bluuebunny/crossref_metadata_2025_split huggingface dataset:
CRLF2 and JAK2 Mutations: Occurrence and Function In Pre-B ALL Cell Lines
Authors: Quentmeier Hilmar , Eberth Sonja , MacLeod Roderick AF , Nagel Stefan , Romani Julia , Zaborski Margarete , Drexler Hans G. 
Published: October 2019

Abstract
Abstract 5109
Thymic stromal lymphopoietin (TSLP), a cytokine produced by epithelial cells promotes early B-cell development and activates dendritic cells. It has recently been reported that a subset of B-cell precursor acute lymphoblastic leukemia (pre-B ALL) overexpresses the TSLP receptor CRLF2. CRLF2 overexpression is linked to translocations between sex chromosomes – localizing CRLF2 – and the immunoglobulin heavy chain locus on chromosome 14, or to an interstitial deletion on the gonosomes. Both events, translocation and deletion juxtapose CRLF2 to a different promoter (IgH or P2RY8). Performing quantitative real-time PCR we tested pre-B ALL and acute myeloid leukemia (AML) cell lines for overexpression of CRLF2. AML cell lines were included in the screening because we knew from an earlier TSLP project that the AML cell line MUTZ-3 is TSLP-responsive, and thus positive for the cytokine receptor. Three of 63 (5%) pre-B ALL cell lines tested (INC, MHH-CALL4, MUTZ-5) overexpressed CRLF2 mRNA. CRLF2-high cell lines carry a t(14;Y). With respect to the 58 AML cell lines tested: some expressed CRLF2 mRNA, but none of them rivalled the aforementioned pre-B cell lines. Pre-B ALL cell lines show the association between chromosomal CRLF2 aberrations and JAK2 pseudokinase domain mutations that has been described for primary pre-B ALL cells: cell lines MHH-CALL4 (JAK2I682F) and MUTZ-5 (JAK2R683G) and – newly described - also the CRLF2-high pre-B ALL cell line INC express a mutated version of Janus kinase 2 (JAK2R683G). We established a PCR based assay system that allowed for the rapid detection of the JAK2R683G mutation: none of the CRLF2-low or –negative pre-B ALL cell lines exhibited this mutation. All three CRLF2-high/JAK2mu cell lines showed high phosphorylation levels of the JAK2 downstream target STAT5. Inhibition of the JAK kinase led to dephosphorylation of STAT5. However, repression of 3H-thymidine uptake and induction of apoptosis by inhibition of the JAK2/STAT5 pathway was weaker in the JAK2mu pre-B ALL cell lines than in the JAK2V617F positive essential thrombocythemia-derived cell line SET-2. Provided that these results reflect the situation in primary cells, mutated JAK2 seems to be of lesser importance for growth and survival of pre-B ALL cells than for cells from myeloproliferative neoplasms. The CRLF2-high/JAK2mu cell lines INC, MHH-CALL4 and MUTZ-5 are promising model systems for the study of the roles of high-level CRLF2 expression and of JAK2 mutations in pre-B ALL.

Disclosures:
No relevant conflicts of interest to declare.
Expression of the Hypoxia-Inducible Factors In the AML and MDS: Is Not Associated with the Von Hippel-Lindau Gene
Authors: Tong Hongyan , Mei Chen , Li Kongfei , Jin Jie 
Published: October 2019

