bluuebunny/crossref_metadata_2025_split huggingface dataset:
Erythropoiesis Is Highly Stimulated in CD34+ Cells in Low-Risk Myelodysplastic Syndromes (MDS) with an Improper Mitochondrial Function.
Authors: Tehranchi Ramin , Invernizzi Rosangela , Zhivotovsky Boris , Fadeel Bengt , Forsblom Ann-Mari , Travaglino Erica , Samuelsson Jan , Hast Robert , Nilsson Lars , Wibom Rolf , Grandien Alf , Cazzola Mario , Hellstrom-Lindberg Eva 
Published: April 2020

Abstract
The apoptosis of early erythroblasts from patients with low-risk MDS, refractory anemia (RA) and RA with ringed sideroblasts (RARS), is mediated through a constitutive cytochrome c (cyt c) release from mitochondria (Tehranchi etal, 2003). Moreover, mature erythroblasts in RARS, but not in RA, show mitochondrial accumulation of aberrant ferritin (MtF) (Cazzola etal, 2003). This study aimed at further describing the pathophysiology of ineffective hematopoiesis in low-risk MDS, by studying cyt c release and MtF expression during erythroid differentiation and mitochondria ATP production in MDS bone marrow cells. We assessed freshly isolated CD34+ cells and day 4–14 erythroblasts from RARS, RA and normal bone marrow (NBM). CD34+ cells from all individuals were negative for MtF. NBM showed only few positive cells (0–4%, d4–14), and RA erythroblasts a median of 3% (0–8%) MtF+ cells. RARS erythroblasts, on the contrary, showed an early increase in MtF+ cells and a continuous increase during the culture period (d4=10%, d7=17%, d14=19%). There was a positive correlation between MtF expression and cyt c at day 14 ( r2=0.8). There was no significant difference in mitochondria ATP production between RARS, RA and NBM (all complexes or cyt -dependent complex IV). We found a significant over-expression (mRNA) of the pro-apoptotic genes for cyt c, Bid and Bax at day 0. Moreover, genes involved in erythroid differentiation were significantly up-regulated in MDS CD34+ cells: 6-fold for GATA-1 and 23-fold for β-globin; p&lt;,0005 for both. GATA-1 and β-globin expression increased during normal erythroid maturation, but in MDS erythroblasts GATA-1 declined and β-globin showed only a weak increase. Comparing RARS with RA, the former showed both higher expression of the β-globin and GATA-1 genes, and a higher degree of cyt c release and MtF expression. This indicates that the cellular abnormalities leading to erythroid apoptosis as well as efforts to compensate for these defects are present at the stem cell level in RARS. G-CSF that reduces cyt c release in MDS erythroblasts (RARS&gt;RA) showed no effect at all on ATP production or cyt c mRNA. Moreover, G-CSF tended to increase MtF expression in some RARS erythroblast cultures, indicating that it allows survival of pro-apoptotic MDS erythroblasts rather than addressing the cause of apoptosis. In conclusion, the aberrant MtF expression of RARS erythroblasts occurs at a very early stage of erythroid differentiation and is paralleled by an up-regulation of genes involved in erythroid differentiation. Alternative mechanisms may be involved in RA pathogenesis, since these cells show cyt c release but only moderate MtF expression, and less gene up-regulation.
Absence of Cytogenetic Abnormalities of PRV-1, TPO and C-MPL Genes Detected by Fluorescence In Situ Hybridization in Essential Thrombocythemia.
