bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Ratio of ADAMTS13 to VWF-Propeptide Can Reflect the Disease Severity and the Extent of Inflammation of the Patients with Severe Sepsis or Septic Shock,
Authors: Nishio Kenji , Fukushima Hidetada , Watanabe Tomoro , Seki Tadahiko , Matsui Hideto , Sugimoto Mitsuhiko , Matsumoto Masanori , Fujimura Yoshihiro , Okuchi Kazuo 
Published: October 2019

Abstract
Abstract 3349

Background:
Sepsis is a critical consequence of self defense to infection, which includes inflammatory and hypercoagulant response. Many stimulatory mediators involved in these responses cause the Weibel-Palade body exocytosis through endothelial activation, leading to surface expression of P-selectin and ultra-large von Willebrand factor (ULVWF), and simultaneously release of VWF-propeptide on a equimolar basis to ULVWF. Where ADAMTS13 is functionally inactivated, ULVWF would not be proteolysed and cause platelet thrombosis in the microcirculation, leading to organ failure. Therefore, the balance of the ULVWF and ADAMTS13 may indicate the prothrombotic state of the patients. However, it is difficult to estimate the amount of ULVWF expressed. It is reasonable to think that plasma VWF propeptide levels will reflect more precisely the extent of expressed ULVWF and P-selectin rather than VWF level. These allow us to speculate that the balance of VWF propeptide and ADAMTS13 can be associated with magnitude of prothrombotic and inflammatory response.

Aim:
To investigate whether the levels of molecules mentioned above are associated with the disease severity of sepsis or inflammation during the clinical course.

Method:
Inclusion criteria are as follows, 1) severe sepsis or septic shock defined by the Society of Critical Care Medicine Consensus Conference Committee 2) Platelets count less than 120,000/μL. SOFA(sequential organ failure assessment)score, APACHEII score, Creatinine, VWF propeptide, VWF, P-selectin, ADAMTS13, TNF-alfa, IL-6 were measured at day 0, 3, 5, 7 (day 0 is the day when the diagnosis as severe sepsis or septic shock was made), and each other's correlation was studied.

Result and Discussion:
VWF propeptide level was increased to 293.8+/−153.8% at day 0, and gradually decreased day by day. VWF was increased to 212.3+/− and slightly increased during the course. P-selectin was not increased during the course. The plasma level of ADAMTS13 at day 0 was decreased to 25+/− 8.7%, and increased during the course in the survivors, however almost not changed in the non-survivors. As we speculated, the value of VWF propeptide/ ADAMTS13 were well correlated to SOFA score, Creatinine, APACHEII, TNF-alfa almost on each study day.

Conclusion:
This result shows that the ratio of VWF propeptide to ADAMTS13 can be used to estimate disease severity of sepsis and magnitude of inflammation. This will improve our understanding of underlying pathophysiology of sepsis.

Disclosures:
Matsumoto: Alexion Pharma: Membership on an entity's Board of Directors or advisory committees. Fujimura:Baxter BioScience: Membership on an entity's Board of Directors or advisory committees; Alexion Pharma: Membership on an entity's Board of Directors or advisory committees.
Inhibition of Platelet Glycoprotein β3 by Chimeric Monoclonal Antibody Prevents Arterial Thrombosis in Beagle Dogs,
Authors: Ji Shundong , Jiang Miao , Dong Ningzheng , Bai Xia , Ruan Changgeng 
Published: October 2019

