bluuebunny/crossref_metadata_2025_split huggingface dataset:
Influence of Valproic Acid on Cytokine Expression of Spleen Cells and Acute Graft-Versus-Host-Disease in a Murine Model.
Authors: Jang Pil-Sang , Lee Dae-Hyung , Park Young-Shil , Chung Nak-Gyun , Cho Bin , Jeong Dae-Chul , Kim Hack-Ki 
Published: April 2020

Abstract
Inhibition of histone deacetylase, one of the mechanisms for the immunologic changes by antiepileptic drugs, induces gene expression which, in turn, alters cytokine secretion. Valproic acid (VPA), the most commonly used antiepileptic drug, inhibits TNF-α and IL-6 secretion induced by PHA in THP-1 cells, a human monocytic leukemia cell line. We examined the changes in cytokine secretion induced by differing concentrations of VPA and whether VPA could prevent lethal GVHD in murine allogeneic bone marrow transplantation (BMT).
For the evaluation of inhibitory effect of VPA, C3H/He (H-2k) splenocytes, irradiated BALB/c (H-2d) splenocytes, and PHA were cocultured with 0.1 mM, 1 mM, and 10 mM VPA. Concentrations of 0.1 mM and 1 mM VPA did not inhibit cell proliferation but 95% was inhibited at 10 mM. Apoptosis was induced more than 99% cells with 10 mM VPA but less than 5% of cells with 0.1 mM, and 1 mM. RT-PCR was performed to evaluate the changes and concentrations of each cytokine. TNF-α and IL-1β secretion, determined by the RNA quantity, was inhibited significantly more at 1 mM VPA than 0.1 mM (P&lt;0.05). Decrease in IFN-γ concentration and increase in TGF-β concentration were significantly more at 1 mM VPA than 0.1 mM (P&lt;0.05). However, differences in IFN-γ and TGF-β secretion, determined by the RNA quantity, were not statistically significant. Lethally irradiated BALB/c mice were transplanted with C3H/He bone marrow cells and splenocytes. Recipients were given 30 mg/kg and 100 mg/kg VPA intraperitoneally once daily from day −3 to day +14, respectively, whereas controls received phosphate buffered saline. There were no significant differences in the GVHD severity and mortality.
These results show that VPA inhibits the secretion of inflammatory cytokines and stimulates the secretion of an antiinflammtory cytokine without inhibition of cell proliferation. Although lethal GVHD was not prevented or controlled in a major and minor histocompatibility complex-mismatched murine allogeneic BMT model, further investigations concerning the effects of VPA on GVHD prevention using a congeneic murine model should be performed.
ATM and ATR Are Not Required for DNA Damage-Dependent Accumulation of Chk1 at the Centrosome.
Authors: Loeffler Harald , Bochtler Tilmann , Ho Anthony D. , Jeggo Penny A. , Bartek Jiri , Kraemer Alwin 
Published: April 2020

Abstract
ATR-Seckel syndrome and ataxia-teleangiectasia are autosomal recessive disorders associated with hematologic malignancies. They share the feature of genetic instability and are caused by defects in the DNA damage response: underlying causes are a hypomorphic ATR mutation and inactivating ATM mutations, respectively. A common substrate of ATM and ATR in the DNA damage response is the nuclear checkpoint kinase Chk1, which we have recently shown to localize to interphase centrosomes and thereby negatively regulate entry into mitosis by preventing premature activation of cyclin B-Cdk1 (Nat Cell Biol6: 884–891, 2004). Here, we demonstrate that DNA damage by ultraviolet radiation or hydroxyurea treatment enhances the centrosomal localization of Chk1. In fibroblasts derived from ATR-Seckel or ataxia-teleangiectasia patients, this DNA damage-dependent centrosomal accumulation of Chk1 was rather more pronounced in comparison with control fibroblasts. Accordingly, this cellular reaction is not dependent on ATR or ATM. These results indicate that centrosomal accumulation of Chk1 might contribute a novel regulatory mechanism to the DNA damage checkpoint. Our study might help to further dissect the contributions of ATR and ATM to the DNA damage response, and to better understand their role in the development of hematologic malignancies.
Both G0/G1 Cell Cycle Arrest and Continuous Activation of MAPK Induce Autophagic Cell Death in bcr-abl-Positive Leukemic Cells.
Authors: Mishima Yuko , Terui Yasuhito , Mishima Yuji , Takizawa Toshihiro , Kimura Shinya , Maekawa Taira , Hatake Kiyohiko 
Published: April 2020

