bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of the Leukemia-Specific Domains of the CALM/AF10 Fusion Gene, a Product of the Leukemia Associated T(10;11) Translocation.
Authors: Deshpande Aniruddha J , Arseni Natalia , Lin Yihui , Xu Guoliang , Hiddemann Wolfgang , Bohlander Stefan , Feuring-Buske Michaela , Buske Christian 
Published: April 2020

Abstract
We have demonstrated that the expression of the CALM/AF10 (C/A) fusion gene in murine BM cells results in an aggressive acute myeloid leukemia (AML). We sought to identify the domains involved in leukemogenesis mediated by the CALM/AF10 fusion gene. For this purpose we employed the CFU-S assay in which it was observed that C/A transduced BM cells generated an average of 75±23 day 12 CFU-S colonies per input 10^5 cells as compared to an average of 2 ±3 colonies with cells expressing the empty GFP vector (~ 37 fold increase; P &lt; 0.0005) in the number of day-12 CFU-S content. Expression of the CALM gene truncated to the breakpoint of CALM/AF10 (CALMdelta3’) or the CALM/ AF10 fusion gene with a deleted octapeptide motif - leucine zipper domain (CALM/AF10 delta OM-LZ) gave 10 (±11) and 12 (±11) day12 CFU-S respectively per input 10^5 bone marrow cells showing that the AF10 portion of C/A, especially the OM-LZ region is necessary for the observed enhancement of d-12 CFU-S. BM cells transduced with a construct harboring the CALM gene fused only to the OM-LZ domain of AF10 (CALM+ OM-LZ) showed 68(±5) d-12 CFUS per input 10^5 cells (~34 fold Vs MIG; P=0.0006) comparable to the C/A fusion gene.
We observed that the C/A fusion gene fails to show leukemic transformation of BM progenitors in CFC or proliferation assays in vitro in contrast to its striking leukemogenic potential in vivo. We tested different mutants of the C/A fusion gene using these assays and observed that the expression of a mutant of the CALM/AF10 fusion gene with a C-terminal portion of CALM (amino acids 400 – 648) fused to the OM-LZ motif of AF10, showed a significant increase in the number of secondary CFCs (32 fold Vs MIG; 15.38 fold Vs C/A), with the appearance of predominantly blast-like colonies. Interestingly, this mutant could also initiate leukemias in mice (n=5) with a latency and phenotype similar to mice injected with C/A transduced BM cells.
Taken together, we demonstrate that the OM-LZ domain of AF10 is necessary for the expansion of early hematopoietic progenitors by CALM/AF10 and also sufficient for in vivo leukemic transformation. We also demonstrate that the deletion of amino acids 1 to 410 of CALM confers in vitro transformation potential to the C/A fusion gene. Our data identify the domains of C/A that are crucial for leukemogenesis and provide insights into the mechanism of transformation in t(10;11) (p13;q14) positive leukemia.
Free Protein S Levels on the Risk of First Venous Thrombosis and Recurrence. Results from a Retrospective Family Cohort Study in 1143 Relatives.