Abstract
Abstract 3357
In this present study in beagle dogs, we evaluated the antithrombotic efficacy of Pulaimab, the chimeric monoclonal antibodies SZ21-F(ab)2 fragments against platelet glycoprotein (GP) β3, in a modified Folts model. The Folts model is widely accepted to be effective and clinically relevant for testing potential anti-thrombotic agents, in this model the cyclic flow reductions (CFRs) are caused by platelet dependent thrombi that form under high-shear conditions at injured stenosed sites of an artery. Thirty beagle dogs of either sex, weighing 7.5 to 14 kg, were randomly divided into five groups of six (three females and three males), the details were following: negative control group (injected normal saline), positive control group (injected 0.2mg/Kg of Reopro), and three experimental groups [injected 0.2, 0.4, 0.8 mg/Kg of SZ21-F(ab)2, respectively]. A dose range from 0.2 to 0.8 mg/kg of Pulaimab significantly reduced the CFRs by 21–73%, without reduction of platelet numbers and prolongation of the bleeding time. Ex vivo ADP-induced platelet aggregation was equally reduced. The present study demonstrates that the inhibition of platelet GP αIIbβ3function by SZ21-F(ab)2 is a powerful intervention to prevent platelet thrombus formation in injured arteries without thrombocytopenia and prolongation of the bleeding time. We therefore conclude that F(ab)2 fragments of inhibitory anti-GPβ3 antibodies may be useful compounds to prevent thrombosis.

Disclosures:
No relevant conflicts of interest to declare.
Storage-Induced Changes Promote Erythrocyte Recognition by Autoantibodies,
Authors: Dinkla Sip , Novotný Vera M.J. , Joosten Irma , Bosman Giel J.C.G.M. 
Published: October 2019

Abstract
Abstract 3371
Physiological erythrocyte removal is associated with a selective increase in the expression of senescent cell antigens related to band 3, and subsequent autologous antibody binding and phagocytosis. Chronic erythrocyte transfusion often leads to immunization and the formation of alloantibodies and autoantibodies. In order to understand the molecular basis for this phenomenon we investigated whether erythrocyte storage leads to increased expression of non-physiological neoantigens.
Immunoprecipitations were performed with erythrocytes and vesicles from blood bank erythrocyte concentrates of increasing storage periods, using plasma containing erythrocyte autoantibodies from selected patients with autoimmune hemolytic anemia (N=6). Patient plasma autoantibody binding to erythrocytes increased with erythrocyte storage time, while the exact opposite was observed for plasma from healthy volunteers, showing that erythrocyte antigenicity changes during storage. SDS polyacrylamide gel analysis combined with proteomics revealed band 3 to be a dominant target of the autoantibodies. From vesicles that are formed during erythrocyte storage, the autoantibodies precipitated protein complexes distinct from those recognized on erythrocytes, indicating that they have a different immunization potential. Soluble immune mediators including complement factors were present in the patient plasma immunoprecipitates, but not in the control immunoprecipitates.
These findings support the hypothesis that disturbed erythrocyte aging during storage of erythrocyte units contributes to transfusion-induced alloantibody and autoantibody formation.

Disclosures:
No relevant conflicts of interest to declare.
The MEIS1 Interactome in Megakaryocytes Reveals a Role in Cell Cycle Regulation,
Authors: Salunkhe Vishal A , Macaulay Iain , Nuernberg Sylvia , McCarthy Cathal , Ouwehand Willem Hendrik , Fitzgerald Desmond J. , Maguire Patricia 
Published: October 2019

