bluuebunny/crossref_metadata_2025_split huggingface dataset:
National Burden of Emergency Department Care for Sickle Disease: Impact of Age, Insurance Status, Income, Hospital Type and Location On Subsequent Hospital Admission.
Authors: Goel Ruchika , Viswanathan Prabhu , Krishnamurti Lakshmanan 
Published: October 2019

Abstract
Abstract 2488
Poster Board II-465

BACKGROUND.
Limited data exist about the national burden of emergency department (ED) care for patients with sickle cell disease (SCD). The proportion of patients being seen in the ED who require hospitalization and factors contributing to the above are poorly understood. We present here analysis of such potential factors from the largest nationally representative ED visit data till date.

METHODS.
The Nationwide Emergency Department Sample (NEDS) is one of a family of databases and software tools developed as part of the Healthcare Cost and Utilization Project (HCUP), a Federal-State-Industry partnership sponsored by the Agency for Healthcare Research and Quality. The NEDS was constructed using records from both the HCUP State Emergency Department Databases and the State Inpatient Databases. The NEDS is the largest all-payer ED database and contains almost 26 million records for ED visits for over 950 hospitals and approximates a 20-percent stratified sample of U.S. hospital-based ED's thus generating weighted estimates of over 120 million ED visits.
The following ICD-9-CM diagnosis codes were used to define SCD related hospitalizations: 28241, 28242, 2825, 28260, 28261, 28262, 28263, 28264, 28268, and 28269 and only ‘primary discharge diagnosis' patients were considered. We analyzed the NEDS data for the year 2006.

RESULTS.
Approximately three-quarters of the total in-patient admissions for SCD in 2006 came through the emergency department. The total number of patients presenting to the ED with SCD as primary diagnosis was 166,043. Of these visits, 68,420 (41.2%) resulted in admission to the hospital.
There was a statistically significant difference (p<0.001) in the proportion of ER visits resulting in hospitalization for the following factors (each studied independently):

CONCLUSION.
SCD is responsible for a significant burden of ED care in the US. A number of factors including of patient age, insurance status, income, hospital type and location may impact the likelihood of subsequent admission to the hospital. High quality care of uncomplicated pain crises in an ED or day hospital is believed to result in a decrease of avoidable hospitalizations. However, the multiplicity of factors independently associated with likelihood of subsequent hospitalization as demonstrated by these data suggest the need for caution in using the proportion of ED visits resulting in hospitalizations as a surrogate marker of quality of care.

Disclosures:
No relevant conflicts of interest to declare.
Ly6C+ Inflammatory Monocytes Are Extrahepatic Precursors of Hepatic Stellate Cells in Liver Injury.
Authors: Masuya Masahiro , Araki Hiroto , Ohishi Kohshi , Katayama Naoyuki 
Published: October 2019

Abstract
Abstract 2518
Poster Board II-495

Objective:
It has been known that hepatic stellate cells (HpSCs) play an important role in the development of liver fibrosis. We previously reported that the HpSCs are hematopoietic origin in carbon tetrachloride (CCl4)-induced liver injury. However, the differentiation pathway from hematopoietic stem cells to HpSCs remains largely unknown. In this study, we tried to elucidate the ability of terminally differentiated cells to transdifferentiate into HpSCs.

Methods:
Male enhanced green fluorescent protein (EGFP)-transgenic mice 10–12 weeks old were used as bone marrow and spleen donors. Bone marrow mononuclear cells were incubated with purified anti-CD45R/B220, anti-CD4, anti-CD8, anti-Ly6G, anti-TER119, anti-c-kit, anti-Sca-1, anti-Thy-1.2, anti-IL-7 receptor α chain, and immunomagnetic beads. The resulting lineage negative cells were stained with anti-Ly6C, anti-c-kit, and anti-CD45.2. CD45.2+Ly6Clowc-kit+ cells (monocytic precursors), CD45.2+Ly6Chighc-kit− cells (mature monocytes), FSClowSSChigh CD45.2+ cells (eosinophils) were enriched by FACSAria. Ly6G+ cells (neutrophils), CD45R/B220+ cells (B cells) and CD3ε+ cells (T cells) were isolated from bone marrow or spleen using magnetic cell sorter device. We transferred these isolated cells into CCl4-injured mice twice a week for two weeks, and analyzed the presence of EGFP+ HpSCs in the injured liver using immunofluorescence and laser confocal scanning microscopy.