Authors: Lijfering Willem M , Mulder René , Kate Min Ki ten , Mulder Andre , van der Meer Jan 
Published: April 2020

Abstract
Conflicting data have been reported on the risk for venous thrombosis in subjects with protein S type III deficiency. As some studies included subjects with protein S type I deficiency, combined with an inadequate cut-off level for defining protein S type III deficiency (i.e. lower limit in healthy volunteers, generally &lt; 65 IU/dL) might explain this discrepancy. We performed a single center retrospective study in a large cohort of families wherein probands had thrombosis and a thrombophilic defect, to define an optimal free protein S level that can identify subjects at risk for venous thrombosis. Probands were excluded as were relatives with protein S type I deficiency. Free protein S antigen levels were measured after precipitation of protein S complexed with C4b-binding protein with polyethylene glycol. 1143 relatives were analyzed. Mean age at onset of the first episode of venous thrombosis was 39 years. Relatives with venous thrombosis had lower free protein S levels than relatives without venous thrombosis (P&lt; 0.001), which was most pronounced in the lowest quartile (Figure). Only relatives with free protein S levels below the 5th percentile (&lt; 41 IU/dL) or below the 2.5th percentile (&lt; 33 IU/dL) were at higher risk of first venous thrombosis compared to relatives with free protein S levels in the upper quartile (³ 91 IU/dL); annual incidence 1.20% (95% CI, 0.72–1.87), and 1.81% (95% CI, 1.01–2.99), respectively; hazard ratio, adjusted for age, sex and thrombophilic defects 6.7, (95% CI, 3.3–13.5) and 11.3 (95% CI, 5.2–22.1). Recurrence rates in these subgroups were 12.12% (95 CI, 5.23–23.88) and 12.73% (95% CI, 5.12–26.22) per year. Relatives with free protein S levels lower than the 5th or 2.5th percentile had an adjusted 2.8 (95% CI, 0.8–8.6) and 3.1 (95% CI, 0.9–10.5) fold higher risk of recurrence compared to relatives with free protein S levels in the upper quartile. We conclude that decreased free protein S levels are a risk factor for venous thrombosis. However, the free protein S level to identify subjects at risk for venous thrombosis due to protein S type III deficiency lies far below the lower limit of the laboratory reference range obtained from healthy volunteers.
Figure. Cumulative distribution curve of free protein S levels in relatives of probands with a thrombophilic defect Figure. Cumulative distribution curve of free protein S levels in relatives of probands with a thrombophilic defect
Gray dashed, dashed dotted and dotted lines display free protein S level for relatives with and without venous thrombosis at cumulative frequency of 25%, 50% and 75%, respectively.
Decreased beta2-Glycoprotein I Plasma Levels as a Risk Factor for Myocardial Infarction in Men.
Authors: De Laat Bas , de Groot Philip G. , Derksen Ronald H.W.M. , Urbanus Rolf T. , Mertens Koen , Rosendaal Frits , Doggen Carine J.M. 
Published: April 2020

Abstract
Background: Several factors influence the occurrence of acute myocardial infarction. One of these factors is thought to be Von Willebrand Factor which serves as adhesive surface for platelets to adhere to the vessel wall. We have recently found that beta2- glycoprotein I is able to inhibit platelet binding to von Willebrand Factor by binding to the A1 domain of Von Willebrand Factor 1. This could indicate that beta2-glycoprotein I possesses antithrombotic properties with respect to arterial thrombosis. In the present study we investigated whether differences in beta2-glycoprotein I plasma levels influence the risk of myocardial infarction.
Methods and Results: We have measured beta2-glycoprotein I and Von Willebrand Factor antigen levels in 539 men with a first myocardial infarction and in 611 control subjects who participated in the case-control Study of Myocardial Infarction Leiden (SMILE). Although we did not find a profound effect of beta2-glycoprotein I plasma levels on myocardial infarction in the overall population (odds ratio 0.93, 95% confidence interval 0.65–1.33), there appeared to be a dose-dependent protective effect of increasing beta2-glycoprotein I plasma levels on myocardial infarction in men of 60 years and older. In this age group we found an odds Ratio of 0.44 (95% confidence interval 0.25–0.77) for high beta2-glycoprotein I levels compared to low levels. Furthermore, high plasma levels of beta2-glycoprotein I remained protective for myocardial infarction despite high levels of Von Willebrand Factor. In addition, we studied a possible association between age and Von Willebrand Factor and beta2-glycoprotein I plasma levels. It appeared that both Von Willebrand Factor and beta2-glycoprotein I plasma levels increased with age, but a larger increase in Von Willebrand Factor plasma levels was observed than in beta2-glycoprotein I plasma levels (13.7 % every 10 years versus 5.7% every 10 years).
