bluuebunny/crossref_metadata_2025_split huggingface dataset:
Complete Eradication of Minimal Residual Disease (MRD) in Patients with Hairy Cell Leukemia (HCL) after Cladribine (2CDA) Followed by an Extended Course of Rituximab.
Authors: Ravandi-Kashani Farhad , O’Brien Susan , Michael Keating , Jones Dan , Faderl Stefan , Ferrajoli Alessandra , Wierda William , Odinga Shirley , Pierce Sherry , Koller Charles , Verstovsek Srdan , Jorgensen Jeffrey , Kantarjian Hagop 
Published: April 2020

Abstract
The nucleoside analogs 2CDA and pentostatin are effective in the initial therapy of patients with HCL, but relapses are common. We have investigated whether an 8 week course of therapy with weekly rituximab after 2CDA can eradicate MRD assessed by flow cytometry as well as by immunoglobulin heavy chain (IgH) polymerase chain reaction (PCR) assay using framework-1, -2 and -3 primer sets. Treatment constituted 2CDA 5.6 mg/m2 IV given over 2 hours daily for 5 days. One month after initiation of 2CDA, bone marrow (BM) MRD was assessed and rituximab 375 mg/m2 IV was given weekly for 8 weeks. BM MRD was reevaluated again after the completion of rituximab. To date, 12 patients have been treated including 10 with newly diagnosed disease and 2 after relapse from prior therapy (2CDA in one patient and chlorambucil in another). The median age of the patients was 55 years (range 31–73). Two patients had variant HCL. Fluorescent in situ hybridization (FISH) was positive for the presence of a clone with p53 deletion/monosomy 17 in 2 patients. One patient with variant HCL had complex cytogenetics. Seven of 10 evaluable patients had mutated IgVH gene whereas IgVH was unmutated in 3 patients. All 12 patients (100%) have achieved CR defined as no hairy cells in BM and blood with normalization of counts (ANC &gt;1.5 x 109/L, Hgb &gt;12.0 g/dL, Plt &gt; 100 x 109/L) after completion of all therapy; 5 patients still had 1% to 50% hairy cells in BM after 2CDA therapy. MRD by flow was positive in 11 patients after 2CDA therapy but became negative in all 12 patients after rituximab. MRD by PCR was positive in 5 of 10 evaluable patients after 2CDA therapy and became negative in 10 of 11 evaluable patients (equivocal in 1) after rituximab. With a median duration of follow-up of 11 months (range 3–13) no patients have relapsed (median response duration 6+ months, range 1–13). One patient has developed a second cancer (pancreatic). We conclude that therapy with an extended course of rituximab is effective in eradicating MRD in HCL. The predictive value of achievement of negative MRD, presence of chromosome 17 abnormalities, or the mutational status of IgVH on the risk of relapse should be evaluated in larger series and with longer follow-up.
Transient Response to Imatinib in an Atypical Chronic Myeloproliferative Disease Associated with ins(9;4)(q34;q21q31) and a CDK5RAP2-PDGFRA Fusion Gene.
Authors: Walz Christoph , Schultheis Beate , Metzgeroth Georgia , Schoch Claudia , Curtis Claire , Lengfelder Eva , Haferlach Torsten , Berger Ute , Hochhaus Andreas , Hehlmann Rüdiger , Cross Nicholas C.P. , Reiter Andreas 
Published: April 2020

Abstract
Chronic myeloproliferative disorders (CMPD) associated with rearrangements of the ‘platelet-derived growth factor receptor A’ (PDGFRA) at chromosome 4q12 are excellent candidates for targeted therapy with imatinib. To date, two fusion genes involving PDGFRA have been described: FIP1L1-PDGFRA and BCR-PDGFRA. Here we report a female patient who presented in advanced phase of an atypical myeloproliferative disease. Routine cytogenetic analysis revealed an ins(9;4)(q34;q21q31). Although no break was visible at 4q12, FISH analysis with flanking BAC probes indicated that PDGFRA was disrupted. To identify the partner gene we employed 5′-RACE PCR, exploiting the fact that all known PDGFRA fusions involve exon 12 of this gene. The resulting PCR-products consisted of 5′-sequences derived from CDK5RAP2 (CDK5 regulatory subunit associated protein 2) located on 9q33 and 3′-sequences derived from PDGFRA. CDK5RAP2 encodes a protein that is believed to be involved in centrosomal regulation. FISH analysis confirmed the co-localization of 5′ CDK5RAP2 and 3′ PDGFRA sequences. RT-PCR confirmed the in-frame mRNA fusion between exon 13 of CDK5RAP2, a 40-bp insert which was partially derived from PDGFRA intron 11 and a truncated PDGFRA exon 12. No reciprocal fusion transcript could be amplified by RT-PCR. The predicted CDK5RAP2-PDGFRA protein consists of 1003 amino acids and retains both tyrosine kinase domains of PDGFRA and several potential dimerisation domains of CDK5RAP2 suggesting a mechanism of tyrosine kinase activation similar to BCR-ABL. Treatment with 400 mg imatinib was initiated and the patient achieved a complete cytogenetic and molecular remission. However, hematological response was only partial with residual blasts repeatedly detectable in the blood and marrow. The patient rapidly developed acute leukemia while still remaining in complete cytogenetic and molecular remission suggesting the outgrowth of a second imatinib-resistant leukemic clone. These findings and the fact that the ins(9;4) was only seen in 24% of metaphases at diagnosis suggests that CDK5RAP2-PDGFRA may have been a secondary abnormality. In summary, we have identified a third fusion gene involving PDGFRA, underlining the fundamental role of activated tyrosine kinases in CMPD’s and their possible response to targeted therapy with imatinib.