Abstract
Abstract 3380
Haematopoiesis is highly coordinated process of fate determination at branch points that is regulated by transcription factors and their cofactors. Our comprehensive catalogue of transcripts in the eight main mature blood cell elements, including erythroblasts and megakaryocytes (MKs) showed that the transcription factor MEIS1 is uniquely transcribed in MKs and the CD34+ haematopoietic stem cell. Gene silencing studies in mice and zebrafish has shown a pivotal role for MEIS1 in haematopoiesis, megakaryopoiesis and vasculogenesis, although its precise hierarchical position and function remain unknown. To gain further insight in the role of MEIS1 in megakaryopoiesis, we used a proteomics approach to search for its nuclear interaction partners.
Co-immunoprecipitation was used to isolate MEIS1 interacting proteins from the nuclear fraction of the MK cell line, CHRF 288–11 and resulting eluates were subjected to proteomics analysis using one-dimensional electrophoresis and liquid chromatography (LC) coupled to tandem mass spectrometry (MS) or GeLC-MS/MS. In total 70 proteins were identified to co-immunoprecipitate with MEIS1 from 3 replicate MS analyses. These included the previously validated MEIS1 interactors PBX1 and HOXB9, as well as numerous novel interactors such as ARID3B and DHX9.
Network analysis of our MEIS1 interactome dataset revealed a strong association with cell cycle regulation. In fact, we had identified a myriad of cell cycle regulators including CDK1, CDK2, CDK9, CUL3, PCNA, CDC5L, ARID3B and MDC1. These interactions are consistent with recent microarray studies in promyelocytic leukemic cell lines that link MEIS1 with cell cycle entry and its regulation of genes such as CDK2, CDK6, CDKN3, CDC7 and Cyclin D3 among others. To confirm the novel interaction of MK MEIS1 with cell cycle regulators we performed reverse immuno-precipitation/immunoblot analysis in CHRF cells and purified MEIS1 containing multiprotein complexes from L8057 murine megakaryoblastic cells. Using a cell cycle specific PCR array, we demonstrate that MEIS1 overexpression in L8057 cells regulates numerous cell cycle regulatory genes. Preliminary analysis using flow cytometry demonstrated that MEIS1 overexpression resulted in an altered cell cycle progression. Furthermore, genome wide ChIP-Seq analysis in CHRF cells for MEIS1 revealed binding sites in Cyclin D3 and CDK6, two known key regulators of the cell cycle and megakaryopoiesis.
Taken together this study provides evidence linking MEIS1 to the cell cycle control of MKs and will help elucidate the role of MEIS1 in cell cycle progression, megakaryopoiesis and associated disorders.

Disclosures:
No relevant conflicts of interest to declare.
PIM Kinases Inhibition, a Rational Strategy in Peripheral T-Cell Lymphomas,
Authors: Martin-Sanchez Esperanza , Rodriguez-Pinilla Socorro M. , Sanchez-Beato Margarita , Dominguez-Gonzalez Beatriz , Wozniak Magdalena B. , Mollejo Manuela , Alves Javier , Rodriguez-Peralto Jose Luis , Menarguez Javier , Cigudosa Juan C. , Ortiz-Romero Pablo L. , Garcia Juan F. , Bischoff James R. , Piris Miguel A. 
Published: October 2019

Abstract
Abstract 3494
The search of an efficient therapy for Peripheral T-cell lymphomas (PTCL) patients is still a challenge, in part due to the very little knowledge about the PTCL pathogenesis, and the absence of appropriate models. This heterogeneous group of very aggressive malignancies can not be cured with conventional therapies; therefore, new therapeutic strategies are needed to improve the poor outcome in these patients. PIM family is composed of 3 kinases (PIM1, PIM2 and PIM3) which play an essential role in cell proliferation and survival. They are mainly activated through JAK/STAT signaling pathway, and are frequently altered in human malignancies by amplification, mutation and overexpression. The aim of this study is to determine the efficiency and the mechanism of action of PIM inhibition in PTCL.
Gene expression profiling of twenty two PTCL cases and seven reactive lymph nodes was performed. We observed a strong overexpression of the three PIM family genes in PTCL cases, especially PIM2. In addition, Gene Set Enrichment Analysis identified an overexpression of STAT3 and IL-2 pathways in PTCL cases, probably responsible for the strong expression of PIMs we found. Furthermore, PIM genes expression was confirmed by quantitative RT-PCR in 6 PTCL-derived cell lines compared to normal T cells from healthy donors, highlighting again the relevance of PIM2.
Genetic inhibition was carried out using small interference RNA to specifically abolish the expression of each PIM1, PIM2 and PIM3 in a panel of 6 PTCL cell lines belonging to different PTCL subgroups. Additionally, pharmacological inhibition with one PIM inhibitor (ETP-39010) was performed.
Surprisingly, genetic inhibition of each of the PIM gene alone did not show any cellular effect, neither cell cycle arrest nor apoptosis. But interestingly, we found that specific inhibition of each of the PIM genes caused an increased expression of the other PIM family members, probably leading to a compensatory mechanism among these kinases balancing the lack of one of them, avoiding pro-apoptotic effects and allowing cell survival.
Accordingly, a simultaneous inhibition of PIM1, PIM2 and PIM3 using the pharmacological pan-PIM inhibitor produced a decrease in cell viability and a strong induction of apoptosis in all cell lines, without cell cycle arrest. Several PIM inhibitor biomarkers have been identified at the mRNA level, involving the DNA damage response signaling.
In conclusion, our results indicate that PIM kinases inhibition could be an effective therapeutic approach for PTCL.