Conclusions: In this study high circulating levels of beta2-glycoprotein I appeared to be associated with a lower risk of myocardial infarction in men over 60 years. In addition we observed a larger increase in Von Willebrand Factor levels with age than beta2- glycoprotein I levels. As beta2-glycoprotein I possesses antithrombotic properties by inhibiting the activity of Von Willebrand Factor in-vitro, this might indicate that during aging the haemostatic balance slowly shifts to a more prothrombotic state 1. Future in-vivo experiments are needed to investigate the exact contribution of beta2-glycoprotein I on the pathophysiology of myocardial infarction and arterial thrombosis in general.
Efficacy of Dasatinib in Patients (pts) with Previously Untreated Chronic Myelogenous Leukemia (CML) in Early Chronic Phase (CML-CP)
Authors: Cortes Jorge , O’Brien Susan , Borthakur Gautam , Jones Dan , Ravandi Farhad , Koller Charles , Mesina Ofelia , Ferrajoli Alessandra , Shan Jianqin , Kantarjian Hagop 
Published: April 2020

Abstract
Background: Dasatinib (BMS-354825) is a multi-targeted kinase inhibitor of BCRABL and SRC with significant activity in pts with CML-CP resistant to or intolerant of imatinib (IM). We initiated a phase II trial to study efficacy and safety of dasatinib in pts with previously untreated CML-CP.
Aims: To investigate the efficacy and safety of dasatinib as initial therapy for patients with CML-CP.
Methods: The primary objective was to estimate the proportion of pts attaining major molecular response (MMR) at 12 months (mo). Pts with previously untreated CML-CP were eligible and received dasatinib 100 mg/day, randomized to either 50 mg-twice-daily (BID) or a 100 mg-once-daily (QD).
Results: Fifty pts have been enrolled (25 on the QD schedule, 25 BID). Median age was 45 years (yrs) (range 18–76 yrs); 75% are Sokal low risk. Median follow-up is 24 months (mo). Overall, 44/45 (98%) evaluable patients achieved complete cytogenetic response [CCyR]. The CCyR rate at 3, 6 and 12 mo compares favorably to that observed in historical controls treated with imatinib 400mg or 800 mg daily:
Percent with CCyR (No. evaluable) Mo on therapy Dasatinib Imatinib 400mg Imatinib 800mg P value 3 78 (45) 37 (49) 62 (202) 0.0003 6 93 (41) 54 (48) 82 (199) &lt;0.0001 12 97 (35) 65 (48) 86 (197) 0.0001 18 88 (33) 68 (38) 89 (179) 0.004 24 80 (25) 70 (40) 88 (173) 0.006
MMR was achieved in 12/35 (34%) at 12 mo and 12/25 (48%) at 18 mo. One of 46 (2%) evaluable pts have achieved confirmed complete molecular response, and 1 other unconfirmed (ie, only achieved on their last assessment). Grade 3–4 non-hematologic toxicity (regardless of causality) included pruritus (13%), fatigue (6%), neuropathy (4%), and memory impairment (4%). Pleural effusion occurred in 21% evaluable pts (grade 3–4 in 2%). Grade 3–4 hematologic toxicity (transient) was thrombocytopenia in 11%, neutropenia in 21%, and anemia in 9%. Twenty-seven (54%) pts required transient treatment interruption. The actual median daily dose for all pts was 100mg. There is no significant difference in grade 3–4 toxicity by treatment schedule. Four pts came off study: 1 pts choice after 1 dose, 1 for toxicity (pleural effusion, QD schedule), and 2 lost response after multiple treatment interruptions (1 myelosuppression, 1 pleural effusion, both BID schedule). Two other pts have lost response because of non-compliance. 24 month EFS (event = loss of CHR, loss of MCyR, AP/BP, death, or off because of toxicity) is 81%.
