bluuebunny/crossref_metadata_2025_split huggingface dataset:
Second Autologous Peripheral Blood Stem Cell Transplantation with High Dose Melphalan (HDM/SCT) in Patients Relapsing After An Initial Course of HDM/SCT for the Treatment of AL Amyloidosis.
Authors: Quillen Karen , Seldin David C. , Finn Kathleen T. , Sanchorawala Vaishali 
Published: October 2019

Abstract
Abstract 2318
Poster Board II-295
High-dose melphalan and autologous stem cell transplant (HDM/SCT) can induce complete hematologic responses (CR), defined as disappearance of the underlying monoclonal gammopathy from serum and urine by immunofixation electrophoresis, and of the clonal plasma cell dyscrasia by bone marrow immunohistochemistry, and extend survival in patients with AL amyloidosis. HDM/SCT results in a CR in 40% of patients, and leads to clinical improvements in organ function in >70% of those who achieve a CR. However, hematologic and clinical relapses occur in ∼8% of patients who initially achieve a CR. Tandem cycles of HDM/SCT, which are typically performed within 12 months of each other, have been shown to achieve a higher ultimate CR rate of >60%. Among patients who do not achieve a CR following a single cycle of HDM/SCT, 30% nonetheless experience improvement in organ function. However, in this latter group, clinical improvement is not durable. We designed a study to explore the feasibility, and efficacy, of a second cycle of HDM/SCT in patients who relapse after initially responding to a first cycle of HDM/SCT.

Results:
Eleven patients, median age 55 (range 39-62), M:F 7:4, who had achieved hematologic and clinical responses after an initial cycle of HDM/SCT, were treated with a second cycle of HDM/SCT when a hematologic and/or clinical relapse occurred after a median time interval of 34 months (range 12-63). Five patients underwent a second course of G-CSF mobilization and a mean of 5.1 million (range 3.4-7.6 million) CD34 cells/kg was collected in a median of 2 days; the other patients had cells saved from the first mobilization. Six patients received 200 mg/m2 HDM; 5 patients received modified high-dose HDM at 140 mg/m2. Engraftment occurred at a median of 10 days for neutrophils, and 12 days for platelets (two days without platelet transfusion support); this engraftment timing is similar to that following the initial transplants (10 days for neutrophils, 13 days for platelets). There was no treatment-related mortality, but toxicity was moderate; almost all patients (except one) experienced grade III/IV non-hematologic toxicities. Of the 11 patients, 3 achieved hematologic CR at one year; these patients are alive and in continuous remission at 2-6 yr after the second transplant, including one patient who received a subsequent renal transplant. Three patients died of progressive disease at 1-2 years after the second transplant. Five patients are alive at 1-3 years post second transplant, in partial remission.

Conclusion:
27% (3/11) of patients with AL amyloidosis who experience a hematologic or clinical relapse after responding to initial HDM/SCT can achieve a hematologic CR with a second course of HDM/SCT.

Disclosures:
No relevant conflicts of interest to declare.
Increased Levels of Histone In Human Plasma In Septic Patients with DIC.
Authors: Kawasugi Kazuo , Yamamoto Tadashi , Shirasaki Ryosuke , Tashiro Haruko , Shirafuji Naoki 
Published: October 2019

Abstract
Abstract 1422
Sepsis is a life-threatening condition that is characterized by a whole-body inflammatory state (called a systemic inflammatory response syndrome, SIRS). Histones are essential for packing DNA into the cell nucleus and also play role in regulating gene expression but the new study (Jun Xu et al, Nature Medicine 15. 1318, 2009) have shown histones released into blood stream at the onset of sepsis can have devastating effects (endothelial damage and organ failure etc). However, about extracellular histones in the plasma from human, they have shown only one patient's data. Therefore, we measured extracellular hisotones in the plasma from septic patients with DIC (n=20). Also, we investigated extracellular hisotones in the plasma from septic patients without DIC (n=5). Extracellular histones in the plasma from human were examined by western blot analysis for histone H3 (H3).
The thrombin antithrombin complexes (TAT) levels were higher in septic patients with DIC as reported by others. We detected high levels of extracellular hisotones (H3) in the plasma of all septic patients with DIC by western blotting. However, we did not find any extracellular histones (H3) in the plasma of fiver septic patients without DIC by western blotting. Moreover, high-mobility group box 1 (HMGB1) concentrations in the human plasma were determined using ELISA kit (Shino-Test, Japan). Plasma levels of HMBG1 were negative for four cases in 20 septic patients with DIC. Also, Plasma levels of HMBG1 were negative in septic patients without DIC.
These results suggest that assess of extracellular hisotones in the plasma may be helpful in the making the diagnosis in septic patients with DIC. It appears that extracellular hisotones in the plasma may contribute to develop DIC in patients with sepsis.

