bluuebunny/crossref_metadata_2025_split huggingface dataset:
L-Gossypol Has Significant In Vitro Activity Against Primary Acute Myeloid Leukaemia Cells, Inducing Apoptosis through the Intrinsic Pathway.
Authors: Osman Hani Y. , Knapper Steven , Burnett Alan K. 
Published: October 2019

Abstract
The potential anti-tumor effect of the polyphenolic cotton oilseed extract gossypol has been known for a number of years. Studies in cell lines and primary tumor material have suggested both antiproliferative effects and the induction of apoptosis (Huang et al. Anticancer research 26:3a) with activity being seen against even multidrug resistant tumors. Although the mechanism of action of gossypol has not yet been well-characterised, there is some evidence that it may block the binding of the apoptotic regulator BCL-2 with the pro-apoptotic proteins BAX and BAD (Prada et al. blood 106:11). Acute myeloid leukaemia (AML) is characterised by accumulation of primitive blasts in bone marrow and peripheral blood. The disease is highly heterogeneous and inherently resistant to therapy, dysregulation of apoptosis frequently leading to a survival advantage in blast cells. We studied the in vitro activity of gossypol against cryopreserved primary blasts obtained from 44 AML patients. Cells were incubated for 48 hours with a range of concentrations of gossypol, this being followed by MTS cytotoxicity assay. IC50 concentrations ranged from 792nM to in excess of 100 mM (median 22.791 mM). Statistical analysis showed no significant variation in drug sensitivity according to sex, age, presenting white cell count, FLT3 mutation status, karyotype or clinical outcome measures. The in vitro combination of gossypol with the cytotoxic agent cytosine arabinoside (AraC) was assessed in 40 primary samples using the median effects principle of Chou and Talalay. The median combination index of 0.946 suggested an overall additive effect when the agents were combined, although numbers were insufficient to establish this conclusively (95% CI for median 0.84–1.31). Flow cytometric annexin V binding assay confirmed apoptosis to be the principal mode of cell death in response to gossypol, cell viability obtained using this method correlating well with that obtained using the MTS assay. Gossypol caused downstream activation of Caspase 3/7 via Caspase 9 as determined by luminometric analysis, indicating that the intrinsic pathway is the main inducer of this cycle. Gossypol is effective in inducing apoptosis in primary AML blasts and there is potential for greater efficacy when used in combination with cytotoxic chemotherapy. Further in vitro development of this agent is now required to further define its future clinical role.
Metalloproteinase Inhibitors Increase the Survival of Long-Term Refrigerated Platelets in Mice.
Authors: Josefsson Emma C. , Rumjantseva Viktoria , Dahlgren Claes , Bergmeier Wolfgang , Wagner Denisa D. , Hartwig John H. , Hoffmeister Karin M. 
Published: October 2019

Abstract
The αMβ2 receptor on macrophages, which recognizes clustered β-N-acetylglucosamine (βGlcNAc) residues on glycoprotein (GP) Ib-V-IX, is one phagocytic receptor involved in the removal of transfused refrigerated platelets. Although coverage of the exposed βGlcNAc residues by galactosylation prevents the clearance of murine platelets refrigerated for less than 4h, it does not impact the clearance of platelets refrigerated for 48h, suggesting that further changes occur in platelets during prolonged refrigeration. We have now found that murine GPIbα and GPV are partially shed from the platelet surface after refrigeration for 48h and re-warming. Other members of the complex, GPIX and GPIbβ, were not affected. Platelets isolated from mice with inactive ADAM17 enzyme, a GPIbα and GPV sheddase, had preserved surface GPIbα and decreased GPV shedding compared to WT mice after long-term refrigeration. 48h refrigerated platelets with inactive ADAM17 or platelets treated with the metalloproteinase inhibitor GM 6001, had increased survival when transfused into WT mice. Metalloproteinase inhibitor-treated and galactosylated, 48h refrigerated and re-warmed mouse platelets exhibited survivals equivalent to fresh platelets. We conclude that 48h long-term refrigeration and re-warming of platelets induces shedding of GPIbα and GPV in murine platelets, and that metalloproteinase inhibitors increase the survival of long-term refrigerated platelets in mice. Further changes in GPIb-V-IX in long-term refrigerated platelets may recruit additional phagocytic receptors to promote clearance, in addition to the αMβ2 receptor.
The Expression and Clinical Significance of PRAME Gene in Acute Leukemia.
Authors: Fu Rong , Ding Kai , Shao Zonghong 
Published: October 2019

