bluuebunny/crossref_metadata_2025_split huggingface dataset:
Matrix Metalloproteinase 9 Levels Are Increased in Pegfilgrastim and Filgrastim Induced Mobilisation but Do Not Predict Efficiency of Mobilisation
Authors: Willis Fenella M , Theti Davinder , Gordon-Smith Edward C. , Pettengell Ruth 
Published: April 2020

Abstract
Release of matrix metalloproteinase 9 (MMP-9), neutrophil elastase and cathespin-G from bone marrow stromal cells and granulocytic precursors in response to cytokines and chemotherapy results in degradation of important retention signals such as SDF-1 and V-CAM and promotes egress of progenitor cells from the bone marrow into the peripheral blood. In this study we investigated whether plasma MMP-9 levels could predict efficiency of haemopoietic progenitor cell mobilization
Methods: Plasma MMP-9 levels were assessed at 0, 24 and 48 hours in chemotherapy naïve patients with solid tumours. Patients were randomised to receive a single administration of 6, 12 or 18mg of pegfilgrastim on day 1 or daily administration of filgrastim 10 μg/kg/day from day 1 until day 7. Eight subjects were studied in each group. Each group was balanced in terms of baseline characteristics. Blood samples for peripheral CD34+ cell count and colony assays were collected on the first 7 days and day 9 and 12 post therapy.
Results: The most common tumour types were non-small cell lung cancer (n = 15; 47%) and ovarian cancer (n = 9; 28%). In the filgrastim and pegfilgrastim 6mg, 12mg and 18mg groups a significant rise in MMP-9 levels were seen between 0 and 24 hours (380 vs. 2020 ng/ml, p=0.002; 220 vs 1920 ng/ml, p=0.015; 610 vs. 2510 ng/ml, p=0.0002 and 250 vs. 2050 ng/ml, p=0.003, respectively). In all groups, there was no significant increase in plasma MMP-9 levels between 24 and 48 hours. No significant differences in MMP-9 levels were observed between the 4 cytokine groups over 0 to 48 hours. Across all the groups no significant correlation was seen between plasma MMP-9 levels and peak colony forming units (CFU) per millilitre of blood and peak CD34 positive cells per millilitre of blood (r2=0.103, p=0.08; r2=0.09, p=0.1, respectively).
Conclusion: In all cytokine groups, a significant increase in plasma MMP-9 levels occurred between day 1 and 2. No association between MMP-9, levels numbers of peripheral blood CD34 positive cells and CFU levels could be established. Plasma MMP-9 levels were not affected by pegfilgrastim dose. MMP-9 does not appear to be a predictor of mobilization efficiency for filgrastim or pegfilgrastim at doses up to 18mg.
Iron Chelation, Blood Transfusion and Splenectomy Decreases Angiogenesis in Patients with Thalassemia Major
Authors: Canatan Duran , Olgar Seref , Kara Aslihan , Hicyilmaz Hicran , Balta Nihal 
Published: April 2020

Abstract
Introduction: Thalassemia major (TM) is an important cause of severe anemia that necessitates regular blood transfusion to prevent the profound weakness and cardiac decompensation caused by the anemia. However, iron overloading is an inevitable consequence of prolonged transfusion therapy. In addition, extramedullary hematopoiesis and hemosiderosis cause spleen, liver and marrow enlargement. In recent years the role of angiogenesis has been investigated in physiologic and pathologic conditions. However, it is known that angiogenetic factors, especially vascular endothelial growth factor (VEGF), cause differentiation of the hemangioblast. The role of angiogenesis has been investigated in different kinds of anemia, such as malignancy related anemia and sickle cell anemia. However, the role of angiogenesis has not been investigated in thalassemia major (TM) patients. In this study the angiogenesis was researched in thalassemic patients by serum VEGF measurement.