Abstract
Abstract 5114
The hypoxia-inducible factor (HIF) is a transcriptional factor with important roles in tumor biology, which activates transcription of genes encoding glucose transporters, glycolytic enzymes, and vascular endothelial growth factor. The VHL (von Hippel-Lindau) gene normally regulates ubiquitin mediated proteolysis of HIF-1a. VHL inactivation blocks HIF-1 proteolysis, resulting in increased HIF-1 expression. In the previous study, the expression of HIF-1a was detected in children ALL and NHL. To clarify the possible involvement of the HIF-1a and VHL gene in the development and progression of acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS), we analyzed the mutation and methylation of the VHL gene in 98 patients (54 AML and 44 MDS). The mutations in VHL gene was examined by direct exons sequence and the methylation status of the promoter region was determined using methylation-specific polymerase chain reaction (MSP). In addition, we examined the expression of HIF-1a in 44 of these patients (25 AML and 18 MDS) through the immunoblotting. VHL mutations were identified only in one patient with AML. However, no mutation was detected in the coding region of the VHL gene in MDS. Meanwhile the methylation of VHL gene was not found in the AML and MDS. But the expression of HIF-1a was found in 10 (66.7%) of 25 patients with AML and 5 (38.5%) of 18 patients with MDS. There is no significant correlation between the expression of HIF-1a and the VHL gene. Our primary date support that the HIF-1a may be involved in the development and progression of AML and MDS, but is not associated with the VHL gene.

Disclosures:
No relevant conflicts of interest to declare.
Are Beta-Receptor and Angiotensin-Blocking Drugs Protective Against Venous Thromboembolism (VTE)? A Population Based Case-Control Study
Authors: Ashrani Aneel A. , Geng Wencheng , Crusan Daniel J. , Petterson Tanya M. , Bailey Kent R. , Leibson Cynthia L. , Heit John A. 
Published: October 2019

Abstract
Abstract 5118

Background:
Increased factor VIII:C (FVIII:C) and hypofibrinolysis are VTE risk factors, and beta-blockers and angiotensin converting enzyme (ACE) inhibitors reduce FVIII:C and enhance fibrinolysis, respectively.

Objective:
To test the hypotheses that beta-blockers and ACE inhibitors reduce VTE risk.

Methods:
Using longitudinal, population-based Rochester Epidemiology Project resources, we identified all Olmsted County, MN residents with objectively-diagnosed incident VTE over the 13-year period, 1988–2000 (n=1306), and one to two Olmsted County residents per case matched on age, event year and duration of prior medical history (n=1500). For cases and controls, we reviewed their complete medical history in the community for previously-identified VTE risk factors (e.g., hospitalization with or without surgery, nursing home confinement, trauma/fracture, leg paresis, active cancer, superficial vein thrombosis and varicose veins), as well as body mass index (BMI), coronary artery disease (CAD), congestive heart failure (CHF), and the use of statins, beta-blockers, ACE inhibitors and angiotensin II receptor antagonist drugs. Using conditional logistic regression, we tested beta-blockers and ACE inhibitors/angiotensin II receptor antagonists for an association with VTE, both individually and after adjusting for age, BMI, previously-identified VTE risk factors, CAD, CHF and the use of statins.

Results:
Among cases and controls respectively, 191 and 173 received beta-blockers, and 171 and 154 received ACE inhibitors/angiotensin II receptor antagonists. Univariately, both beta-blockers (unadjusted OR=1.31; p=0.02) and ACE inhibitors/angiotensin II receptor antagonists (unadjusted OR=1.32; p=0.02) were modestly associated with increased VTE risk. However, after controlling for age, BMI, previously-identified VTE risk factors, CAD, CHF and the use of statins, beta-blockers (OR=1.06; 95% CI: 0.74, 1.51; p=0.75) and ACE inhibitors/angiotensin II receptor antagonists (OR=0.94; 95% CI: 0.65, 1.37; p=0.75) were no longer associated with VTE.

Conclusions:
Beta-blockers and ACE inhibitors/angiotensin II receptor antagonists do not appear to be protective against VTE.

Disclosures:
No relevant conflicts of interest to declare.
The Risk of Vascular and Thromboembolic Events Is Increased In Young Low-Risk Germ Cell Cancer Patients by Cisplatin Containing Chemotherapy
Authors: Solari Leticia M , Dollinger Kathrin , Heinz Jürgen , Reinhardt Heike , Rohdenburg Susanne , Engelhardt Monika , Waller Cornelius F. 
Published: October 2019

Abstract
Abstract 5119

Introduction:
It has been previously reported that both short and long term vascular and thromboembolic events (TEE) occur following cisplatin containing chemotherapy for germ-cell cancer (GCC) (Bokemeyer et al. 1998, Weijl et al. 2000, Diekmann et al. 2010). It has also been shown that patients with GCC are at higher risk of TEE than patients with non-GCC, while on cisplatin-based chemotherapy (Piketty et al. 2005).

