bluuebunny/crossref_metadata_2025_split huggingface dataset:
Investigation of the Safety and Usefulness of Computed Tomography-Guided Percutaneous Needle Biopsy of Paraaortic Lymph Nodes.
Authors: Aramaki Takeshi , Ikeda Takashi , Moriguchi Michihisa , Tsushima Takahiro , Endo Masahiro , Asakura Koiku , Sawada Akihiro , Kimura Fumihiko 
Published: October 2019

Abstract
Abstract 5027

Background
Various factors such as malignant lymphoma cause abdominal lymph node enlargement. A histological diagnosis of malignant lymphoma is important in planning therapy, and although superficial lymph node biopsies are often obtained, target lesions occasionally arise only around the abdominal aorta. These require surgical, or more recently, endoscopic biopsies. To intraoperatively determine whether target lymph nodes are correctly resected during a surgical biopsy can be difficult, and therapy could be delayed due to postoperative complications.

Objective
To assess the safety and effectiveness of percutaneous biopsy of paraaortic lymph nodes under computed tomography (CT) fluoroscopic guidance.

Subjects and Methods
We retrospectively investigated puncture success rates based on findings of diagnostic imaging, size of target lymph nodes, rate of confirmed diagnosis, frequency of changes in therapeutic plans and incidence of complications among 36 patients after percutaneous needle biopsy using an 18G cutting needle between September 2002 and March 2009.

Results
The pre-biopsy diagnoses for the 36 subjects who underwent percutaneous needle biopsy included: malignant lymphoma (n = 26), lymph node metastasis of esophageal cancer (n = 3), lymph node metastasis of lung cancer (n = 3), lymph node metastasis of pancreatic cancer (n = 3) and unknown (n = 3). The median size of the target lymph nodes was 25 mm (range: 9–102 mm). The results of diagnostic imaging indicated that all punctures were successful. The diagnoses were confirmed in 33 of the 36 patients (91.7%). Histopathological findings confirmed malignant lymphoma (n = 21; Follicular Lymphoma in 11 patients, Diffuse Large B cell Lymphoma in 8, T-Cell Lymphoma in 1, Hodgkin Disease in 1), squamous cell carcinoma (n = 2), poorly differentiated adenoma (n = 2), other malignancies (n = 5), benign lesions (n = 3) and unknown in 3. Therapeutic strategies were altered based on the results of CT-guided percutaneous needle biopsies in 12 of the 36 patients (33.3%). Three patients developed 4 adverse events comprising grade 1 radiation dermatitis (n = 1), grade 1 hematoma (n = 2) and grade 1 pneumothorax (n = 1).

Conclusions
Target paraaortic lymph nodes can be accurately punctured using CT-guided percutaneous needle biopsy, and the specimens were too small to examine the flowcytometry or chromosome analysis, however it was enough to confirmed the diagnosis according to WHO classification of malignant lymphoma. Percutaneous biopsy of paraaortic lymph nodes under CT fluoroscopic guidance is useful for confirming diagnoses with a low risk of serious deep organ damage.

Disclosures
No relevant conflicts of interest to declare.
Acquisition of Isochromosome 7 Is a Late Change in the Pathogenesis of Hepatosplenic Lymphoma, Documented On a Case of Adolescent Girl 5 Years After Renal Transplant with Preceding TCR Gamma-Delta Positive LGL Leukemia.
Authors: Mejstrikova Ester , Kabickova Edita , Sumerauer David , Seeman Tomas , Fronkova Eva , Zemanova Zuzana , Lizcova Libuse , Sedláček Petr , Smisek Petr , Muzikova Katerina , Pospisilova Klara , Janda Ales , Kodet Roman , Capkova Linda , Stary Jan , Hrusak Ondrej 
Published: October 2019

Abstract
Abstract 5031

Background
Post-transplant lymphoproliferative disorders (PTLD) occur in 1-20% of recipients receiving solid organ transplantation. We describe a patient who suffered from hepatosplenic T-cell lymphoma occurring after previous PTLD in a renal transplant recipient.

