bluuebunny/crossref_metadata_2025_split huggingface dataset:
Histidine Decarboxylase (HDC) as Novel Marker of Immature Neoplastic Mast Cells in Systemic Mastocytosis.
Authors: Krauth Maria T. , Agis Hermine , Aichberger Karl J. , Simonitsch Ingrid , Muellauer Leonhard , Mayerhofer Matthias , Horny Hans P. , Valent Peter 
Published: April 2020

Abstract
Synthesis of histamine in hematopoietic progenitor cells may be one of the earliest events in mast cell-development from pluripotent hematopoietic progenitor cells. In the present study, we asked whether the key enzyme involved in histamine generation, histidine decarboxylase (HDC), can be employed as an immunohistochemical marker for the detection of (neoplastic) mast cells (MC) in patients with cutaneous (CM) or systemic mastocytosis (SM). To address this question, we examined bone marrow biopsy specimens in a cohort of 101 patients (CM, n=10; indolent SM, n=46; SM with associated clonal non-MC lineage disease, n=31; aggressive SM, n=10; MC leukemia, n=3; MC sarcoma, n=1) using an antibody against HDC. Independent of the maturation stage of MC or subtype of disease, the anti-HDC antibody produced clear diagnostic staining results in all patients with bone marrow involvement examined including those with MC leukemia and MC sarcoma, in which MC are particularly immature and often escape analysis when examined by conventional stains. In these patients (MC leukemia, n=2; MC sarcoma, n=1), expression of HDC was reconfirmed at the mRNA level by RT-PCR analysis performed with RNA of highly enriched sorted CD117+ MC. In patients with CM or normal/reactive bone marrow (n=30), no HDC-positive infiltrates were detected. In these patients, only a few hematopoietic cells, presumably basophils, were found to react with the anti-HDC antibody. In summary, HDC is expressed in neoplastic bone marrow MC in patients with SM independent of the maturation stage of cells or the variant of disease. HDC should therefore be considered as a new MC marker in the screen panel of antigens employed to diagnose high grade MC malignancies.
WT-1 Over-Expression Could Be Related to " High Metaplastic Index" in Myelofibrosis with Myeloid Metaplasia (MMM) Patients. A Preliminary Report.
Authors: Iuliano Francesco , Cilloni Daniela , Gottardi Enrico , Fava Milena , Fabiano Flora , Peta Antonio , Saglio Giuseppe 
Published: April 2020

Abstract
WT1 is a tumor-suppressor gene coding for a zinc-finger transcription factor located on chromosome 11p13, which was originally identified for its involvement in the pathogenesis of the Wilms’ tumor. In normal bone marrow and peripheral blood(PB)WT1 expression is extremely low and often undetectable even by RT-PCR. By contrast WT1 expression has been previously reported to be increased in MDS, AML, ALL and PNH. In CMPDs, little data are available only in CML. With this background we analyze by RQ-PCR the WT1 transcript amount in 13 patients with myelofibrosis with myeloid metaplasia (MMM) in comparison with a group of 32 age- and sex-matched healthy controls. All measurements were carried out only on PB and performed as previous published. All MMM patients were under cytoreductive treatment and at different time from diagnosis ( range 1 – 7 yrs). In MMM WT1 transcript amount is consistently higher (range 23–2518 WT1 copies/104 ABL copies, mean 382±692) than controls ( range 0 – 22, mean 5,6 ± 5,87) ( P &lt; 0.0001 Mann-Whitney U test). To assess the significance of the WT1 expression as a marker for disease features, the following variables were analyzed and compared : age, hemoglobin level, white blood cell count, circulating blasts, LDH serum levels, number of peripheral CD 34 cells, Dupriez score and, according to Barosi published data, myeloproliferation index, myelodepletion index, severity score. We did not found any significant correlation between WT1 and all these variables, probably because the patients were studied during the course of their disease. Moreover, cytoreductive treatment might have modified the disease characteristics. Stricking, when analyzed WT1 expression according to the therapeutic need, we found a subset of patients with a WT1 overexpression ( range 132–2518, mean 758±911; p &lt;0.06) and spleen size between 4 – 7 cm despite receiving &gt; 1,5 gr/daily of hydroxyurea or more than 1 drug. In such of patients WT1 over-expression allowed an almost complete discrimination between MMM in stable phase and MMM with “high metaplastic index”. In conclusion, our results, although based on a relatively small number of patients, provide an interesting profile of WT1 expression in MMM. The possible clinico-therapeutic implications of these observations are worthy of investigation in well-designed studies.
A Phase II Trial of Imatinib Mesylate (Gleevec®) in Myeloproliferative Disorders Other Than CML.
Authors: Kuriakose Philip , Shurafa Muhammad S. 
Published: April 2020