Abstract
Back ground;
Accumulating evidence suggests that programmed cell death is not defined as apoptosis but cells use different pathways for active self-destruction as reflected by different morphology. Autophagic cell death (APCD), which also be designated type II programmed cell death and appears to be a phylogenetically old phenomenon, is observed in physiological and disease state. Indeed, APCD is observed in several neurodegenerative diseases and cancer cells, however, the APCD in leukemic cells has not been reported yet. Here, we found APCD was occurred in bcr-abl-expressing CML cell lines.
Methods;
Bcr-abl-positive cell line, K562, were cultured with 2nM of TPA up to 48hours. Subsequently, we examined the morphological change by light microscopy and electronmicroscopy. To evaluate the APCD, TPA-treated K562 cells were labeled with autofluorescent agent Monodansylcadaverin(MDC), which specifically accumulates in autophagosomes and we measured accumulation of autophagosmes in cytoplasm by fluorescent microplate reader. TPA continuous activated MAPK of K562 cells and induced cell cycle arrest in G0/G1 phase. In order to elucidate the role of activation of MAPK in APCD, MAPK in K562 cells were inactivated by MAPK inhibitor, U0126 or dominant negative MAPK-transfection, and then, the cells were treated with TPA. Furthermore, to evaluate the involvement of cell cycle arrest in APCD, bcr-abl-transfected murine leukemic cell line, BAF3, was treated with cell cycle arrest inducer, mimosine. In addition, we investigated the expression of an autophagy-related molecule, Beclin 1 by immunohistochemical analysis.
Result;
The 48 hours’ treatment with TPA induced cell death in K562 cells, which accumulated plenty of autophagosomes in cytoplasm. MDC labeling assay revealed that accumulation of autophagosomes were increased in time- and dose-dependent manner. On the other hand, MAPK-inactivated K562 cells exhibited resistance to TPA-induced APCD. Furthermore, the bcr-abl-transfected BAF3 cells exhibited continuous activation of MAPK and underwent APCD by cell cycle arrest by mimosine. Immunohistochemical analysis revealed that Beclin 1 shuttled between nuclear and cytoplasms and co-localized with MAPK during APCD progression.
Conclusion:
APCD in bcr-abl-positive leukemia cells were closely involved in G0/G1 cell cycle arrest and continuous activation of MAPK. These findings are useful in developing the novel strategy to treatment of Ph1 positive leukemia via alternative pathways and another type of cell death.
Frequency of Mesenchymal Colony-Forming Cells (CFU-F) from Human Cord Blood and the Umbilical Vein.
Authors: Oostendorp Robert A.J. , Mohaupt Nicole , Kaltz Nikolas , Hippauf Sandra , Peschel Christian 
Published: April 2020

