bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical Significance and Prognosis of Mir-155 and Mir-146a Expression Levels in Formalin-Fixed/Paraffin-Embedded Tissue of Patients with Diffuse Large B-Cell Lymphoma
Authors: Zhong Hua 
Published: October 2019

Abstract
Abstract 5197

Introduction
Diffuse large B cell lymphoma (DLBCL) is an aggressive malignancy that accounts for nearly 40% of all lymphoid tumors. Both miR-155 and miR-146a expression levels are suggested to be regulated by NF-KB expression in DLBCL. In present study, the expression of miR-155 and miR-146a were investigated to evaluate the prognosis in DLBCL patients.

Patients and Methods
The expression levels of miR-155 and miR-146a in formalin-fixed/paraffin-embedded tissue of patients with DLBCL (n=50) have been compared with reactive hyperplasia lymphoid nodes (n=20) by using real-time polymerase chain reaction. Clinical characters of patients such as LDH, β2-MG, International Prognostic Index (IPI), ECOG score and c-myc expression levels have also been investigated.

Results
1) The expression levels of miR-155 and miR-146a were significantly higher in DLBCL patients than control (P<0.001). (Fig 1)
2) The expression levels of miR-155 and miR-146a were different between germinal center B-cell like DLBCL (GCB-DLBCL) and activated B-cell-like DLBCL (ABC-DLBCL), but the difference had no significance (P>0.05).
3) There was correlation between the expression of miR-155 and miR-146a in DLBCL patients (r =0.639, P<0.001).
4) There was positive correlation in the expression of miR-146a with LDH, β2-MG, IPI, ECOG score and c-myc expression levels in DLBCL patients.
5) Lower expression levels of miR-155 and miR-146a were found to be associated with high complete remission (CR) rate and overall response (OR) rate in DLBCL patients (P<0.05).
6) Lower expression levels of miR-146a in DLBCL patients were only associated with better 5 years progression free survival (5y-PFS) (P=0.044), but lower expression levels of miR-155 in DLBCL patients were associated with better 5y-PFS (P=0.013) and 5 years overall survival (5y-OS) (P=0.007).
7) Multivariate analysis using a Cox proportional hazard model confirmed that either miR-155 or miR-146a was the independent predictor. (For miR-155 5y-PFS, Hazards ratio 0.260, 95% CI, 0.058–0.801; for miR-146a 5y-PFS, Hazards ratio 0.251, 95% CI, 0.068–0.922)
8) Compared to CHOP protocol, the higher miR-155 expression level patients who chose R-CHOP protocol for treatment achieved better 3y-PFS and 3y-OS (P<0.05).

Conclusions
The expression levels of miR-155 and miR-146a were higher in DLBCL patients. Moreover, high expressions of these two microRNAs were associated with the disease progression. Patients with higher miR-155 expression levels may be benefit from the protocol including Rituximab.

Disclosures:
No relevant conflicts of interest to declare.
EBV-Negative Primary Pulmonary Lymphoma in a Patient with AIDS
Authors: Storey Raul E. , Mandadi Mounika , Abdolmohammadi Alireza , Natarajan Nagendra , Kloecker Goetz H , Stephen Slone P 
Published: October 2019

Abstract
Abstract 5203
AIDS-related primary pulmonary lymphoma (PPL) is defined as exclusive involvement of the lung by lymphoma with no evidence of extrapulmonary disease at the time of diagnosis and three months thereafter. It is an aggressive disease with a median survival time of four months. A 46 year old male patient with a seven year history of HIV/AIDS and CD4 count of 200/ul was hospitalized because of flank pain, hematuria and hemoptysis. Abdominal CT scan incidentally revealed multiple nodules in the lower lobes of both lungs. Serial chest CT scans demonstrated multiple nodular opacities in all lobes of the lungs that enlarged rapidly and coalesced together. Clinicians favored opportunistic infection due to rapid disease progression but included Wegener's granulomatosis in the differential diagnosis because of hemoptysis. Open lung biopsy showed diffuse large B cell lymphoma (DLBCL), NOS, with an activated B cell phenotype (CD10−/BCL6−/MUM1+) and high proliferative index (Ki67 90%). CD79A expression confirmed B cell lineage. The diagnostic criteria for PPL were fulfilled because staging failed to detect extrapulmonary disease. Infarction due to the angiodestructive nature of the PPL and short doubling time accounted for hemoptysis and rapid disease progression, respectively. The patient died 45 days after admission because of shock, presumably due to superimposed infection. Unlike prior reports of AIDS-related PPL, our case was negative for CD19, CD20 and Epstein-Barr virus (EBV). Although PPL is a rare disease, it deserves consideration in the differential diagnosis of rapidly progressive lung nodules in AIDS patients. To the best of our knowledge, this is the first reported case of EBV-negative PPL.