Material and method: Forty-four consecutive patients with TM were included in this prospective study. TM patients’ findings were compared with those of a healthy control group (n=12). Blood samples were analyzed using commercially available ELISA kits for VEGF
Results: VEGF levels were not affected by hemoglobin levels, ferritin levels, or chelation type (p&gt;0.05). However, VEGF was positively affected by chelation starting age and negatively affected by yearly blood transfusion rate (p&lt;0.05). In addition, VEGF of patients who underwent splenectomy was higher than those who didn’t undergo splenectomy (p&lt;0.05).
In conclusion, VEGF causes differentiation of hemangioblasts, however, early starting transfusion age and regular blood transfusion decrease angiogenesis in thalassemic patients. The besides regular blood transfusion and effective chelation therapy, splenectomy decreases angiogenesis in this group of patients.
Comparing a Novel PET Agent, Yttrium-86 Oxine, with Biopsy and FACS Analysis for Tracking in Vivo Migration Patterns of Different Preparations of DC from a Single Donor
Authors: Harrison Si mon J. , Thompson Mick , Wall Dominic , Loudovaris Maureen , Mackensen Andreas , Bartholeyns Jacques , Ferrantini Maria , Santodonato Laura , Felzmann Thomas , Hicks Rod , Prince H. Miles , Ritchie David 
Published: April 2020

Abstract
Aim: The objective of this study was compare the in vivo distribution of different preparations of dendritic cells (DC) from a single donor after intradermal (ID) administration and correlate FACS analysis of excised tissue and imaging findings.
Method: DC were generated from a single healthy donor apheresis product. The DC types were GM-CSF + IL-13 (IL-13 imDC), then further matured with FMKp+IFNg (IL-13mDC), GM-CSF + IL-4 (IL-4 imDC) and matured (IL-4 mDC) and GM-CSF+IFN-α (IFN-α DC). Antigen naive DC preparations were labelled with Yttrium-86 oxine (prepared on site) by incubation at room temperature for 30 minutes followed by centrifugation and resuspension. 5 × 105 radio-labelled cells in 50_L was injected ID into the footpad and the volar aspect of the foreleg of Balb/c nude mice (n=5 per group). A cohort (n=5) of control mice were similarly injected with Yttrium-86 oxine only. Images were acquired on a small animal PET (SAPET) (Mosaic, Philips Medical Systems) at 2, 24 and 45 hours post administration. Mice were then euthanized and popliteal, axillary and inguinal lymph nodes (LN) harvested. LN were processed into single cell suspension, stained with anti-human HLA-DR and CD11c and analysed by flow cytometry. SAPET images were graded: 0 – no signal, 1 – marginal, 2 – definite nodal signal, 3 – strong signal. DC detection by FACS was graded: 0 – no detection, 1 – &lt;0.5% DC, 2 – 0.5.⋄1.5% DC, 3 – &gt;1.5% DC of all viable cellular events. Results: Pre-injection analysis of DC showed expression of HLA-DR, HLA-ABC, CD11c, CD80 and CD86, low levels of CD83, CCR5 and CCR7 along with down regulation of CD14. Matured DC populations showed increased levels of CD83 and CD86. Similar phenotypic expression on the DC populations were observed prior to cryopreservation and upon thawing. Mean DC radiolabelling efficiency of 72% (54% – 88%). There was a minor decrease in cell numbers and viability after labelling most likely due to oxine related toxicity. There were no substantial differences in tracking between cell types. In 28 of 43 nodes assessed, FACS and SAPET results were congruent (27 FACS and SAPET positive and 1 FACS and SAPET negative). Of the 15 discrepant samples, 13 were SAPET positive and FACS negative, while 2 were FACS positive and PET negative. Average FACS and SAPET grades at 45 hours were higher for popliteal nodes than axillary nodes. No false positive nodal accumulation was visualised at 45 hours in control mice. Conclusion: This novel PET technique provides a robust, reproducible method for quantification of in vivo functional DC tracking to LN. Detection of DC migration using this novel PET agent is a more reliable technique than biopsy and FACS analysis, which has an inherent sampling error.