Abstract
Objective To investigate the expression of PRAME (preferentially expressed antigen of melanoma) gene in acute leukemia and its clinical significance in monitoring prognosis, detecting minimal residual disease (MRD) and gene immunotherapy.
Methods The expression of PRAME gene mRNA in bone marrow mononuclear cells is measured by reverse transcriptase polymerase chain reaction in 34 patients with acute leukemia and 12 bone marrow samples of health donors. The relationships between PRAME gene expressions and some clinical data, such as gender, age, white blood count, leukemic immunophenotype, the percentage of blast cells, and the karyotype of chromosome, were also estimated.
Results PRAME gene was expressed in 38.2% of all the patients, 40.7% of all the AML patients, which was higher than the 28.6% of ALL patients (p &gt;0.05). There was no expression of PRAME gene in healthy donors. In all the sub phenotypes of AML, the expressive rate of PRAME gene in M3 patients is 80%, which is higher than that in M2 (33.3%) and in M5 (28.6%). The expressive rate of PRAME gene was also positively correlated with the expression of CD15, CD33, and the abnormality in the karyotype of chromosome, but not correlated with age, gender, white blood count and percentage of blast cell in bone marrow.
Conclusion PRAME gene is highly expressed in acute leukemia, and could be regarded as a useful tool for monitoring MRD. Differential expression in acute leukemia patients vs. healthy donors suggests that the immunogenic antigens PRAME are potential candidates for immunotherapy in acute leukemia.
Prognostic Impact of Cytogenetic Abnormalities in Elderly Patients with Acute Myeloid Leukemia (AML) Enrolled in the ALFA-9803 Trial.
Authors: Terré Christine , Gardin Claude , Gachard Nathalie , Tigaud Isabelle , Désangles François , Bastard Christian , Maarek Odile , Lai Jean-Luc , Bernheim Alain , Perot Christine , Nguyen-Khac Florence , Eclache Virginie , Plessis Ghislaine , Turlure Pascal , Thomas Xavier , de Revel Thierry , Fenaux Pierre , Dombret Hervé 
Published: October 2019

Abstract
We have recently reported the superiority of prolonged ambulatory versus short intensive post-remission therapy in 416 patients with AML aged 65 years or more (median age, 72 years) selected to be treated intensively in the ALFA-9803 trial (C. Gardin et al. Blood 2007). We report here on the prognostic value of cytogenetic abnormalities in 339 of these patients with available karyotype (27 not done, 50 failures). When classical endpoints (CR rate, DFS, OS) were used, we were only able to identify one very unfavorable subgroup of 14% patients (N=49 with −7, abnormalities of both chromosomes 5 and 7, 3q abnormality, or complex karyotype with 5 abnormalities or more; CR rate, 30%). No significant differences in outcome were found between patients with any other abnormality and the 171 patients with a normal karyotype/−Y (overall CR rate, 62%). When cumulative incidences of AML-related events (primary refractory AML, relapses) were used as endpoints with induction deaths and deaths in first CR as competing events, three distinct subsets were more easily identified:
14% very unfavorable-risk patients, as detailed above; 59% standard-risk patients (N=200 with t(8;21), inv(16), normal karyotype/−Y, or isolated +8; CR rate, 65%); and 27% unfavorable-risk patients (N=90 with other abnormalities, including notably 5q− and 7q−; CR rate, 58%).
Median failure-free interval (Fig 1; P<.001, by the Gray test) and relapse-free interval (Fig 2; P<.001, by the Gray test) were 2.3, 12.4, and 8.1 months and 4.2, 15.0, and 10.5 months in these three subsets, respectively. Interestingly, prolonged ambulatory post-remission therapy was associated with longer DFS and OS in the standard-risk subset only. The results demonstrate that despite their poor general outcome, older patients with AML may be subdivided in three meaningful cytogenetic subgroups. This 3-subgroup classification should help to optimize disease management decisions such as intensive versus non-intensive or investigational therapies for induction or consolidation.
1. Failure-free interval 1. Failure-free interval 2. Relapse-free interval 2. Relapse-free interval
Flow Cytometric Examination of Non-Hematic Body Fluids in Hematologic Malignancies: A Comparison with Cytology.
Authors: Cesana Clara , Scarpati Barbara , Brando Bruno , Barba Claudia , Ferri Ursula , Scampini Linda , Oreste Pierluigi , Veronese Silvio , Nosari Annamaria , Morra Enrica , Cairoli Roberto 
Published: October 2019