Predictable Prognostic Factor of CD56 Expression in Acute Myeloid Leukemia with t(8:21) Including Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Yang Deok-Hwan , Mun Yeung-Chul , Shin Ho-Jin , Kim Yeo-Kyeoung , Lee Je-Jung , Jin Choi-Young , Seong Chu-Myong , Kim Hyeoung-Joon 
Published: April 2020

Abstract
CD56 expression in acute myeloid leukemia (AML) has been associated with extramedullary leukemia and multi-drug resistance, but the clinical and prognostic significances are not yet clearly defined. Recently, some investigators reported that AML patients with t(8;21) showed more frequent CD56 expression rate and the expression of CD56 antigen adversely affected disease-free survival (DFS). It could explain a diverse clinical outcome in AML patients with favorable cytogenetics. This study investigated CD56 expression in 37 adult de novo AML patients with t(8:21) between November 1996 and June 2005 at three institutions. Immunophenotyping was performed with flow cytometry (Coulter EPICS XL) and considered positive if at least 20% of blasts expressed. CD56 was expressed in 25 cases (67.6%). There was no statistically significant differences in age, sex, leukocyte count, the percentage of bone marrow blasts or the presence of additional cytogenetic abnormalities between the CD56+ and the CD56- group. The complete remission (CR) rate to standard dose cytarabine or N4-behenoyl-1-D-arabinofuranosylcytosine (BH-AC) and idarubicin was similar in both groups (91.7% v 88.7%; P=0.73), but the relapse rate to high-dose cytarabine or allogeneic hematopoietic stem cell transplantation (HST) was quite different (60% v 25%; P=0.08). Allogeneic HST was performed from siblings in 15 patients (40.5%) who achieved CR, 8 patients (32.0%) in CD56+ and 7 patients (58.3%) in CD56- group (P=0.16). The median durations of DFS were significantly shorter in CD56+ (median, 12.2 months) than in the CD56- group (median, not reached) (P =0.02). Also, the median durations of survival showed the same results in the CD56+ group (median, 14.9 months) compared with the CD56- group (median, not reached) (P=0.01). Within fifteen transplanted patients, the median durations of DFS in eight CD56+ patients was significantly shorter than seven CD56- patients (median, 24.4 months v not reached; P=0.02)(Fig.1 and 2).We concluded that CD56 expression was associated with reduced DFS and survival for AML patients with t(8:21) including transplanted patients. Although further larger studies are needed, we suggested that CD56 expression at diagnosis is a predictable prognostic factor in AML with t(8:21).
Fig. 1 Disease-free survival (DFS) and Overall survival (OS) for patients with t(8:21) with CD56+ (n−25) and CD56− (n−12) group. Fig. 1. Disease-free survival (DFS) and Overall survival (OS) for patients with t(8:21) with CD56+ (n−25) and CD56− (n−12) group. Fig. 2 Within transplanted patients, decreases from survival (DFS) for patients with t(8:21) with and without CD56 expression. Fig. 2. Within transplanted patients, decreases from survival (DFS) for patients with t(8:21) with and without CD56 expression.
Understanding the Membrane Proteome of Developing Natural Killer Cells.
Authors: Lund Troy , McCullar Valarie , Anderson Lorraine B. , Miller Jeffrey S. 