Patient/Methods
An 11-yr-old girl underwent kidney transplantation for end-stage Fanconi's nephronophthisis in 2002. In October 2006 significant neutropenia (<200/uL) was firstly detected, without any abnormality in bone marrow (BM) aspirate and without hepato- or splenomegaly. Episodes of neutropenia resolved spontaneously or after enhanced immunosuppression and G-CSF. In January 2007 new episodes of neutropenia and newly significant “monocytosis” were detected in peripheral blood (PB) and BM. Percentage of “monocytes” corresponded with immunophenotypically atypical TCR gamma/delta positive T cells (CD7weakposCD5negCD3bright) in PB and BM. Clonal TCR gamma and delta rearrangements were identified which enabled qPCR minimal residual disease (MRD) assessment. No lymphadenopathy was present, slight hepatosplenomegaly was identified by sonography. Conventional and molecular cytogenetic analyses didn't reveal any chromosomal aberration in PB and BM including changes on chromosome 7. No increased levels of EBV and CMV load by PCR were found. Partial increase of granulocytes and slight decrease of atypical TCR gamma/delta T cells were detected after administration of corticosteroid bolus and mercaptopurin. Three months later she presented with fever, rapidly progressive hepatosplenomegaly and pancytopenia, clinically corresponding with hepatosplenic lymphoma. At this time, newly acquired isochromosome 7q was detected by FISH.

Results
Initial therapy with campath and fludarabine was ineffective. She didn't respond to the 2nd line treatment (prednisone, vincristine, daunorubicine,asparaginase) and died 2 weeks later from lymphoma progression. Autopsy identified severe hemophagocytosis in the liver. Retrospectively, we identified identical clonal TCR rearrangements in the PB samples from March 2006 (∼0.03% of lymphoma PB MRD level), when neither changes in PB count nor clinical symptoms were found.

Conclusion
We detected a “pre-lymphoma” phase with clonal expansion of atypical TCR gamma/delta T cells more than 1 year before lymphoma manifestation. The presence of isochromosome 7q was a late change during this lymphoma genesis.

Grant support
IGA NS/9997-4; IGA NR/9531-3, IGA NS 10480-3, Research Projects MZCR 000064203, MSM0021620813

Disclosures
No relevant conflicts of interest to declare.
The Src Kinases Play a Crucial Role in the Growth of Hematopoietic Cells Transformed with Constitutively Activated Stat5 Mutants.
Authors: Nyga Remy , Gouilleux Fabrice , Cartier Flora , Pecquet Christian , Regnier Aline , Marolleau Jean-Pierre , Rochette Jacques , Moriggl Richard , Bouhlal Hicham , Lassoued Kaiss 
Published: October 2019

Abstract
Abstract 5041
Stat5A and Stat5B transcription factors (TF) play an important role in the control of hematopoietic cell proliferation and survival and are constitutively activated in number of hematological malignancies and solid tumours. Studies conducted with constitutively active (ca) Stat5 mutants (Stat51*6 and cS5F) have shown that deregulated Stat5 activity promotes leukemogenesis. In order to evaluate the role of Src kinases in the transforming properties of Stat5 TF, caStat5 expressing Ba/F3 cells and primary murine bone marrow cells were treated with the PP1 Src kinase inhibitor and with its inactive analogue PP3 as a control. We found that PP1 but not PP3 strongly inhibited Stat5A1*6- and Stat5B1*6- expressing Ba/F3 cell growth and survival while no changes were observed in IL-3 stimulated-parental Ba/F3 cells when treated with PP1. Similarly, we were able to demonstrate specific requirement of Src kinases in cS5F-induced primary bone marrow cell growth. We then examined the contribution of Src kinases to the phosphorylation of various signaling molecules involved in cell growth and survival like Stat5, PI3-K/Akt, Ras/Mapk and NF-kB. The treatment of caStat5 (Stat51*6 or cS5F)-expressing Ba/F3 cells with PP1 resulted in a strong decrease of Erk1/2 phosphorylation, but not of Stat5, Akt and IKK species. In addition, caStat5-expressing Ba/F3 cells were found to express a constitutive molecular complex comprising Stat5, Shc, Grb2, Sos, Erk, Gab2, p85, Lyn and Hck. Finally we found that the Lyn Src kinase was overexpressed in the caStat5-expressing cells. Our data suggest a direct implication of Src kinases in the proliferation of caStat5- Ba/F3 transformed cells through the Shc/Erk1/2 signaling pathway and that Src family members are required for caStat5 (Stat51*6 and cS5F)-induced hematopoietic cell growth.