Disclosures:
No relevant conflicts of interest to declare.
TP53 Mutation Is Rare In Primary Myelofibrosis
Authors: Greaves Wesley O. , Verma Shalini , Bisrat Tigist , Rahimi Hamed , Paladugu Abhaya , Bueso-Ramos Carlos E. , Patel Keyur P. , Luthra Rajyalakshmi , Yao Hui , Verstovsek Srdan 
Published: October 2019

Abstract
Abstract 5241

Introduction
TP53 is the most frequently mutated tumor suppressor gene in human cancers and is usually associated with an aggressive disease course. TP53 mutation has been described in a variety of hematopoietic neoplasms, and has been suggested to play a role in leukemic transformation of myeloproliferative neoplasms (MPN). However, the incidence as well as the clinical and pathogenetic implications of TP53 mutation in each sub-category of MPN, including primary myelofibrosis, have not been described. In this study, we investigated the presence and potential clinical significance of TP53 mutations in a large series of primary myelofibrosis cases.

Patients and Methods
We retrieved archival bone marrow DNA from 51 consecutive patients diagnosed with primary myelofibrosis at The University of Texas MD Anderson Cancer Center between the years 2005 and 2007. Diagnosis was based on morphologic, immunophenotypic, cytogenetic and molecular evaluation of bone marrow in conjunction with clinical data. Only 2 patients had blast counts >10% (11 and 14%). Twenty nine of 50 (58%) patients showed JAK2 p.V617F mutation and all patients were negative for BCR-ABL1 translocation on routine clinical testing. DNA samples were assessed for sequence variation in exons 4 through 9 of TP53 by both high resolution melting curve (HRM) analysis using LightCycler® 480 System (Roche, Indiana IN) and bidirectional Sanger sequencing using 3730XL DNA Analyzer (Life technologies, Carlsbad CA).

Results
The mean overall survival was 5.7 years. Five patients developed acute leukemia, all of whom died of disease. By Sanger sequencing, only one (1.9%) case showed an amino acid-altering mutation in TP53: c.707A>G (TAC to TGC) in codon 236 (p.Y236C) of exon 7. In addition, 8 cases showed silent mutations/single nucleotide polymorphisms of unknown significance - c.36G>A (CCG to CCA) in exon 4 (n=3) and c.213A>G (CGA to CGG) in exon 6 (n=6). The p.R72P polymorphism in exon 4 which has been described in other hematopoietic neoplasms was present in 1 patient. All cases with a mutant sequence by Sanger sequencing also showed a variant melting curve pattern by HRM analysis. The patient with TP53 mutation died 2 years after presentation from progressive myelofibrosis without developing acute leukemia.

Conclusion
TP53 mutation is very rare in primary myelofibrosis.

Disclosures:
No relevant conflicts of interest to declare.
Over-Expression of PAI-1 and TAFI Predict Stroke: Two Faces of Ame Coin
Authors: Corinaldesi Giorgio 
Published: October 2019

Abstract 5249
TAFI (thrombin-activated fibrinolysis inhibitor) and PAI-1 play important roles in fibrinolysis, as they both reduce plasmin generation; PAI-1 a 50 KDa glycoprotein is a serine-proteinase inhibitor; TAFI is a 55 KDa plasma zymogen that is activated by either plasmin/thrombin or thrombin/thrombomodulin complex into an active carboxypeptidase, in particular, it is a basic zinc dependent metallo-carboxipeptidase specific for C-terminal lysine and arginine residue inhibiting degradation of the fibrin clot.
We investigated the influence of PAI-1 and TAFI levels on stroke by determining their plasmatic antigen concentration by an electro-immunoassay. We found that PAI-1 levels are increased during the early stage of stroke and that it may also predict clot lysis resistance in patients treated with trombolysis (38.8 ng/ml +/− 14.6, p <0.0001) compared with control subjects (16.2 ng/ml +/−6.8, p <0.0001); TAFI levels were significantly higher in patient with stroke (184.8 nmol/l +/− 62.6 nmol/l), than in healthy control subjects (102.4 nmol/l +/− 36.4 nmol/l, p <0.0001). We have observed that a combination of both PAI-1 higher than 24.6 mg/ml (OR: 12.8. CI: 1.16 to 134.2), and TAFI higher than >180 nmol/l (OR.12.9 CI 1.42 to 116.6), had a sensibility of 92.2%, and a specificity of 98.4% in predicting stroke. The analysis of these results showed that TAFI antigen levels higher than 180nmo/L combined with PAI-1 levels higher than 24 mg/ml are a strong risk factor for clot lysis, stroke (for the evolution, severity, and outcome) and showed a potentially attractive target for fibrinolytic therapies.

