bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enoxaparin Produces Sustained Antithrombotic Effects in Comparison to Fondaparinux and Rivaroxaban: Clinical Implications.
Authors: Jeske Walter , Masse Nicholas , Emanuele Joseph , Zhu He , McGeehan Elizabeth , Fareed Jawed , Bakhos Mamdouh , Walenga Jeanine M. 
Published: October 2019

Abstract
Abstract 3128
Poster Board III-65
Enoxaparin (Lovenox®), fondaparinux (Arixtra®) and rivaroxaban (Xarelto®) have been shown to be effective in mediating antithrombotic effects in post-surgical indications. Because of marked differences in the pharmacokinetic and pharmacodynamic behavior of these drugs we hypothesized that after the last dosage, the duration of the residual antithrombotic activity of the various drugs also differs. In order to compare the duration of effect of these three drugs, a rabbit stasis-thrombosis model (RSTM) utilizing FEIBA as the thrombogenic challenge and a rabbit ear bleeding model (REBM) were employed. Individual groups of rabbits (n=5) were treated with doses to mimic prophylactic, therapeutic and supratherapeutic (5x therapeutic) drug levels. Enoxaparin and fondaparinux were administered subcutaneously and rivaroxaban was administered by oral gavage. Blood samples were collected at baseline and at 16-18 hours post-administration of the last dose on day 1 or after 4 days of repeated dosing in the RSTM or at 3 hours post-administration in the REBM. At the 16-18 hour time point, circulating anti-Xa levels were not observed in any of the treatment groups. Despite this, enoxaparin treated animals exhibited a strong antithrombotic response following administration of the therapeutic dose (clot score = 1.3 ± 0.6 vs. 2.7 ± 0.6 for saline controls). Dosing once daily for four days did not enhance the antithrombotic activity of enoxaparin. In the plasmatic thrombin generation assays, a reduction in thrombin generation was noted in samples from enoxaparin-treated animals. For enoxaparin, no increase in bleeding was observed at prophylactic or therapeutic doses when compared to saline. An increase in bleeding (∼5 fold vs. saline) was observed at 3 and 6 hours post-administration of the ‘overdose’. A dose-dependent increase in anti-Xa activity was observed in blood samples collected 3 hours post-administration. For fondaparinux, no increase in bleeding was observed with any of the doses tested. There was a moderate increase in anti-Xa activity in the samples from rabbits treated with the ‘overdose’ of fondaparinux. This level was comparable to that observed with the therapeutic dose of enoxaparin. These studies suggest that enoxaparin produces a sustained antithrombotic effect in contrast to rivaroxaban and fondaparinux. These observations underscore that residual/sustained antithrombotic effects of enoxaparin may be partly responsible for the prolonged antithrombotic actions associated with the clinical use of enoxaparin.

Disclosures
No relevant conflicts of interest to declare.
Age of Blood as a Limitation for Transfusion: Potential Impact On Blood Inventory and Availability.
Authors: Fontaine Magali J. , Chung Yenho T. , Erhun Feryal , Goodnough Lawrence 
Published: October 2019

Abstract
Abstract 3142
Poster Board III-79

Background
Evolving concerns about storage lesions for red blood cells (RBC) and their effect on patient outcomes have led to ongoing trials evaluating the benefits of transfusing fresher blood (less than 14 days) to acutely ill patients. If improved outcomes with fresh blood are clearly demonstrated, current inventory management strategies may have to be revisited.

Methods
We evaluated several RBC maximum shelf lives (MSLs) and their impact on RBC availability. First, we determined the average age of the RBC units in our inventory based on the ABO/Rh blood type by analyzing the data set of 18,987 RBCs transfused from April 08, 2008 to April 09, 2009. Secondly, we determined the feasibility of issuing RBC of designated age to patients using the same data set.