Results:
Total number of transfused monocytic precursors, mature monocytes, eosinophils, neutrophils, B cells, and T cells per mouse was 1.1 × 106, 6.4 × 106, 4.8 × 106, 22.0 × 106, 29.0 × 106, and 12.5 × 106, respectively. Intravenous infusion of each type of cell showed their ability to localize to the injured liver. In the liver of mice, which were given eosinophils, neutrophils, B cells, and T cells, EGFP+ cells were all positive for CD45. However, in the liver of mice, which received monocyte precursors and mature monocytes, 20% and 39% of EGFP+ cells were negative for CD45. Vimentin and ADAMTS13 are identified in HpSCs but not hematopoietic cells. About 18% of EGFP+ cells in the liver of mice, which received monocytic precursors, were positive for both antigens. In the liver of mice, which received mature monocytes, 35% and 27% of EGFP+ cells were positive for vimentin and ADAMTS13, respectively.

Conclusion:
Both Ly6Clowc-kit+ monocytic precursors and Ly6Chighc-kit− mature monocytes can give rise to HpSCs in CCl4-injured liver.

Disclosures:
No relevant conflicts of interest to declare.
TET Family Oncogene Fus Is Essential for the Maintenance of Self-Renewing Hematopoietic Stem Cells.
Authors: Sugawara Takeaki , Oguro Hideyuki , Iwama Atsushi 
Published: October 2019

Abstract
Abstract 2529
Poster Board II-506
A Proto-oncogene FUS (fusion derived from malignant liposarcoma), also known as TLS (translocated in liposarcoma), was originally identified in chromosomal translocation of human soft tissue sarcoma. FUS is also known to be fused with an ETS family transcription factor ERG in human myeloid leukemia with t(16;21) which is associated with poor prognosis. Based on its protein structure, DNA- and RNA-binding activity and involvement in many human cancers as the fusion with various transcription factors, FUS is now grouped with EWS and TAFII68 into TET (FET) oncogene family. Multiple functions have been postulated for FUS, including non-coding-RNA-mediated transcriptional repression, posttranscriptional RNA processing and the maintenance of genomic integrity. Fus-deficient (Fus−/−) mice showed a non-cell-autonomous defect in B lymphocyte development, defective B cell activation and increased sensitivity to radiation in previous studies. However, its physiological function in hematopoiesis remains unknown. In this study we performed detailed analyses of Fus−/− hematopoietic stem cells (HSCs). Fus−/− fetal livers at embryonic day 14.5 exhibited a mild reduction in numbers of hematopoietic stem and progenitor cells compared with the wild type. Disruption of Fus, however, did not grossly affect proliferation or differentiation of hematopoietic progenitors. Of note, Fus−/− HSCs had significantly reduced repopulating activity of hematopoiesis in competitive repopulation assays, and did not repopulate hematopoiesis at all in tertiary recipients. Moreover, Fus−/− HSCs were highly sensitive to radiation both in vitro and in vivo and showed a drastic reduction in numbers in recipient mice after sublethal irradiation. All these findings implicate Fus in the maintenance and radioprotection of HSCs. Studies of chromosome stability, telomere length, apoptosis and levels of reactive oxigen species (ROS) appeared not accountable for the apparent defect of Fus−/− HSCs. However, gene expression profiling identified changes in expression of several genes in Fus−/− HSCs, and dysregulated expression of some of these genes might be responsible for the defective function of Fus−/− HSCs.

Disclosures:
No relevant conflicts of interest to declare.
Impact of Hydroxyurea On Peri-Operative Management and Outcomes in Children with Sickle Cell Anemia.
Authors: Hayashi Masanori , Calatroni Agustin , Herzberg Brittany , Thornburg Courtney 
Published: October 2019

