bluuebunny/crossref_metadata_2025_split huggingface dataset:
Methylcellulose as a Wetting Agent in Blood Clot: In Vivo Studies
Authors: ROSENFELD MORRIS 
Published: October 2019

Abstract
Methylcellulose, injected intravenously in dogs, altered the surface properties of blood plasma, as measured both in the fluid and clotted state. In fluid plasma, it altered the pattern of the surface tension-time curve, instituting the following changes: a reduction in the dynamic value of surface tension, a progressive rise in value contrasted to the normal decline, an elevation of the final static level above the pre-injection value. In clotted plasma, methylcellulose converted the normal water repellent surface to one that was water attracting, or hydrophilic. There was no change of coagulation time in glass, but possibly some prolongation in siliconed tubes. No surface tension or hydrophilic changes were induced by the other colloids tested; namely, dextran, polyvinylpyrrolidone and carboxymethylcellulose. The physical changes induced by methylcellulose are of a type that might favorably influence clot structure and adhesion.
A Consideration of Myeloid-Erythroid Balance in Man
Authors: PATT HARVEY M. 
Published: October 2019

Abstract
The number of myeloid and erythroid cells in marrow, their turnover times and rates of production have been estimated from consideration of mitotic activities and related data. An approximation to the chronology of development in the steady state with constant population is presented and its implications are discussed. Estimates of myeloid and erythroid balance in man are compared with similarly derived values for the dog and rat.
Elucidation of B Cell Antigen Receptor Signaling by Global Phosphoproteomics Reveals Novel Signaling Pathways and Regulatory Mechanisms
Authors: Oellerich Thomas , Corso Jasmin , Engelke Michael , Stanze Daniel , Hsiao He-Hsuan , Wienands Juergen , Urlaub Henning , Serve Hubert 
Published: October 2019

Abstract
Abstract 1056
B cell antigen receptors (BCR) transmit information via posttranslational protein modifications, most importantly protein phosphorylation. BCR signaling is a complex process that regulates differentiation and activation responses of the B-lineage. Dysregulation of BCR signaling can induce malignant transformation that can result in the onset of leukemia or lymphoma. To date our understanding of normal- versus malignant B cell biology points to differences in properties of BCR signaling that may be amenable to selective therapeutic targeting. However, comprehensive knowledge about BCR signaling pathways is still lacking which makes it difficult to understand the molecular events that drive B cell function and malignant transformation. To globally characterize BCR signaling, we have applied quantitative phosphoproteomic techniques that allowed us to identify and quantify about 10000 phosphorylation sites as a function of BCR stimulus, time, and subcellular location. Our phosphoproteome approach uncovered many novel BCR signaling effectors and their site-specific phosphorylation dynamics. The majority of phosphorylation events occured on serine/threonine residues, while only about one percent was detected at tyrosine residues. Interestingly, most of the identified proteins contain multiple phosphorylation sites showing different kinetics upon BCR stimulation, suggesting that they serve as signal integrators. Moreover, our functional analysis revealed that certain phosphorylation sites in adapter proteins are not only relevant for regulating the onset but also the termination of BCR signaling. Such signal termination is at least partly achieved by coupling the phosphotyrosine-dependent formation of BCR signalosomes to the kinetically flexible cytosolic Ca2+ concentration. In summary, our analysis revealed the complex composition of BCR signaling pathways and their regulation, and hence provides a basis for the identification of potential BCR-associated drug targets in the future.

Disclosures:
No relevant conflicts of interest to declare.
Molecular Basis of Phospholipid Synergy in Promoting Blood Coagulation Reactions
Authors: Davis-Harrison Rebecca L , Clay Mary , Rienstra Chad M , Morrissey James H. 
Published: October 2019

