bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pediatric Reference Intervals for the DxH 520* Hematology Analyzer
Authors: Smit Astrid C , Seidel Aprille Lynn , Rohrbach Justin , Magari Robert , Lo Karen , Martin Javier , Adeli Khosrow , Higgins Victoria , Shabani-Rad Meer-Taher , Cyfra Maureen , Gwilliam Tracey , Michaud Wendy , Tuson Brian , Schmitt Kristy , Couvrette Nicole , Axelson Sandra , Tejidor Liliana M. 
Published: October 2019

Abstract
Introduction: The DxH 520 is a small hematology analyzer capable of performing CBC and 5-part differential in fresh whole blood samples (venous and capillary) collected in K2EDTA and K3EDTA anticoagulants. Medical care of children and adolescents is significantly dependent on reference intervals to properly interpret laboratory test results. A multi-center study was performed to verify or establish pediatric reference intervals for all parameters (White Blood Cells (WBC), Red Blood Cells (RBC), Hemoglobin (Hgb), Hematocrit (Hct), Mean Corpuscular Volume (MCV), Mean Corpuscular Hemoglobin (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC), Red Cell Distribution Width (RDW), RDW-SD, Platelets (PLT), Mean Platelet Volume (MPV), (Lymphocyte) LY%, LY#, Monocyte% (MO%), MO#, Neutrophil % (NE%), NE#, Eosinophil% (EO%), EO#, Basophil % (BA%), and BA#) with combined genders.
Methods: Whole blood samples from healthy children were tested within 8 hours of collection on the DxH 520 analyzer. Samples generating review flags or suspect messages were excluded from the analyses. Even gender distribution was targeted. A total of 208 specimens were enrolled that included 20 neonates (0 to 30 days), 27 infants (31 days to 2 years), 94 children (3 to 12 years) and 67 adolescents (13 to 21 years). Results were analyzed according to CLSI EP20-A3c guidelines.
Results: Reference intervals for the neonate age group were verified from existing published ranges using the transference method described in the guideline (Table 1). The robust method was used to calculate two sets of reference intervals partitioned by combining data from the infant and children age groups, and from the adolescent age group (Table 2).
Conclusion: Reference Intervals for CBC and differential parameters have been established for pediatric age groups (0 to 30 days, 31days to 12 years, 13 to 21 years) on the DxH 520 Hematology analyzer.
*CE marked. Pending 510(k) clearance by the United States Food and Drug Administration; not yet available for in vitro diagnostic use in the U.S.
Beckman Coulter, the stylized logo, and the Beckman Coulter product and service marks mentioned herein are trademarks or registered trademarks of Beckman Coulter, Inc. in the United States and other countries.

Disclosures
Smit: Beckman Coulter: Employment. Seidel:Beckman Coulter: Employment. Rohrbach:Beckman Coulter: Employment. Magari:Beckman Coulter: Employment. Lo:Beckman Coulter: Employment. Martin:Beckman Coulter: Employment. Tejidor:Beckman Coulter: Employment.
Quantitative Proteomic Profiling Identifies Differentially Regulated Pathways in Fanconi Anemia and Aplastic Anemia
Authors: Hou Hui , Lu Qin , Li Dan , Wu Shuiyan , Chu Xinran , Xu Guoqiang , Hu Shaoyan 
Published: October 2019

