bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Effect of von Willebrand Factor (VWF) on Site-Specific Factor VIII (FVIII) Expression in Hemophilia A Mice.
Authors: Shi Qizhen , Fahs Scot A. , Kuether Erin L. , Montgomery Robert R. 
Published: October 2019

Abstract
von Willebrand factor (VWF) is a carrier protein for factor VIII (FVIII) and protects plasma FVIII from protease degradation. Our laboratory has had a longstanding interest in the association of FVIII with VWF both in vitro and in vivo. Our in vitro studies have demonstrated that FVIII stores together with VWF in both endothelial cells and megakaryocytes if FVIII is made in these cells. Furthermore, we demonstrated that FVIII and VWF are both releasable by agonist stimulation. To investigate the association of VWF and FVIII in vivo, we generated two lines of transgenic mice that express FVIII either in endothelial cells or in platelets using either the endothelial cell-specific Tie2 promoter or the platelet-specific αIIb promoter, respectively. When the platelet-specific FVIII (2bF8) transgene is bred into the FVIIInull mouse, FVIII can only be detected in platelets, with a level of 0.76 ± 0.27 mU/108 platelets in heterozygous and 1.53 ± 0.14 mU/108 platelets in homozygous 2bF8 mice. When the endothelial cell-specific FVIII (Tie2F8) transgene is bred into the FVIIInull mouse, homozygous Tie2F8 mice maintained normal plasma FVIII levels (1.15 ± 0.16 U/ml) and 50% levels in heterozygous mice (0.56 ± 0.16 U/ml). Both 2bF8trans and Tie2F8trans phenotypes effectively abrogate the bleeding diathesis in hemophilic mice. When 2bF8 transgene was bred into a FVIII and VWF double knockout background, the level of platelet-FVIII significantly decreased, but this platelet-derived FVIII was still stored in a-granules and still maintained clinical efficacy. In contrast, when the Tie2F8 transgene was bred into the double knockout background, plasma FVIII dropped to undetectable levels. This is in contrast to the situation in VWFnull mice in which normal endogenous murine FVIII is synthesized with about 10% of normal FVIII activity persisting in plasma. This could be due to a difference in survival between human FVIII and murine FVIII. All Tie2F8trans/FVIIInullVWFnull mice (n=15) survived tail clipping even though there is no FVIII:C detected in the plasma. To investigate the effect of murine VWF on the levels of plasma FVIII, plasma from FVIIInull mice was infused into Tie2F8trans/FVIIInullVWFnull mice to restore VWF levels to 25% of normal. As expected, the endothelial cell-derived plasma FVIII was stabilized by the infused VWF and was detected within 1 hour after infusion, with a peak (25% level) at 4 hours. The level of plasma FVIII at 24 hours was still about 20% of normal while the level of remaining VWF was only 5% of normal. These results demonstrate that VWF is important for site-specific FVIII expression. Co-expression with VWF in platelets is important for optimal platelet-specific FVIII expression and endothelial cell-derived plasma FVIII is VWF-dependent.
Melphalan-Prednisone-Thalidomide to Newly Diagnosed Patients with Multiple Myeloma: A Placebo Controlled Randomised Phase 3 Trial.
Authors: Waage Anders , Gimsing Peter , Juliusson Gunnar , Turesson Ingemar , Fayers Peter 
Published: October 2019

Abstract
Previously untreated patients with multiple myeloma were included in this double-blind randomised trial. Patients not eligible for high dose treatment in Norway, Sweden and Denmark were recruited. The study started in 2002 and accrual of patients stopped 1st of May 2007. Date of analysis was 1st of June 2007. The treatment was randomized between melphalan/prednisone/thalidomide and melphalan/prednisone/placebo. Starting dose of thalidomide was 200 mg escalating to 400 mg. There was no recommendation for prophylaxis for venous thromboembolism. Endpoints were overall survival, event free survival, response, time to progression and quality of life. Interim analysis for safety was performed in 2004 by an independent commitee. Altogether 362 patients with mean age 75 (49–92) years were included, 55% males. The incidence of venous thromboembolism in the blinded treatment arms was 7%. The study will be unblinded and results will be available September 2007.
Human AML Stem Cells Remain Quiescent and Exhibit Chemotherapy Resistance within the Bone Marrow Endosteal Region.
Authors: Saito Yoriko , Yoshida Shuro , Saito Noriyuki , Fukata Mitsuhiro , Ozawa Hidetoshi , Doi Takehiko , Miyamoto Toshihiro , Ohshima Koichi , Uchida Naoyuki , Taniguchi Shuichi , Akashi Koichi , Harada Mine , Shultz Leonard D. , Ishikawa Fumihiko 
Published: October 2019

