bluuebunny/crossref_metadata_2025_split huggingface dataset:
Poor Survival Predicted by MDM2 Oncoprotein Expression in Diffuse Large B-Cell Lymphoma (DLBCL) with Wild-Type TP53 Gene
Authors: Rocque Gabrielle L. , Møller Michael B. , Colleoni Gisele W. B. , Sánchez-Beato Margarita , Green Tina M. , Thorborg Trine , Piris Miguel A. , Eickhoff Jens C. , Twohig Matthew , Malik Jeffrey T. , Young Allen H. , Oberley Terry D. , Malter James S. , Ferry Judith , Chan Wing C. , Weisenburger Dennis D. , Greiner Timothy C. , Young Ken H. 
Published: April 2020

Abstract
The p53 pathway is known to play a role in pathogenesis and prognosis of human malignancies including diffuse large B-cell lymphoma (DLBCL). The MDM2 protein has been shown to regulate p53 function and stability via a multi-factorial negative feedback loop. MDM2 is involved in transport of p53 out of the nucleus and its subsequent degradation. Additionally, MDM2 binds p53 protein and inhibits its function as a transcription factor. As an integral part of the regulation of the p53 pathway, we hypothesize MDM2 expression contributes to the pathogenesis of DLBCL and affects survival. The purpose of this study is to investigate the potential role of the MDM2 protein in DLBCL by correlating expression of MDM2 with p53 expression, TP53 mutation status and clinical outcome. Immunostains for MDM2 and TP53 gene-product proteins were performed in 133 cases from 6 medical centers. A positive immunostain was defined as nuclear staining in 10% or more of the tumor cells. The TP53 gene was analyzed for mutations with PCR-based and sequencing methods. Patients were treated with CHOP (cyclophosphamide, doxorubicin, vincristine and prednisone) or CHOP-like regimen. The Kaplan-Meier method was used for survival analysis. MDM2 immunostains were positive in 64 of 133 cases of DLBCL (48%). There was a trend toward poor overall survival (OS) with MDM2 expression in the entire group of patient with DLBCL (p=0.32). Twelve of 24 cases (50%) with TP53 mutations demonstrated MDM2 expression. Within this TP53 mutated group, no significant difference in OS was observed between MDM2-positive and MDM2-negative cases (p=0.32). Of patients with wild-type (WT) TP53, 51 of 107 cases (48%) expressed MDM2. In this subset of patients, the MDM2-positive phenotype predicted for poor 5-year OS (38% vs 67%, p=0.002), and a significantly shorter median disease-free survival (2.3 years vs. &gt;5.0 years; p=0.013). The complete remission rate was 57% in MDM2-positive cases compared to 73% in MDM2-negative cases in the WT-TP53 group (p=0.08). Multivariate analysis confirmed that MDM2 expression was an independent predictor of poor OS in patient with DLBCL with a WT-TP53 gene (HR 2.0, 95% CI 1.15–3.56; p=0.015). This study demonstrates the negative survival impact of MDM2 expression in patients with wild-type TP53, suggesting that MDM2 provides an alternative mechanism of p53 pathway inactivation in DLBCL cases with a WT-TP53 gene.
Diagnostic Role of Flow Cytometry Analysis of Bioptical Samples in the Diagnosis of Lymphoid Tumors
Authors: Cro Lilla , Ferrario Andrea , Zucal Nadia , Gianelli Umberto , Fabris Sonia , Nobili Lucia , Olivero Barbara , Rossi Francesca G , Baldini Luca , Deliliers Giorgio Lambertenghi 
Published: April 2020

Abstract
We reviewed flow cytometric immunophenotyping (FCI) data of 470 tissue suspensions suspected of being involved by lymphoma (371 lymph nodes, 263 surgical biopsies and 108 fine needle aspirations,16 spleens, 6 tonsils, and 77 other tissues) and we have compared corresponding histologic diagnosis. The screening panel of FCI (CD45, CD19, CD3, CD4, CD8, and sIgκ/sIgλ ratio) identified three main groups:
239 cases with demonstrable light chain restriction (sIgκ/sIgλ ratio ≤ 0.5 or ≥4); 37 cases with demonstrable T cell proliferation and polyclonal B cell population; 194 cases with polyclonal B cell and normal T cell populations (146 cases) or with not detectable CD45 reactivity and other leucocyte markers (48 cases).
