bluuebunny/crossref_metadata_2025_split huggingface dataset:
Absolute Lymphocyte Count Assessed During Routine Follow-up Is a Risk Factor for Relapse Post-Autologous Peripheral Blood Hematopoietic Stem Cell Transplantation in Patients with Diffuse Large B-Cell Lymphoma.
Authors: Porrata Luis , Inwards David J , Ansell Stephen M , Micallef Ivana , Johnston Patrick L , Hogan William , Markovic Svetomir 
Published: October 2019

Abstract
Abstract 4359
Routine serial chimerism analysis after allogeneic stem cell transplantation (Allo-SCT) has been used as a risk factor for post-Allo-SCT cancer relapse. In non-Hodgkin lymphoma (NHL) patients treated with autologous peripheral blood hematopoietic stem cell transplantation (APHSCT), a specific predictor during routine follow-up that could be used to ascertain risk for post-APHSCT relapse has not been identified. Thus, we set out to investigate if absolute lymphocyte count (ALC) measured at post APHSCT follow-up could serve as a marker of post-APHSCT NHL relapse. ALC was obtained at the time of confirmed relapse at follow-up and at last no evidence of relapse follow-up. From 1993 until 2005, 269 patients treated with APHSCT for diffuse large B-cell lymphoma (DLBCL), were included in this study. Patients at last follow-up with no relapse (N=137) had a higher ALC compared with those with confirmed relapsed at follow-up post-APHSCT (N= 132) [median ALC x 109/L of 1.66 (range: 0.13-3.69) vs 0.71 (range: 0.03-4.75), p <0.0001, respectively]. ALC at follow-up was a strong predictor for relapse with an area under the curve (AUC) = 0.86 (p<0.0001). An ALC of 1.0 × 109/L was identified as the optimal cut-off point. An ALC < 1.0 × 109/L at the time of confirmed relapse had a positive predictive value of 89% and a positive likelihood ratio of 8.4 to predict relapse post-APHSCT. Patients with an ALC ≥ 1.0 × 109/L (N = 147) had a cumulative incidence of relapse of 19% versus 92% for patients with an ALC < 1.0 × 109/L (N = 122) (p<0.0001) at follow-up post-APHSCT. This study suggests that ALC at follow-up can be used as a marker to assess risk of DLBCL relapse during routine follow-up after APHSCT.

Disclosures:
No relevant conflicts of interest to declare.
Comparison of Array Comparative Genomic Hybridization (aCGH) to FISH and Cytogenetics in Prognostic Evaluation of CLL.
Authors: O'Malley Dennis P. , Giudice Christina , Chang Averee S , Chang Dorothy , Barry Todd S , Hibbard Michele K , Chen Rebecca , Chen Sing-Tsung 
Published: October 2019

Abstract
Abstract 4393

INTRODUCTION
Chronic lymphocytic leukemia (CLL) is a commonly encountered hematologic neoplasm. Evaluation of prognosis in CLL is strongly based on genetic findings and the most commonly used studies are classical cytogenetics and targeted interphase fluorescence in situ hybridization (FISH). High resolution array comparative genomic hybridization (aCGH) is a relatively new and robust method of evaluating chromosomal alterations over the entire genome. We compared aCGH with routine cytogenetics and FISH in detecting genetic alterations in newly-diagnosed CLL cases.

MATERIALS AND METHODS
aCGH testing was performed on 55 cases of CLL in addition to a standard panel of FISH probes (ATM on 11q22, trisomy 12, 13q14, p53 on 17p13 using a standard cutoff for positivity of 10%). These results were compared to a control group of 100 CLL cases upon which routine cytogenetics and FISH were performed. The frequency of detecting abnormalities was compared between the groups and discordant results between methodologies were compared.