Abstract
AML is characterized by the clonal expansion of immature myeloblasts initiating from rare leukemic stem cells (LSCs). We developed a primary human AML xenotransplantation model using newborn NOD/SCID/IL2rg−/ − mice. From 9 patients, 4x106 T cell-depleted BMMNCs were injected iv into sublethally irradiated recipients. At 3 months, newborn NOD/SCID/IL2rg−/ − recipients demonstrated superior engraftment efficiency compared with adult NOD/SCID/IL2rg−/ − and newborn NOD/SCID/b2m−/ − recipients (37.8%, 11.9%, 12.9%, respectively; p<0.005). When sorted primary AML hCD34+CD38− cells from 6 patients were injected iv, as few as 103 cells engrafted dose-dependently. Transplanted hCD34+hCD38− cells gave rise to hCD34+hCD38−, hCD34+hCD38+ and hCD34− cells. In serial transplantation, injection of 103 purified hCD34+hCD38− cells resulted in long-term engraftment, giving rise to hCD34+hCD38− LSCs and hCD34+hCD38+/hCD34− blasts, demonstrating self-renewal and differentiation capacities of primary LSCs in vivo. We next examined homing and localization of LSCs using this model. In recipient femoral sections, 94.0% and 78.4% of hCD34+ cells were found in the endosteal region at day 3 and at 4 months after iv injection, respectively (10,065 cells and 24,973 cells examined, respectively; p<0.001 for each), demonstrating that human primary LSCs preferentially home to and engraft in the BM endosteal region. Dual FISH analysis using mouse and human pan-centromeric probes confirmed the human origin of these cells. When AML-engrafted mice were treated with cytosine arabinoside (Ara-C) to examine the effect of anti-leukemia drugs in vivo, PB AML burden became markedly reduced by day 8, followed by AML relapse by days 16–28. Apoptosis analysis of day 3-post-Ara-C BM demonstrated that hCD34+hCD38− LSCs are more resistant to Ara-C induced apoptosis compared to hCD34+hCD38+ and hCD34− cells (Annexin V-7AAD-: 77.1%, 33.3%, 27.5% respectively; n=3 for each; p<0.05 for each). TUNEL staining using day 3 post-Ara-C femoral sections demonstrated that TUNEL-negative cells were preferentially located at the endosteal surface while the cells in the central BM cavity were apoptotic. The cells abutting the endosteum were hCD34+hCD38− and were adjacent to osteopontin+ osteoblasts. % of cells in the G0 phase of cell cycle was significantly greater in the hCD34+hCD38− compared with the hCD34+hCD38+ and hCD34− fractions (n=5 for each; p<0.005 for each), suggesting a potential mechanism underlying relative chemo-resiatance of LSCs. In summary, primary human LSCs preferentially engraft in the BM endosteal areas where they are protected from AraC-induced apoptosis. Cell cycle quiescence of LSCs may be one of the mechanisms underlying chemoresistance of LSCs.
Donor-Derived Squamous Cell Carcinoma after Allogeneic Bone Marrow Transplantation.
Authors: Janin Anne , Murata Hideyuki , Lebeuf Christophe , Gluckman Eliane , Soulier Jean , de Thé Hugues , Bertheau Philippe , Socié Gerard 
Published: October 2019

Abstract
Long-term survivors after allogeneic hematopoietic stem cell transplantation (HSCT) develop solid tumors. Chronic graft-versus-host disease (GvHD) increases the risk of squamous cell carcinoma (SCC). Five of 24 patients with HSCT and SCC, had sex-mismatched HSCT. In 3 of them, tumor sex karyotype studied by combined XY FISH and double immunostaining on the same SCC section, with 3D Z-stack analysis, was compatible with a cancer originating from donor hematopoietic cells. After control by short tandem repeats analysis (PCR-STR) of laser micro-dissected tumor and normal epidermal cells from the recipient and donor or PCR-STR and FISH analysis of a tumoral cell line from one SCC, a recipient origin was found in one case but in the two other cases the donor genotype of the tumor was confirmed. In a third step, PCR-STR of laser micro-dissected tumor in 2 cases after sex-matched transplant proved SCC to be of the donor origin. We thus obtain conclusive evidence that 4 epithelial tumors among 8 studied arose from the engrafted donor cells. Tissue stem cells were proposed to exhibit an unexpected level of plasticity, although issues on cell fusions have lead to some controversies. Only transplantation experiments using genetically distinct recipients and donors can unequivocally demonstrate these changes in cell fate. We have analyzed a unique set of squamous cell carcinomas arising in long terms survivors of allogeneic stem cell transplantation. In these patients, chronic graft-versus host disease greatly favors development of solid tumors, possibly as a consequence of mucosal inflammation and immunosuppression. Thus, donor-derived hematopoietic stem cells recruited to sites of chronic inflammation yielded epithelial tumors.
MOZ and MOZ-CBP Cooperate with NFκB To Induce Expression of NFκB-Dependent Genes.
Authors: Chan Edward M. , Chan Rebecca J. , Comer Elisha M. , Goulet Robert J. , Crean Colin D. , Brown Zachary D. , Fruehwald Amy M. , Yang Zhenyun , Boswell Howard S. , Nakshatri Harikrishna , Gabig Theodore G. 
Published: October 2019