In group 1, 235 cases were then evaluated by means of the following markers: CD5, CD10, CD11a, CD11c, CD20, CD22, CD23, CD30, CD38, CD43, CD79a–b, CD103, CD138, FMC7, and, in group 2, 30 cases were evaluated by means of the following markers: CD1a, CD2, CD5, CD7, CD16, CD26, CD43, CD56, CD57, CD45RA/RO, anti-TCR-ab gd. The complete analysis identified in group 1 four main diagnostic subsets: A (26 cases) characterized by CD5+, CD23+/±, sIg dim (CLL-like); B (36 cases) characterized by CD5+, sIg bright (MCL/CLLv); C (89 cases) characterized by heterogeneous expression of Ig, CD20 and CD43 (lymphoproliferative syndromes CD5−, excluding hairy cell leukemia); D (84 cases) characterized by CD10+, CD20 bright, CD43− (follicular NHL like). In cluster B, FISH analysis for t(11;14) resulted positive in 15/27 cases; in cluster D, FISH analysis for t(14;18) resulted positive in 60/60 cases. Histologic analysis convalidated FCI data in 23 cases of subset A (88%), in 29 cases of subset B (80%) and in 60 cases of subset D (71%). Subset C included 39 cases of LCL and 15 cases of MZL. In group 2, all cases studied expressed an aberrant T phenotype but, with the exception of 1 case of NK proliferation, 1 case of likely Sezary syndrome/Micosis Fungoides (CD4+CD7−CD26−) and 1 case of T lymphoblastic lymphoma (CD3cy+TdT+), in the other 34 cases was not possible identify other peculiar subset. In this group, histologic analysis included 19 cases of NHL T (63%). Considering NHL vs no NHL, sensibility (SE) and specificity (SP) of FCI analysis resulted 88% and 92%, respectively; positive predictive value (PPV) was 96% and negative predictive value (NPV 78%). Considering B-cell NHL only, SE was 91%, SP 95%, PPV 97% and NPV 84%. Even if histology is the basis for the diagnosis of lymphoid tumors, however our study supports that FCI can play an important role mainly in B-NHL diagnosis, combining rapidity of analysis with a multiparametric analysis, also in presence of size limitated samples.
FDG PET in the Staging and Management of Cutaneous T Cell Lymphoma
Authors: Woodward Catherine S , Halkar Raghuveer , Delman Keith , Parker Sareeta , King Nassoma , Lechowicz Mary Jo 
Published: April 2020

Abstract
We aimed to assess the usefulness of 18F-FDG PET in siting lymphoma involvement for CTCL patients. We retrospectively analyzed CTCL patients who had received a lymph node biopsy within approximately one month of a PET scan. Between 2003 and 2007, 19 CTCL patients (7 males, 12 females, age range: 4 – 70, median: 55) underwent a combined 56 PET scans. All studies were performed on GE Discovery LS 8 slice MDCT PET CT scanner or GE Discovery DST 16 slice MDCT PET CT scanner (Milwaukee, WI) and the images were interpreted on a GE AW or Xeleris workstation. The patients underwent whole body imaging from the head to foot at 4 minutes per bed position. Of these 19 patients, eight patients (four men, four women, mean age: 56.6, range: 49 – 70) received a lymph node biopsy following a PET scan that demonstrated elevated focal uptake (SUVs ranging from 2.2 to 9.9) in at least one lymph node. In four of the cases, the biopsied lymph node directly confirmed CTCL involvement sited in that node by PET scan (SUV 2.2 to 6.1). Three cases represented false-positive PET scans which were attributed to category 1 dermatopic lymphadenopathy and post radiation change. Despite these lack in correlations, in one case, PET sited adenopathy (SUV 9.9) in an inguinal node that biopsy pronounced benign. Further studies proved that a sampling error produced a false-negative biopsy. Despite the small sample, the study demonstrates the importance of PET imaging in staging patients with CTCL. Findings suggest that lymph node SUV values ranging from 2.2 – 9.9 often correlate with definite involvement of the nodes, and consequentially progression of the disease. It was found that false-positive PET scans may result from post radiation change or dermatopathic lymphadenopathy, category 1. The sample size was too small to detect differences in SUV and correlation in lymph node involvement. Our study demonstrates the need for improvement in the specificity of FDG-PET scans possibly by two point imaging and accurate localization of lymph nodes by use of an intraoperative probe to further enhance its usefulness in the staging of CTCL. Further prospective study is needed to determine correlation with SUV and lymph node involvement.
