bluuebunny/crossref_metadata_2025_split huggingface dataset:
Use of “high flow” Totally Implantable Central Venous Access Ports for Myeloablative Conditioning Followed by Allogeneic Stem Cell Transplantation: a Prospective Study.
Authors: Abdelkefi Abderrahman , Gaied Olfa Ben , Ladeb Saloua , Torjman Lamia , Lakhal Amel , Othman Tarek Ben 
Published: October 2019

Abstract
Abstract 4648

Objectives
A central venous access is always necessary for the management of patients receiving myeloablative conditioning followed by allogeneic stem cell transplantation (SCT). Tunnelled, cuffed silastic catheters are the device of choice for these patients. To our knowledge, there are no prospective studies investigating the use of totally implantable central venous access ports (TIAP) in patients receiving myeloablative conditioning followed by allogeneic SCT. The aim of this prospective study was to investigate the usefulness of a new “high flow” single lumen port device in these patients. <>Methods: Between December 2007 and July 2009, patients with haematological malignancies received a TIAP prior to allogeneic SCT. All patients received the same type of port [silicone, 10 French, high flow rate (3100ml/h), Reference 40010, Laboratoires Perouse, Ivry le Temple, France]. All devices were inserted under local anesthesia through direct puncture of the right subclavian vein using the Seldinger technique. A chest X-ray was always obtained to document correct positioning. All infusions, including the graft itself and all blood drawings, were performed via the port. Port-related bloodstream infection was defined according to Infectious Disease Society of America guidelines. All patients were examined by ultrasonography in case of clinical signs of thrombosis and systematically after discharge.

Results
Forty six TIAP were placed in 46 patients [median age: 25 years (20-41 years); 20 female and 26 male], and remained in place for a cumulative duration of 9756 days in situ (range 30-560 days). No pneumothorax occurred. Port-related bloodstream infection occurred in 2 cases (candida parapsilosis and staphylococcus aureus) [2/46, 4.3% ; 0.2 event per 1000 days]. These ports were removed. No port-related thrombosis occurred. No port-related deaths were observed.
In conclusion, the use of “high flow” totally implantable ports has resulted in a good option for long-term access to central veins and delivery of myeloablative conditioning followed by allogeneic SCT, in spite of severe neutropenia and increased risk of sepsis in this category of patients. Although multicentre randomized clinical trials are needed to define the optimal device in this clinical setting, the results of this prospective study support the wider use of TIAP in patients receiving myeloablative conditioning followed by allogeneic SCT.

Disclosures:
No relevant conflicts of interest to declare.
Population Pharmacokinetics Study of Tacrolimus in Chinese Hematopoietic Cell Transplant Patients.
Authors: Xue Ling , Rui Jian-Zhong , Zhang Yan , Miao Li-Yan 
Published: October 2019

Abstract
Abstract 4668

Background
The use of tacrolimus is complicated by its narrow therapeutic index and wide intra- and interpatient variability. The population pharmacokinetics (PPK) of tacrolimus have been reported in solid organ transplantation patients, but few investigation of tacrolimus PPK in hematopoietic cell transplant patients. Our study is to characterize the factors that influence tacrolimus population pharmacokinetics in Chinese hematopoietic cell transplant patients.

Methods
Sixty-eight patients undergoing hematopoietic cell transplant were enrolled. The database (671 data points) collected from the patients was analyzed by using the nonlinear mixed-effect model (NONMEM) program. Patients were administered oral or/and intravenous tacrolimus as part of immunosuppressive regimen that also included corticosteroids. And the robustness and accuracy of the model were assessed using bootstrap. A number of covariates including demographic characteristic, biochemical and hematological index, drug combination, IOV (inter-occasion variability) and other factors (e.g. Cause, POD, the type of transplantation and the sources of donor) were evaluated for their influence on the tacrolimus population pharmacokinetic parameters.