Abstract
Abstract 1109
Most steps in the blood coagulation cascade obligatorily take place on membrane surfaces and are dependent on the exposure of phosphatidylserine (PS). Many coagulation proteins bind to PS-containing membrane bilayers in a calcium-dependent manner via gamma-carboxyglutamate-rich (GLA) domains. Furthermore, certain phospholipids, most notably phosphatidylethanolamine (PE), strongly synergize with PS to promote clotting reactions. We have recently been investigating the molecular basis of lipid-Ca2+ and lipid-GLA interactions in blood clotting using solid-state NMR (SSNMR) studies that employ isotopically labeled PS. Our studies have revealed that Ca2+ induces two predominant conformations of the PS headgroup (Boettcher et al., Biochem. 50:2264–73, 2011). We now report the quantification of the intermolecular and intramolecular atomic distances within Ca2+-induced PS nanoclusters using SSNMR. We have also expanded SSNMR studies to phospholipid bilayers composed of trinary mixtures of PS, PC, and PE as a means to further investigate the impact of PE on PS headgroup conformations in an intact lipid bilayer (PE-PS synergy). Initial results suggest that incorporation of PE into the bilayer alters the Ca2+: PS ratio required to fully saturate the L-serine headgroups of PS. Additionally, PE appears to affect the predominant PS headgroup conformations in the presence of Ca2+, and may induce a third L-serine conformation. SSNMR studies utilizing isotopically labeled PE headgroups are currently underway which should provide novel information concerning PE's interactions with PS, Ca2+, and/or GLA domains.
Previous SSNMR studies from our lab have also indicated that a fraction of L-serine headgroups in PS adopt a new conformation when bovine prothrombin fragment 1 is bound to PS/PC bilayers (Tavoosi et al., J. Biol. Chem. 286:23247–53, 2011). These data support the idea that GLA domains contain an L-serine-specific binding site. We have now bound human prothrombin fragment 1 to membranes containing labeled PS headgroups for use in SSNMR studies. In addition, we have also employed surface plasmon resonance (SPR) experiments to investigate the interactions of both human and bovine prothrombin with bilayers containing PS and PE. These data demonstrate that binding affinities and binding footprints are similar for both prothrombin proteins, and that PE enhances binding affinities and increases the number of GLA binding sites for both proteins. Together, results from SSNMR and SPR studies suggest that different GLA domains use similar molecular mechanisms to interact with the membrane surface.

Disclosures:
No relevant conflicts of interest to declare.
Factor VIIa-CTP, a Novel Long-Acting Coagulation Factor, Displays a Prolonged Hemostatic Effect and Augmented Pharmacokinetics and pharmacodynamics Following IV and SC Administration in Hemophilic Animal Models
Authors: Hart Gili , Hershkovitz Oren , Bar-Ilan Ahuva , Seligsohn Uri , Fima Eyal 
Published: October 2019

Abstract
Abstract 1114

Background:
Prolor Biotech Inc. is a clinical stage public company developing biobetter long acting versions of existing therapeutic proteins utilizing a technology termed CTP. The technology involves fusion of the C terminus peptide of hCG to one or both ends of the target protein. The technology was clinically validated and proven as a safe and efficient way for prolonging the half-lives of several therapeutic proteins while maintaining their biological activity.

Aims:
To determine the pharmacokinetic (PK), pharmacodynamic (PD) and long term hemostatic effects of FVIIa-CTP in murine FVIII−/− mice following IV and SC administrations and to characterize FVIIa-CTP in vitro mechanism of action.

Methods:
FVIIa-CTP was expressed in CHO cells, purified and activated utilizing a purification process involving a CTP specific step. FVIIa-CTP was administered to FVIII−/− mice, and following IV and SC injection the PK and PD profiles were determined. In addition, the long term hemostatic effect was evaluated following bleeding challenge (tail clip assay and in a Tail Vein Transection) as compared to commercial rFVIIa. FVIIa-CTP in-vitro characteristics were also evaluated.

Results:
FVIIa-CTP PK parameters following IV administration, as assessed by a clotting assay, were superior to those of rFVIIa. Its half-life and AUC were 5 and 3.5 fold higher, respectively. A 2 fold improvement in the recovery was observed 15–30 minutes post dosing. Thrombin generation parameters were also superior in these studies. In a Tail Vein Transection study, FVIIa-CTP had a profound effect on survival, which was maintained for more than 24 hours. Reduced duration and intensity of bleeding was also observed in the tail clip study. Following SC administration, FVIIa-CTP's bioavailability, Cmax and half life were superior to commercial rFVIIa in both rats and hemophilic mice models. Correction of thromboelastograply in FVIII −/− mice plasma was maintained longer with FVIIa-CTP than with rFVIIa.

Conclusion:
Attachments of CTP to FVIIa led to a markedly enhanced PK, increased exposure, as reflected by AUC,improved recovery and a prolonged hemostatic effect in hemophilic mice with a comparable specific activity to rFVIIa. In addition, SC administration of FVIIa-CTP resulted in improved bioavailability and exposure was significantly prolonged relative to IV administration. Our data suggest that CTP fused FVIIa has the potential to significantly improve the prophylactic and on demand treatment of hemophilic patients.