Authors: Puigdecanet Eulalia , Espinet Blanca , Villa Olaya , Zamora Lurdes , Besses Carles , Bellosillo Beatriz , Salido Marta , Woessner Soledad , Serrano Sergi , Florensa Lourdes , Sole Francesc 
Published: April 2020

Abstract
Introduction. Essential thrombocythemia (ET) is a chronic myeloproliferative disorder (CMPD) with heterogeneous features and no specific diagnostic markers. Consequently, its diagnosis is based on exclusion of other CMPD and secondary thrombocytosis. The role of PRV-1 (Polycythemia rubra vera-1), TPO (Thrombopoietin) and c-Mpl (Myeloproliferative leukemia virus oncogene) in ET pathogenesis has been studied in order to find new molecular targets which would help in ET diagnosis. PRV-1 gene is overexpressed in granulocytes from polycythemia vera (PV) and in some ET patients. TPO serum levels are not diagnostically useful and c-Mpl expression in megakaryocytes and platelets are generally decreased in ET. Mutations in TPO and c-MPL genes have been detected in familial thrombocythemia, but not in patients with acquired ET. The aim of the present study was to analyse PRV-1, TPO and c-MPL genes status by fluorescence in situ hybridization (FISH) technique in order to find new molecular markers in ET patients.
Patients and Methods. Thirty bone marrow samples of ET patients (7M/23F) diagnozed by PVSG criteria with a normal karyotype and 10 bone marrow samples of normal healthy donors were included in the study. All samples were studied by three locus-specific probes for PRV-1, TPO and c-MPL genes as follows: 1. PRV-1 gene (BAC RP11-160A19, 157 Kb, located at 19q13.12-2) labeled in red cohybridized with the 19p telomeric probe (D19S238E, Vysis) labeled in green. 2. TPO gene (BAC RP11-45NP16, 183 Kb, located at 3q27) labeled in green cohybridized with the centromeric probe for chromosome 3 (D3Z1, Vysis) labeled in red. 3. c-MPL gene (BAC RP11-297L5, 190 Kb, located at 1p34) labeled in green cohybridized with the centromeric probe for chromosome 1 (D1Z5, Vysis) labeled in orange. A minimum of 100 interphase nuclei were analyzed.
Results. FISH study showed no PRV-1, TPO and c-MPL cytogenetic abnormalities in any of the analyzed cases, except for one patient in which 21% of interphase nuclei presented a trisomy for the TPO gene region. The monosomy and trisomy thresholds were 6.1% and 4.7% for PRV-1, 4.9% and 3.4% for TPO, and 5.4% and 3.71% for c-MPL, respectively.
Conclusions. Our results suggest a lack of structural and numerical rearrangements of PRV-1, TPO and c-MPL genes in ET patients. The PRV-1 gene FISH results are in line with the previously reported by Najfeld et al (Exp Hematol2003;31:118–21); regarding TPO and c-MPL results, this is the first FISH study reported in the literature in ET.
Dissecting Growth Factors Production in Patients with Agnogenic Myeloid Metaplasia (AMM).
Authors: Wang Jen-Chin , Hemavathy Kirugaval , Goldberg Amit , Chang Tsong S. , Novetsky Allan D. , Lipton Jeffrey 
Published: April 2020

Abstract
Growth factors including Transforming Growth Factor-β 1 ( TGF- β1), Platelet Derived Growth Factor (PDGF) and Fibroblast Growth Factor (FGF) have been implicated as responsible for bone marrow fibrosis in AMM. Although TGF-β1 and FGF have been demonstrated to be increased in blood hematopoietic progenitor cells, a direct measurement of the production of these growth factors by megakaryocytes has not been performed. The current study was devised to study the production of these growth factors production directly in megakaryocytes and monocytes of AMM patients and correlate these with the clinical features. Twelve patients with AMM and 11 normal healthy volunteers used as controls, were studied. CD 34 + cells and CD 14+ cells were obtained from blood mononuclear cells by MACS Progenitor cell isolation kits ( Miltenyi Biotec, CA). The megakaryocytic (CD41+) cells were then obtained by growing the isolated blood CD34 + for 10 days in Iscove Modified Dulbecco Medium with Stem Cell Growth Factor and Thrombopoietin. The mRNA levels of TGF-β1, FGF and PDGF in the isolated megakaryocytes and blood monocytes were assayed by Real-Time RT-PCR. The results were as shown in Table 1.
Among the AMM patients, a patient with prior history of Polycythemia Vera (PV) and a patient with Essential Throbocythemia (ET) were found to have elevated PDGF and FGF expression in their monocytes but the expression of growth factors was not elevated in their megakaryocytes.. These results demonstrate that megakaryocytes are the main source of growth factors responsible for marrow fibrosis. The study also suggests that growth factors produced by monocytes may be responsible for fibrosis in AMM patients with a prior history of PV or ET.
