBC oxidative stress. Importantly, the transgenic mouse model allows us to obtain purified HbE for crystal growth and dynamic studies, without the presence of human HbA2 (co-expressed in human HbE RBC and difficult to separate). We now present the low [PDB 1YVQ] and high salt [PDB 1YVT] liganded COHbE structures (1.8Å resolution) at or near physiological pH. The known in vitro instability of oxy HbE and its propensity for oxidation may be explained by the substitution of the β26 side-chain with Lys resulting in a positive charge repulsion with consequential changes to β117, and other key stabilizing residues at the α1β1 interface that impart normal association and oxidative stability to HbA. The specific alterations observed in this microenvironment may be the key to HbE instability, given the findings by Adachi et al. (Biochemistry, 42:10252, 2003) that β116 in normal HbA plays a critical role in stabilizing α1β1 interactions and inhibiting oxidation. Additionally, the difference in water structure observed in each of our COHbE structures from crystals generated under different conditions clearly suggests that resolved water structure is dependent simply upon crystallization conditions, and caution must be taken in interpretation of thermal instability. The 1.6 Å structure of deoxy HbE [PDB 3DUT] (crystallization conditions: pH 6.8, 1.8M ammonium phosphate/ammonium sulfate/citrate) exhibits definitive quaternary change compared to the deoxy HbA structure obtained under similar conditions. These structural findings, explain in part, our recent spectroscopic findings of alterations in oxy and deoxy HbE-nitrite interactions.
Pharmacokinetics and Pharamcodynamics of Recombinant Human Thrombopoietin Following Single Injection in Rats
Authors: Krzyzanski Wojciech , Rekic Dinko , Gebreegziabher Asfiha G. 
Published: April 2020

Abstract
Background: Thrombopoietin (TPO) is the primary regulator of platelet production. Recombinant human thrombopoietin (rHuTPO) and its derivative have been reported as potent drugs in treatment of chemotherapy induced thrombocytopenia and idiopathic thrombocytopenic purpura. Due to severe immunogenic adverse affects clinical applications of rHuTPO have been limited. However, new TPO receptor agonists that lack this adverse effect are developed. RHuTPO exhibits nonlinear kinetics and complex dynamics both mediated by binding to TPO receptor. Studies of rHuTPO pharmacokinetics (PK) and pharmacodynamics (PD) will facilitate future use of new TPO receptor agonists.
Purpose: To analyze PK and PD of rHuTPO following a single IV bolus administration in rats.
Methods: Adult male Wistar rats of body weight 300–600 g received a single dose of rHuTPO (ProSpec Ltd) (2.4, 4.8 and 9.6 μg/kg) and a placebo via an intravenous injection in tail. Blood samples (100–200μL) were collected at various hours during 3 days for PK analysis and every day for consecutive 14 days for PD assessment. RHuTPO plasma concentrations were measured using ELISA (R&D Systems Inc). Platelet count was determined using hematology analyzer (BC-2800 VET, Mindray). Noncompartmental parameters such as area under concentration curve (AUC), clearance (CL), volume at steady-state (Vss), and terminal half-life (t1/2) were calculated by WinNonlin 5.2 (Pharsight). Percent increase over the baseline for platelet maximum (%MPC) and area under the platelet response curve (%AUCPLT) were applied as PD markers. Student t-test was used to compare the means between control and treatment groups (SAS 9.1, SAS Institute).
Results: CL decreased with increasing dose (0.185, 0.131, and 0.047 L/h/kg), similarly to Vss (3.28, 1.10, and 0.51 L). The t1/2 values were also dose dependent (13.2, 6.1, and 8.2 h). The %MPC values were 43, 77, and 45% for corresponding doses and the peaks occurred on days 5, 6, and 6 following rHuTPO administration. The %AUCPLT values were relatively insensitive to increasing doses (20, 23, and 13%). Both mean peak platelet counts and mean AUCPLT were significantly different (p &lt; 0.05) from the control values with an exception of AUCPLT for dose 2.4 μg/kg.