Disclosures
No relevant conflicts of interest to declare.
Mdm2 and Mdmx Are Not the Major Negative Regulators of the p53 Pathway in Myeloid Leukemias.
Authors: Abramowitz Julia , Neuman Tzahi , Samman Rania , Perlman Riki , Ben-Yehuda Dina 
Published: October 2019

Abstract
Abstract 5046
Mutations in the p53 tumor suppressor gene are rare in human hematological malignancies, suggesting that aberrant p53 function may be due to alterations in its regulatory pathways. p53 is negatively regulated by MDM2 through ubiquitin-dependent degradation and by Mdmx through inhibition of transcriptional function. There is little information on the expression of Mdm2 and Mdmx in most myeloid leukemias. We determined the gene expression and protein levels of Mdm2 and Mdmx in bone marrow samples of leukemia patients. We first performed quantitative evaluation of Mdm2 and Mdmx gene expression in bone marrow samples of 144 leukemic patients at the time of diagnosis: 29 de novo AML patients (AML, M0-M7, M3 excluded), 30 de novo AML M3 patients (APL), 39 therapy-related AML patients (t-AML, M0-M7, M3 included) and 46 CML chronic phase (CML-CP) patients in comparison to 35 normal bone marrow samples from Hodgkin's disease patients. Quantitative Real-Time PCR analysis showed no global statistically significant over expression of Mdm2 or Mdmx in any of the tested leukemias. However, a number of patients in both de novo and therapy-related AML had elevated levels of Mdm2 or Mdmx. Significant down regulation of Mdm2, Mdmx and a splicing variant of Mdmx lacking exon 6 (Mdmx-S) was observed in CML-CP. We next performed IHC staining, evaluated by semi-quantitative score, to examine the levels of Mdm2 and Mdmx protein expression in 151 leukemic patients at the time of diagnosis: 40 AML patients, 23 APL, 47 t-AML and 41 CML-CP patients in comparison to 58 normal bone marrow samples from Hodgkin's disease patients. Protein expression analysis also showed no global statistically significant over expression of Mdm2 in any of the tested leukemias. Nonetheless, a number of patients in both de novo and therapy-related AML had elevated levels of Mdm2 protein. Specifically, APL patients segregated into 2 groups: while half of the patients did not express the Mdm2 protein, the other half over-expressed Mdm2. A significant down-regulation of Mdmx was observed in APL. In summary, while in some leukemic patients Mdm2 and Mdmx might play a role in the regulation of p53, our quantitative analysis indicates that these negative regulators do not seem to provoke the inactivation of the p53 pathway in most myeloid leukemias patients. These findings have implications on the possible use of Mdm2 antagonists like nutlin-3 in myeloid leukemias. To the best of our knowledge this is the largest group of myeloid leukemia patients studied for Mdm2 and Mdmx expression.