Table-1
Megakaryocytes Monocytes * Denotes number of patients with elevated growth factor levels as compared with controls. ** Denotes numbers of patients tested. TGF- β 1 8*/10* 0/10 PDGF 8/10 3/7 FGF 6/10 2/7
Enumeration and Immunologic Characterization of Basophils in Normal Bone Marrow and Patients with Myeloproliferative Disorders.
Authors: Agis Hermine , Krauth Maria T. , Muellauer Leonhard , Schwartz Lawrence B. , Horny Hans P. , Valent Peter 
Published: April 2020

Abstract
Basophils are highly specialized granulocytes that express a unique profile of antigens and increase in myeloproliferative disorders. In chronic myeloid leukemia (CML), basophilia is an independent prognostic variable. So far, however, no reliable immunohistochemical approach for routine-detection and enumeration of bone marrow (bm) basophils has become available. To overcome this disadvantage, we have applied the anti-basophil antibody 2D7 on formalin-fixed, paraffin-embedded sections of normal bm and bm from patients (pts) with chronic myeloid leukemia (CML; chronic phase, n=21; accelerated phase, n=9), other myeloproliferative disorders (idiopathic myelofibrosis [IMF], n=3; polycythemia vera [PV], n=7; essential thrombocythemia [ET], n=7), and normal / reactive bm (n=32). As assessed by serial section-staining of bm specimens, the 2D7 antibody was found to be a basophil-specific immunohistochemical reagent. In serial bm sections, 2D7+ basophils co-expressed histidine decarboxylase, CD15, and CD43, but did not express B- or T-cell restricted antigens corresponding to the phenotype of normal blood basophils. Bm basophils were found to increase in number in pts with CML and other myeloproliferative disorders compared to normal bm (median 2D7+ cells/mm2 bm: normal bm: 7; CML: 46; IMF: 26; PV: 21; ET: 21, p&lt;.05). The highest numbers of bm basophils were recorded in pts with accelerated phase CML (111 2D7+ cells/mm2). Together, we have established a useful immunohistochemical staining procedure for basophil detection in normal bm and pts with myeloid neoplasms. This approach should enable the quantification of basophils in these pts and the monitoring of bm basophil counts during follow up examinations and anti-leukemic therapies.
Monocyte Activation Studies in Patients with Essential Thrombocythemia.
Authors: Goette Nora P. , Marta Rosana F. , Chazarreta Daniel C. , Molinas Felisa C. 
Published: April 2020

Abstract
Essential thrombocythemia (ET), polycythemia vera and mielofibrosis with mieloid metaplasia (MMM) are myeloproliferative diseases that differ in their potential for developing bone marrow fibrosis. In addition to the role of megakaryocytes, the participation of monocytes in collagen deposits and fibroblast proliferation in the pathophysiology of MMM has already been described. We studied here monocyte activation in ET patients by measuring IL-2 receptor expression (CD25) in ex vivo monocytes and after its adhesion to polyestirene. Besides, we evaluated IL-1-b released by mononuclear cells after 24 hs culture. We evaluated 14 untreated ET patients. Blood samples were obtained under sterility conditions and mononuclear cells were separated by Ficoll-Hypaque gradient. CD25 positivity was evaluated by flow cytometry identifying monocytes as the CD14 + population before and after 18 hs cell culture in IMDM and 2% albumin. IL-1b was measured by ELISA technique in the supernatant of cells cultured in IMDM supplemented with 10% FCS during 18 hs. There were no statistical differences in CD25 expression in ET patients (n=8) neither before (3.1%, 1–11.4%) (median and range) nor after (15.2%, 4.7–39.3%) cell culture of CD14 population compared to normal controls (n=12, 2.0%, 0.2–10.3% and 20.3%, 1.4–34.3%, respectively). Levels of IL-1b were found decreased in patients with ET (n=10), 481 pg/ml, 53–1290 pg/ml, compared to normal controls (n=11), 779 pg/ml, 68.7–2166 pg/ml, although not reaching statistical difference (p=0.057). In conclusion, we did not find monocyte activation in ET patients as demonstrated by CD25 expression and IL-1b release. These results suggest that there is not a constitutive or induced activation of monocytes in ET but a trend towards a decreased capacity of activation. We believe this different monocyte behaviour from ET patients compared to that described in MMM patients could account for the dissimilar natural course of these diseases.