Abstract
Myeloproliferative disorders are a heterogeneous group of disorders characterized by increased cell numbers and variable degrees of organomegaly and myelofibrosis. To date, the standard of care has been phlebotomy followed by Hydroxyurea or anagrelide to control thrombocytosis. Anagrelide has gained popularity in recent years due to its theoretical non leukemogenic potential. Gleevec® is a small molecule inhibitor of several tyrosine kinases including c-kit and platelet derived growth factor receptor (PDGFR). The latter is expressed on platelets and megakaryocytes and may offer an explanation for the megakaryocytic hyperplasia and clustering via an autocrine loop action. In vitro, Gleevec® inhibits spontaneous burst formation in polycythemia vera (PV) probably by inhibiting c-kit. By inhibiting PDGFR, it could inhibit megakaryocytic proliferation by blocking an autocrine effect which may be responsible for megakaryocytic hyperplasia and clustering in essential thrombocytosis (ET) and PV. In 2003 we initiated a study to explore the effectiveness of Gleevec® in these two entities. 11 patients have been enrolled, 5 with PV, and 6 with ET. The starting dose in each case was 400mg, with escalation to 800 mg based on response. Complete response (CR) was defined as therapeutic range hematocrit without phlebotomy, a normal platelet count and discontinuation of Hydrea or Anagrelide. There were two CR’s and one PR in PV. All three patients discontinued treatment because of toxicity at 2, 6, and 10 months, respectively. There was one CR and one PR in ET, but both discontinued treatment due to toxicity at 7 months and non-compliance at 2 months, respectively. The toxicities observed that led to discontinuation of therapy were grade 2–3 diarrhea (3/11), grade 2–3 periorbital and ankle edema (5/11), grade 2 skin rash (1/11), grade 1 blurred vision (1/11). One major post operative bleeding was observed and was probably related to platelet dysfunction. Two patients (one each of PV & ET) had to discontinue therapy for lack of response at maximum tolerated dose. The ending dose was 400 mg (3), 600 mg (5), and 800 mg (3), with two at the 800 mg dose level being non-responders, and all others having discontinued therapy on account of toxicity. This study suggests that Gleevec® has activity in PV and ET, but toxicity at the studied dose escalation schedule could lead to discontinuation of therapy. However, this does not preclude its use in a new clinical trial with a more tolerable dosing schedule over a longer period of time.
Regulation of Berg-36 Gene Expression by Survival and Apoptotic Stimuli in B-Chronic Lymphocytic Leukaemia Cells.
Authors: Jewell Andrew P. , Baou Maria , Yong Kwee L. , Carr Robert , Murphy John 
Published: April 2020

Abstract
Chronic lymphocytic leukemia is characterized by the accumulation of monoclonal malignant CD5+ B cells that show resistance to apoptosis in vivo. Berg 36 (ZFP36L1, TIS11b, BRF1, cMG1, ERF1) is a zinc finger containing early response gene that was cloned from PMA-stimulated B chronic lymphocytic leukemia cells. Induced expression of this gene has been linked to calcium ionophore and anti-CD20-induced apoptosis of human B lymphoma cells. Berg36 protein has, in common with two other members of the small gene family (TIS11, TIS11d), been reported to function as an mRNA-binding protein that may promote instability of cytokine mRNAs. We have therefore studied the regulation of Berg36 expression in B-CLL cells in vitro, in response to cytokines and other signals that regulate apoptosis. B-CLL cells from 12 patients were purified and incubated with the following agents either alone or in combinations for a number of hours; IL-4, CD40 ligand, PMA or anti-CD20 antibody (rituximab). Apoptosis was measured after 24 hours by Annexin/PI staining and Berg-36 expression by Northern blot analysis. Spontaneous apoptosis in unstimulated B-CLL cells was 20.9±5.1%. Co-incubation of B-CLL cells with IL-4 reduced the percentage of apoptotic cells to 6.2±1.2%, but had no effect on Berg-36 expression. In contrast, both PMA and CD40 stimulation reduced the percentage of apoptotic cells (to 14.8±3.4% and 10.2±4.2% respectively), and markedly induced Berg36 expression. On the other hand, anti-CD20 antibody induced apoptosis (36.2±6.6%), but also induced Berg36 expression. Specific inhibitors of various intracellular signalling molecules confirmed that induction of Berg36 by different agents was mediated through different signalling pathways. In conclusion, expression of Berg36 can be induced in B-CLL cells by stimuli that induce either survival or apoptosis in these cells.
Blood Flow Cytometry Allows Us To Dispense with Bone Marrow Biopsy in the Evaluation of Treatment Response in CLL.
Authors: Maloum Karim , Charlotte Frederic , Divine Marine , Cazin Bruno , Lepretre Stephane , Delmer Alain , Sonneville Sylvie , Merle-Beral Helene 
Published: April 2020