Disclosures
No relevant conflicts of interest to declare.
High Plasma Levels of Factor VIII and Factor IX Increased the Incidence of Recurrent VTE.
Authors: Corinaldesi Giorgio , Corinaldesi Christian 
Published: October 2019

Abstract
Abstract 5070
The aim of this study was to evaluate the diagnostic accurancy of the measurement of procoagulant factors FVIII and FIX levels and the risk of recurrence of venous thromboembolism (VTE). We have studied 56 patients (14 male, 42 female) with an objectively diagnosis of first deep vein thrombosis enrolled from February 2003 to February 2008, all patients were <60 years, without known malignancies, heart failure, chronic lung disease, infection, recent surgery, or genetic defects as Factor V Leiden (FV-Arg 506 Glu), prothrombin G20210A, deficiencies of Protein S or C or ATIII, hyperhomocysteinemia, dysfibrinogenemias; with increased levels of FVIII (>218 IU/dl), and FIX (>136 UI/dl). FVIII and FIX antigen levels were measured by ELISA. All patients received subcutaneous low molecular-weight eparin as thromboprophylactic treatment for the first 3 month after primary VTE, color Doppler ultrasonography (DUS) was performed periodically after primary VTE, the venous hemodynamic parameters studied included: blood flow velocity (cm/sec), vein cross sectional area (cm/2) blood volume flow (ml/sec), and microvascular injury was evaluated. Our studies have confirmed that a markedly elevated plasma levels of FVIII >256 IU/dl (range 218-294), and FIX >164IU/dl (range 136-192) among the patients with first episode of VTE are independent predictors of early recurrence about 52-74%: these patients have increased fibrin formation, D-Dimer levels (234.6ng/ml), and blood rheologic variables such as plasma viscosity are still present in all patients: plasma viscosity mPa/s shear rate >1.98 and blood viscosity mPa/s shear rate >3.48 especially under low shear conditions. These factors show to be associated with VTE and above all with the recurrence (54 patients), and emerged as relatively independent of other thrombophilic factors. The higt risk of VTE recurrence in these patients requires strategies for its prevention: secondary prophylaxis with the administration of standard oral anticoagulant at moment the use of coumarins (INR: 2.0-3.0) and possibly the need for prolonged, probably life-long treatment.

Disclosures
No relevant conflicts of interest to declare.
A Prospective Study of Platelets Function During the Process of Stem Cells Mobilisation in Healthy, Allogeneic Blood Stem Cell Donors Using the Method of Impedance Aggregometry.
Authors: Najda Jacek , Wojnar Jerzy , Kyrcz-Krzemien Slawomira 
Published: October 2019

Abstract
Abstract 5079
Potential haemostasis disturbances accompanying the process of peripheral stem cells harvest are still underestimated. Although the last two decades have witnessed an important increase in the use of peripheral blood stem cells (PBSC) for allogeneic transplantations, we are still lacking a satisfactory analysis of mobilization related risk of thrombosis or an unexpected bleeding. It was previously proved, that G-CSF administration may induce a hypercoagulable state by increasing levels of FVIII;C. On the other hand, platelet aggregation is reduced after cytokine application. Numerous, remaining unsolved questions in this matter prompted us to study prospectively 26 PBSC healthy donors. All donors were medically evaluated initially to eliminate any identifiable pathology in their medical history, especially concerning cardiovascular system. Any concomitant medication was excluded. All donors studied were excluded if they had an abnormal platelet count, activated partial thromboplastin time (APTT) or prothrombin time(PT) prior to cytokine administration.
All donors received G-CSF (Neupogen, Amgen) at 5ug/kg of body weight subcutaneously twice daily for nine consecutive doses. We were analyzing the group of donors in the context of hemostasis alteration using the method of impedance aggregometry. The basic device used was “MULTIPLATE' aggregometer produced by DYNABYTE (Munich). We analyzed the primary hemostasis measuring the reaction of platelets to collagen (COL-TEST), ADP (ADP-test) and TRAP –test.
We performed platelets aggregation analysis before G-CSF administration, and subsequently on the first day of stem cells mobilization, three times. The first sample was obtained just before the beginning of separation, the second in the half time point of the procedure and the last sample was drawn immediately after the cessation of stem cell collection procedure. We found reduced platelet aggregation after G-CSF administration, comparing the data before cytokine stimulation and preceding the leukapheresis (p<0,05).
In contrast to this finding, there was a considerable increase in platelet activation parameters in the course of apheresis procedure, concerning both COL-test (p<0,04) nad TRAP-test (p<0,05).
The clinical implications of these findings confirm important and significant coagulation system stimulation during separation procedure.