Disclosures:
No relevant conflicts of interest to declare.
Metabolic and Inflamatory Proteins Differently Expressed in Platelets From Deep Venous Thrombosis Patients
Authors: Flores Nascimento Mariane Cristina , Kleinfelder-Fontanesi Karina , de Andrade Orsi Fernanda Loureiro , Seeholzer Steven H , Ischiropoulos Harry , Annichino-Bizzacchi Joyce M 
Published: October 2019

Abstract
Abstract 5256

BACKGROUND:
Deep venous thrombosis (DVT) is multi-causal disease associated to a high morbi-mortality due to complications as pulmonary embolism and post-flebitic syndrome. The incidence is about 20 to 30%, and 25% of the patients will present recurrence in 5 years. The identification of new risk factors is important in clinical practice to prevent new thrombotic events. The role of the platelets on DVT is still not well defined.

AIM:
The objective of this study was to analyze the hole proteins profile of platelets obtained from DVT patients and compare to the same matherial derived from healthy controls.

PATIENTS:
peripheral blood samples were collected from 3 spontaneous DVT patients and from 1 sibling and 1 neighbor for each patient in order to minimize the genetic and environmental interferences. These patients presented spontaneous and recurrent episodes of lower limbs proximal DVT and all of them mentioned a familiar history of coagulation disorders.

METHODS:
the platelets were washed, lysed, and the proteins were alkylated, reduced, precipitated with acetone and hydrolyzed by trypsin. 100mg of peptides were then separated by hydrophobicity using HPLC, and 8 fractions were obtained and directed to the LTQ-Orbitrap mass spectrometer. The proteins search was performed by Sorcerer-SEQUEST.

RESULTS:
We identified 5 proteins that were present on patients and absent in all the controls: Apolipoprotein A1 Binding-Protein, Coatomer (z1 sub-unit), Estradiol 11–17-b Dehydrogenase, Leucotriene A-4 Hydrolase and Sorbitol Dehydrogenase. Western-Blotting was performed with specific antibodies and validated the results.

CONCLUSIONS:
with this study it was possible to identify proteins up to date non-related to the physiopathology of DVT, that could be involved with metabolic and inflammatory processes.

Disclosures:
No relevant conflicts of interest to declare.
Hemolytic Intravascular Anemia (HIA) Microangiopathic-Like and Deep Venous Thrombosis (DVT) Due to Ozone Exposure: Case Report
Authors: Cortázar-Benítez Luis Fernando , Viveros Pablo Vargas , Alvarez Alfredo Aiza , Monroy Rafael Hurtado 
Published: October 2019

Abstract
Abstract 5278
HIA due to ozone exposure on patients with glucose 6 phosphate dehydrogenase deficiency (G6PDd) is extremely rare. The purpose of this report is to describe in detail a case of HIA and DVT on a woman without G6PDd, with a successful treatment with fresh frozen plasma (FFP) transfusion and Thrombolysis.
The patient is a 36 years old woman, that 3 months before she was diagnosed with multiple sclerosis (MS) because paresthesias in the fingers of her left foot and she received treatment with blood ozone exposure (at unknown dose) in 3 sessions each week for 3 months. The patient attended to our center with severe anemic syndrome during the last 2 weeks and disabling pain of her left leg of 12 hours of evolution. Physical examination showed pale ++++, jaundice ++, functional systolic murmur grade IV, without adenomegaly or splenomegaly, increasing volume, induration, erythema and intense pain from the ankle to the popliteal space of her left leg. The urine was dark. Laboratory data were haemoglobin 5 g/dL, hematocrit 17%, reticulocytes 62%, and platelets 281×109/L. Peripheral blood smear showed esquistocytes +++ and spherocytes ++, suggesting intravascular hemolysis. Total bilirrubin 2.99mg/dL, direct bilirubin 0.57, and LDH 750 U/L. Doppler ultrasound: obstruction of the deep and superficial venous system of tibial, peroneal and left popliteal veins. Four red cells units were transfused and FFP transfusion was started every 6 hours, anticoagulation with enoxaparina sodium (1mg/Kg/day) and thrombolysis with rhTPA 100 mg for 3 hours infusion. The patient successfully improved with increase and maintenance of hemoglobin, decrease of the reticulocytes count and evident clinical improvement of her left leg. She was in-hospital for 8 days at the end of which was achieved ambulation, Doppler showed remission of DVT.
The association between exposure to ozone and HIA has not been informed in the absence of G6PD deficiency, and today, little is known of the ideal treatment. Though plasmapheresis is the treatment of choice in a HIA, the presence of DVT and be in a period appropriate for thrombolysis, determined the use of FFP transfusion as the main treatment. The right clinical evolution observed in the treatment of our patient gave her solving clinical problems. Ozone has been widely used for a variety of off-label purposes. In vitro experiments had demonstrated hemolysis with ozone concentration > 30 mcg/mL, therefore this case must represent an important alert for those ozone users, however the mechanism of hemolysis because ozone exposure remains to be elucidated.