A Part of Acute-Type ATLL Cases Has Genomic Imbalance in Common with Lymphoma-Type ATLL
Authors: Nakagawa Masao , Nakagawa-Oshiro Aya , Tsuzuki Shinobu , Utsunomiya Atae , Seto Masao 
Published: April 2020

Abstract
Acute-type Adult T-cell leukemia/lymphoma (ATLL) is classified into four clinical subtypes (Acute, Lymphoma, Chronic and Smoldering). Acute-type ATLL comprises clinically heterogeneous groups because this subtype is defined as a group not classified into three other subtypes. When lymphoma-type progress to show PBL involvement of more than 1%, it becomes acute-type ATLL. We tentatively designate such cases showing leukemic cells in peripheral blood with prominent nodal involvement as “acute-type ATLL-PNI”. Clarification of the molecular basis for clinical heterogeneity of Acute-type ATLL is an important issue for therapeutic strategy. We previously studied acute-type ATLL with fulminating leukemic manifestation (more than 60% leukemic cells in peripheral blood) by array CGH and identified a part of the molecular basis for the ATLL subtype (Oshiro et al., Blood107:4500,2006). Here we analyzed six such cases of “acute-type ATLL-PNI” by means of array CGH consisting of 2304 artificial chromosome clones that cover the whole genome at a 1.3 mega base resolution. Regions of recurrent gain were detected on 2p16.1, 3p26.3-26.1, 3p24.2-24.1, 3p22.2-22.1, 3p14.3, 3p14.2-3q13.3, 3q25.32-26.2, 3q26.31-26.33, 3q27.3-29, 6p25.3-24.1, 7q32.3-33 and 17q23.3-24.2 and regions of recurrent loss on 4q21.22-21.23, 6q13-16.3 and 13q32.1-33.1. We compared the genomic profile of these six cases of “acute-type ATLL-PNI” with that of 49 lymphoma-type ATLL cases previously reported by us (Oshiro et al., Blood, 2006). They were very similar to each other and thirteen of the 15 regions of recurrent genomic imbalance in “acute-type ATLL-PNI” were also found in lymphoma-type ATLL (86.7%). Our findings clearly showed that “acute-type ATLL-PNI” have genomic imbalances in common with lymphoma-type ATLL rather than acute-type. Our array data also indicated that “clinically defined” acute-type ATLL contained leukemic-type and lymphoma-type with leukemic transformation.