Abstract
Introduction: Flow cytometry is well known to detect malignant cells in peripheral blood and bone marrow of patients with hematologic malignancies. However, its role in evaluating non-hematic body fluid contamination by tumor cells is largely unexplored.
Patients and Methods: Data detected by flow cytometry in non-hematic body fluid samples drawn between 2002 and 2007 from patients with hematologic neoplasms were retrospectively compared with morphological findings obtained from cytospin slides. Immunophenotyping was carried out by using disease-specific multicolor panels of quadruple monoclonal antibodies, conjugated with the fluorochromes FITC, PE, PerCP, and APC, respectively. Acquisition of information on 1x104 to 1x105 stained cells depending on the whole sample cellularity was assessed on a dual-laser FACSCalibur flow cytometer using the CellQUEST software (Becton Dickinson, San José, CA, USA).
Results: Fourty-five samples (bronchoalveolar fluid, n=5; ascites, n=2; hydrocele, n=2; pleural effusion, n=8; and cerebrospinal fluid, n=28) from 32 patients were available for comparison. Diagnoses were as follows: chronic myelomonocitic leukemia (n=1), acute promyelocytic leukaemia (n=2), acute myelomonocytic leukaemia (n=1), B-chronic lymphocitic leukemia (n=4), follicular lymphoma (n=1), acute lymphoblastic leukaemia (n=6), lymphoblastic lymphoma (n=1), high grade non-Hodgkin’s lymphoma (NHL), Burkitt-like (n=3), diffuse large B-cell NHL (n=6), peripheral T-cell NHL (n=3), and NHL, unspecified (n=4). Flow cytometry detected neoplastic cells in 24 cases. Of these cases, only 17 were positive also by morphology. In 7 cases, in which tumor cells were detected by flow cytometry but not by morphology, clinical data confirmed the presence of the disease. Flow cytometry did not show neoplastic cells in 21 cases. Of these cases, only 18 were negative also by morphology. In the remaining 3, the suggestion of diffuse large B-cell NHL contamination by morphology was not confirmed by flow cytometry, demonstrating T-reactive lymphocytes that were clearly negative for disease-specific markers.
Conclusions: Our data suggest that flow cytometry is a useful tool complementary to morphology for the screening of non-hematic body fluid contamination in patients with hematologic neoplasms.
High-Resolution Analyses of Genetic and Epigenetic Aberrations in Infant Leukemia with MLL Rearrangement.
Authors: Takita Junko , Kato Motohiro , Nakamura Fumihiko , Chen Yuyan , Yamamoto Go , Nannya Yasuhito , Sanada Masashi , Taki Tmohiko , Kikuchi Akira , Igarashi Takashi , Hayashi Yasuhide , Ogawa Seishi 
Published: October 2019