Abstract
Aims: To evaluate the diagnostic value of bone marrow biopsy (BMB) with immunohistochemistry studies compared to blood flow cytometry (FC) in response evaluation after treatment in CLL. Patients and methods: 75 previously untreated patients with stage B and C-CLL were treated with oral Fludarabine (30mg/m2/d) + Cyclophosphamide (200mg/m2/d) combination, day 1 through 5, repeated every 28 days for 6 courses. Response evaluation was performed 2 months after the completion of therapy, according to NCI criteria. Sixty patients out of 75 achieved partial or complete remission and underwent BMB. Minimal residual disease (MRD) was simultaneously evaluated on paraffin-embedded BMB by anti-CD5, -CD20 and -CD3 immunostaining and in the peripheral blood using a very high sensitive FC method (Maloum et al, Br J Haematol, 2002, 119:970–5). BMB immunostaining was considered positive for MRD in case of concurrent expression of CD20 and CD5 by the same cells assessed on consecutive sections. Twenty nine patients could have been analyzed so far. The median follow-up was of 49 months [range 19–56].
Results: In all these 29 patients, BMB was considered histologically normal according to the NCI criteria. In 23/29 cases, bone marrow immunostaining did not show expressing CD20 cells. Fifteen out of 23 were in phenotypic remission using blood FC (PhR), of whom only 2 relapsed (at 31 and 37 months). The remaining 8/23 patients did not achieve PhR and all relapsed (at 22, 27, 37, 40, 45 and 51 months). In 6/29 patients, immunochemistry on BMB showed persistent B CD5+ cells : 3 did not achieve PhR of whom 2 relapsed (at 32 and 37 months) while 3 were in complete PhR and displayed a persistent clinical complete remission with a follow-up of 39, 47 and 47 months respectively. Interestingly, in the latter 3 cases, FC analysis showed an expansion of polyclonal BCD5+ cells in blood as assessed, which could explain the presence of such normal BCD5+ cells in the bone marrow. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and diagnostic accuracy (ACC) to predict relapse for BMB and FC are given in the following table:
Sensitivity (%) Specificity (%) PPV (%) NPV (%) ACC (%) BMB 16.7 76.5 33.3 56.3 51.7 FC 83.3 94.1 90.9 88.9 89.7
Conclusion: These results strongly suggest that BMB should not be indicated in the evaluation of treatment response and is not useful for relapse prediction. In contrast, blood flow cytometry constitutes the best method in routine laboratory to predict clinical outcome and should be included in response treatment criteria in CLL.
Is Zap-70 Protein Detection by Flow Cytometry a Reliable Tool in CLL Prognostic Evaluation?.
Authors: Le Garff-Tavernier Magali , Ticchioni Michel , Letestu Rémi , Brissard Martine , Davi Frédéric , Sémichon Monique , Vuillier Françoise , Cymbalista Florence , Merle-Béral Hélène 
Published: April 2020