Abstract
Although monocytic zinc finger protein (MOZ/MYST3) maintains normal hematopoietic stem cells, its fusion to the coactivator CREB-binding protein (CBP/CREBBP) induces acute myeloid leukemia (AML). Since leukemic stem cells in AML often exhibit excessive signal-dependent activity of the transcription factor NFκB, we hypothesized that cooperation between NFκB and MOZ-CBP represents an alternative mechanism for enhancing NFκB transcriptional activity. In reporter assays, MOZ and CBP separately induce transcription from the NFκB-dependent interleukin-8 promoter; however, these two proteins together markedly activate its expression. Although MOZ has less potent transcriptional activity than MOZ-CBP, both proteins cooperate with steroid receptor coactivator-1 (SRC1/NCOA1) to activate transcription. Since cooperation between MOZ-CBP and SRC1 is strongly reminiscent of the interaction between MOZ-TIF2/SRC2 and CBP required to induce AML (Deguchi et al, 2003), these findings suggest that MYST family fusion proteins may assemble a conserved leukemogenic transcriptional complex composed of a MYST protein (MOZ, MORF), molecular integrator (CBP, p300), and nuclear receptor coactivator (SRC1, TIF2/SRC2). MOZ also induces multiple NFκB-dependent viral promoters. Importantly, MOZ associates in a protein complex with the DNA-binding p65/RELA subunit of NFκB in vivo and interacts directly with p65 in vitro. Whereas deletion of the leukemia-associated protein (LAP/PHD) or MYST domains decreases the transcriptional activity of MOZ, deletion of either the acidic domain or C-terminal domain completely abrogates its activity. Since the C-terminal domain is absent from MOZ-CBP, these results indicate that the potent transcriptional activity of MOZ-CBP represents a gain-of-function property derived from the retained portion of CBP. Collectively, these studies not only demonstrate that MOZ and MOZ-CBP cooperate with NFκB to enhance expression of NFκB-dependent promoters but also suggest that aberrant interaction between MOZ-CBP and NFκB may play an important role in the pathogenesis of certain acute myeloid leukemias.
Novel Concentration-Dependent Roles for Heparin in Modifying LPS-Induced Activation of Mononuclear Cells in Whole Blood.
Authors: Hochart Helene , Jenkins Vince , Smith Owen P. , White Barry , O’Donnell James 
Published: October 2019

Abstract
Background: In addition to their etsblished anticoagulant activity, unfractionated heparin (UFH) and low molecular weight heparin (LMWH) are known to possess clinically important immuno-modulatory properties. However different studies have reported conflicting pro- and anti-inflammatory effects in association with heparin. Moreover, the molecular basis for these heparin effects on inflammation remains unclear. In view of the wide and diverse clinical indications for heparin, it is clearly of direct translational relevance to define how UFH and LMWH differentially regulate inflammatory responses to LPS in-vivo.
Objectives: To determine how UFH and LMWH regulate lipopolysaccharide (LPS)-induced activation of human mononuclear cells in whole blood, and define the role of lipopolysaccharide binding protein (LBP) in mediating this effect.
Methods: Whole blood was pre-treated with UFH or LMWH (0.1–200 IU/ml), prior to stimulation with LPS (10ng/ml). After 6 hours, monocyte pro-inflammatory cytokine (interleukin (IL)-1b, IL-6, IL-8, and TNF-a) secretion was determined by plasma ELISA. Parallel experiments using THP-1 cell line and primary monocytes were performed under serum-free conditions, in the presence or absence of varying doses of recombinant human LBP (range: 50–100nM).
Results: Under serum-free conditions, heparin demonstrated dose-dependent anti -inflammatory effects, significantly reducing secretion of pro-inflammatory cytokines (IL-1b, IL-6, IL-8, and TNF-a) in response to LPS-stimulation of THP-1 cells and primary monocytes. In contrast, in the presence of LBP, both UFH and LMWH demonstrated dose-dependent pro-inflammatory effects at all heparin concentrations. In ex-vivo whole blood experiments, pro-inflammatory effects (increased IL-1b and IL-8 following LPS-stimulation) of heparin were also observed, but only at supra-therapeutic doses (10–200IU/ml).
Conclusion: In keeping with previous reports, we have demonstrated that both UFH and LMWH can significantly down-regulate cytokine (TNF-a, IL-1b, IL-6 and IL-8) secretion in response to LPS-activation in-vitro. However our novel data demonstrate that the effect of heparin on monocyte activation by LPS is significantly more complex in the setting of whole blood. Firstly, in contrast to the anti-inflammatory effects observed under serum-free conditions, we found that in whole blood, high concentrations of heparin exerted marked pro-inflammatory effects. Secondly we have also demonstrated that the effects of heparin in whole blood are entirely dependent upon heparin concentration and LBP concentration.
Platelets and Heparin Induced Thrombocytopenia Antibodies Do Not Influence the Inhibitory Activity of Argatroban on Thrombin Generation.
Authors: Gerotziafas Grigoris T. , Roman Marie-Paule , Verdy Elisabeth , Hatmi Mohamed , Samama Meyer M. , Elalamy Ismail 
Published: October 2019