Results
The basic model was selected a one-compartment pharmacokinetic model with first-order absorption and elimination. The typical values of tacrolimus CL, V, F were 12.1 L·h-1, 686 L, 42.2%, respectively. And the inter-individual variations of CL, V, F were 23.5%, 96.4%, 43.8%, respectively. The absorption rate constant was fixed 4.3 h-1. The residual error between observed and model-predicted concentration was 3.03 ng·ml-1. The covariates that influenced the population pharmacokinetics parameters of tacrolimus included inhibitor (INHI), post operation days (POD), age and hemoglobulin (HGB) in the final regression model. The IOV of CL, V, F were 22.2%, 6.23%, 30.3%, respectively.

Conclusion
The covariates included INHI, POD, AGE, HGB and IOV would significantly influence tacrolimus population pharmacokinetics parameters in hematopoietic cell transplant patients.

Disclosures:
No relevant conflicts of interest to declare.
High Id1 Expression Is Associated with Poor Prognosis in 237 Patients with Acute Myeloid Leukemia.
Authors: Hirsch Pierre , Tang R. , Lapusan Simona , Marzac Christophe , Marie Jean-Pierre , Legrand Ollivier 
Published: October 2019

Abstract
Abstract 4687
Inhibitors of differentiation (Id) or DNA binding helix-loop-helix (HLH) proteins are a group of dominant inhibitors of basic HLH transcriptional factors, which act as positive regulators of cell growth or promote excessive proliferation, and also protect cells against drug-induced apoptosis in mammalians. Recently, Id1 has been identified as a common downstream target of several constitutively activated oncogenic tyrosine kinase(TK), such as FLT3/ITD, in leukemia cells. We analyzed Id1 expression as possible prognostic factor in 237 AML patients. High Id1 expression was associated with older age(p=0.009); and with Ftl3/ITD(p=0.003). However, 61% of the patients in the group of FLT3- AML were Id1+, suggesting that other TK are involved. In whole population, high Id1 expression independently predicted shorter disease free survival (DFS)(p=0.05), and overall survival (OS)(p=0.003). In the subgroup of young patients (age'60 years) with normal cytogenetic, Id1+ was, in multivariate analysis, associated with lower CR rates(p=0.02), shorter DFS(p=0.02) and OS(p=0.006).
In conclusion our data provides a new molecular marker for refining the risk classification of AML especially in young patients with normal cytogenetic. Id1- patients with normal cytogenetic should be classified as favourable-risk leukemia. Id1, as a downstream target of constitutively activated TK, could be a suitable candidate for targeted therapy.

Disclosures:
No relevant conflicts of interest to declare.
Targeting Tissue Factor to Tumor Vessels. Experimental Results and First-in-Man Experience.
Authors: Berdel Wolfgang E , Kessler Torsten , Schwöppe Christian , Liersch Ruediger , Persigehl Torsten , Bremer Christoph , Schliemann Christoph , Mesters Rolf 
Published: October 2019

Abstract
Abstract 469
Activation of blood coagulation in tumor vessels with subsequent tumor infarction is an experimental strategy in cancer therapy. We have fused different targeting peptides, including GRGDSP (RGD), GNGRAHA (NGR) and 5 cyclic derivates to the C-terminus of truncated tissue factor (tTF) to preferentially target tumor vessel endothelial cell integrins such as αvβ3 or aminopeptidase (CD13). tTF fusion proteins were expressed in E. coli, purified and refolded. Molecular integrity of the fusion proteins was evaluated by SDS-PAGE, immunoblotting and mass spectrometry. Subsequently, the tTF-fusion proteins were tested for biological activity with the following results: They retained the thrombogenic activity of tTF as measured by factor × activation in vitro. When tested with their respective target molecules either in a purified preparation or present on growing endothelial cells, there was specific binding. In vivo studies with human tumor xenograft models in nude mice showed either significant inhibition of tumor growth or regression of established tumors of different histologies (e.g. lung, breast, melanoma, sarcoma) by systemic application of the tTF-fusion proteins in contrast to controls including non-targeted tTF. Histology of the tumors treated with tTF fusion proteins revealed thrombotic occlusion of vasculature and blood pooling. Contrast-enhanced magnetic resonance imaging (MRI) of the tumors in vivo before and shortly after application of tTF-RGD and tTF-NGR showed a significant reduction of tumor perfusion. Degree of reduction correlated with in vivo tumor response. Toxicity studies showed acceptable therapeutic range and at therapeutic dosage there was no thrombo-embolic event in histology of normal organs, such as lung, heart, liver, and kidney. After upscaling production to amounts sufficient for clinical use, we have treated the first cancer patients with tTF-NGR. MRI studies even at the lowest dose (1 mg/kg i.v.) given showed reduction of tumor perfusion with no side effects. Targeted infarction of tumor vasculature with tTF fusion proteins may be promising as cancer therapy and should be further studied.