Abstract
Background : Expression of ZAP-70 protein has been shown to be correlated with mutational status of immunoglobulin heavy-chain variable region (IgVH) genes, a major prognostic factor in CLL. We investigated whether the detection of ZAP-70 protein by flow cytometric analysis using unconjugated and conjugated monoclonal antibodies (mAbs) could be applied securely in the workup of patients with CLL. Methods: Flow cytometric analysis of ZAP-70 protein was performed using the method described by Crespo et al (N Engl J Med2003;348:1764) with minor modifications. Both fresh and cryopreserved mononuclear cells from CLL patients and healthy donors were fixed and permeabilized using Fix and Perm kit (Caltag Laboratories), incubated with anti-ZAP-70 mAb (clone 2F3.2, Upstate Biotechnology) and then revealed with goat antimouse FITC mAb (Immunotech). Finally cells were incubated with CD3-APC, CD56-APC and CD19-PC5. We also tested 3 mAbs conjugated to various fluorochromes: 2F3.2-FITC (Upstate), 1E7.2-PE (eBioscience), 1E7.2-PE or -Alexa 488 (Caltag). ZAP-70 protein detection in B-cells was expressed either as a percentage of its expression in the T and NK-cells or as a ratio (R) of T-cell mean cell fluorescence (MCF) to B-cell MCF. Western blotting of protein lysates from purified B-cells was carried out to control results obtained by cytometry in 55 samples. Mutational status was defined using a cutoff of 98%. Results: In 13 healthy donors, the mean percentage of ZAP-70 protein expression obtained by flow cytometry with unconjugated mAb (clone 2F3.2) was 4.69% ± 1.93 [range 2–9%] and the R ratio was 6.64 ± 1.54 and &gt; 4.8. In 83 B-CLL samples, ZAP-70 expression was determined using the same method and compared to IgVH mutational status. Results in table below show a 75% concordance between gene mutations and ZAP-70 expression when considering a percentage of positive B-cells &gt; 20%. A better concordance (81%) is obtained with a threshold T-cell MCF/ B-cell MCF at 4 determined by Youden’s index. To note the high concordance (90%) between unmutated status and ZAP-70 + expression (19/21). Comparison with at least 1 of the 3 conjugated mAbs has been performed for 63 samples, with discordant results in our laboratories.
62 mutated IgVH samples 21 unmutated IgVH samples ZAP-70 B-Cells + ≤ 20 % : 43 &gt; 20% : 19 T-cell MCF/B-cell MCF ≥ 4 : 48 &lt; 4 : 19
Conclusions: Our data document the concordance between IgVH gene mutational status and ZAP-70 protein expression measured by flow cytometry, particularly in ZAP-70 negative samples. We found that the indirect method of labelling with unconjugated anti-ZAP-70 mAb remains until now, in our hands, the gold standard method compared to the available dyes conjugate mAbs.
Flow Cytometric Characterization of Genetically Defined Subgroups of Chronic Lymphocytic Leukemia: Basis for the Comprehensive Definition of Prognostic Parameters.
Authors: Kern Wolfgang , Voskova Daniela , Schoch Claudia , Hiddemann Wolfgang , Schnittger Susanne , Haferlach Torsten 
Published: April 2020

Abstract
Chronic lymphocytic leukemia (CLL) is defined based on cytomorphology and immunophenotype. The most important prognostic parameters are cytogenetic aberrations and the expression of CD38 and ZAP70. We analyzed correlations between immunophenotypic patterns and genetic aberrations in 153 peripheral blood samples from patients with newly diagnosed and untreated CLL. Immunophenotyping was performed applying five-fold staining, a comprehensive panel of antibodies, and CD45-SSC-gating. Genetic aberrations were detected by fluorescence in-situ hybridization using probes targeting ATM for the detection of 11q-, D12Z3 for trisomy 12, D13S319 and D13S25 for 13q-, and p53 for 17p-. As compared to cases without trisomy 12, in cases with trisomy 12 the expression levels were higher for CD22 (66% vs. 34%, p=0.001), CD20 (83% vs. 71%, p=0.034), and FMC7 (28% vs. 7%, p=0.000001). This is in accordance with our finding that higher numbers of prolymphocytes have been found in CLL with trisomy 12. As compared to cases with any cytogenetic aberration, cases with normal cytogenetics had lower expression levels of CD5 (82% vs. 89%, p=0.012) and CD19 (79% vs. 85%, p=0.051). Importantly, there were no significant differences in expression levels of CD38 and ZAP70 between cytogenetically defined subgroups of CLL. Thus, in cases with 11q-, trisomy 12, 13q-, homocygous 13q-, 17p-, and normal cytogenetics CD38 was expressed in 51%, 47%, 37%, 33%, 24%, and 38% and ZAP70 together with cyCD79a was expressed in 17%, 11%, 13%, 13%, 22%, and 15%. These data indicates that, with the exception of trisomy 12, the cytogenetic findings are not reflected by specific immunophenotypic features. In particular, this is true for the prognostically and clinically relevant markers CD38 and ZAP70 the expression of which cannot be deduced from cytogenetics. In a second step, correlations between the expression of CD38 and ZAP70 and other immunophenotypic markers have been analyzed. Cases positive for CD38 (more than 30% positive cells) had lower expression levels of CD19 (78% vs. 87%, p=0.005), CD20 (65% vs. 75%, p=0.023), and CD23 (62% vs. 74%, p=0.005). Cases positive for ZAP70 (more than 20% positive cells with coexpression of cyCD79a) had higher expression levels of CD19 (87% vs. 79%, p=0.011), CD22 (40% vs. 26%, p=0.020), and cyCD79a (75% vs. 59%, p=0.009). These findings are in line with both CD38 and ZAP70 being markers that reflect different biologic backgrounds of CLL which also reveal prognostic information. Taken together, these data strongly suggest that studies evaluating different treatment approaches in CLL should provide a detailed characterization of the analyzed patients including both cytogenetic aberrations and immunophenotypic findings as well as novel genetic markers in order to guarantee the detection of subgroup-specific treatment effects.
The Natural Product Honokiol Induces Caspase-Dependent Apoptosis in Chronic Lymphocytic Leukmemia (CLL) Cells.
Authors: Battle Traci E. , Arbiser Jack , Frank David A. 
Published: April 2020