Conclusion: Rapid CCyR occurs in most patients with previously untreated CML-CP treated with dasatinib frontline therapy with a favorable toxicity profile. Accrual to this trial continues.
Adamts-13 Gene Mutations and Transcription Analysis in Primary Antiphospholipid Syndrome.
Authors: Hancer Veysel sabri , Diz-Kucukkaya Reyhan , topal-Sarikaya Aysegul 
Published: April 2020

Abstract
Von Willebrand factor (vWF) is a large multimeric protein produced by endothelial cells and megakaryocytes. vWF mediates adhesion of plateletes at sites of vascular injury, and transports factor VIII. After secretion, vWF multimers are cleaved by ADAMTS-13 (A disintegrin –like and metalloprotease with thrombospondin type 1 motif 13) enzyme to smaller multimers. Decreased activity of ADAMTS-13 by autoantibodies, mutations and other etiologies causes thrombotic microangiopathy (intraluminal platelet thrombosis in the microvasculature, thrombocytopenia, and red blood cell fragmentation). Antiphospholipid syndrome (APS) is an autoimmune disorder characterized by recurrent arterial and venous thrombosis and fetal losses associated with elevated levels of antiphospholipid antibodies. The pathology of small vessel thrombosis in APS patients has been shown to be related with thrombotic microangiopathy. Recently, Austin et al reported that ADAMTS-13 activity was reduced in patients with APS. They also showed an increase of large vWF multimers in plasma of those patients. In this study, we aimed to investigate ADAMTS-13 mutations which decrease the activity of enzyme and the quantitative gene transcrition rates in patients with primary APS and healthy controls. We evaluated 70 primary APS patients (male to female ratio was 25/45), and age and sex matched healthy controls. Exons 10+11,12,13+14 of ADAMTS-13 gene were amplified with the PCR assay for 365del, Q449stop C508Y mutations and P475S polymorphism analysis. PCR products were cleaved with restriction digestion enzymes and genotyped. C508Y mutation was screened by a DNA sequencer system. Total RNA samples were extracted from peripheral blood and cDNA samples were synthesized with reverse transcriptase. Original primers and probes were designed for the target gene ADAMTS-13 and the reference gene HPRT- 1. cDNA amounts of target and reference genes in patient and healthy control groups were determined by real-time PCR. We did not find C365del, Q449stop mutations and P475S polymorphism in the control group and the patients with APS (Table1.) We used Pfaffl method for transcription analysis and found that mRNA amounts of ADAMTS-13 was significantly decreased in APS group compared to helathy controls (Table 1). Our results may suggest that ADAMTS-13 gene is down regulated at the transcription stage in patients with APS. Transcription factors or existence of other mutation(s) at promotor region may explain this effect. This study demonstrated that ADAMTS 13- vWF related mechanisms may have important role in the development of thrombosis in APS patients.
Table 1. Data of transcription analysis and mutations which affect the activity of ADAMTS13.
C365del Q449stop P475S C508Y ADAMTS13 CP* HPRT1 CP* ADAMTS13 mRNA ratio (PAPS/ Control) * Cross Point APS n=70 0 0 0 0 33,71 30,43 0,5 Control n=100 0 0 0 0 32,23 30,05
Thrombin Generation Is Normal in Most Patients with Cirrhosis despite a Prolonged INR.
Authors: Gatt Alexander , Riddell Anne , Calvaruso Vincenza , Makris Michael , Tuddenham Edward , Burroughs Andrew 
Published: April 2020

Abstract
Introduction: Advanced liver disease is associated with prolongation of the prothrombin time (PT). In order to decrease the inter-laboratory variability of PT measurement, the international normalised ratio (INR) calculated as the ratio of patient’s PT to a normal (control) sample, raised to the power of the international sensitivity index (ISI) of the particular thromboplastin used was developed. However, the ISI is derived from PT results of patients on warfarin and results cannot be extrapolated to liver patients. Despite this, the INR is still commonly performed to assess bleeding risk in patients with liver disease worldwide. Furthermore the INR is only affected by factors I, II, V, VII, and X and is not influenced by other factors such as factor VIII which is usually raised in hepatic cirrhosis. Recently it has been reportd that thrombomodulin addition (to take into account the protein C pathway) normalises thrombin generation (TG)1 despite these patients having a low TG if thrombomodulin is not used.