Results
We found that 80% of the RBC inventory was delivered before 14 days of age (DoA), and that 42% versus 65% was issued before 14 DoA versus 21 DoA, respectively. RBC units spent on average 8.6 days on the shelf with a mean age at delivery of 10.2 days and a mean age at issue for transfusion of 18.8 days which varied depending on the ABO blood type as described in Table 1.
We then defined four scenarios where RBCs have different MSL: Scenario #1, #2, and #3 used a MSL of 7, 14, and 21 days, respectively. Scenario #4 used a combination of different MSLs depending on the category of patients, as follows: 7 days for 12% of patients (i.e., neonates), 14 days for 15% of patients (i.e., adult cardiovascular surgical patients), and 42 days for the 73% remaining patients receiving RBCs for transfusion. The simulations were performed using the original 18,987 data sets and results are summarized in Table 2 showing a significant decrease in the availability of RBC units by 51%, 20%, 10% and 0% for Scenario #1, #2, #3, and #4, respectively. Furthermore, the outdate rate would increase to 3.2%, 2.2%,1.0%, and 4.4% for the respective scenario.

Conclusion
Changing the maximum shelf life for red blood cell units will require novel approaches to RBC inventory management in order to meet hospital demands without risking a major shortage and putting patients at risk.

Disclosures
No relevant conflicts of interest to declare.
Covalent Antithrombin-Heparin Complex Catalysis of Activated Protein C Inhibition by Protein C Inhibitor.
Authors: Van Walderveen Maria Christina , Berry Leslie R. , Chan Anthony 
Published: October 2019

Abstract
Abstract 3167
Poster Board III-107

Introduction
Thrombin (IIa), when bound to thrombomodulin (TM), readily converts protein C (PC) into activated PC (APC). APC functions as an anticoagulant by inactivating activated factors V (FVa) and VIII (FVIIIa), and indirectly reduces IIa generation. Once formed, APC activity is controlled through inhibition by PC inhibitor (PCI), a reaction that is catalyzed by heparin (H). Chan et al. developed a covalent antithrombin-heparin (ATH) complex with increased anticoagulant activity compared to H. The current investigation looked at the role and mechanism by which ATH affects the inhibition of APC by PCI.

Methods
Discontinous second order rate constant inhibition assays of APC+PCI, in the presence of ATH or unfractionated H (UFH), were performed. These experiments were repeated in the presence of low molecular weight H (LMWH) or ATH (LMWATH), or a high molecular weight ATH (HMWATH). The affinity of ATH or UFH for APC or PCI was assessed using native electrophoresis and APC or PCI immobilized onto agarose beads.

Results
Second order rate experiments revealed that, at peak values, ATH (k2=2.0 × 107 ± 1.2 × 106) was a significantly slower catalyst of APC inhibition by PCI than UFH (k2= 3.0 × 107 ± 2.0 × 106; p=0.005). The peak reaction rate value for UFH occurred at a higher catalyst concentration (300nM) compared to ATH (60nM). LMWH was a poor catalyst of APC inhibition by PCI (k2= 4.2 × 105 ± 8.2 × 104). However, both LMWATH and HMWATH had high catalytic function (k2= 4.1 × 107 ± 2.1 × 106 and k2= 2.7 × 107 ± 4.8 × 105, respectively). Although UFH and ATH were able to bind to either APC or PCI at a pH of 7.3, increasing the pH to 8.8 eliminated all binding affinity, except that of ATH for APC.

Conclusion
UFH functions mechanistically through a template-mediated effect by bridging APC with PCI. The catalytic function of UFH is dependent upon H chain length, as LMWH was ineffective at enhancing APC inhibition by PCI. On the other hand, ATH may act mainly through conformational APC activation. There was no H chain length dependence for ATH, as LMWATH was able to significantly catalyze PCI inhibition of APC. We speculate that ATH's AT moiety may assist interaction with APC but prevents the H component of ATH from bridging the enzyme with the inhibitor. Compared to UFH, ATH is a more potent inhibitor of IIa, but is less effective at inhibiting APC and thereby decreasing IIa generation. These results add to the growing evidence that ATH is a superior anticoagulant to UFH.