Published: April 2020

Abstract
We are interested in the biological as well as molecular processes involved in natural killer cell differentiation and function based on the growing interest in these cells for cancer therapy and allogeneic transplantation. Determining the proteomic complement is a useful tool in predicting cellular function and fate. iTRAQ is a new tandem mass-spec method which not only allows identification of proteins in the picomole to femtomole range, but also allows one to quantify differences in specific protein amount between two samples. Using this method one can potentially get reliable proteomic information from cells and also compare expression differences between cell populations. For the first time shown here, we have applied the new method of iTRAQ to identify and quantify the expression of membrane localized proteins in two previously characterized distinct but related NK cells populations. One population was derived in vitro from umbilical cord blood stem cells (CD34+Lin−CD38−) differentiated with murine stroma and human cytokines (IL-3, Flt3L, SCF, IL-7. IL-15) and the other from NK cells expanded from CD3-depleted adult peripheral blood and IL-2, both resulting in cultured populations of &gt;90% NK cells. iTRAQ was employed for multiplex peptide labeling followed by 1- and 2-dimensional high performance liquid chromatography (1D- and 2D-HPLC) and then tandem mass spectroscopy was used to analyze the proteomic contingent of membrane associated proteins. We were able to identify well as quantify differences in expression levels of 35–133 proteins in a typical 1D-HPLC experiment. This increased to &gt;400 proteins when 2D-HPLC was used. Over 75% of the proteins were identified with more than 2 peptide fragments. Gene ontogeny analysis showed the majority of proteins to be involved in cell signaling, nucleic acid binding, or cell adhesion. Nearly all proteins were associated with the plasma membrane, membrane bound organelle (lysosome or peroxisome), or nucleus. Several unique proteins were found which previously have not been known to be expressed by NK cells: BASP1, nmb, and AIF-1. More important, we were able to confirm several of our iTRAQ results by RT-PCR, Western blot, and FACS analysis. This is the first demonstration, and verification of using iTRAQ to determine the “membrane proteome” in NK cells and represents a valid and powerful new tool in the science of proteomics. These novel differences between NK cells and their precursors will be studied further to understand NK cell interactions and novel mechanisms which govern NK cell development.
CyclinA2 as a Proliferation Marker for Peripheral Blood T Cell Subsets Measured by Flow Cytometry.
Authors: Smith Cecilia U. , D’Costa Sybil , Rabellino Enrique M. , Shankey Vincent , Wilkinson Julie 
Published: April 2020

Abstract
Proliferation is regarded as a sensitive parameter for assessing immune response to infectious agents, autoimmune conditions, and organ engraftment in transplantation. Proliferation of peripheral blood mononuclear (PBMC) cells has been measured by a number of flow cytometric methods including propidium iodide (PI), incorporation of Bromo-deoxy-uridine (BrdUr), and dilution of Carboxy-fluorescein diacetate, succinimidyl ester (CFSE) intensity. Standardization of these methods however, has been hampered by their technical complexity. The restricted expression of CyclinA2 protein in the cytoplasm during the S and G2 cell cycle phases has provided the basis for a simplified intracellular flow-based proliferation assay. In these studies, ex vivo stimulated human PBMC were tested for the validation of CyclinA2 as a proliferation marker for T cell subsets. Freshly isolated or cryopreserved PBMCs were incubated in vitro with SEB/CD28 for periods up to 72 hours and processed using different methodologies for lymphocyte cell cycle measurement. These included BrdUr /7AAD, PI/CyclinA2; cylinA2 with additional T cell phenotyping markers and a combined CyclinA2 /BrdUr proliferation method. Results for each cell subset were scored for the percentage of positive proliferating cells by each methodology. PBMC co-stained for CyclinA2 and DNA content by PI demonstrated that all cells expressing Cyclin A2 corresponded to cells at S or G2 phase. Furthermore, the time course assays combining BrdUr labeling with CvclinA2 confirmed that CyclinA2 expression corresponded selectively to cells incorporating BrdUr. When PMBC were surface phenotyped for CD3 and CD4/CD8, reproducible populations of CD4 and CD8 T cell subsets incorporating BrdUr were measured for expression of Cyclin A2. In our assay model 11 to 15 % positively stained for Cyclin A2 corresponded to approximately 10% and 5% of CD3/8 and CD3/4 CyclinA2 cells, respectively. The percentage of total BrdUr and total CyclinA2 positive cells that were detected in the dual-labeled specimens corresponded to the percentage detected by each individual method. In conclusion, measurement of in vitro lymphocytes proliferation using expression of CyclinA2 protein expression is highly correlated with BrdUr uptake demonstrating that CyclinA2 is an accurate marker of cell proliferation. Preparation of cells for measuring Cyclin A2 takes only 60 minutes as compared with a total 12 to 16 hours for BrdUr pulsing and staining time. The simple CyclinA2 preparation method described here makes it a practical, fast, and reliable proliferation flow assay that can be easily standardized for enumeration of proliferating lymphocyte subsets.