Demonstration of Biclonal Chronic Lymphocytic Leukemia with Mutated and Unmutated Clones and Concurrent but Clonally Unrelated Myeloma in the Same Patient by Single Cell Immunoglobulin Heavy and Light Chain Gene Analysis.
Authors: Yavuz Selim A. , Hartmann Dan-Paul , Baidas Said , Lipsky Peter E. , Ozdemirli Metin 
Published: April 2020

Abstract
Patients with chronic lymphocytic leukemia (CLL) may develop other B cell malignancies in their clinical course including aggressive diffuse large B-cell lymphomas and rarely myelomas. In a large proportion of cases, the secondary B cell malignancies reflected the emergence of immunophenotypically and genetically different clones. An immature type plasma cell myeloma developed in a 73-year-old female patient in whom CLL was diagnosed four years previously. The CLL expressed CD5, CD19, CD23, CD38 and surface kappa light chain, but were negative for ZAP-70. Trisomy 12 was detected by FISH analysis. PCR analysis of the peripheral blood for immunoglobulin heavy chain genes demonstrated two sharp bands that were initially interpreted as biallelic heavy chain gene rearrangements. Myeloma cells were CD38 and CD138 positive, CD19 negative and expressed cytoplasmic kappa light chain, but not heavy chains. In order to investigate the clonal relationship between these B cell malignancies, a detailed analysis of VHDJH and VκJκ gene rearrangements in individually sorted CD5 and CD19 double-positive CLL cells and also in CD38-positive and CD19-negative myeloma cells by single cell PCR of genomic DNA and direct sequencing was carried out. This technique permitted identification and pairing of both the heavy and light chain immunoglobulin genes from the same individually sorted cell. A total of 17 individual CLL and 23 myeloma cells were successfully analyzed. Our analysis demonstrated (a) the presence of two discrete clones of CLL, one with usage of [VH1-2*04/D3-3*01/J3*02]-[Vκ2-28*01/J1*01] without VH and Vκ hypermutation and the other with usage of [VH1-2*04/D4-11*01/J6*02]-[Vκ1-5*03/J1*01] with VH and Vκhypermutation; (b) no clonal relationship between the CLL and myeloma cells that utilized different VHDJH and VκJκ rearrangements [VH3-66*02/3-10*01/J4*03]-[Vκ1-33*01/J2*02] with VH and Vκ hypermutation. To our knowledge, this is the first demonstration of a biclonal CLL with mutated and unmutated clones in the same patient along with a third clonally unrelated B cell malignancy. This result suggests that single cell analysis may be necessary to detect subtle biclonality of CLL that might be associated with a more aggressive phenotype.
Activation and Proliferation of B Cell Chronic Lymphocytic Leukaemia (B-CLL) Cells through Anti-CD180, Anti-CD40 and IL-4.
Authors: Porakishvili Nino , Manoussaka Maria , Kulikova Nino , Walton James , Nathwani Amit , Jewell Andrew P. , Stevenson Freda , Clark Ed , Lydyard Peter M. 
Published: April 2020

Abstract
Introduction: We have previously shown that Toll-like receptor RP105 (CD180) is heterogeneously expressed on B-CLL cells and that the ligation of CD180 by monoclonal antibodies (mAb) on CD180+ B-CLL cells resulted in delineation of responder and non-responder B-CLL clones [1]. In this study we have examined the role of IL-4 together with CD180 and CD40 as activation signals.
Methods: Blood mononuclear cells were separated from 7 responder B-CLL patients with both mutated and unmutated Ig Vh genes and 11 controls and were cultured for 72 hours in optimum concentrations of anti-CD180 (G28-8) or anti-CD40 mAb or both in presence and absence of 15 ng/ml of IL-4. CD19+ B cells were stained with mAb to the activation marker CD86 or cell cycle protein Ki-67, measured by flow cytometry and expressed as Mean Fluorescence Intensity (MFI) or % of Ki-67+ cells.