Conclusions: RHuTPO exhibits nonlinear pharmacokinetics in rat. CL and Vss dependence on dose suggests receptor mediated drug disposition as a major reason of nonlinear PK. IV bolus doses 2.4, 4.8 and 9.6 μg/kg of rHuTPO moderately increase platelet count in rats that peaks on day 5–6 after drug administration. The observed lack of the PD marker increase with increased doses implicates that the applied doses are in a flat region of the dose-response curve. Compared to similar data reported for humans, rHuTPO is less efficacious in rats than in humans.
Traditional Chinese Whole-Body Acupuncture Mobilizes CD133(+)CD34(−) Cells
Authors: Moldenhauer Sonja , Burgauner Miriam , Hellweg Rainer , Lun Andreas , Kiesewetter Holger , Salama Abdulgabar , Moldenhauer Anja 
Published: April 2020

Abstract
The objective of this study was to analyze the effects of traditional Chinese acupuncture for spinal cord lesions on the mobilization of stem cells. Therefore, 14 healthy study participants were acupunctured and gave blood samples before, immediately after as well as 24 and 48 hours after acupuncture. At these time points, the frequency of CD133, CD34, CD4, CD14, CD19 and CD45 positive cells were determined by flow cytometry. Furthermore, serum concentrations of matrix metalloproteinases (MMP) 8 and 9, brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), stroma-derived factor (SDF) 1 and granulocyte-colony stimulating factor (G-CSF) were measured by enzyme-linked immunosorbent assays. CD133(+)34(−) cells were significantly increased 48 hours after acupuncture (29.1±5.8% versus 11.4±4.8%, p= 0.015), which was paralleled by significant decreases of BDNF (7.5±1 ng/ml versus 10.8±1.6 ng/ml, p= 0.013) and MMP-9 serum levels (46.4±2.6ng/ml versus 42.5±2.4 ng/ml, p= 0.009). Values in non-treated controls were not affected. No changes in SDF-1, NGF, interleukin-6 and G-CSF concentrations occurred. In conclusion, acupuncture does mobilize non-hematopoietic CD133(+) cells.
Soluble Endoglin and Its Interaction with Adhesion Molecules in Patients with Thalassemia Intermedia
Authors: Kattamis Antonios , Lazaropoulou Christina , Siopi Maria , Kanavaki Ino , Goussetis Evgenios , Ladis Vassilios , Papassotiriou Ioannis 
Published: April 2020

Abstract
Endoglin (CD105) is an accessory protein of the transforming growth factor-β receptor system expressed on vascular endothelial cells. Mutations on the endoglin gene are associated with hereditary hemorrhagic telangiectasias (Osler-Weber-Rendu syndrome) and, thus, endoglin has been extensively studied in the context of this disease. Endoglin is highly expressed on endothelial cells in healing wounds, developing embryos, inflammatory tissues and solid tumors. It is a marker of activated endothelium, while its vascular expression is limited to proliferating cells. Previous studies have shown that the endothelial function is impaired in patients with Thalassemia Intermedia (TI). Oxidative damage resulting from hemolysis and iron load, leads to increased expression of the intercellular and vascular adhesion molecules-1 (ICAM-1 and VCAM-1) and impaired nitric oxide (NO) bioavailability. As endoglin plays a critical role in angiogenesis and dysregulation of its expression and/or activity has been implicated in multiple vascular diseases, we aimed to investigate the expression of endoglin and its correlation with factors of endothelial dysfunction in patients with TI. Thirty adult patients with TI were included in the study, while 20 healthy individuals served as controls. Soluble endoglin, soluble forms of adhesion molecules ICAM-1, VCAM-1, E and P-Selectins, thrombomodulin, von Willebrand factor, as well as NO and vascular endothelial growth factor (VEGF), were measured in patients and controls, by immunoenzymatic methods. The main results of the study are:
levels of endoglin, E-selectin, thrombomodulin, VCAM-1, ICAM-1 and VEGF in patients with TI (5.5±0.4 ng/ml, 91.4±20.0 ng/ml, 48.8±16.5 ng/ml, 1413.7±176.1 ng/ml, 658.8±34.6 ng/ml and 619.1±227.8 pg/ml, respectively) were higher compared to controls (4.9±0.3 ng/ml, 10.3±2.2 ng/ml, 4.0±0.6 ng/ml, 328.3±43.8 ng/ml, 107.6±26.5 ng/ml, 81.3±58.1 pg/ml), (p&lt;0.01), endoglin levels in patients with TI correlated positively with concentrations of ICAM-1 (r=0.760, p&lt;0.003), VCAM-1 (r=0.520, p&lt;0.05), E-Selectin (r=0.790, p&lt;0.0020, P-Selectin (r=0.530, p&lt;0.04), while these correlations were absent in normal individuals.