Disclosures:
No relevant conflicts of interest to declare.
Serum Uric Acid As a Predictor Factor of the Response to Deferasirox Therapy for Patients with b-Thalassemia Major
Authors: Vlachaki Efthimia , Perifanis Vasileios , Kondou Antonia , Neokleous Nikolaos , Teli Aikaterini , Oikonomou Marina 
Published: October 2019

Abstract
Abstract 5306
Serum uric acid of patients with beta-thalassemia major (b-MA), despite the hyperuricosuria observed in these patients, is usually in the upper normal levels or increased due to excessive catabolism of the red blood cells (ineffective erythropoiesis). Deferasirox a new oral iron chelator with potential nephrotoxicity is recently used as iron overload treatment in patients with b-MA.

Aim:
Aim of the study is to investigate the effect of deferasirox on uric acid levels of patients with homozygous b-MA.

Material-Method:
53 patients were enrolled to the present study with b-MA major, (aged 22.4 ± 14.7 years, range 4–12 years) 36 adults and 17 children, 32 females and 21 male. All the patients were transfusion-dependent with pretransfusional haemoglobin 9gr/dl and treated with iron chelation. The comparison was made between two different time points' measurements of uric acid and ferritin, at the beginning before Deferasirox, and one year later. The blood was taken from the patients early at mornings before the transfusion. Also uric acid was measured in 24 hour urine of patients under deferasirox, or other iron chelation therapy or after weekly discontinuation of deferasirox. Patients taken allopurinol or thiazide or with abnormal kidney function were excluded.

Results:
There is statistically significant difference (p< 0.001) between the mean annual value of serum uric acid (before 5.2 ± 1.3mg/dl) and ferritin (before 1653,4 ± 1026,3 ng/dl) before and after the start of deferasirox (uric acid after 4.2 ± 1.3 mg/dl and ferritin after 1529,07 ± 1137,44 ng/dl). Also, statistically significant positive correlation between the levels of serum uric acid and ferritin was found during the treatment with deferasirox. However, comparing the uric acid in urine of patients, it was not reported any statistically significant difference between treatment with deferasirox (859,75 ± 122), other iron chelators or without iron chelation for one week (844,32 ± 146).

Conclusion:
The mechanism of uric acid reduction in patients with b-MA major treated with deferasirox is not clear. However, it does not seem to be associated with excess of excretion by urine. The positive correlation between uric acid level and ferritin, allow us to consider uric acid as a predictor factor of the response to deferasirox therapy.

Disclosures:
Vlachaki: Novartis Hellas S.A.C.I.: Research Funding. Oikonomou:Novartis Hellas S.A.C.I.: Research Funding.
Von Willebrand Factor (VWF) Regulates Angiogenesis: Studies in Blood-Derived Endothelial Progenitor Cells From Von Willebrand's Disease Patients and VWF-Deficient Mice
Authors: Starke Richard D , Paschalaki Koralia , Ferraro Francesco , McKinnon Thomas A J , Dryden Nicola H , Payne Elspeth , Haskard Dorian O , Hughes Alun D , Cutler Daniel , Laffan Michael , Randi Anna Maria 
Published: October 2019