Disclosures:
Berdel: private: patent application on targeting tissue factor. Mesters:private: patent application on targeting tissue factor.
SET-NUP214 Fusion Gene at Chromosome 9q34 Contributed to the Development of Not Only T-ALL, but Also Primary and Secondary AML.
Authors: Shin Myung-Geun , Kim Da-Woon , Kim Hye-Ran , Cho Duck , Kee Seung-Jung , Kim Soo-Hyun , Shin Jong-Hee , Suh Soon-Pal , Ryang Dong-Wook 
Published: October 2019

Abstract
Abstract 4696
SET-NUP214 rearrangement at chromosome 9q34 is an extremely rare recurrent cryptic deletion resulting from del(9)(q34.11q34.13) mainly found in T-cell acute lymphoblastic leukemia (T-ALL). A multiplex RT-PCR system (HemaVision; DNA Technology) covering 28 leukemic fusion transcripts was applied to 271 bone marrow (BM) samples obtained from acute leukemia patients at initial diagnosis to investigate new pathophysiologic role of SET-NUP214 fusion gene. Out of 271 samples, we identified three cases (1.1%) harboring SET-NUP214 rearrangement – each case of adult T-ALL, pediatric acute myeloid leukemia (AML) from chronic myeloid leukemia (CML) and adult de novo AML with minimal differentiation (M0). Adult T-ALL occurred in 38-year-old man presented with dizziness and hepatomegaly. Conventional cytogenetic study disclosed normal karyotype and existence of SET-NUP214 fusion gene generated by del(9)(q34.11q34.13) was confirmed by RT-PCR, FISH and direct sequencing. Hematological remission was achieved after treatment of induction chemotherapy but during 7 months follow-up, SET-NUP214 fusion gene was steadily detected by RT-PCR. 13-year-old female with AML M0 converted from CML after treatment of imatinib for 2 months was the second case. Leukemic cells showed negative myeloperoxidase (MPO) and sudan-black B (SBB) staining, positive reaction against CD13, CD33, CD117 and MPO monoclonal antibodies by flowcytometry-based immunophenotyping. Major BCR-ABL1 rearrangement and SET-NUP214 fusion genes were simultaneously detected by RT-PCR, FISH and direct sequencing. She died at 38 days after diagnosis during the chemotherapy. The fusion protein contributed to the leukemogenesis by promoting expression of HOXA cluster genes. Last case was a de novo AML M0 in 39-year-old male. Cytochemical stains of leukemic cells on MPO and SBB were negative and immunophenotyping studies were positive for CD34, HLA-DR, CD13 and CD33. SET-NUP214 rearrangement was detected by multiplex RT-PCR. After treatment of induction and consolidation chemotherapy, he received allogeneous sibling stem cell transplantation (SCT), then complete chimerism was achieved. In clonclusion, SET-NUP214 rearrangement is associated with the development of not only T-ALL, but also primary and secondary AML via aberrant expression of HOXA cluster genes.

Disclosures:
No relevant conflicts of interest to declare.
Interphase Fluoroscence in Situ Hybridization Improves the Detection Rate of CNS Involvement in Childhood Acute Lymphoblastic Leukemia.
Authors: Park Seungman , Lee Sangho , Kang Hyoung Jin , Shin Hee Young , Ahn Hyo Seop , Kim Hyun Kyung , Lee Dong Soon , Cho Han Ik 
Published: October 2019

Abstract
Abstract 4714

Background
Morphologic examination of CSF cytospin is the standard method in the detection of CNS involvement of childhood acute lymphoblastic leukemia (ALL). However, the result widely varies depending on the status of cell degeneration caused by cytospin procedure. We performed interphase fluoroscence in situ hybridization (FISH) on CSF cytospin cells to examine its potential usefulness in detecting malignant cell population in the CSF.