Disclosures
No relevant conflicts of interest to declare.
Poor Potential of Proliferation and Differentiation in Bone Marrow Mesenchymal Stem Cells Derived From Children with Severe Aplastic Anemia.
Authors: Peng Ching-Tien 
Published: October 2019

Abstract 5081

Introduction
Idiopathic severe aplastic anemia (SAA), characterized by failure of hematopoiesis, is rare and potentially life-threatening to children. However, the pathogenesis has not been completely understood, and insufficiency in the hematopoietic microenvironment can be an important factor. Mesenchymal stem cells (MSCs) play an important role in maintaining bone marrow microenvironment. Therefore, we aimed at the intrinsic defects of bone marrow MSCs derived from SAA children.

Materials and Methods
Bone marrow MSCs were obtained from 5 SAA children and 5 controls. The morphology, immunophenotyping, proliferative capacity and differentiation potential of MSCs from SAA children were determined and compared with those of MSCs from controls.

Results
In vitro, MSCs of SAA and control group shared a similar spindle-shaped morphology. Both revealed a consistent immunophenotypic profile which was negative for CD45, CD14 and CD34, and positive for CD105, CD73, and CD44. However, SAA MSCs had slower expansion rate and smaller cumulative population doubling from passage 4 to 6 (1.83± 1.21 vs 3.36± 0.87; p = 0.046), indicating lower proliferative capacity. Besides, only 3 of 5 cultures of SAA group retained the ability to continue expansion till 80%-90% confluent cell layer beyond passage 6, suggesting earlier senescence of SAA MSCs. After osteogenic induction, SAA MSCs showed lower alkaline phosphatase activity (1.46± 0.04 vs 2.27± 0.32; p = 0.013), less intense von Kossa staining and lower gene expression of core binding factora1 (0.0015± 0.0005 vs 0.0056± 0.0017; p = 0.013). Following adipogenic induction, SAA MSCs showed less intense Oil red O staining (0.86± 0.22 vs 1.73± 0.42; p = 0.013) and lower lipoproteinlipase expression (0.0105± 0.0074 vs 0.0527± 0.0254; p = 0.013).The results of real time-PCR analysis for the assessment of lineage-specific genes were consistent with the findings of histochemical stains, and both indicated that SAA MSCs had poor osteogenic and adipogenic potential.

Conclusions
In this study, we demonstrated that bone marrow MSCs from children with SAA had poor potential of proliferation and differentiation. These alterations in MSCs may contribute to the failure of hematopoiesis, and lead to the development of the disease. Further studies are needed to elucidate the relationship between MSCs and SAA.

Disclosures
No relevant conflicts of interest to declare.
Novel Homozygous Mutation (R329X) in the Hemojuvelin Gene Is Associated with Hypogonadotrophic Hypogonadism, Diabetes Mellitus and Heart Failure Duo to Juvenile Hemochromatosis.
Authors: Zhou Rong-Fu , Ouyang Jian , Guo Xue-mei , Chen Bing , Xu Jing-yan , Shao Xiao-Yan , Zhang Qi-Guo 
Published: October 2019

Abstract
Abstract 5102

Juvenile hemochromatosis (JH) is a rare autosomal recessive disorder characterized by the early onset of severe iron overload. A large variety of mutations within the genes encoding hepcidin (HAMP) and hemojuvelin (HJV) have been identified in patients with JH. But in the Chinese population, the prevalence of JH is quite low. No HJV mutation has been reported so far.