Abstract
Purpose: Prognosis and therapeutic treatment of Fanconi anemia (FA) and acquired aplastic anemia (AA) are different. However, delayed and incorrect treatments are frequently occurred in FA because its symptoms are very similar to those of AA. This study aimed to elucidate the differences between FA and AA at the molecular level and to identify biomarkers and pathways of FA through quantitative proteomics and bioinformatics analyses.
Methods: Samples were obtained from the bone marrow of patients with FA and AA followed the institutional guideline. Proteins were extracted, digested with Lys-C and trypsin, and chemically labeled with tandem mass tag (TMT) duplex reagents (TMT126 and 127) for mass spectrometric analyses. Proteins with fold changes of > 1.5 or < 0.667 were considered as differentially expressed proteins (DEPs). Gene ontology analysis was performed using the Database for Annotation, Visualization and Integrated Discovery. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were conducted using the ClueGO plug-in in Cytoscape. A DEP-associated protein-protein interaction (PPI) network was constructed using STRING and visualized in Cytoscape. The degree and topological index of these DEPs were calculated using CentiScape. The PPI network was subjected to modular analysis using the Molecular Complex Detection plug-in in Cytoscape.
Results: A total of 114 DEPs, including 71 upregulated and 43 downregulated proteins, were discovered in the FA sample compared with those in the AA sample. The upregulated proteins were enriched in nucleosome assembly, cell or subcellular component movement, glycolysis, and gluconeogenesis pathway. The downregulated proteins were enriched in immune response, negative regulation of apoptosis, proteolysis, phagosome, and pantothenate and CoA biosynthesis. Five important modules were detected from the PPI network of DEPs. Eight hub proteins, a-enolase (ENO1), phosphoglycerate kinase 1 (PGK1), HSP90AA1, HSP90AB1, ACTC1, ACTBL2, EEF1A1, and CFL1, with a high degree of connectivity were obtained.
Conclusions: Quantitative proteomics coupled with bioinformatics analyses are useful for the identification of protein biomarkers and pathways associated with FA and AA. Some critical DEPs, such as ENO1, PGK1, ACTC1, ACTBL2, EEF1A1, and CFL1, likely play important roles in FA and could be used as serologic markers for its early diagnosis and to distinguish FA from AA.

Disclosures
No relevant conflicts of interest to declare.
Outcome of Lymphocyte Predominant Hodgkin Lymphoma: A Single Institutional Analysis
Authors: Ezekwudo Daniel E , Anusim Nwabundo , Konde Anish S , Gbadamosi Bolanle , Khoury John , Macari David , Zakalik Dana , Blankenship Leann , Gaikazian Susanna , Jaiyesimi Ishmael 
Published: October 2019

Abstract
Background: Nearly 5% of patients diagnosed with Hodgkin lymphoma have the nodular lymphocyte predominant Hodgkin lymphoma (NLPHL), also known as Hodgkin paragranuloma or lymphocytic and histiocytic predominant Hodgkin disease. Given the rarity of this variant of Hodgkin lymphoma, there is no consensus on the standard of care/regimen in this subgroup of patients. Therapeutic approach has spanned from radiotherapy alone, combined chemo-radiotherapy to chemotherapy alone or observation and recently the use of Rituximab anti CD20 monoclonal antibody as these cells expresses CD20 marker. We opt to evaluate the outcome of patients with newly diagnosed NLPHL in our institution. We also reviewed outcome of NLPHL patients treated with various chemotherapy such as ABVD, CVP or RCHOP ± Radiation.
Methods: We retrospectively reviewed medical records of over 300 patients [mean age 45 (13-76)] with Hodgkin Lymphoma at Beaumont Hospital, an academic community center from January 2007 - January 2017. Given the rarity of the disease, a total of 13 patients were found to have pathologically confirmed diagnosis of NLPHL. Data regarding age, sex, stage of disease, treatment and outcome was analyzed.
Results: Total 13 patients with NLPHL with age range from 13 to 80 years were studied. Demographic distribution of studied patients were 8(62%) white, 4 (31%) African Americans and 1 (7.6%) Asian. Out of these, 8 (62%) were males and 5 (38%) females. A total of 9 (69%) patients have stage III disease on diagnosis while 4 (31%) patients had stage II disease on diagnosis. Out of 9 patients with stage III disease, 6 (67%) received chemotherapy alone (ABVD or CVP) whereas 3 (33%) received combined chemotherapy and radiotherapy. One patient in the chemotherapy alone arm, recurred within 24 months of treatment and received local therapy with radiation. Among those with stage II disease on diagnosis, 3 patients received radiation therapy alone while one patient was observed and did not receive any treatment. Overall, out of 13 patients studied, 12 (92%) are still alive and continues to follow with our clinic with no recorded relapse to date. One patient developed astrocytoma grade III and died of intracranial hemorrhage.
CONCLUSION: Although the standard therapeutic management for patients with NLPHL remains controversial. Our data showed that regardless of the regimen selected (chemotherapy alone or combined chemo-Radiotherapy), patients with Lymphocyte predominant Hodgkin Lymphoma treated with ABVD ± Radiation have excellent outcome. Although our study represents a single institutional analysis, it concurs with recent studies that clinical outcome in patients with NLPHL is favorable with chemotherapy.