Disclosures:
No relevant conflicts of interest to declare.
Gas6 Receptors, Tyro3, Axl and Mer, Differentially Participate in Glycoprotein VI-Mediated Platelet activation
Authors: Zhou Junsong , Wang Lu , Wu Yi 
Published: October 2019

Abstract
Abstract 364
Growth arrest-specific gene 6 (Gas6) has three receptors on platelets - Tyro3, Axl and Mer. It has been shown that these three receptors are equally important in platelet activation (J Clin Invest. 2005;115:237–46), and they synergizes with ADP receptor for platelet activation, but not with collagen receptor (J Thromb Haemost. 2010;8:1797–808). Considering the fact that Tyro3, Axl or Mer plays a distinct role in the function of endothelial cells and leukocytes, we hypothesized that these three receptors play specific role in platelet activation, and examined the role of each receptor in Gycoprotein VI (GPVI)-mediated platelet activation using single receptor knockout mice. In response to 100 ng/ml convulxin or 5 ng/ml poly(PHG), a GPVI-specific synthetic peptide, the aggregation of Axl−/− or Tyro3−/− platelets was almost completely prevented. However, the aggregation of Mer−/− platelets was only delayed, and still could reach the maximum level. In the presence of 100 ng/ml convulxin or 5 ng/ml poly(PHG), the spreading of Axl−/− or Tyro3−/− platelets, but not Mer−/− platelets, on fibrinogen-coated surfaces was markedly inhibited. Moreover, convulxin-stimulated P-selectin expression and JON/A binding were both significantly decreased in Axl−/− and Tyro3−/− platelets, but not in Mer−/− platelets. Upon stimulation with 100 ng/ml convulxin, tyrosine phosphorylation of Syk, linker for activation of T cells (LAT), SLP-76(SH2 domain-containing leukocyte protein of 76) and phospholipase C (PLCγ2) was markedly reduced in the Axl−/− or Tyro3−/− platelets, but not in the Mer−/− platelets. Interestingly, anti-Axl or anti-Tyro3 antibody (20 mg/ml) completely blocked convulxin-stimulated human platelet aggregation, however, the neutralizing anti-Gas6 antibody (60 mg/ml) did not have any effect, suggesting the homodimer formation of Axl or Tyro3 is involved in GPVI-mediated platelet activation. As detected by flow cytometry, there was no difference in the expression of GPVI and integrin aIIbb3 on membrane surface among Tyro-3−/−, Axl−/−, Mer−/− and wild type platelets. In conclusion, Gas6 receptors are not equally important in platelet activation, Axl and Tyro3 play more critical role than Mer in GPVI-mediated platelet activation. The trans-interaction of Axl or Tyro3 mediates cell-cell contact-dependent activation, which facilitates the activation of GPVI in platelets. Thus, Axl and Tyro3 may serve as better targets than Mer for the development of new antiplatelet agents.