Abstract
MLL rearrangement-positive leukemia is one of the most aggressive types of leukemia. It is diagnosed predominantly in infants and typically shows a multilineage phenotype. Since current chemotherapy fails in more than 50% of infantile leukemia with MLL rearrangement, a better understanding of biological features of the disease is importantly in order to develop more specific and successful treatment strategies. Thus, to explore both genetic and epigenetic lesions associated with MLL rearrangement-positive infantile leukemia, we performed genome-wide analyses of copy number alterations/allelic imbalances as well as methylation analysis using Affymetrix GeneChip SNP genotyping microarrays and promoter tiling array combined with methylated DNA immunoprecipitation (MeDIP). Combined with newly developed algorithm, CNAG/AsCNAR, SNP array analysis enables accurate copy number analysis at extremely high resolutions. In addition, by sensing subtle distortions in allele-specific signals caused by allelic imbalance using anonymous controls, sensitive detection of LOH is enabled with accurate determination of allele-specific copy numbers even in the face of up to 70–80% normal cell contamination. In total, 25 specimens from MLL rearrangement-positive leukemia were analyzed using high-density SNP-genotyping microarrays (Affymetrix GeneChip 100K/500K arrays). While unbalanced translocation involving chromosome 11q23 is the most frequent genetic alterations, uniparental disomy of 17q is also common genetic alterations in MLL rearrangement-positive leukemia. A number of other genetic changes were also identified but these were mostly found in a single case. On the other hand, epigenetic abnormalities have been implicated in MLL rearrangement-positive leukemia, because MLL is known to be involved in epigenetic regulations. In our assay, fragmented genomic DNA from leukemia specimens was immunoprecipitated with anti-methylcytosine antibody (MeDIP). The immunoprecipitated DNA was amplified by PCR and subjected to hybridization to the promoter tiling array, in which regulatory regions of more than 25,000 genes are tiled by 6.5 millions of oligonucleotide probes to enable sensitive detection of target sequences. This tiling array covers approximate 59% of CpG islands in the human genomes. In addition to the previously described genes, such as FHIT and HOX family, a number of tumor specific methylated sites were identified in the leukemic cells and which were subsequently verified by bisulfate sequencing. These results indicated that high-resolution analyses of genetic and epigenetic aberrations using microarray techniques are powerful and useful for detection of new findings in the pathogenesis of infant leukemia with MLL rearrangement.
Whole Genome Association Study in Acute Myeloid Leukemia with a Normal Karyotype, Using a Single-Nucleotide Polymorphism (SNP) Analysis.
Authors: Cha Yongjun , Ahn Kwang-Sung , Park Juwon , Kim Byung-Su , Bang Soo-Mee , Kim In-Ho , Yoon Sung-Soo , Lee Dongsoon , Lee Jong Seok , Lee Young Yiul , Park Seonyang , Kim Byoung Kook 
Published: October 2019