Disclosures:
Seligsohn: prolor biotech: Consultancy, clinical advisor Other.
Treatment Irrespective of Symptoms Is a Major Predictive Factor for Thrombus Resolution in Patients with Portal Vein Thrombosis
Authors: Yu Yuan-Bin , Yang Chien-Wen , Liu Chun-Yu , Gau Jyh-Pyng , Hong Ying-Chung , Hsiao Liang-Tsai , Liu Jin-Hwang , Chiou Tzeon-Jye , Hsu Hui-Chi , Tzeng Cheng-Hwai 
Published: October 2019

Abstract
Abstract 1166

Background:
Portal vein thrombosis (PVT) typically presents with abdominal pain, ascites, and splenomegaly, but it is frequently asymptomatic. Current guidelines recommend that anticoagulation be used for symptomatic patients with PVT. However, it remains controversial regarding the benefits of treatments for asymptomatic patients with PVT.

Methods:
We retrospectively enrolled 933 patients with suspicious PVT in the Taipei Veterans General Hospital from January 2002 to December 2011. At total of 93 patients were confirmed using either doppler sonography, computed tomography, or magnetic resonance imaging. Response to treatment was defined as the recanalization or cavernous formation of portal veins. Logistic regression was used to investigate the clinic-laboratory parameters that were predictive for the resolution of PVT.

Results:
Among the 93 patients, the median age was 63 years old (range 1–91), and 61 (66%) of the patients were male. Abdominal pain was the most common symptom, occurring in the 53 patients (57%). Twenty-nine (31%) patients were asymptomatic at the time of diagnosis. For 35 of the treated patients, anticoagulation (71%), anti-platelet agents (20%), and catheter-directed urokinase infusion (26%) were the common modalities. Bleeding was noted in 6 (17%) of the patients, and all of the bleeding incidents were in the gastrointestinal tract. In the multivariate analysis, treatment was the only independent factor for thrombus improvement [odds ratio 8.54, 95% confidence interval 2.61–28.0, P < 0.001]. The results were the same when we analyzed the symptomatic and asymptomatic subgroups. The cumulative probability of improvement was higher among the treated patients compared to untreated patients at 2 years (62.8% vs. 26.2%, P< 0.001), and the benefits of the treatment were evident among the symptomatic and asymptomatic patients.

Conclusions:
Asymptomatic PVT patients may benefit from treatment. Further large-scale or prospective studies are necessary.

Disclosures:
No relevant conflicts of interest to declare.
Gfi1 Is Repressed by p53 and Inhibits DNA Damage-Induced Apoptosis
Authors: Du Pei , Tang Fangqiang , Qiu Yaling , Dong Fan 
Published: October 2019

Abstract
Abstract 1228
Gfi1 is a transcriptional repressor that plays a critical role in hematopoiesis. Gfi1 has also been implicated in lymphomagenesis when aberrantly activated as a result of deregulated expression. It is still poorly understood how Gfi1 expression is regulated and how it acts in the hematopoietic system. We show here that Gfi1 transcription was repressed by the tumor suppressor p53 in hematopoietic cells. Treatment of cells with doxorubicin (Doxo), which induces topoisomerase II-mediated DNA double strand breaks (DSB), led to a steady increase in p53 protein level, which was accompanied by a gradual decline in Gfi1 expression at both protein and mRNA levels. Knockdown of p53 resulted in increased Gfi1 expression and also abolished Doxo-induced Gfi1 downregulation. In contrast, Gfi1 expression was reduced and its downregulation in response to DNA damage was restored in p53-deficient cells transfected with the p53/estrogen receptor ligand binding domain fusion protein (p53ERTAM) in the presence of 4-hydroxytamoxifen (4-OHT). In luciferase reporter assays, Doxo treatment inhibited the activity of a ∼2.4-kb Gfi1 promoter fragment in p53+/+ cells, but not in p53−/− cells. Consistent with this, the wild type p53, but not a DNA binding-defective p53 mutant, repressed the Gfi1 promoter fragment. Chromatin immunoprecipitation (ChIP) assays demonstrated that p53 bound to the proximal region of the Gfi1 promoter. Detailed mapping of the Gfi1 promoter indicated that the core promoter region of Gfi1 spanning −33 to +6 bp is sufficient for p53-mediated repression. This core promoter region contains a putative p53 repressive response element (RRE) and, when the RRE was mutated, p53 failed to bind to and repress the Gfi1 promoter. Significantly, apoptosis induction by Doxo treatment was inhibited upon Gfi1 overexpression, but augmented following Gfi1 knockdown. Together, these data establish for the first time that Gfi1 is repressed by p53 and indicate that Gfi1 acts to protect hematopoietic cells from DNA damage-induced apoptosis. Our findings have important implications for understanding the roles of Gfi-1 in normal hematopoiesis and lymphomagenesis.