Disclosures:
No relevant conflicts of interest to declare.
Yttrium-90-Ibritumomab Tiuxetan and BuCyE High-Dose Chemotherapy Compared with BuCyE Alone As Conditioning Regimen in Patients with B-Cell Non-Hodgkin Lymphoma: IPI Matched Case Control Study At a Single Center,
Authors: Jo Jae-Cheol , Yoon Dok Hyun , Kim Shin , Jang Seongsoo , Park Chan-Jeoung , Chi Hyun Sook , Park Chan-Sik , Huh Jooryung , Lee Sang-wook , Suh Cheolwon 
Published: October 2019

Abstract
Abstract 3703

Background:
Currently, there are accumulating data on the role of radioimmunotherapy in autologous stem cell transplantation (ASCT) with promising results. We compared the efficacy and safety of yttrium-90-ibritumomab tiuxetan (90Y-ibritumomab) combined with intravenous busulfan, cyclophosphamide, and etoposide (BuCyE) with those of BuCyE alone followed by ASCT in patients with relapsed or refractory B-cell non-Hodgkin lymphoma (NHL).
Patients and Methods: From October 2005 to May 2011, 71 NHL patients underwent high-dose chemotherapy with BuCyE (n=52) or 90Y-ibritumomab plus BuCyE (n=19) followed by ASCT at the Asan Medical Center. Of 52 patients receiving BuCyE, we found 19 patients who had B-cell NHL and they comprised control group of BuCyE alone (n=19). Retrospective comparison matched by the same IPI was performed for the efficacy and safety between two groups.

Results:
The patient cohort consisted of 38 individuals (19 males), of median age 52 years (range, 27–65 years). The baseline characteristics were well balanced between the two groups. The median time to platelet engraftment (>20,000/mm3) and the median time to neutrophil engraftment (>500/mm3) did not differ between 90Y-ibritumomab plus BuCyE (12 days [range, 3–103 days] and 10 days [range, 8–12 days], respectively) and BuCyE alone (12 days [range, 8–35 days] and 10 days [range, 9–12 days], respectively). The objective overall response rate was 89.5% (17/19): continued CR, 42.1% (8/19); induced CR, 36.8% (7/19); PR, 10.5% (2/19) in 90Y-ibritumomab plus BuCyE group, and 78.9% (15/19): continued CR, 26.3% (5/19); induced CR, 47.4% (9/19); PR, 5.3% (1/19) in BuCyE group, respectively (p =0.649). Median follow-up duration for survivors was 30 months. Event-free survival tended to be better in 90Y-ibritumomab plus BuCyE group with a median event-free survival duration of 12.5 months in 90Y-ibritumomab plus BuCyE group and 6.2 months in BuCyE group (p =0.236) (figure 1-a).There seemed to be no significant difference in overall survival between the two groups (p =0.767, figure 1-b). The toxicities including the incidence of grade 3 or higher stomatitis, nausea, vomiting, diarrhea, hyperbilirubinemia were similar in the two groups.

Conclusion:
Addition of 90Y-ibritumomab to BuCyE high-dose chemotherapy may potentially benefit patients with relapsed or refractory B-cell NHL at no cost of additional toxicity, which warrants further investigation.

Disclosures:
No relevant conflicts of interest to declare.
Angiogenesis and Tumor Growth Are Increased in Kininogen Deficient Mice
Authors: Zhang Wan-Ming , Hsi Linda , McCrae Keith R. 
Published: October 2019