Disclosures
No relevant conflicts of interest to declare.
Tandem P-Selectin Glycoprotein Ligand Immunoglobulin Prevents Lung Vaso-Occlusion in SCD Mice
Authors: Vats Ravi , Tutuncuoglu Egemen , Tejero Jesus , Shaw Gray , Sundd Prithu 
Published: October 2019

Abstract
Introduction: Sickle cell disease (SCD) is an autosomal-recessive-genetic disorder, which leads to red blood cell sickling, hemolysis and vaso-occlusion. Acute systemic painful vaso-occlusive crisis (VOC) is the predominant pathophysiology requiring emergency medical care by SCD patients. 10-20% of SCD patients hospitalized with VOC tend to develop acute chest syndrome (ACS), a type of lung injury within next few days, suggesting a role for pulmonary vaso-occlusion in ACS. This epidemiology also provides a window for therapeutic intervention provided treatments to prevent vaso-occlusion are available. Earlier, we have shown that VOC involves entrapment of large neutrophil-platelet aggregates in lung arterioles of SCD mice, which is inhibited following intravenous administration of P-selectin function blocking antibody. Recently, a tandem-P-selectin-glycoprotein-ligand-immunoglobulin (TSGL-Ig) with two P-selectin binding sites in tandem, has been shown to prevent P-selectin-dependent liver injury in mice. Here, we test the ability of TSGL-Ig in attenuating P-selectin dependent lung vaso-occlusion in SCD mice.
Materials and Methods: Townes knock-in humanized SS (hα/hα:βS/βS) mice were used as SCD mice. SCD mice were intravenously (IV) challenged with 10 µmole/kg Oxy-hemoglobin (Oxy-Hb) without or with 100 µg of TSGL-Ig to trigger lung vaso-occlusion. Fluorescent anti-mouse mAbs against CD49b and Ly6G, and FITC dextran were IV administered for in vivo staining of platelets, neutrophils, and visualizing lung microvasculature, respectively. Presence or absence of lung vaso-occlusion was assessed using multi-photon excitation enabled quantitative fluorescence intravital lung microscopy (qFILM).
Results and Discussion: IV administration of Oxy-Hb led to occlusion of pulmonary arterioles by large neutrophil-platelet aggregates. Remarkably, IV administration of TSGL-Ig significantly attenuated lung vaso-occlusion in SCD mice by reducing the number as well as size of neutrophil-platelet aggregates in the pulmonary arterioles of SCD mice.
Conclusion: Systemic challenge with Oxy-Hb promotes lung vaso-occlusion in SCD mice. TSGL-Ig significantly reduced oxy-Hb induced lung vaso-occlusion in SCD mice, highlighting the potential of TSGL-Ig to prevent ACS.