Comparative Analysis of CD8+ t Cell Priming Efficiencies against a Panel of Leukemia-Associated HLA-A2 Restricted Epitopes Identifies PRAME as a Possible Vaccination Target in AML
Authors: van den Ancker Willemijn , Westers Theresia M , Bontkes Hetty J , Hooijberg Erik , Stam Anita GM , Santegoets Saskia JAM , Ossenkoppele Gert J , de Gruijl Tanja D , van de Loosdrecht Arjan A 
Published: April 2020

Abstract
Background Therapeutic vaccination with dendritic cells (DCs) is regarded as an option for the treatment of minimal residual disease in acute myeloid leukemia (AML). In this respect, defined epitope sequences derived from Leukemia-Associated Antigens (LAA) are an attractive source of antigen. WT1, PRAME, hTERT, and survivin are all LAA that are expressed in various hematological malignancies and that share a potential for the induction of cellular immune responses. Expression rates in AML of each of these LAA are high, ranging from 60 to 90%. To evaluate the feasibility of using these LAA in DC based vaccine formulations in AML, we investigated the relative Cytotoxic T Lymphocyte (CTL) priming efficiencies in response to a panel of LAA-derived HLA-A2 binding epitopes.
Methods CD8+ cells were isolated from peripheral blood of healthy donors (HD) and stimulated with peptide-loaded DCs. DCs were derived from the human AML cell line MUTZ-3 (MUTZ3-DCs).1 The peptides used were WT1 (126–134), PRAME (100–108), hTERT (540–548), and survivin (96–104). Irradiated CD8− cells from the same donor were also added. CD8+ cells were plated at one million per well in 8 to 30 wells. Weekly CD8+ cells were restimulated with peptide-loaded MUTZ3-DCs. HLA-tetramer positivity (Tm+) for WT1, PRAME, hTERT or survivin epitopes was weekly screened by flow cytometry. Wells were scored positive if more than 0.02% Tm+, viable CTLs were present for at least 2 weeks. Functionality of Tm+ cells was studied by analysis of IFN-gamma production after stimulation with the JY cell line loaded with relevant LAA peptides or with an irrelevant control peptide (BCR-ABL).
Results Percentages of the induced peptide-specific CTLs were low as shown in the table below. Priming efficiencies were highest for PRAME. Furthermore, PRAME and WT1 specific CTLs produced IFN-gamma in an epitope specific fashion, confirming functionality of the primed T cells.
Number of HD that were Tm+ after co-culture Number of wells that were Tm+ after co-culture Median % Tm+ cells per positive well (range) Number of restimulations at maximum of Tm+ PRAME 3/4 28/60 0.10% (0.02–1.07) 1–5 WT1 2/5 4/104 0.07% (0.03–0.09) 1–4 hTERT 1/2 1/24 0.25% 5 Survivin 1/1 2/18 0.17% (0.12–0.21) 8
Conclusions On the whole, the detected LAA-specific CD8+ T cell frequencies were low. Most consistently successful priming results were obtained for PRAME, indicating that this peptide may be an attractive candidate for inclusion in AML-specific DC-based vaccine regimens.