Results: B-CLL cells and normal control B cells responded to CD180-ligation by activation and proliferation (Table). Higher levels of CD86 and Ki67 were detected when both anti CD40 mAb and anti CD180 mAb were added (p&lt;0.05) compared with either alone. IL4 alone induced both activation and proliferation of control cells and this was even higher with the leukemic cells (p&lt;0.01) confirming that IL-4 also provides a strong survival/activatory stimulus for B-CLL cells. Addition of IL-4 had no significant enhancing effect on normal B-cells stimulated with both anti-CD180 and anti-CD40, although IL-4 synergised with anti-CD40 in B cell activation (p=0.026) and with CD180 in B cell proliferation (p=0.044).
Conclusion: CD180 had an additive effect with CD40 ligation in activation and proliferation of both B-CLL cells and normal control B cells. IL-4 provides a strong additional stimulus for B-CLL cells.
CD86 and Ki-67 expression by CD19+ cells
CD86 Ki67 B-CLL Control B-CLL control Spontaneous −IL-4 7.2±4.1 4.0±1.0 8.1±2.7 17.4±2.3 +IL-4 22.4±11.8 11.2±4.2 17.0±12.8 23.6±14.8 CD180 −IL-4 14.7±5.8 26.9±13.0 17.1±10.9 28.1±12.0 +IL-4 35.4±14.5 30.0±14.6 26.9±25.6 48.3±9.7 CD40 −IL-4 19.4±8.8 14.8±5.2 14.9±9.7 34.0±5.1 +IL-4 286±145.5 44.0±19.4 52.7±22.8 41.0±19.8 CD180+CD40 −IL-4 32.5±12.6 97.0±26.2 28.0±10.4 65.5±26.6 +IL-4 299±163.2 63.5±27.4 49.3±14.6 55.5±26.4
CD38 Expression Has an Important Value among Prognostic Factors in Chronic Lymphocytic Leukemia (CLL): A Multivariate Analysis in 160 Patients.
Authors: Pavlovsky Carolina , Pavlovsky Miguel A. , Pardo Maria L. , Sapia Sandra , Mountford Pablo , Fernandez Isolda I. , Corrado Claudia S. , Statistics Mariana Juni , Pavlovsky Santiago 
Published: April 2020

Abstract
Objetives: Analyze the prognostic significance of CD38 expression compared to other prognostic factors.
Patients and methods: We studied retrospectively and prospectively at our institution 160 patients (pts) with diagnostic of CLL who met the diagnostic criteria of National Cancer Institute-Working Group (NCI-WG), 97 men and 63 female with an age range of 33 – 90 years (median 62) diagnosed between January 1989 and December 2003 and evaluated up to June 2004. Sixty nine pts showed CD38 positive cells and 91 were CD38 negative. CD38 expression was determined by flow cytometry defining as a higher expression &gt; 7% CD38 positive cells. (Krober A et al. Blood15: 1410–1416.2002). Event Free Survival (EFS) and Overall Survival (OS) were analized according to different characteristics at diagnosis.
No. Patients %EFS 72 mo p= %OS 72 mo p= Binet Stage A /B-C 129/31 65/30 0.000 90/92 0.245 Rai Stage 0 /I – II – III – IV 92/68 72/37 0.000 94/94 0.339 TDLI &lt; / ≥ 12 mo 23/126 41/63 0.007 65/100 0.001 Light Chains kappa / lambda 94/66 68/50 0.473 90/95 0.702 CD38 Positive / Negative 69/91 32/82 0.000 81/100 0.000
Statistical significant difference was observed in Binet stage, Rai stage, TDLI and CD38 expression for EFS and in TDLI and CD38 expression for OS. Considering these variables with p value of &lt;0,05, a multivariable analysis was performed (Cox) in order to asses the adverse effect of each dichotomize group in EFS.