MLL/AF10(OM-LZ) Transformed Cells Induce Myeloproliferative Disease-Like Myeloid Leukemia in the Sublethally Irradiated Mice
Authors: Fu Jen-Fen , Shih Lee-Yung 
Published: April 2020

Abstract
Chromosomal translocation t(10;11)(p12;q23) resulting in the fusion of MLL gene with AF10 is seen in patients with acute leukemias, especially in acute monoblastic leukemia. The OM-LZ domains of AF10 have been shown to be sufficient for immortalization when fused with 5′MLL, however, the leukemogenesis of MLL/AF10(OM-LZ) is not clear. By retrovirally transducing MLL/AF10(OM-LZ) fusion gene into the murine bone marrow cells, we established six MLL/AF10(OM-LZ)-immortalized cell lines in medium containing IL3, IL6, SCF and GM-CSF. The cytological and immunophenotypic studies showed that all the cell lines were consisted of immature blast-like cells and monocyt/macrophage cells, except that line MA6 also contained about 2% mature granulocytes. The percentage of mature granulocytes in line MA6 raised to 20% upon treatment with G-CSF. Virus integration site (VIS) analysis demonstrated that line MA6 had a VIS at PD-L2 gene. Whether interruption of PD-L2 gene plays a role in releasing of the granulocytic differentiation blockage caused by MLL/AF10(OM-LZ) is under investigation. Nevertheless, line MA6 and the other immortalized cell lines all induced a myeloproliferative disease (MPD)-like myeloid leukemia when they were transplanted into the sublethally-irradiated syngeneic mice. Further studies revealed that the excessive mature granulocytes in the peripheral blood of leukemic mice were generated from the recipient hematopoietic stem/progenitor cells. On the other hand, the hyperleukocytosis found in line MA6 leukemic mice may be partly attributed to the expansion and maturation of the immortalized cells. RT-PCR analysis revealed that the MLL/AF10(OM-LZ)-immortalized cells expressed mRNAs encoding colony-stimulating factors and inflammatory cytokines. The latter were known to be able to stimulate or up-regulate endothelial cells, fibroblasts, T-lymphocytes, granulocytes and monocytes to produce GM-CSF, G-CSF or M-CSF, and, therefore, indirectly regulate myelopoiesis. Our results demonstrated that although the MLL/AF10(OM-LZ)-immortalized cells could not terminally differentiated into granulocytes, they could still induce a MPD by expression and secretion of the colony-stimulating factors and cytokines to over-stimulate the myelopoiesis of the recipient hematopoietic stem/progenitor cells.
Bcl-2 Protects against p53-Induced Apoptosis through Hdm2
Authors: Wergeland Line , Spurgers Kevin B. , Oveland Eystein , Høiby Torill , Cascallo Manel , Honda Tsuyoshi , Lorens James B. , Lozano Guillermina , McDonnell Timothy J. , Gjertsen Bjorn T. 
Published: April 2020

Abstract
Hdm2 is up-regulated in several malignancies including sarcomas and acute myeloid leukemia, where it counteracts the anti-proliferative and pro-apoptotic effect of wild type p53. The anti-apoptotic protein Bcl-2 is often elevated in many tumors with wild type p53 and serves to block p53-induced apoptosis. We demonstrate that the protein level of Hdm2 positively correlates with the level of Bcl-2 and follows the Bcl-2 level in different cell systems. Over-expression of Bcl-2 protects Hdm2 from DNA-damage induced degradation in a dose dependant manner. In addition, modulation of Bcl-2 by shRNA knockdown reduced the Hdm2 protein level in parallel. Consequently, treatment of AML cells with the Bcl-2 small inhibitory molecule HA14-1 attenuated the level of Hdm2. The Bcl-2 level, but not the DNA damage induced Hdm2 degradation, was affected by disruption of the E3 ubiquitin ligase activity of Hdm2. In addition, the DNA-damage induced Hdm2 down-regulation was blocked by disrupted E1 ubiquitin-activation, defect polyubiquitination and by proteasome inhibitors. Finally, we show that Bcl-2 protection from p53-induced cell death requires co-expression of Hdm2 in double null p53/mdm2 mouse embryonic fibroblasts. Our results indicate that Bcl-2 regulates the Hdm2 level and that Hdm2 is a key mediator in Bcl-2 inhibition of p53-induced apoptosis. This is of particular therapeutic interest for cancers displaying elevated Hdm2 and Bcl-2, like sarcoma and acute myeloid leukemia.
Role of PI3-K and Mlck during Activation of Thrombin Receptors
Authors: Gao Haili , Sun Aining , Han Yue , Zhang Wei , Dai Lan , Bai Xia , Wang Zhaoyue , Depei Wu , Ruan Changgeng 
Published: April 2020

Abstract
Object In order to compare the influences of wortmannin on platelet aggregation and platelet membrane surface glycoproteins GPIb expression after thrombin receptors activation, then to investigate the role of phosphatidylinositol 3-kinase (PI3-K) and myosin light chain kinase (MLCK) in the course of thrombin receptors activation.
