bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinicopathologic Significance of CD11c, CD16 and FOXP3 Expression in Diffuse Large B-Cell Lymphoma (DLBCL) Patients Receiving Rituximab, Cyclophosphamide, Anthracycline, Vincristine, and Prednisolone (R-CHOP) Combination Chemotherapy
Authors: Oh Sung Yong , Jung Jin Sook , Kim Sung Hyun 
Published: October 2019

Abstract
Abstract 5112

Backgrounds:
The principal objective of this study was to determine whether the expressions of CD11c, CD16 and FOXP3 are predictive of clinical outcomes in diffuse large B-cell lymphoma (DLBCL) patients receiving treatment with rituximab, cyclophosphamide, anthracycline, vincristine, and prednisolone (R-CHOP) combination chemotherapy.

Method:
The study population consisted of 25 patients with DLBCL between January 2006 and December 2007. All the patients had been treated with R-CHOP combination chemotherapy. The R-CHOP regimen was as follows: 375 mg/m2 rituximab, 750 mg/m2 cyclophosphamide, 50 mg/m2 anthracycline, 1. 4 mg/m2vincristine were intravenously administered on day 1, and prednisolone 100 mg was medicated on days 1 to 5. This regimen was repeated every three weeks. The expressions of CD11c, CD16 and FOXP3 of primary tumors were immunohistochemically evaluated.

Result:
The mean age of the patients was 58 years (range, 27–77 years), and the pilot study population included 11 (44%) females and 14 (56%) males. The stages of the patients were I, II, III, and IV in 4 (16%), 5 (20%), 7 (28%), and 9 (36%), respectively. 60% of the patients were categorized as the low (44%) or low-intermediate risk group (16%) according to the International Prognostic Index (IPI). After a median follow-up of 57months, 8 patients (32%) developed disease recurrence or death. The estimated 5-year Progression free survival (PFS) and Overall survival (OS) were 65% (Figure 1) and 74% (Figure 2), respectively. 8 (32%), 18 (72%), and 17 (68%) were positive for CD11c, CD16 and FOXP3 expression, respectively. Univariate analysis of CD11c, CD16, FOXP3 for PFS and OS are shown in Figures 3–5. We noted no significant correlation between immunostaining of CD11c, CD16, FOXP3, and PFS. Patients with a CD11c -expressing DLBCL had a significantly higher 5-year OS rate than those without (p=0. 033, Figure 3). We classified patients according to the risk group. Each one counted for 1 point; In CD11c and FOXP3 stain, positive was counted as 0, negative was 1. In CD16, positive was counted as 1, negative was 0- and they were summed. The points were separated into low risk (0–1) and high risk (2–3) groups (Figure 6). We could found significant differences of PFS (p=0. 015) and OS (p=0. 046) between two groups.

Conclusion:
Even though this result was obtained from small number and retrospective data set, we can get a possibility of clinical significance of CD11c, CD16 and FOXP3 in DLBCL patients receiving R-CHOP therapy. Prospective large scale study is needed for confirming these promising our pilot study results.

Disclosures:
No relevant conflicts of interest to declare.
NPM-RAR Enhances NFκB Transcriptional Activity
Authors: Redner Robert L. , Abecassis Irina , Rush Elizabeth A. , Chattopadhyay Anuja 
Published: October 2019

Abstract
Abstract 5117
Acute Promyelocytic Leukemia (APL) is characterized by recurrent translocations of 17q21, which result in expression of fusion proteins of the retinoic acid receptor alpha (RARα). We have been studying the t(5;17)(q35;q21) variant of APL, which fuses the N-terminal 117 amino acids of nucleophosmin (NPM) to the C-terminus of RARα. NPM-RAR is capable of binding to retinoic acid response elements (RAREs), and we have previously shown that NPM-RAR can recruit both corepressor and coactivator complexes to retinoic acid target promoters, to modulate transcription in a ligand-dependent fashion, though with a different sensitivity to ligand than RARα. The N-terminal NPM-sequence of NPM-RAR contains a dimerization domain capable of mediating not only homodimerization of the fusion protein, but also heterodimerization with wild-type NPM, to potentially modulate NPM functionality. NPM serves as a molecular chaperone, and binds to a diverse set of proteins. Amongst these are the p50 and p65 subunits of NFκB. NPM has been shown to serve as a coactivator for the NFκB transcriptional complex. We hypothesized that NPM-RAR might interact with NPM to modulate NFκB dependent transcription. We focused on NFκB regulation of the Superoxide Dismutase 2 (MnSOD, SOD2) promoter, since NPM regulation of NFκB mediated transcription has been most extensively studied in this model system. We found that NPM-RAR enhances both basal and TNFα-induced transcription of an NFκB-reporter construct (containing a minimal MnSOD promoter/enhancer) in HepG2 and Hela cells. This effect was not dependent upon translocation of NFκB to the nucleus, indicating that the positive effect on NFκB was mediated downstream of the canonical activation pathway of NFκB. The MnSOD promoter does not contain a consensus RARE, and this effect was not seen in cells transfected with RARα, suggesting that this effect is not mediated through the DNA binding motifs of the NPM-RAR C-terminal RARα sequences. NPM-RAR does not directly interact with NFκB; preliminary experiments using siRNA to down-regulate NPM indicate that NPM-RAR enhancement of MnSOD transcription is dependent upon the presence of wild-type NPM. This leads us to propose a model whereby NPM-RAR binds to the NPM/NFκB transcriptional complex through interaction with NPM, to enhance NFκB mediated transcription.