Time to Failure and CD3-Expression Are Strong Prognostic Factors in Relapsed Anaplastic Large Cell Lymphoma of Childhood and Adolescence.
Authors: Woessmann Wilhelm , Lenhard Meike , Peters Christina , Burkhardt Birgit , Ebell Wolgang , Klingebiel Thomas , Mann Georg , Greil Johann , Sykora Karl-Walter , Janssen Dirk , Zimmermann Martin , Schrappe Martin , Reiter Alfred 
Published: April 2020

Abstract
We examined the feasibility and efficacy of a salvage strategy of intensive re-induction chemotherapy followed by autologous or allogeneic hematopoetic stem cell transplantation (HSCT) for children and adolescents suffering from relapsed anaplastic large cell lymphoma after BFM front-line therapy.
From 4/1990 to 2/2003, 75 evaluable patients, median age at relapse 9.8 (range 0.4–22.2) years, were enrolled. The median time from the start of first therapy to failure was 7.1 (range 1–76) months. 17 pts suffered from progressive disease during front-line chemotherapy. ALK staining was positive in 57 pts, negative in 5 pts and not available in 13 pts. CD3 reactivity was positive in 24, negative in 42 pts and not available in 7 pts. 2 pts had B-cell ALCL. Salvage therapy consisted of 2 or 3, 5-day chemotherapy courses based upon dexa, vindesine, cytarabine, etoposide, MTX 5g/m2, cyclophosphamide/ifosfamide, vincristine, doxorubicine, intrathecal therapy followed by HSCT. Conditioning regimen: total body irradiation (2x2 Gy day −7,−6,−5), etoposide 40 mg/kg day -4, cyclophosphamide 60 mg/kg/d days −3,−2. Data were updated as of April 1st, 2004. With a median follow-up of 6.9 [0.8–13.2] years, the probability of survival at 3 years after first relapse is 54+6%. 28 pts died of lymphoma, 6 pts of toxicity. 41 pts are alive in ≥CR2. 21 pts received no salvage or chemotherapy only (2 TRM, 18 death of disease, 1 CR). 39 pts received autologous HSCT (21 CR, 2 TRM, 16 re-relapses [6 &gt;CR2]). 15 pts received allogeneic HSCT (10 CR, 3 TRM, 2 relapses). 3/17 pts with relapse during front-line survived compared to 37/57 pts who relapsed later (p&lt;.001). pSurvival at 3-years was 62+8% for CD3-neg. pts and 45+10% for CD3-pos pts (ns). However, pEFS after autologous HSCT was dependent on the CD3 status: 69+10% for 23 CD3-neg pts (16 CR, 1 TRM, 6 relapse), 9+9% for 11 CD3-pos pts (1 CR, 1 TRM, 9 relapse), p=.002. In contrast, all 7 CD3-pos patients who received allogeneic HSCT (4 of them after relapse subsequent to autoHSCT) remained free of disease thereafter. 5/5 ALK-negative patients relapsed after front-line therapy, were CD3-positive and progressed again, two of them after autologous HSCT.
We conclude that this strategy proved feasible and efficacious for pts who suffered a relapse after front-line therapy while outcome for pts with progressive disease during front-line treatment was poor. Autologous HSCT is an efficient consolidation for pts with relapses of CD3 negative ALCL but not efficacious for those with CD3 positive tumors. These patients may benefit from allogeneic HSCT.
Delivery and Toxicity of Fludarabine-Based Combination Chemotherapy Regimens in Older Patients.
Authors: Polizzotto Mark N. , Tam Constantine S. , Januszewicz Henry , Prince Miles , Wolf Max , Seymour John F. 
Published: April 2020

Abstract
Fludarabine (F)-based combination chemotherapy regimens are highly effective in a range of indolent lymphoproliferative disorders. Despite the prevalence of such disorders in older patients, the deliverability of these regimes in patients aged &gt;= 60 has not been assessed. We analysed the delivery and toxicity of three F-based regimens, all using F 25 mg/m2/dx3 q28d, in 82 adults aged &gt;= 60 years, and compared this with the same regimens in 99 adults aged &lt; 60. The sample comprised 66 patients (32 &gt;= 60) treated with F and cyclophosphamide (C; 250 mg/m2/dx3); 29 with F and mitoxantrone (M; 10 mg/m2 x1; 12 &gt;= 60); and 86 with FC and rituximab (R; 375 mg/m2 x1; 38 &gt;= 60). 349 cycles in older patients were compared with 393 cycles in younger patients for haematologic nadirs, infectious complications and organ toxicity. Both groups received a median of 4 cycles, although older patients were more likely to require dose reduction (4.3% of cycles versus 1.2%, P &lt; 0.001) and growth factor support (3.8% versus 1.8%, P=0.01). The cohorts were well matched for baseline characteristics other than age.