Abstract
CLL remains an incurable disease that requires innovative new approaches to improve therapeutic outcome. Honokiol is a natural product known to possess potent anti-neoplastic and anti-angiogenic properties. We examined whether Honokiol can overcome apoptotic resistance in CLL cells. Honokiol induces caspase-dependent cell death of CLL cells, characterized by an increase in Annexin V positive, propidium iodide negative cells that was blocked by the caspase inhibitor, Z-VAD-fmk. Further evidence for Honokiol-induced apoptosis of CLL cells was confirmed by an increase in caspase 3 activity and cleavage of PARP. In addition, Honokiol demonstrated an LC50 that was lower for CLL cells than for normal mononuclear cells, suggesting that CLL cells are more susceptible to the cytotoxic effects of Honokiol compared to normal hematopoietic cells. To determine whether Honokiol-induced apoptosis of CLL cells was associated with known positive therapeutic effects, we examined mcl-1 expression, a survival protein whose down-regulation is associated with response to treatment in CLL patients. Honokiol treatment of CLL cells resulted in a significant decrease in the expression of mcl-1 within 24 hours. Furthermore, CLL cells pre-treated with IL-4, a cytokine known to support CLL survival, underwent apoptosis when subsequently incubated with Honokiol, indicating that Honokiol could also overcome the pro-survival effects of IL-4. These data indicate that Honokiol is a potent inducer of apoptosis in CLL cells and should be examined for further clinical application.
Telomere Length as a Prognostic Parameter in Chronic Lymphocytic Leukemia with Reference to IGV(H) Gene Mutation Status.
Authors: Roos Goran , Grabowski Pawel , Hultdin Magnus , Karlsson Karin , Thunberg Ulf , Tobin Gerard , Laurell Anna , Rosenquist Richard 
Published: April 2020

Abstract
Recent studies on the immunoglobulin variable heavy chain (IgV(H)) genes have revealed that B-cell chronic lymphocytic leukemia (B-CLL) consists of at least 2 entities with either somatically mutated or unmutated V(H) genes. Cases with unmutated V(H) genes, considered to evolve from pregerminal centre (GC) cells, have a worse outcome compared to cases showing mutated V(H) genes, that is, GC experienced or post-GC derived. Also, telomere length has been reported to be of prognostic significance in CLL. Telomerase becomes activated during the GC reaction with an elongation of the GC-B cell telomeres and a strong association exists between V(H) gene mutation status and telomere length in CLL (ie mutated cases have longer telomeres).
These results encouraged us to study telomere length and VH gene mutations in relation to clinical parameters in a large series of B-CLL cases (n=261). Telomere length was assessed by a PCR based method (Cawthon, 2002) showing very good correlation to telomere length data obtained by Southern blotting. Ig gene rearrangements were determined by DNA sequencing. The prognostic information given by the Ig mutation status (mutated vs unmutated) as well as by telomere length ("long" vs "short", cut off at median value) was statistically highly significant (p&lt;0.0001 for both). Interestingly, the prognostic information given by telomere length seemed to be restricted to the Ig mutated CLL cases.
Our data indicate that telomere length must be considered as a relevant prognostic factor of importance for B-CLL. Telomere length determination using the here applied PCR technique is fast and accurate and can easily be established in routine diagnostics.