Disclosures:
No relevant conflicts of interest to declare.
Myeloid Lineage-Associated Mutations Are Detected in Blastic Plasmacytoid Dendritic Cell Neoplasm
Authors: Wang Yongbao , Windham Justin , Billouin-Frazier Shere , Jones Dan 
Published: October 2019

Abstract
Abstract 5123

Background/Purpose:
Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare but highly aggressive hematologic malignancy that frequently presents with skin, lymph node, and leukemic disease. Since BPDCN can evolve into acute myelomonocytic leukemia (AMML) with other cases associated with monocytosis, a close relationship of BPDCN to the myelomonocytic lineage is suspected. Here we investigate the frequency of myeloid-associated DNA mutations in BPDCN using a targeted exon sequencing panel.

Methods:
Nine cases of BPDCN (5 skin, 1 lymph node, 3 bone marrow clot sections) were studied. All showed blastic cytomorphology; characteristic expression of CD56 and CD123; CD4 and/or TCL1 expression; and absence of definitive lymphoid, myeloid, and monocytic marker expression. AMML recurrence at 18 months, seen in 1 patient, was also assessed. Genomic DNA was extracted from formalin-fixed paraffin-embedded sections and subjected to a 161-amplicon PCR-based DNA sequencing assay that includes the commonly mutated areas of KRAS, NRAS, IDH1, IDH2, TET2, EZH2, JAK2, and ASXL1. Limited multiplex PCR and product harvesting was performed on an Access Array chip (Fluidigm). Ion adaptors were added and DNA sequencing was performed on a PGM sequencer using the 316 chip with 200 base-pair sequencing chemistry (Life Technologies). Sensitivity of mutation detection varied between 5–10%. Sequencing data were analyzed by SeqNext software (JSI MedSystems). Detected mutations were confirmed using conventional Sanger sequencing, with an approximate sensitivity of 15%, or pyrosequencing (5%).

Results:
Mutations in one or more of the tested genes were found in 6 of 9 patients, with unconfirmed mutations in 2 additional cases due to low-level mutations in samples with poor quality DNA. One patient with a typical BPDCN showed an IDH2 mutation (R140Q) with recurrence as AMML showing this mutation as well as a double mutation in KRAS (G12R, Q61R). Other mutations identified included 3 in ASXL1 including W608C, NRAS G12D, EZH2 C-terminal truncation, and 2 TET2indels.

Conclusions:
The presence of myeloid-associated mutations particularly in IDH2, TET2 and ASXL1 in cases of BPDCN is support for a relationship to the myelomonocytic lineage. The presence of shared and new mutations in the AMML recurrence in 1 patient with BPDCN further indicates a pathogenetic relationship between the two entities.

Disclosures:
Wang: Quest Diagnostics: Employment. Windham:Quest Diagnostics: Employment. Billouin-Frazier:Quest Diagnostics: Employment. Jones:Quest Diagnostics: Employment.
Frequency and Distribution of Asymptomatic Brain Lesions in Patients with β-Thalassemia Intermedia
Authors: Karimi Mehran , Haghpanah Sezaneh , Bagheri Mohammad Hadi , Bordbar Mohammad Reza , Pishdad Parisa , Rachmilewitz Eliezer A. 
Published: October 2019