Disclosures:
No relevant conflicts of interest to declare.
Daily Pain Is Unrelated to Admission Rate in Adult Patients with Sickle Cell Disease
Authors: van Tuijn Charlotte F.J. , Biemond Bart J. 
Published: October 2019

Abstract
Abstract 1016

Background and aims
Sickle cell disease is characterised by frequent episodes of vaso-occlusive pain resulting in frequent health care utilisation and cumulating organ damage. Disease severity is often quantified by the number of hospital admissions. However, sickle cell patients frequently experience episodes of pain without health care utilisation. In the present study, we analysed whether daily self-reported sickle cell pain in adult patients is related to disease severity quantified by frequency of hospital admission or disease severity defined by genotype.

Methods
We conducted a prospective cohort study in order to analyse daily pain in adult sickle cell patients in a tertiary teaching hospital in the Netherlands. Patients were divided in having severe (HbSS/HbS-β0) or milder (HbSC/HbS-β+) disease according to genotype. Daily pain was assessed for a period of 6 months by the use of pain diaries. Pain intensity was measured with different validated pain assessment instruments (Visual Analog Scale (VAS), McGill Pain Questionnaire and Brief Pain Inventory). Daily pain scores were correlated with genotype and frequency of hospital admission.

Results
110 consecutive outpatients with sickle cell disease were approached. 16 patients were excluded for different reasons (pregnancy, unable to fill in a diary or declined participation), 19 patients missed their appointments in the outpatient clinic and 26 patients completed less than a month of their diaries. 49 patients (mean age 33 years (SD 12.6); 67% female) completed at least two month of the diaries (response rate: 65%). No differences in disease severity according to genotype or age were observed between responders/non-responders.
Pain was reported in 18% of the total 5016 patient-days. Overall median pain intensity was 3.6 (VAS). 41% of the patients reported pain in 5% or fewer diary days and 5% of all patients reported pain in more than 95% of the days. The frequency of hospital admission was unrelated to self-reported pain and no difference in daily pain was observed between patients with severe or milder disease according to genotype. Furthermore, lab results (hemoglobin and fetal hemoglobin levels, leukocyte counts, lactate dehydrogenase (LDH) and the presence of alfa-thalassemia) were not associated with daily pain.

Conclusions
Using daily pain diaries, adult sickle cell patients report pain in 18% of their total diary days. Interestingly, our patient population reported less daily pain in comparison to daily pain in adults reported previously (over 50%). Daily pain appeared to be unrelated to disease severity according to genotype or defined by frequency of hospital admission. Our results further stress that daily pain is an important problem in sickle cell patients and appears to be unrelated to the traditional definitions of disease severity.

Disclosures:
No relevant conflicts of interest to declare.
Critical Role for Histone Deacetylase 6 (HDAC6) in the Regulation of IL-6, and the JAK/STA3 Signaling Cascade in Macrophages
Authors: Lienlaf Maritza , Perez-Villarroel Patricio , Cheng Fengdong , Wang Hongwei , Marante Danay , Yoder Sean , V. Woan Karrune , Knox Tessa , Lagor Stephen , Pinilla-Ibarz Javier , Seto Edward , Sotomayor Eduardo M. , Villagra Alejandro 
Published: October 2019

Abstract
Abstract 1039

Background
Histone Deacetylases (HDACs) have divergent effects over the production of anti- and pro-inflammatory cytokines in Antigen Presenting Cells (APCs). We have previously shown that modulation of specific HDACs can alter the immunogenicity of APCs, either to an activating or tolerogenic phenotype. We recently identified HDAC6 to positively regulate IL-10 production. However, the participation of this HDAC in other immune related cellular processes remains unknown. In this work we are presenting evidence of the important role of HDAC6 in the regulation of IL-6 via activation of the JAK/STAT3 pathway.