Disclosures:
No relevant conflicts of interest to declare.
Origin and Characterization of CD166+ Cells During Human Marrow Ontogeny
Authors: Mokhtari Saloomeh , Colletti Evan , Porada Christopher D , Almeida-Porada Graca 
Published: October 2019

Abstract
Abstract 1252
The development of hematopoiesis during embryonic and fetal life has served as a valuable model for characterizing putative HSC markers, as well as for studying the ontogeny of cells comprising the hematopoietic niche, and defining the nature of the interactions of hematopoietic stem/progenitor cells with these microenvironmental cells that drive HSC maintenance and/or differentiation. Alcam (CD166) has been shown to be expressed during ontogeny by the endothelium within the yolk sac and dorsal aorta, playing a key role in capillary tube formation and differentiation, but the adult aorta is devoid of this molecule. In the adult bone marrow, CD166 has been shown to be expressed by both highly primitive HSC and mesenchymal cells of the microenvironment, and to mediate homophilic adhesion between these two cell types. Here, we investigated the origin of the CD166+ cells during ontogeny and extensively characterized their phenotype as the marrow developed to better understand their function. To this end, we used flow cytometry and confocal microscopy to analyze human bone marrow (BM) during the period of development from 10 to 20 gestational weeks. CD166 expression was not detected on any cell population within the BM until week 18 of gestation, a time point when hematopoiesis is already well established within this tissue, as determined by the presence of CD34+CD45+ cells. CD166+ cells were rare at week 18, localized with other circulating cells in hematopoietic areas/spaces, and co-expressed FLK-1, CD44, N-cadherin, and Stro-1. By 20 weeks of gestation, the CD166+ population had increased significantly in number, and could be separated based on phenotype and spacial localization. CD166+/Stro-1+/CD44+/N-cadherin+/FLK-1+ cells increased approximately 4-fold in number between weeks 18 and 20, and they continued to be localized within the hematopoietic zones. In contrast, the more numerous CD166+/Stro-1+/CD44+/FLK-1- population was confined to the perivascular niche, with some of these cells integrating within the forming/maturing vasculature. In conclusion, we found that CD166 expression defines two distinct populations during development; one that seemingly associates with the perivascular niche, and another that localized with circulating hematopoietic cells. Interestingly, even though some CD166+ cells co-expressed N-cadherin, these cells did not localize to the osteoblastic niche. Studies are currently underway to further characterize these two populations with respect to their ability to support and/or give rise to hematopoiesis.

Disclosures:
No relevant conflicts of interest to declare.
Integrated RNA-Seq-Mirnome Analysis Reveal Interplay of Multiple Cancer Signaling Pathways Involved in B Lymphomagenesis
Authors: Muniandy Parameswary A , Haas Brian , Csizmadia Eva , Purushotham Jyothi , Washington Cyrus , Balter Barbara , Yan Catherine T. 
Published: October 2019

Abstract
Abstract 1294
Conditional inactivation of both the essential non-homologous DNA end-joining repair gene XRCC4 and the tumor suppressor gene p53 in mature B cells in mice frequently leads to mature B cell lymphomas. The majority of B lymphomas recurrently harbor reciprocal IgH/c-Myc translocations, activating c-Myc, a hallmark characteristic of Burkitt's lymphoma. We designed an integrated tumor RNA-Seq transcriptome profiling and miRnome based approach to dissect the interplay of signaling pathways and oncogenic signatures that lead to the development of the tumors in our model. Differential expression analysis of mRNA and 709 miRNAs reveal specific alterations in distinct signaling pathways, including upregulation of both Wnt and PI3K pathway signaling. For example, we found that Wnt10b, the wnt ligand expressed in B cells is upregulated. Correspondingly, frizzled receptors and it's downstream effector disheveled are also upregulated, factors that contribute to beta-catenin stabilization. Our studies also reveal increased expression PIK3CD, a PI3K catalytic isoform expressed in B cells, with corresponding downregulation of miRNAs with seed matches to 3'UTR of PIK3CD in the tumor; a factor that likely contribute to the stabilization of PIK3CD mRNA. Our analyses reveal an interplay of multiple pathway proteins and genes that serve as novel therapeutic targets for human B lymphoma.