Methods Peptide SFLLRN (PAR1-AP) and AYPGKF (PAR4-AP)were used for stimulating platelet, then the alterations of platelet aggregation and GPIb were analyzed in the involvement of 100nM wortmannin (inhibitor of PI3-K) and 10μM wortmannin (inhibitor of MLCK).
Results Platelet activation was influenced by either concentration of wortmannin in response to PARs stimulation. Platelet aggregation was apparently inhibited by 10μM wortmannin following both peptides, and slightly inhibited by 100nM wortmannin only upon PAR1-AP activation. In addition, GPIbα interalisation is partly inhibited by 100nM wortmannin in response to PAR1 (P &lt;0.05 at 1, 2, 5min) and PAR4 (P &lt;0.05 at 2, 5, 10min) activation. Meanwhile, 10μM wortmannin induces little alteration for GPIbα centralisation in the course of PAR activation, with a delayed restoration of surface GPIbα observed upon PAR1-AP activation, and no change of GPIbα redistribution exists upon PAR4-AP activation.
Conclusion All the results confirm different roles of PI3-K and MLCK in the course of thrombin receptors activation. PI3-K accelerates the short course of GPIb centralisation for two PARs signal pathways, while MLCK inhibites the restoration of GPIbα in PAR1 pathway
Evaluation of the Role of Flow Cytometry as a Diagnostic Tool for Hereditary Spherocytosis
Authors: Pati Nalini K , Sartor Mary M , Wong Sue , Nemet Adrienne , Bradstock Kenneth Francis 
Published: April 2020

Abstract
Flow cytometric analysis of eosin-5′-maleimide-labeled red blood cells has been proposed as a method of identifying hereditary spherocytosis (HS). The flow cytometric test measures the fluorescence intensity of intact red cells labelled with the dye eosin-5-maleimide (EMA), which reacts covalently with Lys-430 on the first extra cellular loop of band 3 protein. Patients with HS have reduced fluorescence compared to other patient groups and normal controls. The aim of the present study was to assess the utility of flow cytometry in the diagnosis of hereditary spherocytosis. Fresh peripheral blood from 40 normal controls was collected in Lithium Heparin, stained with the dye EMA and analysed by flow cytometry. The normal range was established as 55–74 channel numbers. On 7 known cases of Hereditary Spherocytosis the MFI range was 35.6–44.6 channel numbers, and therefore samples falling into this range were considered to be positive for a diagnosis of HS by EMA. Equivocal results were defined when the MFI was in the range of 45–54 channel numbers. A total of 98 samples were sent for investigation or exclusion of HS. Clinical problems were grouped into the following categories: peripheral blood spherocytosis with negative Coomb’s test and no family history of HS, positive family history of HS, Neonatal hyperbilirubinaemia, haemolytic anaemia of unknown origin.
Indications Coomb’s −ve, Spherocytosis, no FH (n=50) (51%) Positive family history (FH) (n=18) (17%) Neonatal hyperbilirubinemia (NNH) (n=8) (8%) Haemolytic anaemia (HA) (n=16) (17%) Results of EMA Pos Neg Equi Pos Neg Equi Pos Neg Equi Pos Neg Equi Number 19 17 13 9 7 2 2 4 2 0 14 2 Percentage 38 34 26 50 39 11 25 50 25 0 87.5 12.5
Within this cohort the group with FH of HS had the highest positive and least equivocal results. The group with suspected HS (peripheral blood spherocytosis with negative coomb’s test) resulted in positive in 38% and equivocal in 26% cases. The group with NNH had positive results in 25% of cases and the group with HA had no positive results. Equivocal results occurred in 20% of cases suggesting further investigations are required to confirm or exclude HS. The EMA dye method by flow cytometry is a useful test for diagnosing red cell membrane abnormalities due to band 3 defects, but the assay produces a substantial number of equivocal results, and has poor utility in the investigation of haemolytic anaemia of unknown origin.