Method:
We reviewed all GCC patients treated in our department from 2008 until 2010 with cisplatin-based chemotherapy and assessed the incidence of TEE. Cases were analyzed with respect to risk factors for vascular events (obesity, diabetes, nicotine abuse, prior cardiovascular disease, prior TEE), tumor characteristics (low- vs. intermediate/high-risk GCC) and age. In addition, further analysis of hemostasis laboratory tests of these patients is being performed.

Result:
Eight patients (8/50=16%) treated with cisplatin-containing chemotherapy developed TEE. Of these TEE, two were myocardial infarctions, four pulmonary embolisms, one patient showed thrombosis of the jugular vein and one a peripheral (femoral) arterial thrombosis. TE-risk factors included: nicotine abuse in two patients, obesity and nicotine abuse in one patient and no identifiable factors in the others (hemostasis laboratory tests are currently being performed). The median age of these 8 patients was 42 years (range 16–61). Patients were treated with PEB (cisplatin, etoposide, bleomycin) or PIV (cisplatin, ifosfamide, etoposide) initially applying continuous heparin prophylaxis with 15000IE/day (d), d1-5 only during PEB or PIV, but no low-molecular weight heparin (LMWH-) and/or aspirin (ASS-) medication for venous and arterial TEE prophylaxis thereafter. With these clinically very relevant and severe TEEs, we have adapted our PEB- and PIV protocols, adding a LMWH prophylaxis (enoxaparin 40mg d6-21 after chemotherapy).

Conclusion:
We observed an increased risk of major thromboembolic and vascular events with an incidence rate of 16% in GCC patients after cisplatin-containing chemotherapy. Prior studies have identified TEE-risks in GCC and with cisplatin-chemotherapy, albeit LMWH- and/or ASS-prophylaxis - without clinical trials - has not been recommended as yet. Prospective studies investigating thrombosis prophylaxis measures are currently performed in order to achieve the best standard of care for these patients. Currently an extensive hemostasis diagnostic testing from all patients with TEE is being performed.

Disclosures:
No relevant conflicts of interest to declare.
Simultaneous Occurrence of 2009 H1N1 Influenza and Server Acute Hemolytic Anemia —Three Cases
Authors: Wei Ju-ying , Jin Jie , Lou Yinjun 
Published: October 2019

Abstract
Abstract 5139
Most symptoms of 2009 H1N1 virus patients are mild, and most patients could recover well. Lymphopenia and thrombocytopenia are common in 2009 H1N1 patients. We observed that three patients who suffered from acute severe hemolytic anemia (Hemoglobin<60g/L), two of which(Case 1 and 3) also accompany with thrombocytopenia while suffered from H1N1. Two patients(Case 2and 3)got high fever and dark urine at the same time, and another patient(case1)suffered dark urine two days after cough. All patients were confirmed of H1N1 positive by RT-PCR test, while other lab tests and symptoms were consistent with hemolytic anemia. Two of them (case 2 and 3) progressed into hemolytic uremic syndrome and were treated with hemodialysis and methylprednisolone. Another patient (case 3) had been diagnosed with Paroxysmal nocturnal hemoglobinuria (PNH) for one year, and her hemoglobin had maintained about 110g/L, but lower to 45 g/L when she suffered from 2009 H1N1. All three patients were administrated with oseltamivir (75 mg, twice per day) for 5 days by oral and supportive therapy. The hemoglobin level of two patients (Case 1 and 2) increased to 100 g/L after three weeks gradually, and another PNH patient's hemoglobin returned to between 80 and 90g/L after two months. Our three cases had high fever and dark urine simultaneously, which indicated that 2009 H1N1 influenza may induce acute severe hemolytic anemia. Physicians need to be aware of this complication.