Abstract
As a result of numerous genetic aberrations, acute myeloid leukemia (AML) present with very heterogeneous clinical features. Moreover, structural and numerical chromosome aberrations currently comprise the most important basis for predicting these heterogeneous outcomes. However, 30 to 50 percent of AML patients have cytogenetically normal karyotypes. Though submicroscopic genetic alterations (such as NPM1, CEBPA, MLL-PTD) are increasingly being used for clinical purposes, additional markers with prognostic or predictive value are still lacking in this group. In this study, we performed a genome-wide, single-nucleotide polymorphism (SNP) study in order to identify novel genomic regions of interest in normal karyotype AML. 54 untreated AML patients with normal karyotypes were analyzed with an Illumina infinium 317K SNP chip assay. SNP genotype call rate was 99.8 percent, resulting in a resolution of 1 SNP/Mb. In a genome-wide SNP analysis, 317 SNP loci, having a significant difference between AML and normal control, were found. In addition, about 300 loci were also identified, showing a significant difference between patients who achieved CR and those who did not. Some of these were found to be related with drug metabolism and MDR family. Also we used the SNP assay to screen a loss of heterozygosity (LOH), suggesting possible involvement of tumor suppressor genes. In summary, 38 LOH regions larger than 1Mb were observed in 23 cases (43%) among 54 AML patients having normal karyotypes. Most (55%) of them were copy-neutral events and the most frequently identified alterations were located at 3p, 8p, 13q and 22q. 11 (29%) out of 38 LOH regions had overlapping parts among them. Some LOHs were correlated with response to chemotherapy. Various genetic changes were discovered by use of an SNP-based chip array in normal karyotype AML patients. Moreover, some of these were related with CR acquisition and drug metabolism. These results can be used to differentiate normal karyotype AML and warrant further study.
Essential Role of Pro-Apoptotic Mechanisms for Production of Normal Erythrocytes and Prevention of Hemolysis.
Authors: Chen Min , Prchal Josef T. , Hector Sandoval , Thiagarajan Perumal , Dasgupta Swapan K. , Schumacher Armin , Wang Jin 
Published: October 2019

Abstract
Prevention of apoptosis is a well-recognized mechanism essential for productive early erythropoiesis and preventing anemia; however, the importance of pro-apoptotic processes for productive terminal erythropoiesis is not defined. Erythroid cells undergo enucleation and the removal of organelles during terminal differentiation. Although autophagy has been shown to mediate the elimination of organelles for erythroid maturation, the molecular mechanisms underlying this process remain unknown. Here we report a novel role for Bnip3L, a pro-apoptotic Bcl-2 family member, in the regulation of erythroid maturation through autophagy. Bnip3L−/− mice developed hemolytic anemia and expansion of erythroid precursors (Fig.1A). Moreover, erythrocytes in the peripheral blood of Bnip3L−/− mice exhibited mitochondrial retention associated with reduced lifespan (Fig. 1B). While the clearance of ribosomes proceeded normally in the absence of Bnip3L, the entry of mitochondria into autophagosomes was defective (Fig. 1C). Overexpression of Bnip3L led to the loss of mitochondrial membrane potential (ΔΨm). Further, disrupting Δ Ψ m with an uncoupling chemical restored the sequestration of mitochondria into autophagosomes in Bnip3L−/− erythrocytes (Fig. 1C). Our study indicates that Bnip3L-dependent loss of Δ Ψ m is important for targeting the mitochondria into autophagosomes for degradation during erythroid maturation, and interference with this function impairs terminal erythroid maturation and results in hemolysis. These data underlie the importance of pro-apoptotic mechanisms for productive terminal erythropoiesis through autophagy.
Figure Figure
Emodin Induces Leukemic HL-60 Cells Apoptosis by Inhibiting PI3K/Akt Signal Pathway.
Authors: Hu Jian Da , Zheng He-yong , Zheng Zhi-hong , Huang Lv-ye , Lv Lian-huang , Chen Ying-yu 
Published: October 2019