Abstract
Abstract 371
High molecular weight kininogen (HK) is an abundant plasma protein and member of the contact phase of coagulation that is involved in a number of important biological processes. We have previously reported that kallikrein-cleaved HK (HKa) induces rapid apoptosis of proliferating endothelial cells and inhibits angiogenesis. We have recently produced a kininogen deficient mouse by deleting the murine gene for kininogen that is expressed in liver, mKng1. These mice lack detectable kininogen, despite persistent expression of mRNA from the mKng2 gene, primarily expressed in kidney. To further determine the importance of kininogen in the regulation of angiogenesis, we assessed angiogenesis in mKng1−/− mice. These mice demonstrated increased angiogenesis in subcutaneously-implanted Matrigel plugs containing vascular endothelial cell growth factor (VEGF) or basic fibroblast growth factor (bFGF), compared to wild-type littermates. We also assessed the growth of B16F10 melanoma cells and Lewis lung carcinomas planted subcutaneously in mKng1−/− mice. In both cases, in particular with the B16F10 implants, tumor growth occurred substantially faster in the setting of kininogen deficiency (mean tumor volume of mKng1−/− mice 4820 ± 728 mm3 vs 1990 ± 412 mm3 in wild type mice; P = 0.003). Differences in growth rates were apparent approximately 8 days after tumor implantation, and were associated with increases in tumor microvascular density. Several reports suggest that the urokinase receptor (uPAR), which binds HKa, mediates its antiangiogenic activity. However, using Matrigel plug assays we observed that HKa inhibited angiogenesis equally well in wild type and uPAR−/− mice. These studies demonstrate that kininogen serves as an endogenous regulator of angiogenesis. Current studies are focused on better defining the mechanisms of this activity.

Disclosures:
No relevant conflicts of interest to declare.
Phospho-SFKs (Y416) As a Potential Predictive Marker for Dasatinib Therapy in B-Cell Non-Hodgkin Lymphoma and Chronic Lymphocytic Leukemia,
Authors: Zhao Xiaoxian , Hsi Eric D. 
Published: October 2019

Abstract
Abstract 3727
The Src family of protein tyrosine kinases (SFKs) plays an important role in regulating multiple signaling networks including B-cell receptors (BCR) mediated pathways and abnormal SFK kinase activation promotes B lymphoma cell survival. Dasatinib is an oral BCR/ABL1 and SKF inhibitor useful in the treatment of imatinib-resistant CML and Ph+ALL. Given its broad inhibitory activity, dasatinib may be useful in the treatment of other hematologic malignancies and having a biologic predictor of response would be helpful in rational selection of this targeted therapeutic. We hypothesized this agent could have therapeutic potential against lymphoma patients with p-SFK (Y416) expression.
Constitutive p-SFK (Y416) expression (indicating active SFK signaling) was detected in both B-lymphoma cell lines and a subset of primary lymphoma tissues including diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), Burkitt lymphoma and small lymphocytic leukemia (SLL). Dasatinib induced apoptosis of B-lymphoma Raji cells correlated with high level expression of constitutive p-SFK (Y416) and dasatinib rapidly reduced the global level of tyrosine phosphorylations including p-SFK (Y416) in Raji cells. 19 of 28 lymphoma cases (67.9%) were positive for p-SFK (Y416) by Western blot analysis. Dasatinib displayed in vitro dose-dependent (10–200 nM) killing activity against 17 of those 19 p-SFK (Y416) cases (89.5%). In contrast, only 2 of 9 p-SFK (Y416) negative cases (22.2%) had response to dasatinib exposure. Thus presence of p-SFK (Y416) was associated with in vitro response to dasatinib (p <0.0001). Similar to tested Raji cells, dasatinib induced apoptosis of primary B-cell lymphoma cells was accompanied with de-phosphorylation of p-SFK (Y416) and cleavage of caspase-3. 6 of 9 tested CLL cases were p-SFK (Y416) positive. Dasatinib displayed in vitro killing activities against 5 of 6 positive cases with a range of killing from 12% to 53% (mean 26.5%) of malignant B-cells. Meanwhile, one of three negative cases showed response to dasatinib (17% killing).
We conclude that p-SFK (Y416) may be a useful predictive marker of response to dasatinib. Potential uses include pharmacodynamic monitoring or integral biomarker for selecting appropriate patients with B-cell malignancies for clinical trials.

Disclosures:
No relevant conflicts of interest to declare.