Abstract
Abstract 5141
We aimed to determine the frequency of asymptomatic brain lesions in a group of patients with β-thalassemia intermedia (β-TI) and to evaluate correlation of asymptomatic brain lesions with splenectomy, thrombocytosis, blood transfusions and clinical parameters. Ninety five neurologically intact patients with β-TI were randomly enrolled in this cross-sectional study. Diffusion Weighted Imaging (DWI) brain MRI was performed in every patient to detect cerebral white matter lesions (WML). We found an overall frequency of 15 (15. 8%) for WMLs, 14 (23. 7%) in splenectomized and 1 (2. 8%) in nonsplenectomized patients. The presence of WML was significantly associated with splenectomy (P= 0. 008) and thrombocytosis (P=0. 009). However, after adjustment for splenectomy, thrombocytosis was not significantly associated with the presence of WML (P>0. 05). The number of patients with regular blood transfusions and normal MRI was not significantly higher compared to those with abnormal findings (52. 5% vs 26. 7%; P=0. 092). In untransfused patients, hydroxyurea (HU) administration was associated with a lower incidence of WML (P<0. 001). Although in univariate analysis either splenectomy or thrombocytosis showed significant correlation with the presence of single or multiple WMLs, thrombocytosis by itself did not significantly contribute in developing asymptomatic brain lesions. The lack of significant correlation between lesions and regular blood transfusions could be related to the treatment with HU in untransfused patients, which increased fetal hemoglobin levels and improved the morphology and the pathological indices of the red blood cells. Larger prospective studies are suggested for evaluation of the correlation of these factors with developing asymptomatic brain lesions accurately.

Disclosures:
No relevant conflicts of interest to declare.
Aspirin Induces Platelet Apoptosis
Authors: Zhao Lili , Zhang Weilin , Chen Mengxing , Zhang Jiansheng , Zhang Mingyi , Ruan Changgeng , Dai Kesheng 
Published: October 2019

Abstract
Abstract 5144
Aspirin is widely used in the treatment of a number of clinical conditions, such as fever, pain, strokes, and heart attacks. Although aspirin is being thought of a relatively “safety” medicine, it also has some side effects, particularly the risk of bleeding which may be severe and lead to death. The mechanisms, however, are not totally understood. The effectiveness of aspirin has been attributed to its ability to inhibit prostaglandin production by inhibiting the cyclooxygenase (COX) enzyme. Interestingly, it has been reported recently that aspirin induces apoptosis in many cell types. Platelet apoptosis induced by either physical or chemical compounds or platelet storage occurs widely. We have reported that the glycoprotein Ibα-von Willebrand factor interaction, hyperthermia, and calmodulin antagonists induce platelet apoptosis, suggesting that platelet apoptosis might play important roles in controlling the number and function of circulated platelet under physiological conditions, or in the development of platelet-related hemorrhagic diseases. Thus, the aim of the current study is to explore whether aspirin induces platelet apoptosis. Washed platelets (3×108/ml) were incubated with various concentrations of aspirin (2. 5 mM, 5 mM, 10 mM, 20 mM) at 37 °C for 2 hours. The effect of aspirin on platelet mitochondrial membrane potential (ΔΨm) depolarization and phosphatidylserine (PS) exposure were analyzed by cell-permeable lipophilic cationic dye tetramethylrhodamine ethyl ester (TMRE) and annexin V binding, respectively. The data show that ΔΨms depolarization and PS exposures were dose-dependently induced by aspirin in platelets. In order to further confirm that aspirin incurs platelet apoptosis, caspase-3 activation was measured in platelets incubated with aspirin by detecting 32 kD pro-caspase-3 and 17 kD cleaved caspase-3 fragment, and the result shows that aspirin induced caspase-3 activation. Furthermore, platelet shrinkage appeared in platelets incubated with aspirin (20 mM). Caspase inhibitor z-VAD-fmk inhibited apoptotic platelet shrinkage, and blocked ΔΨm depolarization, but had no effect on PS exposure, suggesting that caspase-3 acts upstream of ΔΨm depolarization, and plays a key role in aspirin-induced platelet apoptosis. Whereas, protein kinase C (PKC), protein kinase B (PKB), p38 mitogen-activated protein kinases (MAPK), protein 53 (p53), reactive oxygen species (ROS), and mitogen-activated protein kinase kinase (MAPKK) were not involved in aspirin-induced platelet apoptosis. In addition, platelets incubated with another COX inhibitor indomethacin did not incur ΔΨm depolarazation and PS exposure in platelets. Taken together, the data indicate that aspirin induces platelet apoptosis via caspase-3 activation, and independent of the COX signaling.