Beta S.E. H.R. 95% C.I. p CD38 Positive (1) /CD38 Negative(0) 1.70 0.32 5.5 2.89–10.53 0.000 Binet B-C /Binet A(0) (1) 1.46 0.29 4.33 2.42–7.7 0.000 RAI I-II-III-IV /RAI 0 (0) (1) 1.45 0.30 4.29 2.37–7.78 0.000 TDLI ≥ 12 (1) /TDLI&lt;12 (0) 0.03 0.00 1.03 1.01–1.04 0.000
This analysis shows that the presence of CD38 positive cells at diagnosis conferred an adverse effect on EFS with a HR 5.5 (95% confidence interval 2.89–10.53) over the other variables included.
Conclusion:In our population, patients with CD38 positive expression showed a EFS and OS at 72 months of 32% and 82% ( p=0.000) versus 81% and 100% (p=0.000) in patients with CD38 negative expression. We can conlude that CD38 positive cells will identify a subgroup of patients with aggresive clinical outcome and worse prognosis.
ZAP-70 Versus CD38 Expression as the Prognostic Factors in B-Cell Chronic Lymphocytic Leukemia.
Authors: Hus Iwona , Bojarska-Junak Agnieszka , Podhorecka Monika , Wasik-Szczepanek Ewa , Sieklucka Malgorzata , Skorzynska Katarzyna , Rolinski Jacek , Dmoszynska Anna 
Published: April 2020

Abstract
B-CLL is a heterogeneous disease with a highly variable clinical course. The mutational status of immunoglobulin heavy-chain variable region (IgVH) genes in the leukemic cells seems to be of great importance in assessing the prognosis in B-CLL. Recent studies have shown that both CD38 and ZAP-70 might be surrogate markers for IgVH mutation identifying patients with a more aggressive clinical course. The aim of our study was to estimate the clinical value of both ZAP-70 and CD38 as B-CLL prognostic factors.
The analysis with use of flow cytometry technique was performed at diagnosis in the group of 100 B-CLL patients. 39/100 persons were defined as ZAP-70 positive and 61/100 persons as ZAP-70 negative (based on 20% cut off value in the CD19+/CD5+ population). ZAP-70+ patients had significantly higher lymphocytosis (p=0.003) and LDH level (p=0.009), while hemoglobin (p=0.049) and platelet levels (p=0.002) were significantly lower than in ZAP-70− patients. The proportion of CD19+/CD5+ cells expressing ZAP-70 correlated significantly with stage of disease (p=0.035). Patients with Rai stage III–IV (32/100 patients) had higher percentage of CD19+/CD5+/ZAP-70+ cells than patients with stage 0-II (68/100 patients) (24.81±17.40% and 14.89±13.54%, respectively, p=0.008). The mean treatment-free interval was longer in the ZAP-70 negative (8.46±4.63 months) than in the ZAP-70 positive patients (1.16±0.28 months). Furthermore, patients requiring treatment (57/100) had significantly higher percentage of CD19+/CD5+ cells expressing ZAP-70 protein than untreated patients (20.18±15.09 and 13.44±11.84, respectively, p=0.034). In the midst of patients requiring treatment, percentage of CD19+/CD5+/ZAP-70+ cells was significantly lower in patients achieving remission after first line therapy (26/57) than in patients with progressive disease (31/57) (p=0.004).
According to CD38 analysis the patients with ≥ 20% of CD38 expression were considered positive (CD38+) and those &lt; 20% were considered negative (CD38−). Patients CD38 positive, like the ZAP-70 positive patients, had more advanced Rai stage of disease, significantly higher lymphocytosis and LDH serum level. Patients requiring treatment had higher percentage of CD19+/CD5+CD38+ cells than untreated patients, however no statistical significance was detected. Percentage of CD19+/CD5+CD38+ was not a dicriminating factor between patients with good response to the first line therapy and those with progressive disease.
The obtained results indicate that both ZAP-70 and CD38 expression may be prognostic markers identifying a subgroup of B-CLL patients with aggressive clinical presentation. However, the ZAP-70 expression seems to have more prognostic power, especially in predicting response to the first line therapy.