Abstract
Argatroban is a synthetic, reversible, direct thrombin inhibitor (DTI) used in patients with heparin induced thrombocytopenia (HIT). Argatroban as other DTIs prolongs PT, aPTT and ecarin clotting time. Argatroban added in vitro in normal platelet poor plasma (PPP) inhibits tissue factor (TF) triggered thrombin generation (TG) in a concentration dependent manner. We studied the influence of platelets, HIT antibodies and residual heparin, on the inhibitory effect of argatroban on TG. Argatroban (0 to 2 μM) was added in normal platelet rich plasma (PRP) and PPP, in pool PPP from three consecutive HIT patients (HIT-PPP) and in HIT-PRP prepared by mixing normal PRP with HIT-PPP (v/v). HIT-PRP and HIT-PPP were containing residual heparin (0,035 and 0,07 anti-Xa IU/ml respectively). All experiments were repeated 3 times. TG was triggered in the presence of TF (Dade-Behring Innovin; 1/1000 final dilution in plasma) and assessed with Calibrated Automated Thrombinoscope®. TG in PPP was triggered by adding PPP reagent (Thrombogram-Thrombinoscope®). Lag time (LT), time to peak (ttP), peak (P), endogenous thrombin potential (ETP) and mean velocity index (MVI) of thrombin generation were measured. The concentrations of argatroban which prolonged 2-fold the LT and the ttP and which inhibited 50% (IC50) the P, ETP and MVI were calculated. In the presence of low argatroban concentrations (0,1 an 0,2 μM) an artifactual increase of TG was observed. This is probably due to the interference of alpha2macroglobulin-bound thrombin with the fluorogenic substrate (as shown by Hemker’s group for other reversible DTIs). Argatroban at 1 μM inhibited TG by 50% in both normal PRP and PPP. Argatroban at 1 μM induced a 2-fold prolongation of aPTT (Table 1). HIT antibodies did not modify the inhibitory activity of argatroban on TG in HIT-PRP and HIT-PPP. The presence of traces of heparin in plasma from HIT patients had a synergistic effect with argatroban on the inhibition of TG (Table 1). Our in vitro study shows that argatroban at concentration achieved in clinical practice (about 1 μM) induces 50% inhibition of TG. The inhibitory activity of argatroban on TG is not modified by the presence of platelets or HIT antibodies. In contrast, traces of residual heparin in plasma from HIT patients amplify the inhibition of thrombin generation induced by argatroban. This finding has to be confirmed in a prospective clinical study since it implicates an increased vigilance during the switch from heparin to argatroban in acute HIT patients.
Table 1. Inhibitory activity of argatroban on thrombin generation. normal PRP normal PPP HIT-PRP (0,03 aXa/ml) HIT-PPP (0,07 aXa/ml) Lag-time x2 0,7 μM 0,7 μM 0,9 μM 0,1 μM ttPeak x2 1 μM 1 μM 1 μM 0,2 μM Peak IC50 1 μM 1 μM 0,7 μM 0,3 μM ETP IC50 1 μM 1 μM 0,7 μM 0,3 μM MRI IC50 1 μM 1 μM 0,5 μM 0,3 μM PTx2 - 1 μM - - aPTT x2 - 1 μM - -
The Reliability of Point-of-Care (POC) Prothrombin Time Testing - A Comparison of CoaguChek XS® INR Measurements with Hospital Laboratory Monitoring.
Authors: Ryan Fiona , O’Shea Susan , Ortel Thomas L. , Byrne Stephen 
Published: October 2019