Methods
Stable knockdown clones of HDAC6 (KDHDAC6) and Non-target (NT) cells were generated in RAW264.7 murine macrophages using lentiviral shRNA for HDAC6 or non-target (NT) respectively. Two KDHADC6 and two NT clones were treated with LPS or untreated and then analyzed by microarray using the Affymetrix GeneChip Mouse 430 2.0. Significantly down- or up-regulated genes were analyzed by their ontology distribution and selected genes were validated by quantitative real-time RT-PCR, ELISA, or immunoblots. Additionally, changes in the expression of these selected genes were tested in cells treated with selective HDAC6 inhibitors.

Results
1542 genes were down-regulated and 775 up-regulated in KDHDAC6 cells. Their ontology distribution revealed significant changes in immune-related (632) and apoptosis/cell cycle control (47) genes. Importantly, IL-6 was one of the most highly down-regulated genes in KDHDAC6 cells. Therefore, we next analyzed the relevance of these findings by studying the tolerogenic JAK/STAT3 signaling pathway which is known to be activated by IL-6 and critical in the final outcome of APCs in response to stimuli. We observed a complete abrogation in the phosphorylation of JAK2 and STAT3 proteins in KDHDAC6 cells in response to LPS, which was reverted when these cells were treated with exogenous IL-6.

Conclusions
These results demonstrate the requirement of HDAC6 in the production of IL-6 in response to LPS, and therefore positions this deacetylase as an important regulator of the JAK/STAT3 pathway. These findings provide insight into the molecular mechanisms controlling the immunogenicity of APCs, supporting the use of HDAC6 inhibitors to enhance immune activation.

Disclosures:
No relevant conflicts of interest to declare.
A GM-CSF and IL-4 Fusion Cytokine Triggers Conversion of B-Cells to Tumoricidal Effectors
Authors: Deng Jiusheng , Li Pingxin , Pennati Andrea , Yuan Shala , Hsieh Hsiang-Chuan (Jeremy) , Wu Jian Hui , Galipeau Jacques 
Published: October 2019

Abstract
Abstract 1048
We have previously demonstrated that coupling of GMCSF at the N-terminus of common γ-chain interleukins IL2, IL15 and IL21 leads to meaningful gain-of function activity in interleukin-responsive lymphomyeloid cells. Considering the physiological and immunological importance of IL4 that is a member of γ-chain interleukins, we tested the bioactivity of a novel fusion cytokine consisting of a fusion between GM-CSF and IL-4 (GIFT4). We observed that GIFT4 leads to a pan-STAT hyper-phosphorylation response in resting splenic B-cells distinct from IL4 only and that treated B-cells up-regulated expression of MHCI/II, CD80 and CD86, secreted IL-12, IL-1a, IL-6, and substantial amounts of CCL3 and GM-CSF, akin to recently described innate response activator (IRA) B-cells (Science 335, 597, 2012). In vivo delivery of recombinant GIFT4 protein to normal mice leads to homeostatic expansion of splenic B cells and plasma cells as well as humoral hyper-responsiveness to antigenic challenge. We further showed that B16F0 melanoma cells engineered to secrete GIFT4 are immune-rejected in a B-cell dependent manner. The clinical effect was abolished when B16F0-GIFT4 cells were implanted in B-cell deficient μMT, CD4−/−, CD8−/− or FcγR−/− mice consistent with a pivotal for B cells, their T-cell helper function and antibody-dependent cell-mediated cytotoxicity for the observed melanoma-specific therapeutic effect. Thus, GIFT4 defines a novel engineered cytokine that mediates endogenous expansion of B-cells with potent immune helper and antigen-specific effector function. We propose that GIFT4 protein could serve as a novel immunotherapeutic agent and defines a previously unrecognized potential of B-cells as tumoricidal effectors.

Disclosures:
No relevant conflicts of interest to declare.
Is a Pbac-Score a Good Tool to Quantify Menorrhagia in Women with Von Willebrand Disease and Rare Bleeding Disorders?
Authors: Halimeh Susan , Rott Hannelore , Siebert Manuela , Kappert Guenther 
Published: October 2019

Abstract
Abstract 1133

Introduction:
Von Willebrand disease (VWD) is the most common inherited bleeding disorder. VWD and other autosomal inherited bleeding disorders equally affect women and men.
Menorrhagia or severe menstrual bleeding (HMB) is the most common symptom of women with bleeding disorders. HMB is defined as bleeding that lasts for more than seven days or as the loss of more than 80 mL of blood per menstrual cycle.
The menstrual blood loss can be quantified by the use of a pictorial bleeding assessement chart (PBAC).