Active Von Willebrand Factor in Plasma of Sickle Cell Patients Is An Indicator for Severity of Sickle Cell Anemia
Authors: Chen Junmei , Hobbs William , Le Jennie , Lenting Peter J , Lopez Jose A 
Published: April 2020

Abstract
Vaso-occlusion, hemolysis and oxidative stress are hallmarks of sickle cell anemia. We and others have shown that oxidants generated in sickle cell anemia can stimulate endothelial cells to release ultra-large von Willebrand factor (ULVWF), which is hyperactive compared to the normal plasma VWF in binding platelets and red blood cells, and is capable of binding sickle erythrocytes especially well. For this reason, we surveyed several parameters of VWF quantity and function in plasma from patients with sickle cell anemia (9 individuals, some with multiple determinations). These parameters included VWF antigen levels, VWF multimer composition, ADAMTS13 (the VWF-cleaving metalloprotease) activity, VWF activation factor, and total active VWF. VWF activation factor was measured using a llama antibody (nanobody) that detects an active conformation in the VWF A1 domain that correlates with an enhanced ability to bind platelets. The activation factor is defined as the ratio of nanobody binding of patient plasma to that of pooled normal plasma at a given VWF antigen level. Thus, the activation factor of pooled normal plasma is 1. Elevated VWF activation factor (≥ 1.5, Curr. Opin. Hematol. 2007 14:284–289) has been demonstrated in the plasma of patients with type 2B von Willebrand disease, thrombotic thrombocytopenic purpura, HELLP syndrome, and other prothrombotic disorders. In this study, plasma was collected from patients with sickle cell anemia at baseline during routine outpatient visits. In all of the patients, the VWF antigen level was higher than in the pooled normal plasma (1.5–3.6 fold) or an ethnically matched control. Activation factors were in the range of 0.2 to 6.8. The total quantity of active VWF in plasma was then determined by multiplying VWF antigen level and activation factor. The total active VWF in plasma correlated with the severity of the patient’s disease, as determined by the number of hospital admissions over the previous year and elevated plasma lactate dehydrogenase (LDH). Elevated LDH is a risk factor for hemolysis associated pulmonary hypertension, leg ulcers, priapism and death in sickle cell anemia. In addition, we found that a sub-set of patents had reduced ADAMTS13 activity, which was correlated with higher VWF multimer composition on agarose gels. This indicates that accumulation of active VWF in patients with sickle cell anemia arises not only from increased endothelial secretion but also from decreased ADAMTS13 activity. These findings suggest that high levels of active VWF predispose patients with sickle cell anemia to vasoocclusive complications and correlate with increased hemolysis.
Glycoprotein IIb/Iiia Antagonist Could Reduce the Inflammatory Factors IL-6, IL-1β, TNF-α of Atherosclerosis Rabbits in Vivo
Authors: Yin Songmei , Xie Shuangfeng , Li Yiqing , Nie Danian , Ma Liping , Wang Xiuju , Xiao Jie 
Published: April 2020

Abstract
Background: Glycoprotein (GP) IIb/IIIa antagonists have been widely used in clinical practice. It had been demonstrated that although GPIIb/IIIa antagonists could reduce 30%~ 50% ischemic events around coronary artery intervention therapy, it had only limited effects on the long term survival. We want to explore the effects of GPIIb/IIIa antagonists besides inhibiting platelet aggregation. So we observed the effects of tirofiban (a GP IIb/IIIa antagonist) in vivo.
Design: the atherosclerosis model rabbits were divided into 4 groups. Each group got a 1.7ml/h nature saline (control group), 3.125mg/L, 12.5 mg/L or 50mg/L tirofiban continuous intravenous injection for 48 hours respectively. Platelet aggregation maximum (PAG(M)), plasma interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α were measured at 0 hour, 12th hour, 24th hour and 48th hour time point.
Results: In vivo, compare to the value at 0 hour, the PAG(M) didn’t change significantly at the 12th hour, 24th hour and 48th hour time point in the control group and 3.125mg/L tirofiban group. While the PAG(M) was reduced at the 12th hour, 24th hour and 48th hour time point in the 12.5mg/L and 3.125mg/L tirofiban group. About the inflammatory factors, in control group, compare to the index at 0 hour, plasma IL-1β, IL-6 and TNF-α concentration were all increased significantly at the 12th hour time point. In the 3.125mg/L tirofiban group, the plasma IL-1β and IL-6 concentration elevated significantly at the 24th hour time point. In the 12.5mg/L tirofiban group, the plasma IL-6 and TNF-α concentration at 24th hour time point, the plasma IL-1β, IL-6 and TNF-α concentration at 48th hour time point were all decreased significantly. In 50mg/L tirofiban group, the plasma IL-6 concentration at 12th hour time point, the plasma IL-1β, IL-6 and TNF-α concentration at 24th hour and 48th hour time point were all decreased. At 0 hour, plasma IL-1β, IL-6 and TNF-α concentration had no difference among the 4 groups (IL-1β PANOVA=0.954, IL-6 PANOVA=0.954, TNF-α PANOVA=0.954. n=5). While at 12 hour, 24 hour or 48 hour time points, among the 4 groups, the index value of the IL-1β, IL-6 and TNF-α decreased along with the increase of the tirofiban concentration (PANOVA&lt;0.01. n=5).