RESULTS
In the control group (n=100), the mean age was 71 (52-86) with a male to female ratio of 1.6:1. Genetic abnormalities were detected by classical cytogenetics in 19% (19/100) of cases as compared to FISH which detected abnormalities in 66% (66/100) of cases (Table 1). An additional group of 55 CLL cases [male to female ratio of 2.2:1 and a mean age of 71 (52-90)] was analyzed by both aCGH and FISH. This additional group of CLL cases showed a similar frequency of genetic abnormalities by FISH (60%; 27/45). In contrast to FISH, aCGH detected genetic abnormalities in 82% (45/55) of CLL cases (Table 1).
aCGH identified genetic abnormalities not detected by FISH studies in 16% (7/45) of cases whereas FISH identified abnormalities not detected by aCGH in only 7% (3/45) of cases. Rare recurring genetic alterations were detected by aCGH, which would not have been detected by a standard FISH panel, and included losses in 6q, 8p, 10q, 14q32, and 18q, and gains in 10q.

DISCUSSION
Classical cytogenetics is often performed in cases of CLL, but is not particularly useful as CLL cells are often difficult to grow in culture and because of the low rate of detecting common genetic alterations. Our findings suggest that aCGH is an effective and robust technique for evaluating recurring genetic abnormalities in CLL and is better than classical cytogenetics and standard FISH in detecting common genetic abnormalities in CLL.

Disclosures:
No relevant conflicts of interest to declare.
Mechanism Research of Magnetic Nanoparticle Fe3O4 and 5-Bromotetrandrine On Reversal of Multidrug Resistance in K562/A02 Leukemic Cells.
Authors: Chen Bao-An , Wu Wei-Wei , Cheng Jian , Gao Feng , Xu Wen-Lin , Gao Chong , Ding Jia-Hua , Sun Xin-Chen , Xu Cui-Rong , Chen Ning-Na , Xia Guo-Hua , Wang Xue-Mei , Chen Wen-Ji , Liu Li-Jie , Li Xiao-Mao 
Published: October 2019

Abstract
Abstract 4827

Purpose
The present study aimed to research the machenism of magnetic nanoparticles Fe3O4 (Fe3O4-MNPs) and 5-bromotetrandrine (BrTet) on multidrug resistance cell line K562/A02 solitarily or symphysially.

Mothed
The proliferation of K562 and K562/A02 cells, and the cytotoxicity of PMBCs which were cultured with daunomycin (DNR) alone or in combination with Fe3O4-MNPs (0.1, V/V), BrTet (0.5μM) or both for 48 h, were evaluated by MTT assay. DNR accumulation of K562, K562/A02 cells and PMBCs were analyzed by fluorospectrophotometry after incubated with 2μM DNR in the absence or presence with Fe3O4-MNPs (0.1 V/V), BrTet (0.5μM) or both for 48 h. Real time–PCR analyses and western blotting were performed to examine the mRNA and proteins level, respectively.

Result
The results showed that the combination of Fe3O4-MNPs and BrTet with effective concentration could exaggerate cytotoxicity against MDR cell line K562/A02 significantly. Flow cytometry assay showed that 0.5μM BrTet in combination with Fe3O4-MNPs (0.1, V/V) significantly enhanced the intracellular accumulation of DNR in K562/A02 cells and its potency was greater than that of BrTet or Fe3O4-MNPs alone at the same concentrations. While both Fe3O4-MNPs and BrTet in company with DNR did not increase the cytotoxicity to PMBCs. Both BrTet and Fe3O4-MNPs inhibited the overexpression of MRP1, MRP2, MRP4 and MRP5, and down regulated, mrps mRNA expression in K562/A02 cells to some extent.

Conclusion
We propose that Fe3O4-MNPs loaded with DNR and BrTet probably have synergetic effect on reversal in multidrug resistance. However, this combination shows less cytotoxicity to normal cells and reveals better target. The results may provide evidence for clinic application of them as reversal agents of drug resistance.