Abstract
Emodin (1.3.8-trihydroxy-6-methylanthraquinone), a traditional Chinese herbal medicine, which is a natural anthraquinone compound from the root and rhizome of Rheum palmatum, has been shown to display a number of biological activities such as antiviral, antimicrobial, immunosuppressive, anti-inflammatory, and anticancer. Recent studies showed that it can inhibit proliferation of multiple cancer cells such as breast, hepatocellular, and lung cancer cells in vitro. The PI3K signal transduction pathway has been investigated extensively for its role in oncogenic transformation and in the prevention of apoptosis. The aim of the present study was to investigate the effects of Emodin on inhibiting cell growth and inducing apoptosis in human myeloid leukemia cell line HL-60, and to examine how emodin affects the PI3K/AKT signalling pathway, leading to cell apoptosis. HL-60 cells were exposed to various dosages of Emodin. MTT assay was used to detect HL-60 cell proliferation. Distribution of HL-60 cells in cell cycle was analyzed by flow cytometry and cell apoptosis was observed by MitoCapture apoptosis detection. The protein expressions of Akt signal pathway were detected by Western-Blot. The results showed that Emodin remarkably inhibited the cell proliferation, with 20μmol/L of the IC50 value for 48h. Apoptosis in HL-60 cells could be efficiently induced by Emodin in a dose dependent manner and cells were arrested at G0/G1. The expressions of Akt,p-Akt,IκB-α,p-IκB-α,p65,p-p65,mTOR and p-mTOR in PI3K/Akt signal pathway were downregulated after Emodin treatment. It could be concluded that Emodin can efficiently induce growth inhibition and apoptosis in HL-60 cells .Akt signal pathway may be involved in this process.
High INDO (Indoleamine 2,3-Dioxygenase) mRNA Level in Blasts of Acute Myeloid Leukemic Patients Predicts Poor Clinical Outcome.
Authors: Chamuleau Martine E.D. , Van de Loosdrecht Arjan A. , Hess Corine J. , Janssen Jeroen J.W.M. , Zevenbergen Adri , Delwel Ruud , Valk Peter J.M. , Lowenberg Bob , Ossenkoppele Gert J. 
Published: October 2019

Abstract
Indoleamine 2,3-dioxygenase (IDO) degrades the amino acid tryptophan which is essential for T cells. Tryptophan depletion causes T cell cycle arrest and solid tumors that express high levels of IDO can create immune suppression. Recently, blasts of acute myeloid leukemic (AML) patients were shown to express the IDO protein. We now determined INDO (the encoding gene for IDO) mRNA expression in myeloid leukemic blasts of 285 AML patients by gene expression profiling. Results were validated by quantitative PCR (qPCR) analysis in blasts of an independent cohort of 71 patients. Correlation of INDO expression to relevant known prognostic factors and survival identified high INDO expression as a strong negative independent predicting variable for overall and relapse free survival of AML patients. Inhibition of IDO may result in breaking tumor induced immune tolerance and offers new therapeutic options for AML patients.
Figure Figure
A Study of the Protein Phosphatase 2A Involved in the Induction of NB4 and MR2 Cell Differentiation by ATRA.
Authors: Ouyang Jian , Xu Xihui , Wu De Pei 
Published: October 2019

Abstract
Objective: To study whether protein phosphatases type 2A (PP2A) plays an important role in signal transduction of differentiation of NB4 induced by all-trans retinoic acid, furthermore, to evaluate whether PP2A plays an important role in MR2 resistance to ATRA.
Methods: ATRA, Okadaic acid (OKA) and both chemicals were incubated with NB4 and MR2 cells respectively. Wright’ staining and NBT reduction test were employed to evaluate the changes of cell phenotype. The expression of CD11b was measured by flow cytometer, and the proliferation of cells was measured by MTT method. At last the activity of PP2A was evaluated by Serine/Threonine phosphatase assay system.
Results: Results showed ATRA induced differentiation of NB4 significantly, but no effect on MR2 cell through Wright’ staining and NBT reduction test. OKA, the specific inhibitor of PP2A, could augment the differentiation induced by ATRA, and it combined with ATRA showed the slight induction of differentiation of MR2 cell. MTT showed the inhibitory rate of proliferation of ATRA, OKA and ATRA combined with OKA to NB4 were 25.1%, 10.1% and 38.4% respectively, and to MR2 were 4.8%, 7.2% and 19.3%. When ATRA, and ATRA combined with OKA were incubated with NB4 and MR2 respectively, the expression of CD11b on the surface of NB4 cell were 43.1% and 65.4% and on the surface of MR2 were from 0 to 9.3%. When OKA combined with ATRA were incubated with MR2, MR2 have showed the decreased activity of PPA. Conclusion PP2A plays an important role in the mechanism of differentiation of NB4 induced by ATRA and of resistance of MR2 to ATRA.