Disclosures
No relevant conflicts of interest to declare.
Thalassemia Gene Therapy By In Vivo Transduction of Mobilized Hematopoietic Stem Cells (HSCs) with an Integrating Hybrid Adenovirus Vector System
Authors: Georgakopoulou Afrodite , Wang Hongjie , Kapsali Chrysi , Psatha Nikoleta , Koufali Angeliki , Li Chang , Izsvak Zsuzsanna , Anagnostopoulos Achilles , Papayannopoulou Thalia , Yannaki Evangelia , Lieber Andre 
Published: October 2019

Abstract
To overcome the cost and complexity of current thalassemia ex vivogene therapy protocols, we developed a minimally invasive and readily translatable approach for in vivo HSC gene delivery which abrogates the need for HSC leukapheresis, CD34+ cell selection, ex vivo HSC culture, myeloablation and ultimately, transplantation. Our approach involves HSC mobilization with G-CSF/AMD3100 andintravenous injections of a hybrid vector system consisting ofa CD46-targeting, helper-dependent adenoviral vector and the hyperactive Sleeping Beauty transposase (SB100x) that mediates integration of thevector-encoded γ-globin and mgmtP140K genes. Pretreatment with glucocorticoids before virus injectionsis used to blockthe release of pro-inflammatory cytokines andimmunosuppression is applied in order to avoid responses against human g-globin- and MGMT protein-expressing cells. We tested our approach in a mouse model recapitulating the phenotypeof human β-thalassemia intermedia (Hbbth-3/hCD46++ mice). At week 8 post transduction, hCD46+/+/Hbbth-3 mice expressed HbF in 31.2±2.7% of circulating erythrocytes. Due to a significant drop in HbF expression by week 16 (11.9±3.0%), a 4-dose O6BG/BCNU treatment was administeredin order to in vivo select forgene corrected hematopoietic progenitors, thus recovering the HbF expression in76.0±5.7% of the circulating erythrocytes, by week 29 post in vivo transduction. With an average vector copy number of 1.4/cell, the human γ-globin to mouse α-globin expression was ~10% by HPLC and the human γ-globin to mouse β-globin mRNA ratio ~10%, by qRT-PCR. Hematological parameters (RBCs, Ht, Hb, MCV, RDW, Reticulocytes) at week 29 post in vivo transduction, were significantly improved over baseline or were indistinguishable from normal values, suggesting near to complete phenotypic correction. Treated mice showed significant reduction of spleen size, extramedullary erythropoiesis and parenchymal hemosiderosis. After secondary transplantation and without in vivo selection, more than 90% of donor-derived erythrocytes (CD46+) were g-globin-positive, up to 20 weeks post-transplant. Safety was demonstrated by the good tolerability of treatment, the absence of alterations in hematopoiesis, the normal colony-forming potential of bone marrow cells and the random integration pattern of our vector system. Overall, we present a simplified platform for gene therapy of thalassemia, which can serve as a cost-efficient and "portable" approach to make gene therapy accessible even to resource-poor regions where thalassemia major is endemic but only minimally complex strategies could be adopted.

Disclosures
No relevant conflicts of interest to declare.
Novel Antibodies Against Glycoprotein Ibα Inhibit Pulmonary Metastasis By Affecting Vwf-Gpibα Interaction
Authors: Yingxue Qi , Chen Wenchun , Xu Ke , Wu Fengying , Fan Xuemei , Ren Shengxiang , Liu Junling , Deng Wei , Zhang Jun , Li Renhao , Liu Jianwen , Liang Xin 
Published: October 2019