Disclosures
No relevant conflicts of interest to declare.
Diagnostic Utility of Flow Cytometry in Myelodysplastic Syndrome: A Retrospective Validation From a Single Centre.
Authors: Topcuoglu Pervin , Dalva Klara , Bakanay Sule Mine , Bozdag Sinem Civriz , Arslan Onder , Ozcan Muhit , Arat Mutlu , Beksac Meral , Gurman Gunhan 
Published: October 2019

Abstract
Abstract 4846
Myelodysplastic syndrome (MDS) is heterogeneous clonal hematopoietic stem cell disorder characterized by cytopenia(s) and dysplasia in one or more cell lineage. Though flow cytometry (FCM) is an important diagnostic tool in hematopoietic cell disorders, a prominent immunophenotyping feature in MDS may not be determined. In this study, we retrospectively evaluated flow cytometric features of bone marrow samples diagnosed as MDS with clinical and hematological findings.

Patients-Method
Between Feb 2004 and March 2009, flow cytometric parameters of 73 patients (M/F: 50/23) with MDS were re-analyzed. Median age was 59 years (17-89 ys). Our general principles are to evaluate quality of bone marrow samples, to determine proportion of the cells and features of their light scatter, and to give percentage of the blast. When detected a finding of dysplasia in the first analysis, the second step includes the determination of the maturation of the cells and the presence of the aberrant antigen expression.

Results
The samples were interpreted as MDS in % 76.7, MDS-RAEB-1 or RAEB-2 in %16.4, myeloproliferative disorder in %1.4 and non-diagnostic in %6.8 of the cases by flow cytometric examination. We detected variable degrees of hypogranulation in myeloid lineage in %82.2 of the samples by the light scatter features of the cells: 85% of severe and 15% of moderate or mild hypogranulation. The ratio of myeloid and lymphoid was changing from 0.3 to 17.5 (median 2). The decreasing of this ratio (<1) was observed in 19.4% of the samples. We detected altered expression of mature granulocyte. These included decreasing or lack of expression in CD15 45/73 (61.4%), CD13 38/70 (54.3%), CD16 53/67 (79.1%), CD11b 51/71 (71.8%), CD24 44/69 (65.2%), CD10 23/72 (31.9%) and MPO 14/72 (19.4%). Besides, bright expression of CD33 in 53.5% of the samples was observed. CD36 and CD56 in myeloid lineage were co-expressed in about 50 % of the samples. In 80.8 % of the samples dysplasia in erythroid compartment could be evaluated: Expression of CD71 according to glycophorin A (ratio <1) was decreased in 23.7 %. When we made similar analysis in the samples without RAEB-1 and -2 as pathological examination of bone marrow, 13.4 % of the samples could not be evaluated in favor of dysplasia. Of the samples with dysplasia hypogranulation, aberrant antigen expression of myeloid lineage and eryhtroid dysplasia were observed in 92.1%, 34.1% and 31.5%, respectively. In conclusion, FCM events may help to the differantial diagnosis of MDS especially when combining with clinical events. Improving of the analysis by focusing on the blast characteristics may be a standard approach to evaluate for low risk MDS.

Disclosures
No relevant conflicts of interest to declare.
Differential Diagnosis of Myelodisplastic Syndrome, with Flow Cytometry.
Authors: De Salvo Cardullo Luigi , Duerto Mirian C , De Salvo Dalila DV , Salas-A Rodolfo J , Weir-M Jesus E , Nuñez Ana M , Nuñez Alonso A , Plumacher Zaida A , Fernandez Olga Ch , Avila Estela BEATRIZ , Rios Mariela J , Paz Luis S , Salas Dalida DJ , Fuenmayor Alba M 
Published: October 2019