Angiogenesis is a highly coordinated process in which VEGF, endoglin and hypoxia inducible factor 1 (HIF-1) play a pivotal role by coordinating interaction between endothelial cells, extracellular matrix and the surrounding cells. Taking into assumption that endoglin is a protective-repair tissue protein, our findings support the hypothesis that patients with TI exhibit increased degree of angiogenesis and endothelial regeneration, which are probably compensatory mechanisms in response to tissue hypoxia and damage.
CD44 Signaling Via PI3K/AKT and MAPK/ERK Pathways Protects CLL Cells from Spontaneous and Drug Induced Apoptosis
Authors: Herishanu Yair , Gibellini Federica , Njuguna Ndegwa , Keyvanfar Keyvan , Wiestner Adrian 
Published: April 2020

Abstract
Chronic lymphocytic leukemia (CLL) is an incurable malignancy of mature B-cells. CLL cell proliferation and survival depends on host factors that are present in the tissue microenvironment. In–vitro these cells rapidly undergo apoptosis but can be supported by culture on stroma cells. The extracellular matrix is an important component of the microenvironment. Interactions between tumor cells and the extracellular matrix are, in part, mediated by CD44, whose principle ligand is hyaluronic acid. To explore a possible role of this adhesion receptor in CLL, we evaluated the effect of CD44 engagement on the survival of CLL cells and the induction of drug resistance. Dimerization of CD44 activated the PI3K/AKT and MAPK/ERK pathways and resulted in increased levels of MCL-1. There was no increase in MCL mRNA in CD44 stimulated cells, consistent with an effect on protein stabilization. Consistent with the induction of these anti-apoptotic mechanisms, CD44 protected CLL cells from spontaneous apoptosis: CD44 stimulated CLL cells had a 46% increase (range 7% – 181%) in viability over the corresponding unstimulated control cells (n=20, p&lt;0.0001). Furthermore, CD44 activation also protected from fludarabine-induced apoptosis: cell viability for isotype treated cells 76 ±6%, for isotype and fludarabine treated cells 47 ±18% and for CD44 stimulated and fludarabine treated cells 69 ±16% (p=0.005). CLL cells of the IgVH unmutated subtype expressed CD44 more highly than IgVH mutated CLL cells (MFI ratio 224 ±43 to 122 ±44, respectively, p&lt;0.0001) and derived a more pronounced survival advantage from CD44 activation: % live cells (CD44 stimulated - control) 21 ±9% vs. 13 ±6%, respectively (p=0.04), which could contribute to the more progressive nature of this subtype. PI3K or MEK inhibitors as well as obatoclax, an antagonist of MCL-1, blocked the pro-survival effect of CD44 activation. Furthermore, obatoclax was able to sensitize CLL cells to fludarabine. In conclusion, CD44 engagement protects CLL cells from spontaneous and fludarabine induced apoptosis. This prosurvival effect is mediated through PI3K/Akt and MAPK/ERK pathways and increased MCL-1 levels. Targeting either signaling pathway or MCL-1 with specific inhibitors could overcome the protective effect of the microenvironment in CLL and provides a rationale for combination therapy.