Disclosures
No relevant conflicts of interest to declare.
Stat5 Is Essential for BCR-ABL-Transformed Chronic Myeloid Leukemia (CML) Associated with Increased CCN3 Gene Expression.
Authors: Zhu Bing-Mei , Wickre Mark , Na Risu , Klover Peter , Martin Cyril , Kimura Akiko , Hennighausen Lothar 
Published: October 2019

Abstract
Abstract 3271
Poster Board III-1

Introduction:
Signal transducers and activators of transcription5 (Stat5) proteins are involved in many cellular processes through mediated cytokine, hormone, and growth factor signaling. But its role in disease pathogenesis has not been fully elucidated. Recently, activation of BCR-ABL has been reported to regulate a novel gene, CCN3 in cell lines and primary cells derived from chronic myeloid leukemia (CML) patients. To investigate the function of Stat5 in CML initiation and maintenance and determine the downstream target genes on Jak-Stat5 pathway, we developed a BCR-ABL - induced CML - like disease model by using retro-viral infection in Cre-mediated Stat5 knockout transgenic mice and analyzed the progress of CML. We also used Stat5 knockout (Stat5 KO) mice to perform gene profiling. Results: Our study showed that loss of Stat5 resulted in increased survival rate and remission of CML. Microarray analysis showed that CCN3 expression was down-regulated in KL cells derived from Stat5 KO mice. BCR-ABL-activated Stat5 increased expression level of CCN3 in CML cells. We further determined that Stat5 binds to CCN3 promoter region in IL-3 stimulated 32D cells and BCR-ABL-induced CML cells. Conclusions: Our study suggested that Stat5 is essential for BCR-ABL transformed CML and that CCN3 is involved in normal hematopoiesis and CML development. Further study will be necessary to uncover the function of CCN3 and more targets of Stat5 pathway in CML development and discover the therapeutic significance.

Disclosures:
No relevant conflicts of interest to declare.
T/NK Lymphocytosis in CML Ph+ Patients During Dasatinib Therapy.
Authors: Rossi Antonella Russo , Ricco Alessandra , Carluccio Paola , Delia Mario , Leo Manuela , Casieri Paola , Stagno Fabio , Minervini Angela , Liso Vincenzo , Specchia Giorgina 
Published: October 2019

Abstract
Abstract 3279
Poster Board III-1

Background.
Some authors reported that Natural Killer (NK) cells from CML patients are defective in NK cell activity and NK cell number decrease as the disease progresses to the advanced phase and probably the abnormal BCR/ABL gene causes abnormal NK cell differentiation.TK inhibitors reduce BCR/ABL transcription and could restore NK cell numbers and/or function.Moreover Dasatinib,by the blockade of SRC Kinases,could affect the development of NK cells as well as T-lymphocytes.Our aim was to verify the impact of Dasatinib treatment on T CD8+ and NK cells modulation.

Methods.
We evaluated 24 patients with CML resistant/intollerant to Imatinib and treated with Dasatinib at a starting dose of 70 mg/BID or 100 mg/QD.Blood count were monitored; lymphocytosis has been definited by an increased number of peripheral blood lymphocyte counts ≥ 3.0×10(e)9/L and by the predominance of LGLs in peripheral blood smear. Immunophenotyping was done with flow-cytometry using antibodies against the following antigens: CD2, CD3, CD4, CD5, CD7, CD8, CD16, and CD56.