Abstract
Abstract 4860
Our flow cytometry laboratory processed 38 bone marrow (BM) samples (24 males and 14 females) from patients with Myelodisplasic Syndrome diagnosis, from January 2008 to May 2009. Upon examination, most BM smears corresponded with an hypercellular bone marrow and increased intermediate and immature cells. All patients classified according to OMS for Myelodisplasic Syndromes. Twenty-four (n=24) patients (65%) presented RA, five (n=5) patients (13%) RAEB-1, one (n=1) 1% presented RARS, and eight (n=8) patients (21%) classified as RAEB-II. Cytogenetic result analyses for all patients classified according with the Index Prognostic Standard Score (IPSS). Twenty-five (n=25) patients or 66% corresponded with a good cytogenetic risk; seven (n=7) or 20% corresponded with an intermediate risk and six (n=6) or 14% with poor risk. Cell cytometry results corresponded with increased immature cells. In 20 patients, the DR/CD34 average 27,29% (range 0-67); in 24 patients, the DR/CD33 average 31,80% (range 6-100); in 29 patients, the DR/CD117 average 18,89% (range 0-43); in 7 patients, the CD33/CD34 average 22,14% (range 0-50); in 5 patients, the CD38 average 47,60% (range 32-61); in 4 patients, the BCL2 average 21,75% (14-26 and in 3 patients, the DR/CD64 average 44,33% (range 30-60). All patients with high average of DR/CD64 classified as CMML. Patients with RAEB-II diagnosis classified as MDS in transformation. Most patients displayed 10% CD34+ values higher than normal human average. We conclude the cytometry is an important tool in the differential diagnosis of MDS.

Disclosures
No relevant conflicts of interest to declare.
Arsenic Trioxide-Mediated Growth Inhibition of Myeloma Cells Is Associated with An Extrinsic Signaling Pathway Involved in Caspase 3 and Caspase 8.
Authors: Shi Jumei , Wu Yi , Wang Siqing , Meng Xiuqin , Wei Rong , Hu Xiaojing , Zangari Maurizio , Tricot Guido J. , Zhan Fenghuang 
Published: October 2019

Abstract
Abstract 4899
Arsenic trioxide (ATO) is a well-known inhibitor of cell proliferation in certain forms of malignancy and has been successfully used in the treatment of acute promyelocytic leukemia. Preclinical and clinical studies showed that ATO has anti-myeloma effects both as a single agent and in the combination therapy; however, the underlying molecular mechanism remains elusive. This study was performed to evaluate the molecular mechanism underlying its anti-myeloma activities. Cells from OPM2, U266, RPMI8226 myeloma cell lines and patients diagnosed with myeloma (n=6) were cultured with various concentrations of ATO for 4 days. Cell growth and viability were assayed by trypan blue dye exclusion. Cell cycle and apoptosis were analyzed by flow cytometry using CellQuest software and Vybrant Apoptosis Assay Kit. Alterations of the signaling pathways induced by ATO were tested by real-time PCR and western blot. ATO induced potent inhibition of myeloma cell growth compared with untreated control cells. Further investigation showed that ATO down-regulated c-Myc and phosphorylated (p)-Rb, while it up-regulated p53, p21Clip1, and p27Kip1 proteins, resulting in G2/M cell cycle arrest and cell growth inhibition. ATO treatment increased mRNA levels of interferon regulatory factor-1 (IRF-1) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), as well as protein levels of caspase 8 and cleaved caspase 3, indicating involvement of the extrinsic apoptotic pathway. No significant change was detected in the expression levels of Bax, Bcl-xL caspase 9 and Bcl-2, indicating that the intrinsic signaling pathway was not involved. A pan-caspase inhibitor abrogated ATO-induced apoptosis of myeloma cells. Our data suggest that ATO induces apoptosis in MM cells most likely through an extracellular signaling pathway.