Materials and Methods
The study used 19 CSF cytospin specimens of the childhood ALL patients under treatment. All of them had cytogenetic abnormalities in the bone marrow nuclear cells at the initial diagnosis. None of the patients showed malignant cells in morphologic examination of the cytospin. Interphase FISH was performed to detect recurrent genetic abnormalities including rearrangement of BCR/ABL, TEL/AML1, MLL and IgH rearrangement, and p16 deletion using LSI BCR/ABL dual color, dual fusion translocation probe, LSI TEL/AML1 ES dual color translocation probe, LSI MLL dual color, break apart rearrangement probe, LSI IGH dual color, break apart rearrangement probe, and LSI p16/CEP 9 dual-color probe (Abbot Molecular Diagnostics). FISH signals were interpreted using automated slide scanning system (Metafer 4 system) and manually read for confirmation.

Result
Two patients had the same cytogenetic abnormalities in CSF cytospin as observed in the bone marrow mononuclear cells: one patient had 4 copies of AML1 gene, suggesting tetrasomy 21 or multiple fusion partner gene other than chromosome 12, both in bone marrow and in CSF; the other had 3 copies of p16 gene including one with low signal intensity both in bone marrow and in CSF. Ten patients did not have any cytogenetic abnormalities. We could not interpret the result in the other 7 patients due to low cell count and/or low signal intensity.

Conclusion
Out of 19 who were diagnosed not to have malignant cell in CSF in morphologic examination, 2 patients had cells with the same clonal abnormalities in the CSF as bone marrow. Our result indicates that FISH study would be helpful in evaluating CSF involvement of hematologic malignancies including ALL. In order to improve the performance, it is required to prevent degeneration of the cells by prompt specimen delivery and slide preparation.

Disclosures:
No relevant conflicts of interest to declare.
The Use of ABL and GUS as Control Genes for Normalizing Quantification of Fusion Transcripts by Real Time PCR: Our Experience of More Than 1000 Samples.
Authors: Lassoued Mohamed Amin Ben , Beaufils Nathalie , Loundou Anderson Dieudonné , Arnaud Christiane , Gabert Jean 
Published: October 2019

Abstract
Abstract 4727

Introduction
Real-time quantitative PCR (RT-PCR) is a sensitive and accurate tool for monitoring leukemic patients by amplification of fusion gene (FG) transcripts.
In order to correct variations in RNA quality and quantity and to calculate the sensitivity of each measurement, a control gene (CG) transcript should be amplified in parallel to the FG transcript.
In our laboratory two CG are used: Abelson (ABL) and beta-glucuronidase (GUS).
Clinical application for monitoring the treatment of chronic myelogenous leukemia (CML) patients with tyrosine kinase inhibitor makes the quantification of which CG to use very important into the clinical practice today.

Objective
The aim of this study was to assess the degree of agreement between the GC, compare the expression of GC (type and time of sampling and pathology), try to redefine the limits between the 4 groups of quality (very good, good, poor and very poor quality samples) and to focus on CML patients.

Materials and methods
A total of 1087 samples (peripheral blood and bone marrow) were obtained from 475 patients from January 2005 to December 2007. This study includes patients with acute myeloid leukaemia (AML), acute lymphoblastic leukemia (ALL), chronic myeloid leukaemia (CML) and Hypereosinophilic syndrome (HES).
We collected 366 samples of 41 CML patients between 2002 and March 2009 including 32 CML at diagnosis.
RT-PCR analysis was performed using the EAC protocols.

Results
Comparison between CML, AML, ALL and HES patients showed that ABL and GUS expression did not differ significantly (Bland and Altman method).
ABL and GUS gene expressions did not differ significantly between CML patients at diagnosis and the others.

Conclusion
Based on the literature and our local experience of more than 1000 samples, we propose to use only ABL as CG in routine practice in onco-hematology particularly for CML patients at diagnosis.

