bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical and Molecular Findings in Childhood Polycythemia Vera.
Authors: Cario Holger , Pahl Heike L. , Milen Minkov , Harald Reinhard , Kohne Elisabeth , Debatin Klaus-Michael 
Published: April 2020

Abstract
Background: Polycythemia vera (PV) and congenital erythrocytoses constitute extremely rare diseases in pediatric and juvenile patients. Systematic data on clinical and laboratory evaluations as well as on treatment modalities are sparse.
Aims/Methods: We developed a protocol (PV-ERY-KA 03) for the systematic collection of clinical, hematological, biological as well as treatment data of children and adolescents with PV or congenital erythrocytoses. The collaborative trial was initiated in March 2004.
Results: To date, 5 (2 male / 3 female) pediatric patients with PV have been enrolled. The median age at onset was 14 years (7–18.5 years). 4/5 patients are of German, 1/5 of Turkish origin. Only 3 patients presented with PV-related symptoms (Budd-Chiari syndrome, isolated headache, or headache together with tinnitus, dizziness and weakness). At first presentation, the median hemoglobin content was 18 g/dl (15.5–21.1 g/dl), the median hematocrit 0.60 (0.59–0.73). Thrombocytosis was present in 4/5, leukocytosis in 2/5 patients. All patients presented with mild to marked splenomegaly. Granulocytic PRV-1-mRNA expression and JAK2V617F mutation were assessed in 4/5 patients. PRV-1-mRNA expression was significantly elevated in 3 of them and borderline the 4th patient. 3 patients including the patient with borderline PRV-1-mRNA expression presented with a heterozygous JAK2V617F mutation. Treatment strategies included phlebotomy (4/5), hydroxyurea (1/5), IFN-alpha (1/5), and aspirin (3/5). Stem cell transplantation from a matched unrelated donor was performed in one patient (now in complete remission). Because of progressive liver failure, the patient with Budd-Chiari syndrome underwent orthotopic liver transplantation.
Conclusions: The activating JAK2V617F mutation previously reported to be present in up to 90 percent of adult PV patients, is also involved in childhood PV. The age of onset and clinical presentations in the rare cases of childhood PV are varied. Different treatment strategies are applied. A collaborative assessment of children with PV may help to optimise treatment in these patients, but may in addition contribute to a better general understanding of the disease’s pathogenesis.
Bone Marrow Evaluation According to the PVSG and WHO Criteria in 90 Essential Thrombocythemia (ET) Patients Treated with PEG Interferon alpha-2b. Preliminary Results.
Authors: Gugliotta Luigi , Bulgarelli S. , Asioli S. , Tieghi A. , Gardini G. , Sabattini E. , Franco V. , Boveri E. , Kvasnicka H. M. , Vianelli N. , Candoni A. , Latagliata R. , Passamonti F. , Rupoli S. , Martinelli V. , Zaccaria A. , Lauria F. , De Biasi E. , Sacchi S. , Gobbi M. , Tabilio A. , Cacciola E. , Cacciola R. , Grossi A. , Mazza P. , Bonifazi F. , Favini P. , Mandelli F. , Baccarani M. , Thiele J. 