Samples and methods:
In 195 women with menorrhagia and in 45 controls menstrual blood loss was quantified using pictorial blood assesment charts (PBAC) and results were compared.

Results:
In 169 of 195 women (86%) a bleeding disorder could be detected. In those with a bleeding disorder, the distribution was as followed: 62% had a von Willebrand disease, 14,4% had a factor-VII-deficiency (F7D), 5% had a factor-XIII-deficiency (F13D) and the remaining 18,6% had other beedling disorders (e. g. hypofibrinogenaemia and other mild factor deficiencies). The median PBAC-Score of all patients was 268 (range: 10–4212). In our controi group of 45 women the median PBAC-Score was 46,5 (3- 137).ROC-Analysation shows that the PBAC (AUC=0.977) is much more useful than the number of bleeding days (AUC=0.855) in order to distingish controls from patients suffering from menorrhagia due to a coagulation disorder. We found that the best cutoff for the PBAC is 100 with an sensitifity of 88% and a specifity of 97%.

Discussion:
Attempts to measure the quantity of menstrual blood loss can be useful in clinic practice. One study found that variables predicting a blood loss higher than 80ml per menses were clots greater than one inch, low ferritin levels, or changing a pad or tampon more than hourly (flooding). A prospective method of quantifying menstrual blood loss includes the use of a pictorial bleeding assessement calendar (PBAC). We are of the opinion, that we would have detected more bleeding disorder also in the patients, where we did not find any diagnosis until now, if we would have controlled them more than one time during the cycle period.

Conclusions:
Women with hyermenorrhagia frequently suffer from a bleeding disorder, in 86% of our patients an abnormal coagulation was found.
The PBAC-Score is an easy tool to quantify menstrual blood loss in women.
In our study a PBAC-Score above 100 was suspicious of having a bleeding disorder.

Disclosures:
No relevant conflicts of interest to declare.
Risk of Venous Thrombosis Associated with White Cell Count On Peripheral Blood and Its Interrelationship with Other Environmental Risk Factors
Authors: Rezende Suely M , Lijfering Willem M. , Rosendaal Frits R. , Cannegieter Suzanne C. 
Published: October 2019

Abstract
Abstract 1149

Background:
Antithrombotic prophylaxis needs to be balanced against bleeding risk. Therefore, risk stratification is useful to identify patients who would benefit most. In this study, we analyzed how VT risk relates to different leukocyte count. We also analyzed the effect of high leukocyte counts in groups with high risk of VT.

Methods:
In the MEGA case-control study, blood leukocyte count and information on environmental risk factors were collected from 2443 patients with VT and 1459 partner controls. Logistic regression analyses were adjusted for age and sex.

Results:
Population characteristics are detailed in Table 1. Risk of VT increased for measurements above the 97.5th percentile for total white cells, granulocytes, lymphocytes and monocytes. Adjusted odds ratio, [OR] for 99th percentile was 1.37 (95% confidence interval [CI], 0.73–2.56), 1.48 (95% CI, 0.79–2.77), 1.20 (95% CI, 0.63–2.31) and 1.88 (95% CI, 1.02–3.46), for total white cells, granulocytes lymphocytes and monocytes, respectively, compared to the reference percentile (5th-95th) (Table 2). We also analysed the effect of high leukocyte counts on VT in high risk groups, including surgical patients, hospitalized patients, and patients with cancer. Adjusted OR for high leukocyte counts (white cell, lymphocyte, monocyte or granulocyte counts > 97.5th percentile) was 6.3 (95% CI, 0.84–47.1) for groups who had surgery or were hospitalized and 2.2 (95% CI, 0.6–8.2) for patients with cancer, respectively, compared with cell counts within the percentile between 5th and 97.5th.

Conclusions:
High blood leukocyte count is associated with increased risk of VT. The risk is further increased in high risk groups. These results may assist in individually tailored thromboprophylaxis in high risk groups.