Disclosures:
No relevant conflicts of interest to declare.
High Dose Cyclophosphamide for Moderate to Severe Refractory Multiple Sclerosis: 2-Year Follow-up (investigational new drug No. 65863).
Authors: Gladstone Douglas 
Published: October 2019

Abstract
Abstract 4744

Background
High dose cyclophosphamide (HDC) is a chemotherapy treatment designed to eradicate autoreative B- and T-cells responsible for lymphocyte-mediated autoimmune illness, while sparing the pluripotent blood stem cell of any ill effects. Multiple sclerosis (MS) is the most common inflammatory and demyelinating immune-mediated disorder of the central nervous system in young adults.

Methods
Patients with moderate to severe, refractory MS, defined as an Expanded Disability Status Scale (EDSS) score of 3.5 or higher after 2 or more Food and Drug Administration-approved disease-modifying agents, received 200 mg/kg of cyclophosphamide over 4 days. For the following 2 years, quarterly EDSS score evaluations and biannual brain magnetic resonance imaging and neuro-ophthalmologic evaluations were obtained.

Results
15 patients were evaluated for clinical response. During follow-up, no patients increased their baseline EDSS score by more than 1.0. EDSS score stability or decrease was realized in 5 of 7 (71%) patients with relapsing remitting MS and 5 of 8 (62%) patients with secondary progressive MS patients. 4 patients required additional immunomodulatory treatment after treatment. Neurologic improvement involved changes in gait, bladder control, and visual function. Treatment response was seen regardless of the baseline presence or absence of contrast lesion activity.

Conclusions
HDC can effectively decrease symptoms, stop disease progression, and allow for disability regression in RR and SPMS patients. The most appropriate candidates for HDC, its duration of benefit and the potential need for prophylactic preventative immune manipulation after HDC all require further investigation.

Disclosures:
Off Label Use: cyclophosphamide: IND# 65863.
Elevated Heat Shock Protein 70 (HSP70) Serum Levels After Tumor Radiofrequency Ablation Might Represent a Favorable Prognostic Marker.
Authors: Haen Sebastian P. , Pereira Philippe L. , Clasen Stephan , Schmidt Diethard , Boss Andreas , von Herbay Alexandra , Kanz Lothar , Rammensee Hans-Georg , Gouttefangeas Cecile 
Published: October 2019

Abstract
Abstract 4748

Introduction
Radiofrequency (RF) ablation is a safe and effective method to locally treat malignancies of the liver, kidney and more recently lung. For the treatment of small tumors, long term results are comparable with outcome after surgical resection and better than current chemotherapies. RF-ablation has now been reported to induce heat shock protein (HSP) expression on sublethally damaged hepatocytes. HSPs are potent alarm signals for the immune system causing i.e. activation of T cells and dendritic cells. In addition to triggering immune responses directed against HSPs themselves, HSPs can carry antigens derived from those cells of which HSPs originated and thereby elicit specific immune reactions against these cellular epitopes.

Patients and Methods
We here analyzed serum samples of 22 patients to evaluate the systemic release of HSP70 using a specific HSP70 ELISA. Patients were followed for a median of 30 months until death or three years after the last RF-treatment. As a control group, sera from 20 patients undergoing diagnostic liver biopsy were screened.

Results
The median increase of HSP70 in serum comparing levels before and one day after the procedures was 1.6-fold in the RF-ablation group, while no soluble HSP-70 could be detected in the control group before and after the biopsy. Nine patients showed a higher than two-fold increase of HSP70 in the serum after ten sessions of RF-ablation compared to levels prior to treatment. No correlation of HSP70 levels with tumor volume, the ablated non-malignant tissue or the total size of the necrosis was observed. In the subgroup of patients with elevated HSP-70 serum levels, 67% (6 out of 9) were alive at the end of follow up with only one suffering from progressive disease. In contrast, 61% (8 out of 13) of the other patients suffered from progressive disease or had deceased at the end of follow-up.

Conclusion
RF-ablation not only induces cellular HSP expression but also can lead to a systemic release of serum HSP70. Patients with strong increases in HSP-70 serum levels tended to have a better survival than patients without induction of HSP-70 in the serum. Thus, immunomodulation might represent an additional beneficial effect of RF-ablation besides local tumor destruction, since HSPs are strong stimuli for the immune system and can mediate tumor-specific T cell immunity.

Disclosures:
No relevant conflicts of interest to declare.