Published: April 2020

Abstract
Ninety ET patients diagnosed according to the PVSG criteria were enrolled in a phase II study (sponsored by the Schering-Plough Company) designed to evaluate the efficacy, safety and tolerability of a two years treatment with PEG Interferon alpha-2 b (PEG Intron). The patients, observed in 16 Italian Centres belonging to the GIMEMA Cooperative Group and judged at high risk, had been previously treated with cytoreductive (97%) and antiplatelet (91%) drugs. At the study start the patients, 60 F and 30 M, mean age 45 years, showed splenomegaly in 22% of cases. The Hematological Response (HR: PLT&lt;500 x109/L) was observed in 64/81 (79%) and 48/55 (87%) of the patients on PEG Intron treatment at the end of the first and second year, respectively; the spleen enlargement disappeared in 75% of cases. The Bone Marrow data reported in the study CRFs document that the baseline increase of cellularity, of granulopoyesis, of megakaryocytes (MK) number, size and ploidy significantly decreased during PEG Intron treatment, while the MK displasia and fibrosis rate globally increased. In these patients a revision of the Bone Marrow biopsy slides was blindly performed by an Expert Pathologist Panel by applying the WHO criteria. The evaluation of two-thirds of cases showed the following distribution at the baseline: true ET (23%), IMF-0 (pre-MF 17%), IMF-1 (early MF 40%), IMF-2/3 (classical MF 3%), MPD-U (17%). Interestingly, the true ET patients never showed spleen enlargement that, otherwise, was present in IMF-0 (20%) and IMF-1/2/3 (42%) patients. The patients with true ET did not show any bone marrow evolution during the PEG Intron treatment, while the IMF-1/2/3 picture significantly increased. Moreover, the rate of the HR at the end of the first year was higher in true ET(93%) respect to the IMF-0 (60%) and IMF-1/2(71%) patients. These preliminary data show that the patients classified as true ET (WHO Criteria) are not splenomegalic, have no myelofibrotic evolution and reach a better response to the IFN treatment.
Megakaryocyte-Active Chemokines: Dysregulation in the SDF-1a/CXCR4 Axis in Patients with Essential Thrombocythemia.
Authors: Salim Juan P. , Marta Rosana F. , Molinas Felisa C. 
Published: April 2020

Abstract
Chemokines belong to a large family of molecules that are implicated in the localization and production of blood cells. Some of them, such as Interleukin 8 (IL-8) and GRO-a, participate in the regulation of megakaryopoiesis, mostly by exerting an inhibitory action. Recently, the involvement of stromal derived factor 1 (SDF-1) in synergizing the stimulatory effect of thrombopoietin on megakaryopoiesis and its participation in megakaryocyte transendothelial migration has been described. The aim of the present study was the evaluation of the plasma levels of IL-8, GRO- a and SDF-1 in patients with essential thrombocythemia (ET), a myeloproliferative disorder characterized by megakaryocytic hyperproliferation with increased circulating platelet count. Besides, the corresponding chemokine receptors were assayed on platelet membrane from ET patients. A cohort of 27 patients diagnosed according to the Polycitemia Vera Study Group were enrolled in the study (mean age, 45; 21 women). Twenty-seven normal subjects matched by sex and age were taken as the control group. The Ethic Committee from IDIM A. Lanari approved the study and all patients and normal controls signed the informed consent. Plasma levels of the chemokines were measured by ELISA technique (R&D Systems) according to the manufacturer. Expression levels of IL-8 receptors (CXCR1 and CXCR2), GRO-a receptors (CXCR2) and SDF-1 receptors (CXCR4) on platelet membrane were evaluated by flow cytometry using specific MoAbs and the corresponding isotype controls (B-D Pharmingen). Flow cytometry results were expressed as relative fluorescence intensity (RFI, the relationship between the mean fluorescence intensity from the specific antibody and the isotype control). Results were expressed as median and range. Statistic analysis was carried out using Mann-Whitney Wilcoxon rank sum test and Wilcoxon signed rank test. Plasma levels of the chemokines measured in 19 ET patients were similar to that found in normal controls, IL-8 2.5, pg/ml (0.8–28.2) and 2.8 pg/ml (1.1–16.5), GRO-a, 30.0 pg/ml (7.4–463.1) and 23.9 pg/ml (9.6–148.0), SDF-1, 1895.0 pg/ml (1246.0–2719.0) and 1915.0 pg/ml (822–2424.0), respectively. The expression levels of CXCR4 receptor was found diminished in platelets from ET patients, RIF 16.94 (1.3–31.3) compared to normal controls, 27.4 (2.4–58.4); p=0.0059, n=10. However, the expression of CXCR1 and CXCR2 in platelets from ET patients was normal. In conclusion, although plasma levels of the chemoquines IL-8, GRO-a and SDF-1 were normal in these patients, the decreased level of CXCR4 on platelet membrane suggests a dysregulation in the SDF-1a/CXCR4 axis in patients with essential thrombocythemia.