Conclusion: In vivo, full dosage tirofiban could inhibit PAG(M), decrease the production of rabbit plasma IL-6, IL-1β and TNF-α. While little dosage tirofiban (≤3.125mg/L) had no effect on PAG(M), could not decrease the production of rabbit plasma IL-6, IL-1β and TNF-α. GP IIb/IIIa antagonist could reduce the inflammatory factors in full dosage.
Disequilibrium of Human Platelet Antigen 1 (HPA-1) in U.S. Population
Authors: Hutchinson Tyler Davi d , Song Yuhua , Trainor Kevin , Hashmi Ghazala 
Published: April 2020

Abstract
Background: Alloimmunization against Human Platelet Antigens (HPA) is associated with Neonatal Alloimmune Thrombocytopenia (NAIT), post-transfusion purpura and refractoriness for platelet transfusion. A flexible BeadChip™ design was developed to simultaneously detect 22 platelet antigens, including HPA-1, and used to assay over 1,000 random blood donors from across the United States.
Methods: Samples from 19 labs/centers from across the country were assayed for 11 HPA loci (HPA-1 through 9, 11 and 15) using the BioArray Solutions HPA Assay. Each locus was independently assessed for Hardy-Weinberg Equilibrium.
Results: Allele and genotype frequencies for each locus were reported. Platelet antigens HPA-2 through HPA-9, HPA-11 and HPA-15 were all found to be in Hardy-Weinberg Equilibrium with a Chi-Squared value of &lt;3.84 (1 degree of freedom, 5% confidence interval). HPA-1, however, did not exhibit Hardy-Weinberg Equilibrium yielding a Chi-Squared value of 43.4.
Conclusions: After reaffirming there was no sampling preference by inclusion of a second blinded random group, it was acknowledged that HPA-1 did not conform to a Mendelian distribution of alleles. The lower incidence of heterozygote HPA-1 individuals may lend credence to the recent finding by Ivanov et al (Akush Ginekol, 2007) linking the polymorphism in GPIIIa that is responsible for the HPA-1 antigen with embryo implantation failure. Further research may help elucidate the causes behind the HPA-1 disequilibrium and how much implantation failure impacts HPA-1 frequencies.
Distinct Gab2-Mediated Signaling Pathways Are Essential for Myeloid or Lymphoid Transformation and Leukemogenesis by BCR-ABL
Authors: Mohi M. Golam , Chan Wayne W. , Li Shaoguang , Neel Benjamin , Van Etten Richard A. 
Published: April 2020

Abstract
The BCR-ABL oncogene encodes an activated fusion tyrosine kinase that causes chronic myelogenous leukemia (CML) and B-lymphoid acute lymphoblastic leukemia (B-ALL) in humans. An autophosphorylation site at Tyr 177 of BCR-ABL recruits Grb2 via its SH2 domain, and is required for efficient induction of CML-like myeloproliferative disease by BCR-ABL in a mouse BM retroviral transduction/transplantation model. We showed previously (Sattler et al., Cancer Cell2002;1:479) that the scaffolding/adapter protein Gab2 is recruited to Y177 of BCR-ABL via a Grb2/Gab2 complex, and in vitro transformation of primary myeloid and lymphoid progenitors by BCR-ABL was impaired in bone marrow from mice with homozygous null mutations in the Gab2 gene (Gab2−/− mice), coincident with decreased activation of the ERK and Akt signaling pathways. Here, we demonstrate an essential requirement for Gab2 in myeloid and lymphoid leukemogenesis by BCRABL. Whereas recipients of BCR-ABL-transduced Gab2+/+ BM develop fatal CML-like myeloproliferative disease within 4 weeks of transplantation, recipients of BCR-ABLtransduced Gab2−/− BM fail to develop CML but succumb after a long latent period to T-cell acute lymphoblastic leukemia, phenocopying the disease induced by the BCR-ABL Y177F mutant. These results suggest that the Y177F and Gab2 mutations have an epistatic relationship, and that the critical transforming signals from Tyr177 of BCR-ABL are transmitted through