Disclosures
No relevant conflicts of interest to declare.
Compassionate Use of Combined Anti-Inflammatory and Angiostatic Treatment as Third-Line Therapy in Eight Patients with Multiple Myeloma – a Combined Treatment Setting of Lenalidomide with Pioglitazone, Dexamethasone and Low-Dose Treosulfan.
Authors: Reichle Albrecht , Vogelhuber Martin , Holler Ernst , Hahn Joachim , Andreesen Reinhard 
Published: October 2019

Abstract
Abstract 4958

Introduction
Therapeutic options for patients with refractory and multifold pretreated multiple myeloma (MM) are limited, as patients have already received the novel targeted therapy approaches at this stage. Continuous production and release of pro-inflammatory cytokines may account for its resistance towards cytotoxic agents. Therefore, a multi-targeted approach consisting of anti-angiogenic, anti-inflammatory and anti-tumor components was implemented for response induction prior to and after allogeneic transplantation.

Methods
Within a compassionate-use program eight patients suffering from refractory or relapsed osteolytic multiple myeloma (MM) or plasma cell leukemia (PCL) and proven progression of serum paraprotein levels were treated in third-line continuously with daily lenalidomide 10 mg, pioglitazone 60 mg, treosulfan 250 mg once daily, and dexamethasone 1 mg.

Results
Here we report results of eight patients (48 to 63 years old) treated for refractory (n= 7) or relapsed (n= 1) multiple myeloma (n= 7) or PCL (n= 1). Prior therapy consisted of double autologous transplantation with high-dose treosulfan/melphalan conditioning, lenalidomide and/or bortezomib. One patient suffered from progressive disease following allogeneic stem cell transplantation (allo-SCT). No side effects &gt; WHO 2 occurred on treatment. In one patient, bortezomib-induced motoric polyneuropathia partially resolved on study medication. Biomodulatory therapy induced clinically meaningful and very rapid responses, stable disease, n= 2, partial remission (PR), n= 3, and very good PR (VPR, n= 3). Therapy-induced PR and VPR facilitated allo-SCT in 2 refractory patients. A third patient with refractory disease after allo-SCT received donor lymphocytes. Two patients with allo-SCT survived in VPR for 10+ and 3+ months, respectively. One patient remained in ongoing VPR since 9 months after donor lymphocyte infusion. Patients not proceeding to allo-SCT had SD for 3 (PLC) and 4 months, VPR and PR for 3+, 4+ and 5+ months.

Conclusions
Together, biomodulatory therapy has remarkable clinical activity in pretreated and refractory patients with MM. The eight cases show comparatively high efficacy at low toxicity rates of a biomodulatory acting therapy approach: Although the substances have limited or no efficacy at the respective dose levels, and were already administered previously at higher dose-levels (i.e. treosulfan and/or lenalidomide), the combined treatment setting is promising since it targets the myeloma cells itself as well as the surrounding stroma. Therefore, this treatment schedule deserves further investigation and discharges now in a novel multi-center phase II trial for third-line therapy in multiple myeloma.

Disclosures
No relevant conflicts of interest to declare.
Prospective Exploratory Phase II Studies of A Rotating Regime of Nilotinib and Imatinib for Frontline Treatment of Philadelphia POSITIVE (Ph+) Chronic Myeloid Leukemia (CML) and Acute Lymphoblastic Leukemia (ALL).
Authors: Saglio Giuseppe , Abruzzese Elisabetta , Alimena Giuliana , Bocchia Monica , Carella Angelo Michele , Fanin Renato , Martinelli Giovanni , Rosti Gianantonio , Russo Domenico , Specchia Giorgina , Baccarani Michele 
Published: October 2019