Disclosures:
No relevant conflicts of interest to declare.
Interleukin-9 Promotes Survival of Diffuse Large B Cell Lymphoma Cells Through Upregulation of P21cip1
Authors: Lv Xiao , Wang Xin , Feng Lili , Ge Xueling , Fang Xiaosheng , Sui Xiaohui , Liu Xin 
Published: October 2019

Abstract
Abstract 1544

Introduction:
Diffuse large-B-cell lymphoma (DLBCL) is known as an aggressive malignancy arising from B lymphocytes. Despite its greatly improved prognosis as a result of chemotherapy and immunotherapy with monoclonal antibodies, the exact molecular etiology of DLBCL is not well understood. Interleukin-9 (IL-9) is initially described as a growth factor secreted by activated Th2 cells. Various observations have demonstrated its diverse actions in immune disorders. Recent years, the determination of its growth-proliferative and anti-apoptotic activities on multiple transformed cells implies a potential role of this cytokine in tumorigenesis but there are still no reports about its oncogenic activities in DLBCL. Our study is aimed to test the expression of IL-9 and its receptor (IL-9R) in DLBCL patients and illustrate its pathogenic effect on DLBCL cell lines in vitro.

Methods:
Blood samples and araffin-embedded tissues from twenty DLBCL patients were collected prior to therapeutic interventions. Serums from healthy volunteers served as normal control. IL-9 levels in sera were quantified using human ELISA kits. The expression of IL-9R protein in DLBCL tissues and lymphoma cell lines (LY1, LY8, SP53, Mino and Jurket) was determined by immunohistochemical staining and western-blot, respectively. IL-9R genes were knocked down in DLBCL cell lines LY1 and LY8 by lentivirus-mediated gene silencing (interference sequence 5'- GCTCGTGCCATCTGACAATTT -3'). LY1, LY8 and the stable transfected cells were treated with IL-9 alone and in synergy with rituximab (10ug/ml).

Disclosures:
No relevant conflicts of interest to declare.
Chronic Myeloid Leukemia Philadelphia Chromosome Positive (CML Ph+) with Imatinib (IM) Intolerance or Resistant Disease. Response to Thalidomide (TALI) Treatment. Preliminary Report of a Serial of Cases with Encouraging Results
Authors: Hurtado Rafael , Viveros Jose Pablo Vargas , Silvia Carrillo Muñoz , Cruz-Velazquez Judith , Candelaria Myrna , Cervera Eduardo 
Published: October 2019

Abstract
Abstract 1689
The increase dose of Imatinib, the shift to a Tyrosine Kinase (TKI) inhibitor of second generation or hematopoietic stem cell transplantation are treatment options when there is loss or lack of response to IM in CML; However, in patients without access to these therapeutic alternatives, the use of alkylating drugs or investigational agents is indicated. NF Kappa B pathway is constitutively activated in CML patients therefore the combination of IM with a NF Kappa B inhibitor (as thalidomide) contributes a new very interesting field of research. We treated 21 patients (11 female) with CML Ph +− resistant (n=13) or intolerant (n=8) to IM, mean +− SD age was 44 + 11 years. At diagnosis 16 (79%) were classified as high risk, according with Eutos scale. Mean +− SD time of IM treatment before adding thalidomide (100mg daily) was 47.8 +− 31 months. At the time of this preliminary analysis, mean follow up since thalidomide addition was 5 (1–18 months) and only 13 patients are evaluable for response (Global response rate was 76%): Six had hematological response & Four achieved complete cytogenetic response (CCgR). The first case was a 53 year old female, who started treatment with IM in accelerated phase, was intolerant to IM and neither achieve CCgR with IM, but reached it after 6 months of adding thalidomide and remains with this treatment after 18 months. The second case was a 38 year old female, who achieved CCgR after 6 months within IM treatment, lasted 53 months, and finally thalidomide was added after losing CCgR; she achieved a 2nd CCgR after 7 months with this combination, and continues with thalidomide at 11 months. The third one is a 44 year old female, who only achieved partial CCgR (pCGR), and after indicating thalidomide on accelerated phase, she had CCgR after 4 months of this drug and remains at 6 months of treatment. Finally, the fourth case is a 26 year old female that received thalidomide because of a suboptimal response to IM; she achieved CCgR after 2 months within thalidomide. Eight patients are not evaluable for response yet, but are ongoing and all will be detailed at ASH meeting. Toxicity has been acceptable, mainly grade 1 neurotoxicity & none severe adverse events have been documented. This is the first communication of the use of Thalidomide with Imatinib with promising results which may represent an alternative therapeutic in CML with antiproliferative enhancer and/or synergistic effect in patients with resistance to IM without opportunity of access to other treatments. This cases report open a future very interesting field of research in intolerance and resistance CML.

Disclosures:
No relevant conflicts of interest to declare.