Disclosures:
No relevant conflicts of interest to declare.
Burr Red Blood Cells (BC) In Patients with Disseminated Intravascular Coagulation (DIC)
Authors: Patel Vipul R. , Khokhar Jatinder , Murthy Hemant S , Braverman Albert S. 
Published: October 2019

Abstract
Abstract 5141
Burr red blood cells (BC) in patients with disseminated intravascular coagulation (DIC).

Background:
Angiopathic hemolysis with schistocytes occurs in a minority of DIC patients. Burr cells (BC), are morphologically homogeneous, with a serrated membrane resembling that of red cells (RBC) in a hypertonic medium. They are associated with acute renal insufficiency, and have been observed in DIC.

Methods:
Criteria for patient inclusion were sepsis, often with multi-organ failure and hypotension. Of the 39 patients studied, 20 had DIC, based on platelet counts <100,000, and an International Society of Hemostasis and Thrombosis score of ≥5, derived from D-dimer or fibrin split product levels, prothrombin time elevations, and fibrinogen levels. The percentages of BC and schistocytes were determined by 1000 RBC counts, without knowledge of data concerning DIC. Patients' [Hb]'s, serum creatinine and blood urea nitrogen (BUN), were recorded.

Results:
Five of the 20 DIC patients succumbed to their acute illness, while 15 recovered without developing renal insufficiency. The BC%'s of the 20 DIC patients ranged from 0–80%, with a median of 18% and a mean of 27%; in the 19 patients without DIC the range was 0–45%, the median 6.4% and the mean 11% (p=0.046). The schistocyte percentages in patients with and without DIC ranged from 0.20–17, and 0–78; the respective medians and means were 1.1% and 1.0%, and 3.0–5.8%. The median and mean [Hb]'s of the patients with and without DIC were 9.3 and 11, and 9.6 and 10.3, respectively. BC% did not correlate with the [Hb]. The median creatinine levels of patients with and without DIC were 0.92 and 0.93 mg/dl, though the DIC patients' BUN's were higher (medians 31 vs 15). The BC% correlated with neither the BUN nor the BUN/creatinine ratio. The median and mean serum sodium levels in patients with and without DIC were 137.

Conclusion:
BC's are significantly more frequent in septic patients with DIC than in those without it. BC% did not correlate with anemia, implying that BC do not cause significant hemolysis, though there was a trend to lower [Hb] in the DIC patients. BC% was much higher than that of schistocytes in both groups, and schistocyte percentages did not correlate with DIC or BC%. Because of the DIC patients' higher BUN's and BUN/creatinine ratio's, we cannot exclude a role for the factors, which caused pre-renal azotemia in BC formation. BC formation may prove to be a criterion for DIC.

Disclosures:
No relevant conflicts of interest to declare.
Too Much Ferritin, Too Little Overload
Authors: Crisostomo Ricardo H. , Docena Christian P. 
Published: October 2019

Abstract
Abstract 5154
A healthy 42y/o, White, male, without any systemic illnesses or complaints presents with a serum ferritin of 1800ng/mL; the discrepancy between his optimal physical status and the laboratory result, poses a challenge to the most astute clinician. The fundamental goal of this abstract is to provide recognition and prevention to torment that patients with Hereditary Hyperferritinemia Cataract Syndrome (HHCS) are exposed to. The history will offer an important clue, of early onset cataracts. Our patient was guided by an older sibling who herself for many years underwent unnecessary clinical management for hemochromatosis. The presence of hereditary hyperferritinemia cataract syndrome (HHCS) has been defined in families all across the world and likely is under-diagnosed. Such a rare syndrome may never make it into a hematology practice and be missed while the focus is on the “panic” ferritin result. Our patient is part of a family who has had the syndrome since at least the mid 1800's. The scope of this disorder is bringing to reality that confirmation of genetic testing doesn't change management and that the only intervention will be ophthalmologic. The abundant testing for ferritin across medical specialties may cause a rise in HHCS cases, recognizing this syndrome early on will lead to protect patients from insistent and costly interventions. These patients tolerate poorly phlebotomies and will develop iron deficiency anemia. Although appropriate data is available regarding HHCS there is no relevant clinical data to support a poor clinical outcome, and the precise recognition this triad (cataract, asymptomatic patient and hyperferritinemia) will avoid worthless and costly investigations. In this case, a family member who was victimized by excessive workup, was able to research, discover the family mutation and offer this practitioner the tools towards practicing better patient care. In summary, Hereditary Hyperferritinemia Cataract Syndrome is a benign condition that requires avoidance of increased healthcare costs for the proband.