Abstract
Abstract 4972
The introduction of Imatinib (IM) and subsequently of Dasatinib and Nilotinib (NI) has made a substantial contribution to the therapy of all Ph+ leukemias. With IM alone, a complete cytogenetic response (CCgR) is achieved in about 75% of CML patients who are treated frontline, but about 15% of them loose the response in the first 3 years. A complete hematologic response (CHR) is achieved in almost all ALL patients, but about 50% of them relapse with BCR-ABL mutated clones within one year. With NI alone, a complete cytogenetic response is obtained in about 40% of Ph+ leukemia patients who are resistant to IM, and in more than 90% of early chronic phase, previously untreated, CML patients. NI is a derivative of IM, but has different pharmacokinetic and pharmacodynamic properties, and inhibits most of BCR-ABL mutants which are resistant to IM. Moreover, the toxicity profile of the two drugs is different. The administration of two tyrosine kinase inhibitors (TKIs) may increase the strength of initial therapy, so increasing the rate and the solidity of the response, and reducing the rate of failures. To test the feasibility and the validity of this hypothesis the GIMEMA CML WP designed a rotating regime of NI and IM to be tested front-line in patients with Ph+ CML, and in the patients with Ph+ ALL who are more than 60 years old. The first course is with NI 400 mg twice daily. The second course is with IM 400 mg once daily. A course lasts 3 months in CML, and 6 weeks in ALL. Treatment duration (study core) is 2 years in CML (8 courses), and 36 weeks in ALL (6 courses). The primary endpoint is event-free survival at 24 months in CML, and disease-free survival at 6 months in ALL. CML patients enrollment (n = 120) has been completed between February and August 2009. The results of the first interim analysis at 3 and 6 months will be presented. The enrollment of ALL patients will begin September 2009.

Disclosures
No relevant conflicts of interest to declare.
Effect of CD20 Expression Levels On B-Cell Lymphoma Patients' Background and Prognosis.
Authors: Suzuki Yuhko , Yoshida Tsutomu , Nakamura Naoya , Togano Tomiteru , Miyazaki Koji , Ishii Ryuuji , Ohsaka Manabu , Danbara Mikio , Horie Ryouichi , Nakayama Meijin , Okayasu Isao , Higashihara Masaaki 
Published: October 2019

Abstract
Abstract 5000

AIM
The anti-CD20 antibody Rituximab has improved the disease-free survival and overall survival of patients with several types of B-cell lymphoma, in which CD20 is expressed on the cell surface and in the cytoplasm. Flow cytometry indicated that CD20 expression varies from null, weak to strong. Despite its clinical importance, the influence of CD20 expression levels on lymphoma therapy has not been well elucidated. Patients, Material and

Methods
214 cases were analyzed, all newly diagnosed with B-cell lymphoma at our institute from January in 2002 to April in 2009. All were biopsied and analyzed by flow cytometry. The biopsy specimens were fixed in formalin and stained with Hematoxylin-Eosin; they were also immunostained. Histological subtypes were defined according to the World Health Organization Classification Ver. 3. The mean florescence intensities of CD20 and CD19 were determined by flow cytometry, and cytoplasmic expression of CD20 was determined by immunohistochemistry. 1) The cases were categorized as follows: A: CD20-negative, B: CD20/19 less than 20%, C: CD20/19 20-50% and D: CD20/19 more than 50%. Patients' backgrounds, pathological diagnoses, primary lesions, etc. were also analyzed. 2) Among DLBCL cases, 76 treated with R-chemo were selected and analyzed with respect to treatment response and overall survival.

Results
1) Diagnoses: DLBCL 128, Follicular lymphoma 58, MALT 7, CLL 4 and so on. Among the DLBCL cases, immunohistochemistry indicated six (4%) were CD20-negative and three were CD20-positive; the ages in the CD20-negative DLBCL group tended to be high (74.28 vs 64.36, p=0.06, t-test). Weak CD20 expression was observed in 15 cases (B: 4, C:11). No statistically significant differences were seen with respect to gender, IPI, clinical stage, biopsy lesion, CD5 expression or Karyotype. No FL cases were CD20-negative, but two ( 3.4%) showed low CD20 expression. 2) Kaplan-Meier analysis of 76 cases of DLBCL treated with R-Chemo showed a significantly lower overall survival for Group A, (CD20-negative) while there was no significant difference in overall survival of Groups B+C(CD20-weak) and D(CD20-normal). Our results indicate that even patients with lower levels of CD20 expression in B-cell lymphoma may respond favorably to anti-CD20 therapy. Further studies will be necessary to explore this possibility.

Disclosures
No relevant conflicts of interest to declare.