Results.
With a median of 19 mo. of Dasatinib therapy (range 3–43), 15/24 cases (62.5%) developed peripheral blood lymphocytosis. Median onset of lymphocytosis was 3 months after the initiation of Dasatinib therapy (range 1–12) and duration was 14 months (range 6–40).Lymphocytosis was CD3+/CD8+/Cytotoxic T Cell in 9 patients (60%) and CD3-/CD16+/CD56+/NK Cell in 6 patients (40%).In all 15 patients no symptoms or signs suggestive of LGL leukemia or viral infections were documented.There was no significant difference in terms of the frequency of severe adverse events, including pleural effusion between patients with and without lymphocytosis. 11(73%) of the 15 patients who developed lymphocytosis achieved MMolR and 4/11 presented Bcr/Abl mutation at the time of imatinib treatment (F317L, E255K, F359V, E255K),whereas only 3(33%) of the 9 patients without lymphocytosis achieved MMolR and 1/3 presented Bcr/Abl mutation (F359V). Moreover molecular response was earlier in the group of patients with lymphocytosis (8vs12 mo.).

Conclusions.
The development of lymphocytosis in our patients seems to be associated to an improved response to dasatinib in terms of molecular response and time to response. The assessment of higher frequency lymphocytosis requires further analysis; a larger patients'cohort should be needed to explore the biological role of lymphocytosis and the impact on the long term outcome in patients treated with dasatinib.

Disclosures:
No relevant conflicts of interest to declare.
High CD34 Yield Not Impaired by Melphalan 10mg/m2 Test-Dosing for Pharmacogenomics in M-VTD-PACE as Induction Therapy in Total Therapy 4 (TT4) for Low-Risk and Total Therapy 5 (TT5) for High-Risk Newly Diagnosed Multiple Myeloma (MM).
Authors: van Rhee Frits , Alsayed Yazan , Cottler-Fox Michele H. , Haessler Jeff , Waheed Sarah , Nair Bijay , Crowley John , Anaissie Elias , Barlogie Bart 
Published: October 2019

Abstract
Abstract 3411
Poster Board III-299

Introduction:
Based on profound outcome differences in Total Therapy 3 (TT3) among patients with gene expression profiling (GEP)-defined low- and high-risk groups, TT4 for low-risk MM randomizes patients between standard and light version of TT3, whereas TT5 for high-risk MM applies dose-dense treatment sequences. Both trials employ M-VTD-PACE combination therapy for induction followed by HPC collection with the objective of collecting a minimum of 20 × 106 CD34 cells per kg body weight. The M-VTD-PACE regimen includes M (melphalan, 10mg/m2) as a test-dose after a preceding bortezomib test-dose (1.0mg/m2), each of which is followed 48hr later by GEP studies to examine prognostically relevant and drug-unique GEP alterations. There was obvious concern that the M test-dose might interfere with HPC collection. Here, we examined the CD34 yield and the days required in the first 84 patients in comparison to predecessor TT3 protocols.

Methods:
Box-plots were used to display the total CD34 collections and the CD34 collection days. Comparisons were made using the Wilcoxon Rank Sum Test.

Results:
As is evident from the appended Figure, the addition of M did not adversely affect CD34 total yield or the kinetics of procurement. Results will be presented on the engraftment kinetics and early outcome results for this first personalized medicine trial in MM.

Conclusion:
We were able to introduce M pharmaco-genomic investigations in this first-in-kind risk-directed front-line protocol for MM without compromising HPC mobilization.

Disclosures:
van Rhee: Genzyme Corporation: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.
Bryostatin Enhances the Cytotoxic Effects of Anti-CD22 Immunotoxins in CLL by Two Distinct Mechanisms: Implications for a Sequential Therapy.
Authors: Ringshausen Ingo , Decker Thomas , Biberacher Viola , Meyer zum Buschenfelde Christian , Oelsner Madlen , Wagner Michaela , Bogner Christian , Peschel Christian 
Published: October 2019