A Cautionary Tale.
Authors: Pearlman Eugene S. , Mitchell Farzine , Carloss Harry 
Published: April 2020

Abstract
RV, a 62 year old caucasian male was admitted to LH from the ER for prolonged epistaxis. Ten months prior to admission he had appeared at the ER with the same complaint. At that time the patient’s Hb, Hmct, MCH and MCHC were 12.3 g/dL, 29.2 percent, 35.8 pg and 42 percent respectively. All hematological and chemical assays used instrumentation from Beckman-Coulter Inc.( Brea, CA). The patient’s epistaxis was attributed to anti-platelet therapy with aspirin and clopidogrel for coronary artery disease and he was subsequently lost to follow-up. At his most recent ER appearance his Hb, Hmct, MCH and MCHC were 11.8 g/dL, 22.9 percent, 42.5 pg and 51.6 percent respectively. The patient was admitted to allow evaluation by ENT because of difficulty controlling his epistaxis. Shortly after admission, a CBC revealed nearly identical hematological parameters to those seen in the ER and the pathologist was telephoned. Attempts at warming the patient’s specimen and even acquiring another specimen brought to the lab in a water bath resulted in only trivial changes in the patient’s CBC. If however, patient plasma was replaced by an equal volume of isotonic diluent supplied by the manufacturer, the Hb, Hmct, MCH and MCHC dropped to 6.5 g/dL, 19.4 percent, 23.1 pg and 33.5 percent respectively. Examination of a smear revealed numerous rouleaux. Serum protein and albumin assays revealed concentrations of 9.7 and 2.7 g/dL respectively suggesting an increase in the globulin fraction. Subsequent SPE, IFE and quantitative nephelometry assays performed at our reference lab (ARUP; Salt Lake City, UT) indicated the presence of an IgM-kappa monoclonal protein [6.99 g/dL]. A presumptive diagnosis of Waldenstrom’s macroglobulinemia was confirmed by bone marow biopsy. It has long been known that an MCHC greater than 36 percent should prompt a careful investigation of the data [Am J Clin Pathlol.83:218 (1985)]. In this case the elevated concentration of IgM resulted in increased turbidity and a spuriously elevated Hb [Arch Pathol Lab Med.124:616 (2000)]. Yet the clue went unrecoginized by laboratory personnel as well as ordering physicians leading to a probable delay of nearly one year in the diagnosis of the patient’s underlying hematologiclal disorder.
CD4+ Perforin+ T Cells in B-CLL: MHC Class II Restriction and Anti-Cytomegalovirus Reactivity.
Authors: Walton James , Lawson Keirissa , Manoussaka Maria S. , Nathwani Amit , Emory Vincent , Lydyard Peter M. , Porakishvili Nino 
Published: April 2020

Abstract
Introduction: An expansion of CD4+ T cells expressing perforin (PF) has recently been described in B-CLL and we have previously demonstrated anti-cytomegalovirus (CMV) reactivity within this population (Walton et al; 2004; Blood [ASH annual meeting abstracts] 104:4787). Here we further characterise the anti-CMV response of CD4+PF+ T cells in B-CLL and investigate the role of CMV in CD4+PF+ T cell expansion.