Fluctuation of Cystatin C Concentration during Deferasirox Treatment in Thalassemic Patients May Reflect Hemodynamic Changes
Authors: Papassotiriou Ioannis , Kattamis Antonios , Hantzi Eugenia , Delaporta Polyxeni , Sergounioti Athanassia , Margeli Alexandra , Ladis Vassilios 
Published: April 2020

Abstract
Deferasirox (Exjade®, ICL670) is an once-daily, oral iron chelator for the treatment of transfusional iron overload and its efficacy and tolerability have been established in adults and children with a range of transfusion-dependent anemias, such as thalassemia. We observed alterations in cystatin C plasma concentration during longterm treatment of thalassemia patients with deferasirox. Cystatin-C is a non-glycosylated protein that belongs to the cystatin superfamily of cysteine protease inhibitors. Plasma cystatin-C has been suggested to be an ideal endogenous marker of glomerular filtration rate (GFR). We aimed to investigate if these changes in cystatin C concentration were solely due to deferasirox treatment, or whether other factors contribute to these changes. 150 b-thalassemia patients treated with deferasirox at doses between 20–40 mg/Kg/day were followed longitudinally. Cystatin C concentrations were measured by an immunonephelometric technique (Dade Behring, Siemens Healthcare Diagnostics) at regular monthly intervals (5–12 monthly serial measurements per patient), while GFR was calculated according to the recently proposed cystatin C–based prediction equation using only its concentration in mg/L: GFR [mL/min/1.73m2] = 76.7 x Cystatin C−1.18. To further evaluate the effect of deferasirox on renal function we measured: a) plasma levels of Neutrophil-Gelatinase-Associated-Lipocalin (NGAL) (R&D Systems) a protein expressed on tubular cells and which production is markedly increased in response to harmful stimuli, such as ischemia or toxicity, b) N-terminal pro-B-type natriuretic peptide (NT-proBNP) (Roche Diagnostics), in order to correlate cystatin C fluctuation with left ventricular ejection fraction (LVEF) and c) ferritin level, as a marker of iron overload/mobilization. The main results of the study showed that cystatin C concentration fluctuated during deferasirox treatment (ANOVA repeat measures p&gt;0.850). Baseline mean values were 0.97±0.27 mg/L and reached a maximum of 1.01±0.29 mg/L at 4 months of treatment. According to the guidelines and as no significant increase of creatinine levels was observed, the drug dose was not reduced. Analysis of data showed that: a) there is no correlation between cystatin C fluctuation and NGAL levels, p &gt; 0.6745, evidence that the tubuloglomerular feedback is not activated b) cystatin C and NT-proBNP levels correlated positively with a binomial equation (p&lt;0.004) and c) cystatin C and ferritin levels correlated positively with also a binomial equation (p&lt;0.001). These findings suggest that cystatin C fluctuation during deferasirox treatment do not reflect renal injury. Hemodynamic signals, LVEF alterations and iron mobilization seem to play central role on changes of cystatin C.
Induction of Adenovirus Specific T-Cells in Response to 15-Mer Peptides in Cord Blood
Authors: Haveman Lianne Margreet , Bulatovic Maja , Prakken Berent , Bierings Marc 
Published: April 2020

Abstract
Human adenovirus (HAdV) infections are a serious life-threat in haematopoietic stem cell transplantation (HSCT) patients, particularly in children. In recent years, umbilical cord blood transplants (UCBT) emerged as an alternative stem cell source. The advantages are the rapid availability of donor cells, the possibility of using HLA-mismatched transplants and a lower risk of graft versus host disease. However, the immune response after UCBT is often delayed, leading to a higher risk of viral infections. Because no effective antiviral medication exists for severe HAdV infection adoptively transfer of HAdV specific T-cells from cord blood may be a promising treatment. The aim of this study was to see if it was possible to induce HAdV specific T-cells in response to five recently detected 15-mer peptides ( Haveman LM, Int. Imm. 2006), a pool of these five peptides or complete inactivated HAdV (MOI 10). Mitogen Concovalin A ( ConA) was used as control. Cord blood mononuclear cells (CBMC) derived from 34 uncomplicated full term deliveries were isolated and cultured with the different antigens. Proliferation expressed as Stimulation Index (SI) was determined. A SI&gt;1.7 was seen as positive. In 14 neonates a response to in total 36 of the 5 different HAdV peptides could be seen and in 7 of them also a response to the pool of the peptides. Only in 3 neonates a response to complete HAdV could be detected (Figure 1). To induce HAdV specific T-cells CBMCs of 20 neonates were cultured with the different HAdV antigens in addition of interleukin 2 (40IU/ml) for 7 days. By using FACS analysis the upregulation of the activation markers CD25 and CD69 could be detected in response to the HAdV peptides (n=41) or to the complete HAdV (n=26). Induced HAdV specific T-cells expressed the cytotoxins IFN□, TNFα, perforin and granzyme B, produced by CD8+ T-cells, but in higher extent by CD4+ T-cells. This indicates that induced HAdV specific T-cells play an essential role in killing HAdV infections. No clear correlation could be seen between proliferation and the production of the cytotoxins. The induction of a specific immune response to HAdV peptides in cord blood is an important step towards adoptive therapy.