Abstract
Abstract 3429
Poster Board III-317
Recently, treatment of CLL has been improved by the addition of Rituximab, a monoclonal antibody directed against CD20. Although Rituximab displays modest activity when used as monotherapy, the combination of Rituximab and chemotherapy has demonstrated impressive efficacy in CLL. However, even though remissions can be achieved in the majority of patients, CLL still remains an incurable disease for most patients. Therefore, alternative treatment options are needed. BL22 is a recombinant protein composed of the variable region of a monoclonal antibody, which binds to CD22 on the surface of normal and malignant B-cells and of PE38, a truncated Pseudomonas exotoxin. Previously, we demonstrated that BL22 induced cell death in CLL, involving the intrinsic apoptotic pathway. However, apoptosis induction correlates with expression of CD22 on the surface of CLL cells and was only moderate in CD22low expressing cells. The aim of this study was to increase BL22 cytotoxicity by modulating the surface expression of CD22 on CLL cells. Bryostatin is a macrocyclic lactone which structurally mimics the PKC-activating second messenger diacylglycerol. This PKC modulator has demonstrated anti-leukemic effects in CLL by itself in vivo. Here we show that Bryostatin up-regulates the expression of CD22 on CLL cells in a PKC dependent pathway, thereby increasing the cytotoxic effects of BL22. However, PKC-mediated up-regulation of Mcl-1 attenuated the anti-leukemic effects of BL22. We unravel that CD22 and Mcl-1 up-regulation by Bryostatin occurs in a dose dependent manner. Importantly, higher doses of Bryostatin prevented the undesirable up-regulation of Mcl-1 by inhibition of PKC in spite of CD22 up-regulation. Therefore, combined treatment of BL22 and Bryostatin seems to be a promising approach to enhance the cytotoxic effects of the immunotoxin by modulating PKC activity and CD22 up-regulation. In addition, our data provide evidence for a sequential therapy with both drugs, since CLL cells can be primed with Bryostatin. After a single dose of Bryostatin, PKC-modulation lasts for several days, allowing to administer BL22 at later time points, thereby decreasing the risk of potential drug interactions. Finally we demonstrate that stromal cell mediated drug resistance in CLL can be overcome by Bryostatin/ BL22 therapies. Conclusively, Bryostatin enhances the anti-leukemic effects of BL22 in CLL. Clinical trails are needed to prove if such an approach has clinical impact on the treatment and prognosis of B-CLL.

Disclosures
No relevant conflicts of interest to declare.
Enhanced In Vivo activation of Adoptively Transferred Genetically Targeted T Cells Following Cyclophosphamide Chemotherapy: Initial Results From a Phase I Clinical Trial Treating CLL Patients with Autologous CD19-Targeted T Cells.
Authors: Brentjens Renier J. , Hollyman Daniel , Park Jae , Santos Elmer , Yeh Raymond , Stefanski Jolanta , Taylor Clare , Weiss Mark A. , Filippa Daniel , Riviere Isabelle , Sadelain Michel W 
Published: October 2019

Abstract
Abstract 3436
Poster Board III-324
Patient T cells may be genetically modified to express chimeric antigen receptors (CARs) targeted to antigens expressed on tumor cells. We have initiated a clinical trial treating chemotherapy-refractory chronic lymphocytic leukemia (CLL) patients with autologous T cells modified to express the 19-28z CAR targeted to the CD19 antigen expressed on most B cell malignancies. In the first cohort of this trial, patients were infused with the lowest planned dose of modified T cells alone. All patients treated in this cohort experienced low-grade fevers following modified T cell infusion, and 2 of 3 treated patients exhibited subjective and laboratory evidence of transient reductions in tumor burden. The first patient treated on the second cohort of this study received prior cyclophophamide chemotherapy followed by the same dose of modified T cells administered to the first cohort of patients. This patient experienced persistent fevers, dyspnea, hypotension, renal failure, and died 44 hours following modified T cell infusion, likely secondary to sepsis. Modified T cells were not detectable in the peripheral blood of treated patients at 1 hour following completion of T cell infusion. However, post mortem analyses revealed a rapid infiltration of targeted T cells into anatomical sites of tumor involvement. Serum levels of the inflammatory cytokines IL-5, IL-8, and GM-CSF, but not TNFα, markedly and rapidly increased following infusion of genetically targeted T cells in this patient, mirroring the in vitro cytokine secretion profile of this patient's T cells, and consistent with marked in vivo activation of the modified T cells. Similar cytokine signatures were not found in patients from the first cohort. Significantly, serum cytokine analyses from the second cohort patient revealed a marked increase in the pro-proliferative cytokines IL-2, IL-7, IL-12, and IL-15 following cyclophosphamide therapy, in contrast to the baseline levels found in the first cohort. This report demonstrates the high efficiency trafficking of CD19-targeted T cells and in vivo activation of T cells encoding a second generation CD28/zeta chain-based chimeric antigen receptor. Furthermore, these data highlight mechanisms whereby cyclophosphamide may generate an in vivo milieu that enhances the anti-tumor efficacy of autologous tumor targeted T cells.