Disclosures:
No relevant conflicts of interest to declare.
Erythropoietin Induced Erythroid Precursor Pool Depletion Causes Erythropoietin Hyporesponsiveness
Authors: Yan Xiaoyu , Ait-Oudhia Sihem , Krzyzanski Wojciech 
Published: October 2019

Abstract
Abstract 5156

Background:
Erythropoietin (EPO) hyporesponsiveness is demonstrated by a persistence of anemia despite high dose of recombinant human erythropoietin (rHuEPO) or requirement of large doses to maintain the target hemoglobin concentration. The purpose of this study is to demonstrate that the bone marrow depletion induced by rHuEPO treatment may another contributing factor to hyporesponsiveness.

Methods:
Healthy Wistar rats were given single dose (SD) or multiple doses (MD) of rHuEPO. In MD study, animals were challenged with thrice-weekly (100 IU/kg) over two weeks and the pharamcodynamic biomarkers included reticulocyte (RET) and red blood cells (RBC) counts, and hemoglobin (HGB) concentrations. In SD study, in addition to the biomarkers in the peripheral blood, the erythropoietic responses in bone marrow and spleen were also quantified using a flow cytometric immunophenotyping technique. The total marrow and spleen cellularity was quantitatively assessed using flow cytometry. A mathematical approach involving measuring RET age distribution was developed to evaluate the RET loss due to neocytolysis. The pharmacokinetics of rHuEPO was assessed by measuring serum concentrations over time using ELISA.

Result:
In MD study, a slow and oscillatory decline of RET response was observed before the administration of the last dose on day 11. The RET kept decreasing below the baseline and reached a nadir on day 22, followed by a slow return to the baseline on day 28. These observations demonstrated the tolerance and rebound phenomena. In SD study, the RET decreased below the baseline after day 6. A depletion of the bone marrow erythroid precursor cells was also observed. Meanwhile, neocytolysis of RET was only observed from day 3–5 after rHuEPO injection. These results suggested that the reduced levels of RET after day 6 resulted from the depletion in the erythroid precursor cells in bone marrow induced by rHuEPO treatment.

Conclusion:
The EPO hyporesponsiveness is of clinical importance because it is associated with an increased risk of death or cardiovascular events. The findings in this study demonstrate that EPO-induced erythroid precursor depletion may contribute to the rHuEPO hyporesponsiveness. When the erythroid precursor cells are depleted, an increase of rHuEPO dose would not show much benefit, due to the smaller number of cells present to initiate the erythropoiesis. These findings provide additional justification for reducing the doses of erythropoietin stimulating agents in anemic patients demonstrating hyporesponsiveness.

Disclosures:
No relevant conflicts of interest to declare.
Favorable and Sustained Response of Primary Autoimmune Hemolytic Anemia to Rituximab
Authors: Al-Momen Abdul Kareem M , Aleem Aamer , Gahtani Farjah Al , Hasanato Rana , Anjum Farhan Ali , Blecana Sri-Halima , AlSaleh Khalid 
Published: October 2019

Abstract
Abstract 5160

Background:
Primary autoimmune hemolytic anemia (PAIHA) is a rare autoimmune disorder that males and females of all age groups, but more frequent in young adults. Most patients respond initially to steroids and intravenous immunoglobulin's followed by Splenectomy for non-responders. Around one third of the patients do not respond to any standard treatment and another one third show only transient response and therefore labeled as refractory PAIHA. In this refractory group, various immune modulators and suppressants are used with variable response. These include azathioprine, vincristine, cyclophosphamide and mycophenolate.

The Aim:
The aim of our study is to explore the safety and efficacy of Rituximab in the treatment of refractory, PAIHA.

Patients and methods:
Thirteen patients (seven males and six females, aged 33. 7 +/− 15 years) with refractory PAIHA (Hb 73. 1 +/− 13. 9 g/l) were given Rituximab 375 mg/m2 intravenously weekly for four weeks, then monthly for four months.

Results:
All patients showed a rapid and favorable response with significant rise in Hb level to 128 +/− 10. 8 g/l within four months. Rituximab was tolerated very well and the response was sustained in all patients during the follow up to 25. 6 +/− 21. 8 months

Conclusion:
Rituximab is probably highly effective in the treatment of refractory PAIHA with sustainable response. This has been shown in refractory autoimmune hemolytic anemia in adults and children as well as in chronic lymphocytic leukemia associated autoimmune hemolytic anemia (1–3)

Disclosures:
Off Label Use: Rituximab was used in Primary Autoimmune Hemolytic Anemia compassionately.
Hereditary Spherocytosis: How the New Information Provided by the Routine Hematology Analysers May Help in Its Differential Diagnosis or Flagging
Authors: Barella Susanna , Simon-Lopez Ramon , Gaetano Nicola Di , Galanello Renzo 
Published: October 2019

Abstract
Abstract 5163

Introduction:
Hereditary Spherocytosis (HS) is one of the most common inherited hemolytic anemias. Many of them are autosomal dominant, being about 25% of the cases transmitted recessively.