Abstract
Background The development of Point of Care (POC) testing devices enables patients to test their own International Normalised Ratio (INR) at home, and has lead to the development of patient self- testing (PST) programmes. This facilitates more frequent testing of anticoagulation levels which in turn improves time in therapeutic range (TTR) and reduces bleeding and thrombotic complications in patients on oral anticoagulation therapy (OAT).1 External quality control of POC devices is essential to ensure that results are robust. One approach is to collect a venous sample at the same time as POC testing, which is analysed using an appropriate hospital laboratory analyzer. It has been recommended that INR results should be within 0.5 of each other when measured by each method2. However, previous studies have shown that when compared with clinical laboratory values, POC devices may show statistically significant differences between the INR values.3 Aim The aim of this study is primarily to compare TTR and adverse events between two different approaches to OAT management and more specifically in this presentation to examine the intra patient variability of INR measurements performed by two different techniques.
Methods 160 patients routinely attending the anticoagulant clinic of Cork University Hospital (CUH) were enrolled to take part in a prospective randomized controlled, cross-over design trial of supervised PST of warfarin therapy using an internet based expert system (Coagcare®, ZyCare, Inc) and POC testing device (CoaguChek XS®, Roche, UK) vs. rountine medical care in CUH. During the 6 months PST arm of the study a venous blood sample was taken from each patient for laboratory determination of the INR on 3 occasions; at enrolment, 2 months and 4 months. This measurement was then compared with the INR measured on the Coaguchek XS® machine using capillary blood from the patient’s fingertip.
Results 88 patients have being enrolled in the PST arm of the study to date and 454 POC measurements have been compared with laboratory values. Good correlation has been shown between the two methods of INR determination (r2 = 0.8314), however, statistically significant differences did occur (p < 0.005). A Bland and Altman plot showed good agreement between the dual measurements at values between 2.0–3.5 INR units. However there seemed to be increasing disagreement between the two methods as the INR rose above 3.5. Four patients were excluded from the study because the differences in the INR measurements were greater than 0.5 INR units. Conclusion POC testing devices for OAT are reliable and safe. External quality assessment should be inherent to any programme involving these devices. Clinicians involved in the use of POC anticoagulation monitoring should be aware that POC and laboratory values may differ and intervene if there is not adequate correlation between the two INR measurements.
Acquired Genetic Abnormalities in Acute Lymphoblastic Leukaemia in Patients with Down Syndrome.
Authors: Kearney Lyndal , Horsley Sharon W. , Bateman Caroline M. , De Castro David Gonzalez , Young Bryan D. , Chaplin Tracy , Eguchi Minenori , Greaves Mel 
Published: October 2019

Abstract
Children with Down syndrome (DS) have a 30 fold increased risk of developing leukaemia compared to their non-DS counterparts. While it is now known that cooperating mutations in the haemopoietic transcription factor GATA1 occur in all cases of acute megakaryoblastic leukaemia in Down syndrome patients, the additional genetic events which confer an increased risk of acute lymphoblastic leukaemia in Down syndrome (ALL-DS) are unknown. We initiated a search for mutations in the coding region of candidate genes including RAS, B-RAF, FLT3, KIT and JAK2 in a series of ALL-DS cases. No mutations were identified. We then carried out high resolution (Affymetrix 250K Nsp and 250K Sty) SNP array analysis of leukaemic blast cell DNA from 9 cases of ALL-DS, the majority of which had matched remission DNA. Overall, the whole and partial chromosome gains and losses were in agreement with the karyotype, but in all cases these were updated and refined. There were between 1 and 12 additional copy number alterations per case, with small focal deletions comprising 1 or 2 genes being more frequent than gains. The most common of these was a focal deletion of the CDKN2A gene (4 cases), all of which had either a partial deletion or copy number neutral LOH of the whole of 9p. The other most common focal deletions were of 12p13.3 (ETV6 gene) and 9p13.2 (PAX5), found in 2 cases each. Other regions affected included 3q13.2 (BTLA), 5q33.3 (EBF1), 13q14.2 (RB1) and 20p12.2 (including c20orf94), identified in 1 case each. These results indicate that the secondary genetic events in ALL-DS are similar to those for ALL overall (Mullighan et al., Nature446: 758–764, 2007). The number and pattern of submicroscopic genetic abnormalities more closely resembles that of ETV6-RUNX1 positive ALL than high hyperdiploid ALL (which includes acquired trisomy 21).