Aim: We speculated that TG, which is a global assay of coagulation and sensitive to all coagulation factors, when triggered by a low tissue factor (TF) concentration might not correlate with the INR in patients with liver disease and that contact inhibition with corn trypsin inhibitor (CTI) might better reflect the coagulation potential in this patient group.
Results: 73 unselected patients with liver cirrhosis due to various diseases and 25 normal subjects were studied. INR and TG using the calibrated automated thrombogram (CAT) at 1pM tissue factor (TF) with CTI, 5pM without CTI and with and without Protac (a Protein C activator) were performed using platelet poor plasma (PPP). The INR range was 0.8–4.0 (mean 1.6). At 5pM TF without Protac, the patient group had a significantly lower endogenous thrombin potential (ETP) than the controls (mean ETP difference 752nM/min; P &lt;0.0001). With Protac, no significant differences could be detected between the 2 groups. However, if the ETP without Protac was divided by the ETP with Protac x 100, the liver group showed more resistance to PC activation (mean % difference 25.4; P 0.0002). At 1pM TF, the mean ETP in the cirrhosis cohort was slightly lower than the normal group (difference between means 216nM/min; P 0.03). However, only 7 (9.6%) patients had ETP values less than the normal range (mean±2SD). No correlation was found between the ETP at 1pM and the INR. The mean FVIII:C was raised at 185.6 (78–420U/dl).
Conclusion: TG measured at low TF with CTI is normal in the majority of patients with cirrhosis. These patients are also more resistant to PC activation and have supranormal FVIII:C. Thus most patients have a normal or high thrombin potential despite an abnormal INR. These findings have important implications as in the absence of bleeding, “prophylactic” plasma and clotting factors are unnecessary.
Clinical and Laboratory Risk Factors for Venous Thromboembolism in Patients with High-Grade Gliomas: Results of a Prospective Study of 107 Newly-Diagnosed Patients.
Authors: Streiff Michael B. , Ye Xiaobu , Desideri Serena , Kickler Thomas , Jani Jayesh , Grossman Stuart 
Published: April 2020

Abstract
Introduction: Patients with malignant gliomas are at high risk for venous thromboembolism (VTE). The reason for this association is unclear. We sought to identify clinical and laboratory risk factors for VTE in adult patients with high-grade gliomas.
Methods: The NABTT CNS Consortium prospectively enrolled patients with newly-diagnosed grade 3 or 4 malignant glioma prior to anti-neoplastic therapy. Patients with a previous history of VTE, anti-neoplastic therapy or on chronic anticoagulation were excluded. At enrollment, we collected demographic and clinical information (age, gender, ethnicity, tumor histopathology and grade, Karnofsky Performance Status (KPS), and ABO blood group) and blood samples for measurement of factor VIII activity (FVIII), fibrinogen, and quantitative D dimer using standard laboratory assays. Endogenous thrombin potential (ETP) was measured using Innovin® and a synthetic chromagenic thrombin substrate on a BCS® coagulation analyzer (Dade Behring Inc. Newark, DE). Fisher’s exact test and the Student’s-T test were used for individual comparison of categorical data and continuous data, respectively. Cox regression modeling was used to examine the association of factors with VTE. The probability of thrombosis-free survival was estimated using the product-limit method of Kaplan and Meier.