Gab2. Co-expression of Gab2 with BCR-ABL in Gab2−/− BM restored efficient induction of CML-like leukemia, but mutants of Gab2 that lacked either the pleckstrin homology domain or Tyr binding sites for the SH2 domains of the downstream Gab2 effector molecules SHP2 or p85 PI3K failed to rescue myeloid leukemogenesis by BCR-ABL, although the mutant Gab2 proteins were expressed in circulating myeloid cells. Gab2 deficiency attenuated B-lymphoid transformation by BCR-ABL in vitro, and significantly prolonged the latency of B-ALL induced by BCR-ABL in mice. In contrast to CML, induction of B-ALL in Gab2−/− BM was rescued by either WT Gab2 or the Gab2 mutant defective in p85 binding. These results demonstrate that BCR-ABL absolutely requires signaling via Gab2 to both SHP2 and PI3K to cause CML, while a Gab2-SHP2 signaling pathway contributes to the pathogenesis of BCR-ABL+ B-ALL.
GSK-3β Inhibition Activates WNT, Delays Division and Preserves the Function in Hematopoietic Stem Cell
Authors: Dolnikov Alla , Ko Kap-hyoun , Song Emma , Holmes Tiffany , O’Brien Tracey A 
Published: April 2020

Abstract
Wingless (Wnt) signaling appears to play an integral role in expansion and quiescence of stem and progenitor hematopoietic cell populations. To better understand this dichotomous role and aid in the translation of therapeutic goals including cellular therapies, greater understanding of the molecular mechanisms that regulate Wnt/β-catenin in the bone marrow niche are needed. We explored the impact of Wnt signalling in the context of the human HSC niche by using co-culture of human cord blood or bone marrow derived HSC with mesenchymal stem cells (MSCs) together with pharmacological inhibition of GSK-3β (iGSK-3β). We have previously shown that activation of the Wnt/β-catenin pathway using pharmacological inhibition of GSK-3β (iGSK-3β) in human cord blood derived HSCs acts to preserve their primitive status, often lost during the ex-vivo expansion process (1). Here we show using cell division tracking that iGSK-3β delays stem cell divisions induced by cytokines in co-culture with MSCs. Functional analysis of iGSK-3β-treated cells revealed an increased proportion of primitive cells producing mixed Colony Forming Units (CFU-GEMMs) formed in the methylcellulose cultures and Cobblestone Area Forming Cells identified in long-term co-cultures with the bone marrow stroma MS5 cells. In addition, iGSK-3β-treated cells exhibited a higher frequency of SCID Repopulating Cells. We also show that the iGSK-3β effect, at least in part, is mediated through VLA4 integrin engagement by fibronectin. Importantly, iGSK-3β also induced niche-independent delay in stem cell divisions seen in cytokine supported suspension cultures. Up-regulation of HoxB4 and p21Waf1 and down-regulation of cyclin D1 was observed in HSCs treated with iGSK-3β. These findings confirm that iGSK-3β– regulated Wnt/β-catenin activity is crucial for the maintenance of HSC quiescence, both in the MSC niche and in a niche-independent environment and that iGSK-3β can circumvent the requirement of exogenous Wnt ligand for the induction of Wnt/β-catenin signalling in MSC niche. Our results are consistent recent data showing that blocking of canonical Wnt signaling, by over-expression of dickkopf1 (Dkk1) increases the number of proliferating HSCs in the murine marrow microenvironment but reduces the ability to reconstitute the hematopoietic system of irradiated recipient transgenic mice (2). These findings contribute to a better understanding of the mechanisms regulating the balance between self renewal and quiescence and help to facilitate progress towards clinical application of Wnts as mediators of ex-vivo stem cell expansion or interruption of the Wnt pathway as potential anti-leukemic therapeutic strategy.