Abstract
Abstract 5311
Dysregulation of angiogenesis is implicated in many diseases. Von Willebrand factor (VWF), a large plasma glycoprotein essential for normal haemostasis is synthesized by endothelial cells (EC) and megakaryocytes. Raised VWF plasma levels are a risk factor for arterial thrombosis, whilst deficiency of VWF causes Von Willebrand disease (VWD), the most common congenital bleeding disorder in man. VWD can be associated with angiodysplasia, vascular malformations linked to defective angiogenesis which are responsible for intractable bleeding. We recently showed that VWF is involved in angiogenesis. Inhibition of VWF expression in human umbilical vein EC (HUVEC) with specific siRNA resulted in increased in vitro angiogenesis on Matrigel, proliferation and migration. Mechanism studies implicated the endothelial VWF receptor, integrin αvβ3 and the angiogenesis regulator angiopoietin-2. The findings were confirmed in EC from VWD patients and in VWF-deficient mice. Blood outgrowth endothelial cells (BOEC) isolated from peripheral blood of patients with VWD showed decreased VWF release, compared to control BOEC, and increased in vitro angiogenesis, migration and proliferation, similar to what observed with VWF siRNA-treated HUVEC. In vivo studies using the matrigel model and imaging of blood vessels in the ear showed increased angiogenesis and vascular network in VWF-deficient mice compared to controls. Recent studies in patients BOEC and in VWF deficient mice provide new insight into the complexity of this phenotype. We have used the mouse model of post-natal angiogenesis in the retina to carry out detailed analysis of angiogenic networks in the VWF-deficient mouse, and found increased vascular density and defective vascular network. Moreover, using BOEC from patients with type I and type II VWD, we have studied VWF intracellular distribution by immunofluorescence confocal analysis and found patterns of expression that point to a variety of defects in synthesis, storage and secretion. These studies define a new function for VWF, which may have clinical implications for VWD and for patients at risk of CV disease. Moreover, studies with BOEC from VWF patients provide a novel understanding of the physiopathology of this disease.

Disclosures:
No relevant conflicts of interest to declare.
First Case of Platelet-Type Von Willebrand Disease (PT-VWD) Associated with Type 2B Von Willebrand Disease (2B VWD)
Authors: Dreyfus Marie , Desconclois Celine , Guitton Corinne , Baas Marie-Jeanne , Mandard Helene , Boisseau Pierre , Goudemand Jenny , Caron Claudine , Itzhar Nathalie , Veyradier Agnes , Lanza Francois , Fressinaud Edith 
Published: October 2019

Abstract
Abstract 5313

Introduction
VWD 2B and PT-VWD are rare diseases, due to mutations inducing a gain of function respectively of von Willebrand factor (VWF) and of its platelet receptor, Glycoprotein (GP)1bα

Case history
We report here the case of a young girl, born with an extensive purpura and a severe thrombocytopenia: platelet count: 16G/L. There was no associated biological nor clinical abnormality. A high dose of 1g/kg of immunoglobulin G infused on day 1 was unsuccessful, and a HPA-1a (−) platelet concentrate infusion led to a partial and transient increase of the platelet count up to 60G/L. Thrombocytopenia then resolved spontaneously.
Biological study showed no sign of materno-fetal allo- or auto-immunity, parents were not consanguineous.
The diagnosis of type 2B VWD was performed when she was 5 months old: VWF:RCo < 13 IU/dl, VWF:Ag 60 IU/dl, positive ristocetin induced platelet aggregation (RIPA) at a low ristocetin concentration (0.5 mg/ml). RIPA mixing studies were unconclusive.
The same biological abnormalities were found in the father, whereas the mother had normal hemostasis tests. The biological phenotype also included a study of the multimeric VWF structure, showing a marked decrease in percentage of VWF high and intermediate molecular multimers. Genetic analysis performed on VWF gene showed the heterozygous p.Pro1266Leu missense mutation in the VWF A1 domain. This mutation ( o ) is only slightly deleterious, and induces usually a mild disease, without thrombocytopenia, even in stress situations, with normal VWF multimeric distribution; therefore, it could not explain the biological phenotype severity in this family.
GPIBA was then analysed, and a candidate point mutation p.Met239Ile was evidenced. This mutation had not been described yet, but p.Met255Val had already been found in diagnosed cases of PT-VWD.

Conclusion
This case underlines the utmost importance to characterize precisely neonatal thrombocytopenia mechanism. Furthermore, it points out the difficulties to performing PT-VWD diagnosis, which incidence is most probably underestimated. In our case, it was the systematic and extensive biological workout performed in this case of isolated neonatal thrombocytopenia, without any obvious cause, which led to the diagnosis of a PT-VWD, inducing a severe biological phenotype, associated with type 2B VWD characterized by a mild expression. It is, to our knowledge, the first case described to date of such a morbid association.

Disclosures:
No relevant conflicts of interest to declare.