Results: One hundred seven patients (49% male) with a median age 57 years (range 28–85) were enrolled between 6/05 and 4/08. Ninety patients (84%) had glioblastoma multiforme. Median KPS at enrollment was 90. After a median follow up of 324 days, twenty two patients (21%) have suffered VTE and 45 patients (42%) have died. Median time to VTE was 67 days post-operation (95% Confidence Interval 33–128). No fatal VTE have occurred. VTE was associated with a lower KPS (p=0.008), higher baseline FVIII (178% versus 151%, p=0.04) and greater ETP (473nmol/L versus 438nmol/L, p=0.04) but not with ABO blood group. Patients suffering VTE were more likely to die than patients without VTE (68% versus 35%, p&lt;0.005).
Conclusions: In a prospective cohort study of newly-diagnosed patients with high-grade gliomas, we have identified a substantial incidence of VTE. Patients with VTE had a lower initial KPS, higher factor VIII activity, greater ETP and were more likely to die than patients without VTE. With additional follow up, we hope to identify additional clinical and laboratory risk factors for VTE. These data should lead to greater insight into the pathogenesis of VTE in patients with high-grade gliomas and facilitate identification of patients in whom primary long-term VTE prophylaxis may be beneficial.
Lactadherin and Clearance of Platelet-Derived Microvesicles.
Authors: Dasgupta Swapan Kumar , Abdel-Monem Hanan , Niravath Polly , Le Anhquyen , Bellera Ricardo , Langlois Kimberly , Nagata Shigekazu , Rumbaut Rolando , Thiagarajan Perumal 
Published: April 2020

Abstract
Objective— In platelets, as in most mammalian cells, the anionic phospholipids such as phosphatidylserine are present only in the inner leaflet of the membrane bilayer. During platelet activation, phosphatidylserine moves from the inner to the outer leaflet of the membrane bilayer. The transbilayer movement of phosphatidylserine is responsible for platelet procoagulant activity as the exposed phosphatidylserine provides high affinity binding sites for the assembly of the prothrombinase and tenase complex. Externalization of anionic phospholipids in platelet is accompanied by the release of phosphatidylserine-rich microvesicles. These microvesicles account for the procoagulant activity of plasma by providing an efficient catalytic surface.
Lactadherin, also known milk fat globule-EGF 8, is a 45 kDa glycoprotein secreted by macrophages. Lactadherin contains EGF-like domains at the amino terminus and two C-domains at the carboxy terminus that share homology to the phosphatidylserinebinding domains of blood coagulation factors V and VIII. Lactadherin binds to apoptotic lymphocytes and phosphatidylserine-expressing red blood cells via the C-domains and anchors them to macrophage integrins via its RGD sequence in the EGF domain. We have examined the role of lactadherin in clearance of phosphatidylserine-rich platelet-derived microvesicles.
Methods and Results—Platelet-derived microvesicles were labeled with the fluorophore BODIPY-maleimide and incubated with THP-1 cell derived macrophages. The extent of phagocytosis was quantified by measuring the intracellular fluorescence by flow cytometry. Lactadherin promoted phagocytosis in a concentration-dependent manner with a half-maximal effect at ~ 5 ng/ml. A monoclonal antibody to lactadherin and a carboxy terminal fragment of lactadherin inhibited lactadherin-dependent phagocytosis. Lactadherin-deficient mice had increased number of microvesicles in their plasma and generated more thrombin compared to their wild type littermates. In addition, splenic macrophages from lactadherin-deficient mice showed decreased capacity to phagocytose platelet microvesicles. Finally, in a in vivo model of light/dye-induced endothelial injury/ thrombosis model, lactadherin-deficient mice, showed enhanced thrombus formation (5.93 ± 0.43 min) compared to their wild-type littermates ( 9.80 ± 1.14 min; P=0.01, n=9 in each group) in the cremastric venules.
Conclusion— Our studies show that lactadherin mediates the clearance of PS expressing platelet-derived microvesicles from the circulation and that a defective clearance can induce a hypercoagulable state.