Figure 1. Proliferation of CBMC in response to HAdV peptides, complete HAdV en ConA Figure 1. Proliferation of CBMC in response to HAdV peptides, complete HAdV en ConA
Leukemia, Solid Tumors and Methylarginines: Is There Any Relation?
Authors: Doroszko Adrian , Niedzielska Ewa , Wojcik Dorota , Szuba Andrzej , Andrzejak Ryszard 
Published: April 2020

Abstract
Background: Despite improvements achieving complete remission and progress in supportive care of children with acute leukemias or solid tumors during the last decade, there are no optimal prognostic markers in stratification of risk of adverse events leading to poor outcome. The endogenous nitric oxide synthesis inhibitor asymmetrical dimethyl-L-arginine (ADMA) is associated with atherosclerosis and represents an independent marker for cardiovascular morbidity and mortality.
Aim: To investigate the clinical significance of methylarginines in leukemia and solid tumors in children.
Material and method: Concentration of metabolites of nitric-oxide pathway in children during therapy of acute leukemia (ALL) – N 12 and solid tumors N 15 (osteosarcoma, RMS, PNET, Wilms’ tumor) was determined. Correlations of biochemical markers, blood count parameters and plasma level of methylarginines were investigated. In addition, the influence of cytostatics on NO-pathway metabolites was analyzed.
Results: Table 1: Differences in mean plasma concentration of arginine, ADMA and SDMA in pts with leukemia vs pts with solid tumor
Leukemia N=12 Mean ±SD Median±SEM Solid tumor N=15 Mean ±SD Median±SEM p Arg 31,28±8,63 30,82±2,49 49,59±10,81 49,18±3,00 0,0003 ADMA 1,17±1,10 0,53±0,32 0,98±1,10 0,46±0,31 NS SDMA 1,18±1,10 0,80±0,32 1,56±1,02 1,09±0,28 NS Arg/ADMA 67,40±54,31 69,48±15,68 228,82±451,44 117,35±125,21 NS
Table.2. Differences in mean plasma concentrations of arginine, ADMA and SDMA depending on gender, treatment, etc.
N=27 Arginine ADMA SDMA Arg/ADMA Sex 1. Mail; 2. Femail 1. 0.NS NS NS NS HSCT (1-yes; 0-no) 1. 0.NS NS NS NS Remission (1-yes; 0-no) 1. 0.NS NS NS NS Currently infection (1-yes; 0-no) 1. 0.NS NS NS NS Glucocoricoids (1-yes; 0-no) 1. 31,9±9,30. 51,0±11,2 p=0,0011 NS NS NS Cytostatic – any (1-yes; 0-no) 1. 0. NS NS NS NS Doxorubicine (1-yes; 0-no) 1. 0. NS NS NS NS Vincristine (1-yes; 0-no) 1. 0. NS NS NS NS Cytarabine (1-yes; 0-no) 1. 32,0±10,50. 45,6±10,8 p=0,022 NS NS NS Methotrexate (1-yes; 0-no) 1.0 NS NS NS NS Thioguanine/6-merkaptopurine(1-yes; 0-no) 1. 0. NS NS NS NS Cyclophosphamide/Iphosphamide(1-yes; 0-no) 1. 0. NS NS NS NS
Conclusions: Plasma level of L-Arg was significantly higher in patients with solid tumor The L-Arg/ADMA ratio was not significantly higher in patients with solid tumors and lower plasma level of ADMA was found. Treatment with glucocorticoids and cytarabine was associated with significantly lower level of L-Arg. No significant treatment dependent difference was found in level of nitric oxide metabolic pathway.