Disclosures:
No relevant conflicts of interest to declare.
Hematopoietic Stem Cells Are Dependent On Homologous Recombination Only During Active Cell Cycle in Nuclease-Mutant Exonuclease1mut mice
Authors: Desai Amar , Gerson Stanton L. , Qing Yulan 
Published: October 2019

Abstract
Abstract 1225
Hematopoietic stem cell (HSC) maintenance and self-renewal is crucial for long term tissue repopulation and immune function. HSC populations require functional DNA repair pathways in order to maintain their reconstitution capabilities but the pathways involved and the mechanisms of regulation are still being elucidated. It has been proposed that quiescent HSCs rely on the error prone non homologous end joining pathway for DNA double strand break (DSB) repair while HSCs in cycle use both NHEJ and the high fidelity homologous recombination (HR), but functional in vivo studies have not yet been completed. Exonuclease 1 participates in homologous recombination. We used Exo1mut fibroblasts to demonstrate that loss of Exo1 function results in a defective HR response, increased sensitivity to DSB inducing agents, and aberrant DNA damage signaling. However, Exo1mut mice did not appear to require HR to maintain quiescent HSCs at steady state or to respond to DNA damage. Exo1mutmice were able to sustain long term serial repopulation, displayed no defect in competitive repopulation or quiescence maintenance, and did not display increased sensitivity to whole body ionizing radiation (IR). In contrast, when Exo1mut HSCs were pushed into cell cycle with 5-Fluorouracil, the hematopoietic population and HSCs became hypersensitive to IR stress relative to WT B6 mice, as shown by decreased bone marrow cellularity, colony forming unit defects, loss of the HSC population, and finally animal death. Thus, loss of Exo1, and in turn fully functional HR, in quiescent HSC is not critical to stem cell function, survival, or recovery after DNA damage, whereas HR mediated repair of DNA damage is essential for HSC maintenance after cell cycle entry. In HSCs, DNA damage repair response, and sensitivity is dependent on cell cycle.

Disclosures:
No relevant conflicts of interest to declare.
Elevation of Mitochondrial DNA Copy Number in Bone Marrow Cells Associated with Increased Risk of Childhood Acute Myeloid Leukemia
Authors: Kim Yu-Na , Kim Hye-Ran , Kim Hwan-Young , Dai Trang Nguyen Thi , Han Dong-Kyun , Baek Hee-Jo , Kook Hoon , Shin Myung-Geun 
Published: October 2019

Abstract
Abstract 1423

Background:
Alteration of mitochondrial DNA (mtDNA) is now implicated in tumorigenesis as well as poor clinical outcomes. mtDNA copy number is altered in patients with various hematological malignancies such as lymphocytic leukemia, lymphoma and other disorders. Moreover, recent studies has reported that mtDNA copy number is associated with increased risk of non-hodgkin lymphoma (Blood 2008;112:4247–9). Therefore, the present study was fist to validate the hypothesis that alteration of mtDNA copy number in bone marrow cell would be associated with risk of childhood acute myeloid leukemia (AML).

Patients and Methods:
Direct sequencing of mtDNA control region, analysis of mtDNA minisatellites and mtDNA copy number were performed in 48 patients with initially diagnosed childhood AML cases and 48 matched case controls from Environmental Health Center for Childhood Leukemia and Cancer cohort. Molecular analysis of mtDNA including quantitation of mtDNA copy number was determined using our published protocol (Int J Cancer 2012;131:1332–41).

Results:
Numerous sequence alterations of mtDNA were only found in patient's bone marrow cells. Instability of mtDNA minisatellites was mainly occurred in the 303 polyC and 16184 polyC minisatellite. mtDNA copy number was significantly elevated in patients with childhood AML compared with those of control group (p=0.000075, Fig. 1). There was a dose-response relationship between tertiles of mtDNA copy number and risk of childhood AML (odds ratio [OR], 95% confidence interval [CI]: 1.0; 1.145 [0.412–3.183]; and 6.818 [2.244–20.712], respectively; p(trend) =.000). The effect was most pronounced for childhood AML harboring normal karyotype (OR: 1.0; 1.75[0.369–8.302]; 9.00 [1.888–42.904]; p(trend) =.008) (Table 1).

Conclusion:
mtDNA minisatellite inability was frequently observed in bone marrow cells of childhood AML. Elevation of mtDNA copy number could be associated with increased risk of childhood AML, particularly normal karyotype childhood AML.

Disclosures:
No relevant conflicts of interest to declare.