Abstract
Abstract 2567
Poster Board II-544

Surgical procedures in children with sickle cell anemia (SCA) can be complicated by vasoocclusive events (VOE) such as acute chest syndrome (ACS) and pain. Peri-operative management requires a multidisciplinary approach to provide appropriate pre-operative intravenous hydration and intra- and post-operative monitoring. Transfusion therapy has been controversial. Our institution previously described a low incidence of complications in children who received serial transfusions over 3-4 weeks prior to surgery. Subsequently, an increasing number of children have been prescribed hydroxyurea (HU) to prevent SCA complications. In general, children on HU at our institution only receive a single top-off transfusion the day prior to surgery if their hemoglobin is less than 10 g/dL. We hypothesized that children in the HU group would have a lower number of serial transfusion compared to the non-HU group and that there would be no difference in complications or days to discharge between the two groups. We conducted a single-institution retrospective cohort study of children with SCA, who were age less than 18 years and underwent at least one surgical procedure at Duke University Medical Center between January 1, 2003 and April 30, 2008. Data were abstracted from electronic and written medical records. Descriptive statistics were used to characterize the cohort. Wilcoxon test was used to compare continuous variables and Pearson test was used to compare categorical variables between the non-HU and HU groups. Fifty-three subjects were included (Table 1). The non-HU group was significantly younger than the HU group, but children in the non-HU group were significantly more likely to be transfused pre-operatively, primarily with serial transfusions or erythrocytopheresis, compared to the HU group. One subject in the non-HU group developed a pre-operative delayed hyperhemolytic transfusion reaction. Post-operative complications are detailed in Table 1; the overall rate was low. Two subjects in the HU group developed acute chest syndrome despite pre-operative transfusion; one episode was likely related to underlying asthma and poor response to hydroxyurea; the second was likely related to pain and hypoventilation after laparoscopic splenectomy and tonsillectomy/adenoidectomy. Overall, there were no significant differences in complications and no significant difference in days to discharge between the two groups. In summary, children with SCA on HU may safely undergo surgery without significantly reducing their percent HbS. Nonetheless, attention should still be made towards multidisciplinary effort to reduce intra- and post-operative complications, and clinicians should consider response to HU, pulmonary status and type of surgery when planning peri-operative management in children with SCA on HU.

Disclosures:
Off Label Use: hydroxyurea in young children.
Role of the MAPK and PI3-Kinase/Akt Pathways in the Pre-B Cell Receptor (pre-BCR)-Induced NF-κb Activation and Rag1 and Rag2 Down Regulation.
Authors: Lassoued Kaiss , Fuentes Vincent , Gamlouch Hussein , Bissac Eliane , Marolleau Jean-Pierre , Rochette Jacques , Bouhlal Hicham 
Published: October 2019

Abstract
Abstract 2667
Poster Board II-643
The pre-BCR acts as a critical checkpoint in pre-B cell development and might be also involved in leukemogenesis. Using the 697 and Nalm6 human pre-B cell lines, we have previously shown that pre-BCR stimulation resulted in cell cycle progression associated with activation of number of adaptors and signaling pathways including the PI3-Kinase/Akt, Ras/MAPK, AP1 and the canonical NFkB pathway. We have also demonstrated that Src kinases together with Syk played a crucial role in controlling the pre-BCR-associated functions, acting upstream the above-mentioned signaling pathways. Pre-BCR crosslinking also induced down regulation of Rag1 and Rag2 transcription. In this study we aimed to evaluate the role of MAPK and Akt in the pre-BCR-induced NF-kB activation and Rag1/2 down modulation. For this purpose the 697 pre-B cells and normal bone marrow primary pre-B cells were treated with the U0126 and LY294002, and with MEK1/2 and Akt inhibitors, respectively. A dominant negative form of Akt fused to the HIV1 Tat peptide was also used to inhibit the PI3-Kinase/Akt pathway. We bring evidence that LY294002 could alter the pre-BCR-induced NF-kB activation by inhibiting : i) p105 degradation, ii) p50 NF-kB1 nuclear translocation and, iii) the binding of p50 to an oligonucleotide containing a specific consensus sequence. On the contrary, U0126 significantly enhanced p105 degradation, indicating that MAPK and Akt exerted antagonistic effects on the pre-BCR-induced NF-kB activation. Strikingly the baseline levels of Rag1 and Rag2 transcripts were increased in the LY294002 but not the U0126-treated pre-B cells. Futhermore, both inhibitors were shown to induce a strong increase in the expression of Rag1 and Rag2 transcripts upon pre-BCR crosslinking, suggesting that this receptor exerts dual effects on Rag1/2 expression with a predominant negative regulatory component mediated by both PI3-K and MAPK. No changes in the levels of Pax5, E2A, EBF, IFR4, IRF8, FOXO1, FOXO3, Myb, MAZ, LEF1 and SP1 (transcription factors implied in the regulation of Rag1 and Rag2 transcription) were observed in the pre-BCR stimulated or unstimulated-697 cells, treated or not with the MAPK and Akt inhibitors. Our results suggest that the pre-BCR signaling is a complex and tightly self-controlled process, which deregulation might alter cell growth and survival pathways via NF-kB as well as genomic stability trough Rag1/2 expression.