Methods: Peripheral blood mononuclear cells (PBMC’s) from 24 untreated B-CLL patients (17 CMV seropositive [SP], 7 CMV seronegative [SN]), 2 SP treated (Campath) patients and 12 healthy age-matched control individuals (8 SP, 4 SN) were fixed, permeabilised and stained with anti-CD4PerCP, anti-IFN-γ-APC and anti-PF-FITC monoclonal antibodies (mABs) (BD). PBMC were cultured for 18 hrs with DOWNE cell lysate (Dade Behring) containing CMV-antigen or lysate alone and with anti-CD28 and anti-CD49d mAbs (BD), in the presence of Brefeldin A (eBiosciences). In blocking experiments PBMC were pre-incubated with anti-HLA DR,DP,DQ mAb (BD) for 1 hour. The CMV specific response was assessed by flow cytometry (Dako Cyan, Summit software) as the percentage of IFN-γ+ cells in PF+ and PF− CD4+ T cell populations. Statistical analysis was performed using the Mann-Whitney U test and Spearman rank correlation.
Results: The proportion of CD4+ T cells expressing PF directly ex vivo was significantly higher in SP B-CLL patients (17.5±18.6%) compared to SN patients (2.0±2.3%, p=0.019). In seropositive aged matched controls the percentage of CD4+ cells expressing perforin was positively correlated with the percentage of CMV-reactive CD4+ cells (r=0.976, p&lt;0.01). In contrast, there was no significant correlation in the patient group. However, two patients with relatively large expansions of CD4+PF+ cells (37.7±3.39%) post-Campath treatment had high percentages of CMV-reactive CD4+ cells (10.93±0.62%) compared to SP B-CLL patients (1.34±1.19%) and SP controls (1.31±1.14%), implying Campath related CMV reactivation. The addition of anti-HLA-DR,DP,DQ mAb to patients’ PBMCs, prior to CMV stimulation, led to an 80% (from 3.26% to 0.79%) and 90% (from 3.9% to 0.45%) reduction in the proportion of antigen reactive CD4+ and CD4+PF+ cells respectively.
Conclusions: A population of major histocompatibility complex (MHC) class II restricted, CMV reactive, CD4+PF+ T cells exists peripherally, in a large group of CMV SP B-CLL patients. Furthermore, CMV is associated with CD4+PF+ T cell expansion in patients and controls. Our data implies that high numbers of B-CLL cells inhibit anti-viral effector function, leading to increased viral activity and chronic antigenic exposure, potentially driving CD4+PF+ T cell expansion.
Sphingosine 1-Phosphate (S1P) Induces Migration and ERK/MAP-Kinase-Dependent Proliferation in Chronic Lymphocytic Leukemia (B-CLL) Due to Expression of the G Protein-Coupled Receptors S1P1/4.
Authors: Seitz Gabriele , Yildirim Sedat , Boehmler Andreas M. , Kanz Lothar , Möhle Robert 
Published: April 2020

Abstract
Egress of lymphocytes from lymphoid organs into the circulation has been shown to depend on the presence of the lipid mediator sphingosine 1-phosphate (S1P) in the peripheral blood, and expression of corresponding S1P receptors (i.e., S1P1), that belong to the family of 7-transmembrane G protein-coupled receptors (GPCR). As circulating lymphocytic lymphoma cells are a hallmark of chronic lymphocytic leukemia, we analyzed expression of different S1P receptors and the effects of S1P on B-CLL cells. By qualitative and quantitative (TaqMan) RT-PCR, significant mRNA expression of S1P1 and S1P4 was found in CLL cell lines (EHEB, MEC-1) and in most samples (S1P1 in 88%, S1P4 in 100%) of primary CD19+ cells isolated from the peripheral blood of untreated B-CLL patients. mRNA of other S1P receptors (S1P2, S1P3, S1P5) was less consistently detected. Normal, nonmalignant B cells were strongly positive for S1P1, while other S1P receptors were weakly expressed or negative. S1P induced typical effects of chemotactic GPCR, such as actin polymerization (analyzed by flow cytometry) and chemotaxis (measured in a modified Boyden chamber assay) in CLL cell lines and primary B-CLL cells. After serum deprivation in vitro, S1P induced phosphorylation of ERK/MAP-kinase as analyzed by Western blot, demonstrating that S1P receptors expressed in CLL were able to activate signaling pathways of GPCR not only related to cell migration and chemotaxis, but also to cell proliferation. Of note, the S1P1 ligand FTY720, which induces receptor internalization after prolonged exposure and acts as an antagonist, resulted in apoptosis in CLL cell lines and primary CLL cells in vitro, as measured by MTT-test and staining with Annexin-FITC, respectively. We conclude that sphingosine 1-phosphate, which is present in the peripheral blood in considerable amounts, contributes to the trafficking of B-CLL cells expressing the GPCRs S1P1/4, and to their prolonged survival.