Overall, older patients had a slightly higher rate of infections (18%/cycle versus 15%/cycle), though this was not statistically significant (P = 0.28). For severe (grade &gt;=3) infections the difference was minimal: 6% versus 7% (P&lt; 0.5). The rates of neutropenia &lt; 1.0 and 0.5 were 13% and 22% versus 11% and 20% for older and younger patients, respectively (all P values&gt;0.1). The rates of thrombocytopenia &lt; 100 and &lt; 50 were 21% and 5% versus 16% and 5% for older and younger cohorts (all P values &lt; 0.1). Other organ toxicities were uncommon, and showed no difference between age groups. Treatment-related mortality in both cohorts was &lt;1% (P &gt; 0.5).
Comparison within the cohort aged over 60 showed that those aged 70 and over were at higher risk of haematological and infectious toxicity. 82 cycles delivered to 23 patients aged &gt;= 70 were compared with 267 cycles delivered to 61 patients aged 60 to 69. The rate of infection for those over 70 was 25% versus 16% in those aged 60 to 69, though this was not statistically significant (P=0.07). For severe infections (grade &gt;=3), the rates were 13% versus 6% (P=0.03), while rates of neutropenia &lt; 0.5 and thrombocytopenia &lt; 50 were 32% and 15% versus 8% and 3% for those &gt;= 70 and 60 to 69 respectively (all P values &gt; 0.001).
These results demonstrate that F-based regimens are well tolerated and can safely be delivered to most older patients, with a modestly increased rate of infectious morbidity, but no increased treatment-related mortality. However, for patients aged &gt;=70 the increased rate of toxicity mandates careful patient selection and monitoring.
Systematic Review of 18F-Fluorodeoxyglucose Positron Emission Tomography for Post-Treatment Evaluation of Malignant Lymphoma.
Authors: Zijlstra Josée M. , Hoekstra Otto S. , Lindauer-van der Werf Gerda , Hooft Lotty , Riphagen Ingrid I. , Huijgens Peter C. 
Published: April 2020

Abstract
Despite the increasing number of publications concerning 18F-Fluorodeoxyglucose Positron Emission Tomography (FDG-PET) for post-treatment evaluation of lymphoma and the increasing availability of this novel diagnostic modality, its exact role in response assessment after therapy is still unknown. The aim of this study was to systematically review the literature regarding the diagnostic performance of dedicated FDG-PET in evaluation of first line therapy of Hodgkin’s disease and (aggressive) non-Hodgkin’s lymphoma, and to calculate summary estimates of its sensitivity and specificity. The databases of PubMed and Embase were searched for relevant studies, up to January 2004. Criteria for inclusion of studies were 1) histologically proven Hodgkin’s disease (HD) or aggressive non-Hodgkin’s lymphoma (NHL), 2) evaluation of post-treatment patients following first line therapy, 3) the use of dedicated (ring) PET using 18FDG, and 4) study population of at least ten patients. Two reviewers independently assessed the methodological quality of each study. As a valid reference test, histology or follow-up of at least 12 months were accepted. A meta-analysis of the reported sensitivity, specificity, and positive Likelihood Ratio (LR+) and negative Likelihood Ratio (LR−) of each study were performed.
Fifteen studies, involving 705 patients, met the inclusion criteria. The studies had several design deficiencies. The majority of studies did not describe whether the reference test was interpreted without knowledge of the FDG-PET findings. Furthermore, in only two studies, patients entered the study consecutively. In all studies, there was a description of the spectrum of patients included, i.e. all patients for post-treatment evaluation or only patients with substantial residual masses post-treatment. Pooled sensitivity, specificity, LR+ and LR- for detection of residual disease in Hodgkin’s lymphoma were 84% (95% CI 71–92%), 90% (95% CI 84–94%), 5.5 (95% CI 3.4–8.7) and 0.25 (95% CI 0.1–0.55), respectively. For non-Hodgkin’s lymphoma, pooled sensitivity, specificity, LR+ and LR− were 72% (95% CI 61–82%), 100% (95% CI 97–100%), 36 (95% CI 11–125) and 0.30 (95% CI 0.21–0.42), respectively.
In conclusion, FDG-PET showed reasonable sensitivity and high specificity for evaluation of first line therapy in Hodgkin’s and in non-Hodgkin’s lymphoma.