Abstract
Background:
Platelet glycoprotein Ibα (GPIbα) extracellular domain, which is part of the receptor complex GPIb-IX-V, plays an important role in tumor metastasis. However, the mechanism through which GPIbα participates in the metastatic process remains unclear. In addition, potential bleeding complication remains an obstacle for the clinical use of anti-platelet agents in cancer therapy.
Methods and Results:
To generate antibodies that bind to mouse platelet GPIbα, washed mouse platelet lysate was used as the antigen for rat immunization. Obtained hybridoma clones were screened in ELISA for binding affinity to the GPIb-IX complex. Positive clones were further screened for their abilities to inhibit platelet-cancer cell adhesion. Finally, at static condition, two antibodies, 2B4 and 1D12, had virtually no effect on the activation of integrin αIIbβ3, which is used to indicate platelet activation. Then, we characterized the binding sites of 2B4 and 1D12 by 20 purified recombinant GPIbα fragments binding. Results showed that 2B4 and 1D12 shared the same binding sites with vWF.
To determine whether 2B4 and 1D12 affect vWF binding, we tested the binding by flow cytometry using recombined mouse vWF, and then, we investigated platelet aggregation induced by several agonists, including vWF binding agonist ristocetin. Our data demonstrated clearly that 2B4 and 1D12 could inhibit vWF binding.
To investigate whether the inhibition of vWF-GPIbα interaction was associated with tumor metastasis, we examined the effect of 2B4 and 1D12 in each of the following interactions in vitro: between activated platelets and tumor cells, platelets and endothelial cells. Meanwhile, We further investigated the inhibitory effect of these antibodies in vivo using the experimental metastasis model and the spontaneous metastasis model. Results showed that 2B4 and 1D12 could potently inhibit the adhesion of cancer cells in vitro, and metastasisin vivo.
We next investigated whether 2B4 and 1D12 could affect platelet activation and/or induce thrombocytopenia in vivo. Results showed that the addition of 2B4 or 1D12 to PRP did not induce platelet aggregation and injection of 2B4 or 1D12 Fab at appropriate dose did not affect tail-bleeding time and platelet count.
Based on the above findings, we obtained anti-human platelet GPIbα monoclonal antibody YQ3 using the same approach to explore the role of human GPIbα in cancer metastasis. As expected, YQ3 inhibited lung cancer adhesion and demonstrated similar value in metastasis. More importantly, for all three mAbs in our study, none of their Fabs induced thrombocytopenia.
Conclusion:
Our results therefore supported the hypothesis that GPIbα contributes to tumor metastasis, and suggested potential value of using anti-GPIbα mAb to suppress cancer metastasis.

Disclosures
Li: Neoletix: Consultancy, Equity Ownership.
Regulatory T Cell Selective IL-2-Fc Fusion Proteins for the Treatment of Autoimmune Diseases
Authors: Varma Rajat , Liu Ke , Avery Kendra , Rashid Rumana , Schubbert Suzanne , Leung Irene , Bonzon Christine , Bogaert Liz , Desjarlais John , Bernett Matthew J. 
Published: October 2019

Abstract
Regulatory T cells (Tregs) characterized as CD4 positive cells, expressing CD25 (IL-2Rα) and the forkhead transcription factor FoxP3 maintain immune tolerance in tissues by suppressing the function of both CD4 and CD8 effector T cells. It is well documented that there is a paucity and/or dysfunction of Tregs in most autoimmune diseases and therefore a promising therapeutic approach has been to restore their numbers and function. Treg homeostasis depends on IL-2 derived signals and Tregs are sensitive to IL-2 stimulation in part due to the high abundance of CD25. As a result, encouraging results have been seen in patients with autoimmune diseases who were treated with low doses of IL-2. Single or repeat low doses of IL-2 led to selective expansion of Tregs over effector cells and provided clinical benefit. Such IL-2 based therapies can be further improved by enhancing the selectivity of IL-2 for Tregs and improving the pharmacokinetics of the drug. Here we have developed an IL-2-Fc fusion protein that selectively acts on Tregs in vitro and in vivo. IL-2 mediated signal transduction occurs through the formation of a high affinity tetrameric complex consisting of IL-2, CD25, CD122(IL-2Rβ) and CD132(IL-2Rγ). IL-2-Fc fusion proteins were designed to be monomeric by fusing variant IL-2 molecules to one side of our heterodimeric Fc-region. To achieve Treg selectivity, we designed variants of IL-2 that have weakened interaction with CD122 and/or enhanced interaction with CD25 such that signaling is only observed in CD25 high cells. These IL-2-Fc variants were produced with good yield and purity. We identified novel variants that selectively led to phosphorylation of STAT5 transcription factor in Tregs from human peripheral blood mononuclear cells (PBMCs). These Treg-selective IL-2-Fc variants stimulated Tregs with varying potencies. Further, they selectively enhanced and sustained the proliferation of Tregs in vivo in non-human primates. These promising preclinical results suggest that clinical development of Treg selective IL-2-Fc fusion proteins for autoimmune diseases is warranted.