Disclosures:
No relevant conflicts of interest to declare.
Smarca5 Regulates Ctcf Recruitment to Chromatin, Including to Regulatory Loci Involved In Control of Globin Gene Expression In Erythroleukemia
Authors: Kapalova Martina , Kokavec Juraj , Curik Nikola , Burda Pavel , Skoultchi Arthur I. , Stopka Tomas 
Published: October 2019

Abstract
Abstract 5159
Transcription factor Ctcf (CCCTC-binding factor) represents a major regulatory component of epigenetic regulation by recognizing its unmethyled DNA binding sites, resulting in changes in expression of neighboring genes. Ctcf plays an important role in transgenerational genetic imprinting. Very little is known about its role in hematologic malignancies. Ctcf has been described to promote differentiation of human erythroleukemia K562 cells (Torano 2005). We studied Ctcf in mouse erythroleukemia (MEL) cells and found it is expressed at both the mRNA and protein levels. Using chromatin immunoprecipitation (ChIP), we found that Ctcf is recruited to the H19/Igf2 imprinting control region (ICR) and also to the promoters of the alpha globin genes (Hba-a1, Hba-a2) as well as the beta globin locus control region (LCR) in MEL cells. To determine the mechanism by which Ctcf interacts with chromatin, we tested its interaction with chromatin remodeling proteins that associate with these DNA targets, including the well known Imitation Switch (ISWI class) ATPase Smarca5 (Snf2h). Using coimmunopreciptiation and ChIP experiments we found that Smarca5 and Ctcf interact on DNA. Next, we used MEL cells expressing an inducible Smarca5 shRNA. Doxycycline induction of Smarca5 shRNA led to a 5-fold decrease in Smarca5 mRNA and protein levels within 48hrs. ChIP experiments demonstrated that depletion of Smarca5 was accompanied by loss of Ctcf from the aforementioned loci indicating Ctcf requires Smarca5 for its association with chromatin. Furthermore, this was followed by significantly decreased levels H19 RNA. Our data provide evidence that Smarca5 regulates Ctcf recruitment to chromatin, including to regulatory loci involved in controlling globin gene expression. (Grants # IGA 10310-3, MSMT 2B06077, GAUK 251070 45410, SVV-2010-254260507, NIH R01CA154239).

Disclosures:
No relevant conflicts of interest to declare.
Beta Thalassemia In Different Tribal Population In West Bengal, India
Authors: Majumdar Deboshree , Basak Jayasri , Chakraborty Abhijit , Mukhopadhyay Soma , Pal Nabamita , Mukhopadhyay Ashis 
Published: October 2019

Abstract
Abstract 5170

Background:
The Thalassemias are a group of anemia that results from a genetic defect whch reduces the rate of synthesis of normal globin chains. Beta Thalassemia results in severe microcytic, hypochromic, haemolytic anaemia among affected patients. It has emerged as one of the most common health problems among the tribal populations in West Bengal. A project entitled, “Status of Prevalence and Awarness Among Thalassaemia Population: Special reference to various tribes belonging in different Districts of West Bengal” has been sanctioned by the Department of Biotechnology, Govt. of India(BT/PR10336/SPD/24/293/2007). This study is aimed to observe the spectrum of various beta mutations among the tribes. In the screening schedule thalassemia awarness programme was conducted in various rural areas of West Bengal. We mainly confronted with the Sardar tribes in extreme south of West Bengal. In North Bengal there were Toto, Rabha, Oraon, Munda, Toppo, Baraik, Nagbanshi, Khalko, Kheria, Kerketta and Soren.