Methods and results
The proband was a 25-year-old young man of Asia decent presented with hypogonadotrophic hypogonadism, diabetes mellitus and heart failure but no family history of iron disorders. His serum iron level was 34μmol/L, with a transferrin concentration of 8.5g/L, serum ferritin concentration was 8140 μg/L. Echocardiography revealed that he had generalized cardiac enlargement, cardiac dysfunction, and severe mitral and tricuspidal insufficiency, pulmonary hypertension. Ultra-sonography showed diffuse hepatomegaly and splenomegaly, seroperitoneum and right hydrothorax. Liver biopsy showed severe diffuse hepatocellular siderosis and cirrhosis,hemosiderin pigmentation. To search for possible variants in the HJV gene, we performed PCR and direct sequencing in proband and his family. A homozygous nonsense mutation in exon 4 of HJV (R329X) was identified in the JH patient and heterozygous mutation of R329X in his father and mother. No mutations were found in other genes associated with adult or juvenile hemochromatosis including HFE, transferrin receptor-2 (TFR2), ferroportin (SLC40A1), hepcidin (HAMP).

Conclusions
A novel nonsense mutation (R329X) has been identified in the HJV gene for the first time in China. This mutation elevates ferritin levels and leads to JH associated with hypogonadotrophic hypogonadism, diabetes mellitus and severe cardiomyopathy.

Disclosures
No relevant conflicts of interest to declare.
Hematologic Manifestations and Changes of Cytokines or Hematopoietic Growth Factors in Mice with Iron Overlaod.
Authors: Jeong Dae-Chul , Hwang Hui Seung , Chung Nack Gyun , Cho Bin , Shin Hyun Jung , Kim Hack Ki , Kim Yoo Jin , Han Chi Wha 
Published: October 2019

Abstract
Abstract 5103

Background
Iron overload by repeated transfusions induced organ toxicity including liver, heart. We investigated hematologic manifestations and cytokines or hematopoietic growth factors in murine secondary hemochromatosis.

Materials and methods
We established murine secondary hemochromatosis model using 6 week-old male C57/BL6 (H-2b) with iron dextran. Mice (n=10∼12) were intraperitoneally injected with 10 mg of iron dextran for 2 or 4 weeks. We divided five groups: control (PBS injection), iron 100mg, iron 200mg, iron 200mg with deferasirox (DFX) 300mg, and only DFX 300mg. We examined hematocrit, platelet counts and plasma iron concentration (PIC) in peripheral blood, and liver iron contents (LIC) by atomic absorption spectrophotometer. We evaluated colony forming capacity from bone marrow according to experimental group. For cytokines and hematopoietic growth factors, we performed real-time PCR for IL-1b, iNOS, IFN-g, TNF-a, TGF-b, SCF, TPO, GM-CSF, and IL-11 in bone marrow. We compared each values of relative ratio with b-actin.

Results
There was no difference of hematocrit among experimental groups. The platelet counts were significantly decreased in iron 200mg among groups (P<0.05), and showed increased trends after administration of DFX. The levels of LIC and PIC were dependent on cumulative dose of iron loaded, and decreased by DFX (P<0.01). This findings showed positive correlation between PIC and LIC (P<0.01, R2=0.726). The CFU-GEMM and CFU-GM decreased in iron 200mg, iron 200mg+DFX300mg, and DFX300mg compared with control and iron 100mg (P<0.01). Most colonies in DFX300mg were not observed except CFU-GM. In cytokines, there was shown no difference for IL-1b, iNOS, IFN-g, TNF-a, TGF-b according to experiments (P>0.05). However, SCF was shown diminished expressions for treated mice compared with control (P=0.02). The levels of TPO were increased in hemochromatosis, and decreased after administration of DFX (P=0.05). The GM-CSF was observed significantly lower in iron 200mg, iron 200mg plus DFX, DFX than control and iron 100mg (P<0.01).

Conclusions
Our results suggested that iron overload might affect hematopiesis and these findings were due to effects of hematopoietic growth factors including SCF, TPO, GM-CSF, not inhibitory cytokines. Also, we need further study for DFX in hematopoiesis.

Disclosures
No relevant conflicts of interest to declare.