Diagnostic of HS:
Classic testing for HS includes: Hematologic testing of red blood cell indices (RDW, MCHC, Reticulocyte count), peripheral blood smear (presence of spherocytes), osmotic fragility and Eosin-5-maleimide binding to band 3 and Rh-related proteins forms that may be used as screening tests for hereditary spherocytosis.

Objective:
Recently have been developed new parameters and information in the new automated hematology analyzer called DxH8008™ from Beckman Coulter as @MSCV (@Mean Sphered Cell Volume), @RSF, @MAF, @ LHD%. All this parameters may be used to create flagging for laboratory use only (LUO) or Research use only (RUO). The purpose of this study is to investigate the possible use or utility of this new information for the screening/flagging of Hereditary Spherocytosis. There are previous studies showing the possible benefit of using MCV minus @MSCV for the detection/flagging of cases with spherocytes.

Patient and Methods:
We have collected 28 patients with Hereditary Spherocytosis. All of them were confirmed by red cell morphology, osmotic fragility and Eosin-5-maleimide binding to band 3 and Rh-related proteins forms.

Results:
Using ROC analysis, the best parameters differentiating the Hereditary Spherocytosis from the normals were: RET% (AUC 0. 996), MCV - @MSCV (AUC 0. 996), @MSCV (AUC 0. 969), RDW(AUC 0. 892), MCHC (AUC 0. 860), HGB (AUC 0. 787).
Using ROC analysis, the best parameters differentiating the Hereditary Spherocytosis from other anemias (excluding normals)were: MCV - @MSCV (AUC 0. 991), MCHC (AUC 0. 987), RET% (AUC 0. 857).

Disclosures:
Simon-Lopez: Beckman Coulter: @LHD, @MAF, @RSF, @LHD, @MAF, @RSF Patents & Royalties, Employment. Di Gaetano:Instrumentation Laboratory spa: Work for a distributor of Beckman Coulter Instruments in Italy Other. Galanello:Novartis: Research Funding, Speakers Bureau; Apopharma: Research Funding, Speakers Bureau; Ferrokin: Research Funding.
Iron Chelation Therapy and Mobilization of Hematopoietic Peripheral Progenitors in Patients with β-Thalassemia Major
Authors: Forni Gian Luca , Podestà Marina , Musso Marco , Piaggio Giovanna , Musallam Khaled M , Balocco Manuela , Pozzi Sarah , Frassoni Francesco 
Published: October 2019

Abstract
Abstract 5178
Several reports established an association between iron chelation therapy with deferasirox and hematopoietic improvement in patients with myelodysplastic syndromes. Data in β-thalassemia major (TM) patients is absent. We evaluated levels (frequency and aboslute number) of several hematopoietic peripheral progenitors (HPP: Colony Forming Unit-Granulocyte/Macrophage [CFU-GM], Erythroid Burst-Forming Unit [BFU-E], Colony Forming Unit-Granulocyte, Erythrocyte, Macrophage, Megakaryocyte [CFU-GEMM], and Long Term Culture-Initiating Cells [LTC-IC]) in 26 TM patients (median age 28. 5 years, 42. 3% males) and 12 age-matched controls. All TM patients had to be using the same iron chelator for at least 6 months with >80% compliance. The levels of all HPP were significantly higher in TM patients than controls, and varied between splenectomized and nonsplenectomized patients (lower CFU-GM and BFU-E, and higher LTC-IC in splenectomized patients). Patients using deferasirox (n=9) showed significantly higher levels of BFU-E compared with both deferoxamine (n=10) and deferiprone (n=7) treated patients (p<0. 01). After adjusting for age, sex, splenectomy status, serum ferritin changes, the association between higher BFU-E levels and deferasirox compared with deferoxamine or deferiprone therapy remained statistically significant (R2=0. 698, p=0. 006). Mobilization of BFU-E in TM patients receiving regular iron chelation therapy depends on the type of chelator used, independent of iron status.

Disclosures:
Forni: Novartis Pharmaceuticals: Research Funding; Ferrokin: Research Funding. Musallam:Novartis Pharmaceuticals: Honoraria.