CD38 Expression in Chronic Lymphocytic Leukemia (CLL) Patients Is Associated with Family History of Hematological Neoplasms.
Authors: Domingo-Domenech E. , Domingo-Claros A. , Gonzalez-Barca E. , Benavente Y. , Alvaro T. , Bellas C. , Font R. , de Sevilla A. Fernandez , de Sanjose S. 
Published: April 2020

Abstract
Introduction: Family clustering of hematological tumours has largely been reported in chronic lymphocytic leukemia (CLL) patients.
Objectives: To identify clinical and biological parameters at diagnosis that could be associated to family history of haematological neoplasms in CLL patients.
Methods: Eighty-five unselected consecutive CLL patients were included in the case-control multicenter study Epilymph_Spain. Immunophenotype data, including CD38 expression, genetics and clinical presentation were performed at diagnosis. Personnel interviews were conducted to collect data on family history of cancer, including site, age at diagnosis, and status of the affected relative. Statistical differences between each group were analysed using Chi-Square tests.
Results: Out of the 85 CLL patients included in the study, 50 (59%) reported a first-degree relative with cancer, from which 7 (8%) were haematological tumours. In the following table, the most relevant variables related with the family history of hematological neoplasms in CLL patients are presented.
Conclusions: Family history of haematological neoplasms, CD38 expression and younger age at presentation of the disease seems to identify a subgroup of CLL patients that could have a genetic origin.
Family history of Haematological neoplasm p NO (n=78) YES (n=7) Median age (range) 71 (45–87) 62 (53–74) 0,061 Gender (M/F) 51 /27 4 /3 0,476 CD38 + 21 (27%) 5 (71%) 0,026 Rai 0–1/2–4 67 /11 7 /0 0,369
Comparison of Genomic Aberrations and Expression of ZAP-70 and CD38.
Authors: Yañez Lucrecia , Cuadrado Maria Angeles , Insunza Andres , Bermudez Arancha , Mesones Belen Gonzalez , Conde Eulogio , Bureo Encarna , Iriondo Arturo 
Published: April 2020

Abstract
Expression of CD38 and /or ZAP 70 and chromosome abnormalities such as deletions of 11q23 and 17p13 have been previously identified by several groups as predictors of disease progression and decrease overall survival in Chronic Lymphocytic Leukemia (CLL). Moreover, few are described about the relationship between ZAP70/CD38 status and their genomic aberrations associated.
We analysed 86 patients (median age: 70 yrs; female/male: 28/58) diagnosed of CLL between 1980 and 2005. Cytoplasmic ZAP-70 expression and cell surface expression of CD38, was determined by flow cytometry in CD19+ CD5+ B-CLL cells. Trisomy 12 and deletions in 13q14, 11q22–23 and 17p13, were detected by the interphase cytogenetic fluorescence in situ hybridization (FISH). Biological study in patients diagnosed before 2001 was done on cryopreserved blood cells.
Concordant expression of ZAP-70 and CD38 was seen in 66 of 86 patients (76%). Fifty-one patients were ZAP-70- and CD38-, and 15 patients were ZAP-70+ and CD38+. Discordant ZAP70/CD38 status was present in 20 patients. Cytogenetic study identified chromosome aberrations in 85% of patients. Deletions in band 13q14 (43%), followed by trisomy 12 (21%), deletions in 11q22–23 (11%) and in 17p13 (10%) were the most frequent abnormalities. Normal karyotype was presented in 28% of patients and complex karyotype in 11%.