Disclosures:
No relevant conflicts of interest to declare.
Platelet Derived CD154 Induced by Tat Contributes to HIV-1 Associated Autoimmune Thrombocytopenia.
Authors: Wang Jianhui , Zhang Wei , Li Zongdong 
Published: October 2019

Abstract
Abstract 2669
Poster Board II-645
Enhanced platelet activation was reported in HIV-1 infected patients and strongly correlated with plasma viral load. The HIV-1 Tat protein is able to serve as a ligand of the integrin αvβ3 and several chemokine receptors (CCR2 and CCR3) and to induce downstream signaling in cells express those receptors but the effect of Tat on platelet activation has not been fully investigated yet. Activated platelets are known to express and release a variety of proteins that can modulate the immune system, and platelet derived CD154 is reportedly involved in the development of autoimmune thrombocytopenia. However, the full mechanism underlying HIV-1 induced platelet activation and the biological consequences are not completely understood. In this study, we demonstrate that Tat is able to interact with platelets and β3 integrin by S35 label Tat-platelet binding assay and GST-Tat protein pull-down. We then show that Tat is able to induce platelet activation, up-regulates both CD62P and CD154 in both mouse and human platelets, and results in micro-particle release (as demonstrated by electron microscopy). Tat induces greatly diminished activation in integrin β3 knock out platelets or in platelets pretreated with CCR3 or calcium flux inhibitors, suggesting the requirement of chemokine receptor CCR3, integrin β3 and calcium flux for Tat induced platelet activation. The effect of Tat induced platelet activation on the immune response was studied both in vitro and in vivo. An enhanced immunoglobulin class switch was found in mouse spleen B cells co-cultured with platelets treated with Tat in vitro. In addition, an early antibody response against adenovirus was found in Tat injected mouse immunized with adenovirus suggesting an enhanced immune response in vivo. Thus, we have described a new mechanism in which Tat is able to induce platelet activation and have generated a model in which platelet activation can contribute to the development of HIV-1 associated thrombocytopenia.

Disclosures:
No relevant conflicts of interest to declare.
Granzyme B Produced by Human Plasmacytoid Dendritic Cells Suppresses T Cell Expansion.
Authors: Fabricius Dorit , Vollmer Angelika , Blackwell Sue , Maier Julia , Sontheimer Kai , Beyer Thamara , Mandel Birgit , Lunov Oleg , Tron Kyrylo , Nienhaus G. Ulrich , Simmet Thomas , Debatin Klaus-Michael , Weiner George J , Jahrsdoerfer Bernd 
Published: October 2019

Abstract
Abstract 2674
Poster Board II-650
Human plasmacytoid dendritic cells (pDC) play a central role in regulating adaptive T cell responses in the course of neoplastic, viral and autoimmune disorders. In several of these diseases, elevated extracellular levels of the serine protease granzyme B (GrB) are observed. We found that human pDC can be an abundant source of GrB based on FACS analysis, ELISpot, ELISA, Sensizyme, Western immunoblotting, RT-PCR, and fluorescence microscopy. GrB is actively secreted by pDCs and reaches maximal levels up to two logs higher than those produced by classical GrB producers such as CTL or NK cells. However, pDC GrB production is not accompanied by perforin secretion. Spinning disk confocal microscopy revealed that GrB+ pDC bind to and transfer active GrB to T cells. Importantly, this GrB transfer induces a suppression of T cell proliferation in a GrB-dependent, perforin-independent manner, a process reminiscent of regulatory T cells. GrB expression in pDC is regulated on a transcriptional level by JAK1, STAT3 and STAT5. IL-3 and IL-10 enhance GrB production by pDCs while GrB production is inhibited by toll-like-receptor agonists and CD40 ligand. These findings suggest that GrB production by pDCs is involved in the complex interactions between pDC and T cells and that GrB-secreting pDC may play a regulatory role related to anti-tumor immunity, anti-viral immune responses, and autoimmune processes.