The Antioxidant Effects of Capparis Ovata and Deferasirox in Patients with Thalassemia Major
Authors: Canatan Duran , Duman Handan 
Published: April 2020

Abstract
Introduction: Iron overload and auto-oxidation of unpaired globin chains is the main cause of oxidative stress in thalassemia. The high levels of MDA in iron overloaded thalassemic patients is the best sample of oxidative stres. Generally decreased iron burden was associated with decreased oxidant damage. In vitro studies were shown that iron chelators such as deferoxamine and deferipron neutrolyse intraselluler free iron and inhibits oxidation. We aimed to show the antioxidant effects of capparis ovatta and deferasirox in thalassemic patients.
Material-Method: A total number of 40 thalassemia major patient aged between 7–30 years, who has been taken regular transfusion (15 cc/kg/month to maintain Hb &gt;10 gr/dl) and chelation therapy (30 mg/kg/day ICL-670) are involved. They were separated into two groups as control and study group randomly. Both study and control group were followed by transfusion and chelation therapy. In study In addition study group have been taken capparis marmelade at the breakfast with a dose of a dessert-spoon (12.5 gr) younger than 10 years and a soup-spoon (25 gr) older than ten years for 6 months. Hematological and biochemical parameters, ferritin at every month and oxidative-antioxidant status (MDA, CAT, Gpx, SOD) were measured at the beginning and at the end of the study.
Results: Serum ferritin and MDA levels declined significiantly in both groups (for ferritin; control group p= 0.00; study group p=0.00) during the study but a much more decrease occured at MDA levels in the cappari given group (p=0.02). There was no statistically significant difference between the groups at the initial and last SOD CAT, GPX, SOD levels. Further more in the study group a significant decrease in liver function tests was occured (AST p= 0.05, ALT p= 0.01).
Conclusion: Our findings suggest that combination of capparis with deferasirox may be usefull for decreasing the effect of oxidative damage.
Vaccination with Leukemia Cell Expression Membrane Bound GM-CSF Overcomes the Host Immunosuppression Induced by Leukemia
Authors: Ling Di , Arlinghaus Ralph B. , Ling Xiaoyang 
Published: April 2020

Abstract
Immunotherapy of leukemia involves stimulating host-cell mediated immunity by facilitating immune recognition of leukemia cells, which are normally weakly immunogenic. We previously showed that vaccination with membrane bound GM-CSF leukemic cells protects mice from leukemia challenge (Ling et al., Oncogene, 2007). In these studies, after addition of a transmembrane domain to the original GM-CSF coding sequence (tmGM-CSF), the construct was transduced into murine leukemia cells (WEHI-3B), which was shown to be more than 98% on the cell surface. Vaccination with lethally irradiated tmGM-CSF expressing murine leukemia cells prevents leukemia in immunocompetent mice (BALB/c), as 100% of vaccinated BALB/c mice were protected from leukemia (Ling et al, Oncogene 2007). No protection was observed by vaccination of nude mice, indicating that adaptive immunity is involved in the protective response. In the present studies, we extended our original observation and provided evidence to show that leukemic mice undergo immunosuppression and that vaccination with leukemia cells expressing cell surface tmGM-CSF overcomes immunosuppression. Vaccination with lethally irradiated leukemia cells expressing cell surface tmGM-CSF overcame the immunosuppression induced by leukemia development, as normal levels of CD4+/CD25+/Foxp3+ T-regulatory (Treg) cells were maintained in spleens and thymus after challenge with leukemia cells. In contrast, the Treg population was significantly increased in leukemic mice vaccinated with leukemia cells lacking cell surface tmGM-CSF (p&lt;0.001) after leukemia challenge, and these mice had a lower CD8+/Treg cell ratio (p&lt;0.01). The ratio of CD8+/Treg cells was higher in tmGM-CSF/GFP vaccinated mice than in GFP vaccinated mice (p&lt;0.001), which in-turn leads to a more effective CD8+ T-cell response. DC levels were also increased from normal levels in mice vaccinated with tmGM-CSF+ leukemia cells compared to control vaccinations. These results suggest that vaccination with leukemia cells expressing GM-CSF on their cell surface leads to an effective cell-mediated immune response in the vaccinated host by overcoming an impaired host cellular immunity induced up-regulation of Treg cells caused by the leukemia process. This strategy has potential for use in the treatment of various human leukemias.