In the concordant ZAP70-CD38- cases and ZAP70+ CD38+, 13q14 aberration (47, 9%) and normal karyotype (42%) respectively, were more frequent than in the other groups but in our study this differences were not statistically significant. Moreover deletion in 11q22–23 (25%) was seen statistically significant, p = 0, 05, in patients with concordant expression of ZAP70+ and CD38+. Patients with discordant status in the expression of ZAP70 and CD38, were statistically associated with trisomy 12 (38%) p = 0,002.
In conclusion, in our study we find that expression in ZAP70 and CD38 is associated with a different genomic aberration. Deletion in 13q14 appears more frequent in patients with concordant ZAP70- and CD38- expression. Trisomy 12 is associated with discordant status expression. Finally, the worse prognostic group ZAP70+ and CD38+ is associated with a normal karyotype or deletions in 11q22–23.
Expression of ZAP 70 and CD38 and genomic aberrations ZAP70+ CD38+ ZAP70- CD38- Discordant status p-value Del 11q22-23 25% 4,2% 12,5% 0,05 Del 17p13 8,3% 4,2% ---- 0,52 Trisomy12 8,3% 10,4% 37,5% 0,002 Del 13q14 16,7% 47,9% 31,3% 0,10 Normal 41,7% 33,3% 18,8% 0,39 Complex Karyotype 20% 5,9% 20% 0,13
Inhibition of Bortezomib-Induced Apoptosis in Chronic Lymphocytic Leukemia by Red Blood Cell Uptake.
Authors: Wheat Luise M.C. , Kohlhaas Susan L. , Monbaliu Johan , De Coster Roland , Majid Aneela , Walewska Renata J. , Cohen Gerald M. , Dyer Martin J.S. 
Published: April 2020

Abstract
Bortezomib (PS-341/Velcade™) is a reversible inhibitor of the proteasome that has shown promising activity in clinical trials in several malignancies including multiple myeloma, mantle cell lymphoma and follicular lymphoma, including those with refractory disease. However, results have been less encouraging in chronic lymphocytic leukemia (CLL) and we have, therefore, sought to determine the barriers to effective therapy with bortezomib in this disease. Patients with CLL were eligible but were required to have received no therapy in the six months prior to the study. In a panel of 26 patients with CLL, both purified mononuclear cells and whole blood were tested for their apoptotic response to bortezomib (1–100 nM) up to 24 h by flow cytometry and western blotting. In all cases, purified CLL cells were sensitive to bortezomib-induced apoptosis in a concentration and time-dependent fashion, irrespective of stage of disease, resistance to prior therapy, IGHV mutational status or the presence of TP53 mutations. Apoptosis was induced at low (&gt;10 nM) nanomolar concentrations of bortezomib by activation of the intrinsic apoptotic pathway. Bortezomib-induced apoptosis correlated with levels of ubiquitination, Bax activation, and caspase cleavage. Apoptosis of CLL cells was obtained at drug levels readily obtained in vivo using currently-used dosing protocols. However, in vitro, it was necessary to maintain these concentrations for 16–24 hours to obtain maximal apoptosis. Apoptosis measured in a whole blood apoptosis assay was markedly less than in isolated lymphocytes at comparable time points and concentrations. Activity of bortezomib in purified cells was not diminished by addition of exogenous plasma but was abrogated by addition of autologous red blood cells (RBC), suggesting preferential active uptake of the drug by these cells. These data were confirmed in animal models showing preferential distribution of bortezomib to the RBC fraction. RBC uptake may therefore account for the low serum levels of bortezomib attained in vivo during terminal half-life and thus the lack of activity against cells in the peripheral blood. Together with pharmacokinetic and in vivo data, these studies suggest that different dosing schedules of bortezomib other than bolus injections may be more effective in patients with CLL.