Method:
In the year 2009–2010, 1289 tribes were screened. Mass awarness programme were followed by collection of peripheral blood samples in EDTA vials and transported to the laboratory in ice packs. NESTROFT was done on spot using 0.36% Saline Buffer solution (Sodium chloride, Sodium dihydrogen phosphate, Disodium hydrogen Phosphate). Complete Blood Count was performed within 24 hours of collection. HPLC (High Performance Liquid Chromatography) was done to identify the beta samples. DNA was extracted from the blood of beta carrier and beta major and subjected to ARMS (Amplification Refractory Mutation System) PCR (Polymerase Chain Reaction) to detect the point mutations. Result:

Conclusion:
Molecular Characterization of Beta globin gene mutations among these tribes have confirmed the presence of the following mutations: IVS-1 nt5 (G>C), IVS-1 nt1(G>T), codon 15 (G>A), codon 26 (G>A), the mutation which leads to HbE, codon 19 (A>G), codon 17 (A>T), -28 (A>G). The most common mutation observed among Totos and Rabhas were codon 26 (G>A) of North Bengal. The prevalence of Beta carrier among them is negligible. The mutation IVS-1 nt5 (G>C) is prevalent among the Oraon, Nagbanshi and Sardar tribes of Bengal. The other mutations which are present among them are codon 15 (G>A) and codon 30 (G>C). There are two codon 17 (A>T) and one -28 (A>G) mutations which are present among Mundas. The rest of them are mutation IVS-1 nt5 (G>C). In few cases presence of sickle cell anemia was observed. Our results have added to the existing data on the common beta globin gene defects which are prevalent among the general population of West Bengal, India.

Disclosures:
No relevant conflicts of interest to declare.
A Novel Dendritic Cell Population Generated by Fusing Granulocyte-Macrophage Colony-Stimulating Factor and Interleukin-21 Induces Tumor-Antigen Specific Immunity
Authors: Williams Patrick , Bouchentouf Manaf , Rafei Moutih , Romieu-Mourez Raphaelle , Hsieh Hsiang-Chuan (Jeremy) , Boivin Marie-Noelle , Yuan Shala , Forner Kathy-Ann , Birman Elena , Galipeau Jacques 
Published: October 2019

Abstract
Abstract 5177

Introduction:
Cancer recruits the immune system to promote its growth and to inhibit reactions against itself. We generated a fusion of GMCSF and IL-21 (GIFT-21) with the aim of stimulating distinct, but complementary, elements of the innate and adaptive immune system against cancer. In a previous study, GIFT-21's aberrant interactions with its cognate receptors on macrophages resulted in an unanticipated pro-inflammatory response and tumor rejection in mice.

Results:
We further explored this phenomenon by treating mice with dendritic cells (DC) derived by treating monocytes with GIFT-21. B16 melanoma and D2F2/neu breast cancer growth was inhibited only in mice treated with a single injection of antigen naïve GIFT-21 DCs. This effect was lost in CD8-/- and CCR2-/- mice and when mice were treated with β2 microglobulin deficient GIFT-21 DCs, and we confirmed that GIFT-21 DCs migrated to and sampled from the tumors to present tumor antigens to CCL2 recruited CD8+ T cells via MHCI.

Conclusion:
When stimulated with GIFT-21 DCs, the immune system can identify cancer specifically, independently of cancer type. We conclude that GIFT-21 and its associated cellular products may serve as novel therapeutic platforms for the treatment of cancer.

Disclosures:
No relevant conflicts of interest to declare.