Disclosures
Varma: Xencor: Employment, Equity Ownership. Liu:Xencor: Employment, Equity Ownership. Avery:Xencor: Employment, Equity Ownership. Rashid:Xencor: Employment, Equity Ownership. Schubbert:Xencor: Employment, Equity Ownership. Leung:Xencor: Employment, Equity Ownership. Bonzon:Xencor: Employment, Equity Ownership. Bogaert:Xencor: Employment, Equity Ownership. Desjarlais:Xencor: Employment, Equity Ownership. Bernett:Xencor: Employment, Equity Ownership.
A Population-Based Study of Prognosis and Survival in Primary Hepatobiliary Lymphoma
Authors: Sindel Ariel , Al-Juhaishi Taha , Sabo Roy , Yazbeck Victor Youssef 
Published: October 2019

Abstract
Background:
Primary Hepatobiliary Lymphoma (PHBL) is a rare disease. In review of the literature, there are only few retrospective studies. On prior studies, the median age at presentation was 62 years and was associated with a median overall survival of 162 months. In this study, we sought to characterize standard prognostic factors and their effects on overall survival utilizing the Surveillance, Epidemiology and End Results (SEER) database.
Methods
The Surveillance, Epidemiology and End Results (SEER) database was used to identify patients diagnosed with PHBL between 1973 and 2014. B-Cell and T-Cell lymphomas of all stages were included. Patient characteristics and demographics are summarized. Overall survival (OS) was estimated using the Kaplan-Meier method for all patients and for sub-groups. Comparisons are made between the survival functions between the levels of these categorical measurements using the log-rank test, where we adjust the significance level to account for multiple comparisons using the Bonferroni correction.
Results:
A total of 1751 patients were included. The mean age was 64 years (2-97), with the majority of patients being male (n=1081, 62%) and Caucasian (n=1278, 73%). B-cell Non-Hodgkin's Lymphoma was the predominant lymphoma (n=1297, 75%), followed by Not Otherwise Specified (n=338, 19%), and T-cell NHL and Hodgkin's Lymphoma were nearly identical at n=54 and 50 respectively. Staging at presentation varied with the majority being 1 and 4 (n=703, 40% and n=698, 40%), this was followed by stage 2 (n=196, 11%). The median overall survival time was 16 months (95% CI; 12, 23). By diagnoses, the median survival time (Figure 1) for B-Cell NHL was 23 months (95% CI; 17, 33) and was significantly longer survival than patients with T-Cell NHL (1 month, 95% CI; 0, 8, p=0.0014) and NOS (9 months, 95% CI; 6, 12, p=0.0076) but was not significantly different than patients with Hodgkin's Lymphoma (12 months, 95% CI; 1, 71 p=0.15). By staging, patients with stage 1 (38 months, 95% CI; 24, 52) did not have a statistically significant difference in survival in comparison to stage 2 (15 months, 95% CI; 9, 48, p=0.85) or 3 (10 months, 95% CI; 2, 34, p=0.0173). Results were not statistically significant regarding gender nor ethnicity. On a multivariant analysis, surgery, chemotherapy, or radiation were all associated with better survival when adjusted for patient age, sex, race, disease type and cancer stage. However, there did not appear to be a statistical significance when compared to each other.
Conclusion
PHBL is a rare lymphoma affecting mostly elderly, Caucasian males. B-Cell NHL subtype and stage 1 were associated with a more favorable prognosis. However, compared to DLBCL, PHBL seem to have a poorer outcome. While treatment alone conferred a longer survival, the type of treatment option did not have an overall change to prognosis.

Disclosures
No relevant conflicts of interest to declare.