Disclosures
No relevant conflicts of interest to declare.
Rituximab Combining with Fresh Frozen Plasma for the Treatment of Patients with Refractory Advanced CLL.
Authors: Xu Wei , Li Jianyong , Miao Kourong , Hong Ming , Qiao Chun , Wu Yu-Jie , Fan Lei , Liu Peng 
Published: October 2019

Abstract
Abstract 3450
Poster Board III-338
Chronic lymphocytic leukaemia (CLL) is characterized by a progressive accumulation of monoclonal B lymphocytes in the bone marrow, lymphatic organs and blood. Even with the use of newer therapies, many patients with CLL develop progressive, treatment-resistant disease. Rituximab (RTX), a monoclonal antibody, targets CD20 on B lymphocytes and widely used in indolent B cell neoplasms. The lower density of CD20 on CLL cells may play a role, however, the causes of the lesser susceptibility of CLL to RTX remain poorly understood. Complement deficiencies have been identified in many CLL patients and appear to be more pronounced in patients with more advanced disease. Since complement mediated cell lysis is one of the major mechanisms of RTX action, we hypothesized that the therapeutic effect of RTX in CLL may be enhanced by the provision of complement through concurrent administration of fresh frozen plasma (FFP). Between August 2008 and April 2009, twelve patients with refractory advanced CLL were enrolled in this open clinical trial. All had been previously treated with fludarabine and two also failed treatment with RTX. The median age patients was 57 years (range, 48-82 years). According to the Binet staging system, 2 patients were in Binet B, 10 patients in Binet C. Florescence in situ hybridization (FISH) studies identified del(13q14) in 8 patients, and 4 patients with del(13q14) as the sole abnormality. Trisomy 12 was found in 3 patients, del(11q22.3) in 2 paitents, del(17p13) in 2 patients, del(6q23) in 1 patient, and IgH translocation in 1 patient, respectively. By flow cytometry, CD38 was positive in 9 patients, and ZAP-70 positive in 8 patients. Immunoglobulin heavy chain variable gene (IgVH) mutational status was detected by multiplex PCR, and six patients had unmutated VH. Two units of FFP followed with RTX 375 mg/m2 as a single agent, repeated every 1-2 weeks up to a total of 2-5 cycles. Complete response (CR) was achieved in 4 patients (33.3%), partial remission (PR) in 6 patients (50.0%). A major improvement of clinical signs and symptoms was achieved in all patients. Lymphocyte counts dropped markedly followed by shrinkage of lymph nodes and spleen and improvement of the anaemia and thrombocytopenia. This could be maintained over 9 months (range, 3-11 months) without progression. Toxicity was minimal and the treatment was well tolerated in all cases. It was show that RTX combining with FFP may provide a useful therapeutic option in patients with refractory advanced CLL.

Disclosures
No relevant conflicts of interest to declare.