Disclosures:
No relevant conflicts of interest to declare.
Tumor-Derived Microvesicles From Sera of Patients with Acute Myeloid Leukemia Suppresses NK Cell Cytotoxicity Via Transforming Growth Factor Beta-1.
Authors: Szczepanski Miroslaw J , Szajnik Marta E , Czystowska Malgorzata , Mandapathil Magis , Welsh Ann , Whiteside Theresa L , Boyiadzis Michael 
Published: October 2019

Abstract
Abstract 2678
Poster Board II-654
Natural killer (NK) cell cytotoxicity in patients with acute myeloid leukemia (AML) is significantly decreased relative to that in normal controls (NC). However, the mechanisms responsible for low NK cell activity in AML are not known. We considered the possibility that tumor-cell-derived microvesicles (MV) mediate suppression of NK cells. MV originate from the endosomal compartment of activated normal and neoplastic cells. Evidence suggests that tumor-derived MV exert detrimental effects on cells of the immune system and may play a role in tumor progression. To determine their contribution to immune suppression in AML, MV were isolated from sera of patients newly diagnosed with AML prior to any treatment and used to evaluate MV-mediated NK cell suppression. The protein content of MV isolated using exclusion chromatography and ultracentrifugation from sera of 19 AML patients was significantly higher than that of MV isolated from sera of 25 NC (75μg±12/mL vs 1.2μg±0.4/mL, p<0.001 ). MV from AML patients were positive for membrane-associated TGFb-1 and FasL in Western blots, whereas no TGFb-1 or FasL was detected in MV from NC. For functional assays, NK cells sorted from peripheral blood of NC were cultured with MV isolated from sera of the AML patients. A significant decrease in NK cell cytotoxicity was observed after co-incubation with MV (2412 LU before vs 1640 LU after, p<0.002). Concomitantly, a decrease in the expression of the NK cell activating receptor, NKG2D, was observed (57% before vs 38% after, p<0.001). The addition of TGFb1-neutralizing antibody abrogated the effects of MV on the NK cell cytotoxicity and receptor expression. The increased levels in sera of AML patients of MV mediating potent NK cell suppression is likely to compromise anti-tumor immune responses. Therefore, modulation of the levels and functions of MV might provide new immunotherapeutic approaches in AML.

Disclosures:
No relevant conflicts of interest to declare.
Comprehensive Geriatric Assessment-Adapted Chemotherapy in Elderly Patients (>70 years) with Diffuse Large B-Cell Non-Hodgkin's Lymphoma (DLBCL): Final Results and Long Term Follow-up.
Authors: Tirelli Umberto , Fratino Lucia , Balzarotti Monica , Uziel Lilj , Giacalone Annalisa , Ferreri Andres , Santoro Armando , Spina Michele 
Published: October 2019

Abstract
Abstract 2684
Poster Board II-660

Background:
R-CHOP is the standard treatment for elderly patients (pts) with DLBCL. Many pts aged 70 years (yrs) or more are unable to receive R-CHOP and the majority of them are excluded from clinical trials. Comprehensive geriatric assessment (CGA) is an useful instrument to predict the clinical outcome of elderly pts with cancer. Within the GOL (Gruppo Oncoematologico Linfomi) we started a phase II study aiming to evaluate feasibility and activity of a CGA-driven chemotherapy for elderly pts with DLBCL

Material and methods:
Pts with no comorbidity received CHOP/R-CHOP; pts with mild cardiopathy received epirubicin instead of doxorubicin; in pts with moderate/severe cardiopathy the use of anthracyclines was omitted; pts with diabetes did not receive prednisone; in pts with neuropathy vincristine was omitted. The dosage of chemotherapy was decided according to CGA: pts with a good score (ADL=6 and IADL>6) received full doses of CT; pts with an intermediate score (ADL=5 and IADL>4) received 75% of the dose; pts with a poor score (ADL<5 and IADL<5) received 50% of the dose.

Results:
One hundred pts (41 males and 59 females) have been treated. The median age was 75 yrs and stages III-IV were diagnosed in 51% of pts. 61% of pts received full doses of CT; 25% received 75% of dose and 14% received 50% reduced dose; 86% of pts received an anthracycline and 54% rituximab. Toxicity was quite acceptable. Grade 3–4 neutropenia was observed in 30% of pts, mucositis in 12%, and peripheral neuropathy in 9%. Four toxic deaths were observed. Overall, 81% of pts achieved complete remission; with a median follow-up of 50 months, 20% of them have relapsed. The 5 yr-OS, DFS, EFS are 58%, 78% and 50%. It is remarkable that the 5-year specific survival is 72%.

Conclusions:
Our results demonstrate that a CGA-driven approach is feasible in elderly pts with DLBCL. This strategy allows to offer a curative approach to all pts with aggressive NHL, avoiding to under treat pts with a potentially cured disease or over treat pts with severe comorbidities.

Disclosures:
No relevant conflicts of interest to declare.