Abstract
Umbilical cord blood (UCB) is frequently collected for the purpose of hematopoietic stem cell therapy. However, the use of these cells is infrequent. To investigate other possible applications we recently showed that UCB is an excellent source of endothelial colony-forming cells which improve vessel density and heart function ofter myocardial infarction (Ott, Keller, et al., FASEB J, 2005;19:992–4). We here investigated whether UCB or the umbilical cord would be suitable as a routine source of mesenchymal colony-forming cells (CFU-F). CD34+ and AC133+ cells were selected by MACS (Miltenyi) from mononuclear UCB cells. The resulting cells were cultured at limiting dilutions on fibronectin in medium with 2% FCS (BioWhittaker: EGM-2). CFU-F were isolated from 3 of 7 (42%) UCB donors investigated. However, the CFU-F frequency is extremely low: only 1.3 in 100 ml of UCB. Similraly to the endothelial cells we previously described, these few UCB-derived stromal cells (CBSC) could easily be expanded. In Parallel, we also investigated CFU-F from the human umbilical vein: human umbilical vein stromal cells (HuvSC). Here, CFU-F were isolated from all donors when veins were prepared within 6 hours of collection. The CFU-F frequency was estimated to be 3 to 5 per 10 cm of umblical vein. Like the UCB-derived CFU-F, HuvSC were easily expandable. To find out whether these stromal cells showed characteristics of immature cells, we investigated the presence of embryonic markers by RT- and realtime PCR. We confirmed the expression of POU5F1 (Oct4), SSEA-4 and Stella-related (DPPA3). Immature HuvSC were shown to express mRNA of many genes associated with mesenchymal lineages but did not express the hematopoietic markers CD45 or CD34. HuvSC were shown to differentiate into adipogenic, osteogenic and myogenic lineages, though their differentiation is not as pronounced as bone marrow derived MSC. HuvSC did also support long-term production of immature cobblestone area-forming cells week 6. In conclusion, UCB and the umbilical vein contain CFU-F with characteristics of immature mesenchymal stem cells. However, not all UCB collections contained CFU-F and the CFU-frequency is very low. Thus, UCB can, in our view, not be used as routine MSC source. On the other hand, CFU-F grew from small stretches of umbilical vein (HuvSC), making these cells excellent source of mesenchymal progenitors for tissue engineering purposes.
Hematopoietic Differentiation of ES Cells from P230 Bcr/Abl Transgenic Mouse.
Authors: Tanosaki Sakae , Inokuchi Koiti , Sawaguchi Kazuhiro , Takeuchi Jyunko , Tamai Masato , Uchida Naoya , Watanabe Ayako , Inami Mituharu , Yamaguchi Hiroki , Dan Kazuo 
Published: April 2020

Abstract
Chronic neutrophilic leukemia (CNL) is a distinct disease form with less aggressive phenotype mimicking chronic myelogenous leukemia (CML). The patients present lower white blood cell counts consisting principally of neutrophilia, more severe thrombocytosis, less severe anemia, and delayed or absent transformation to blastic crisis. CNL sometimes has the translocation (9; 22) results in the transcription of an e19/a2 type Bcr/Abl mRNA that codes for a 230-kD Bcr/Abl protein (P230). Recently, we generated the P230 Bcr/Abl-transgenic mice, whose disease phenotype is myeloproliferative disese (MPD) with thrombocytosis. To investigate the disease phenotype of p230 Bcr/Abl-expressing MPD, we derived embryonic stem (ES) cells from the P230 Bcr/Abl transgenic mouse and non-transgenic control mouse. We differentiated ES cells on OP9 stromal cells, which supportive for hematopoietic differentiation. After day 5, ES cells were differentiated into megkaryocytes with TPO, erythrocytes with EPO and myeloid cells without cytokines. At day 13, floating cells were analyzed with lineage specific markers of megakaryocyte (CD61), erythroid (TER119) and myeloid (CD11b) cells by FACS analysis. P230 Bcr/Abl ES cell had more tendency of myeloid differentiation than control ES cell. With methylcellulose colony assays, we found that P230 Bcr/Abl ES cell produced more colony forming units. Intriguingly, the number of myeloid colonies was significantly increased and that of erythroid colonies was slightly decreased with no significance. It is reported that P210 Bcr/Abl markedly suppressed erythroid colony development. These data suggested that P230 Bcr/Abl enhances myeloid development in hematopoiesis and that it suppresses erythropoiesis not as strongly as P210 Bcr/Abl.
Overexpression of MEL1 as a Novel Fusion Partner of AML1 in the Blastic Phase of Chronic Myeloid Leukemia with the Recurrent Cryptic Translocation t(1;21)(p36.3;q22).
Authors: Hazourli Sawcene , Chagnon Pierre , Fetni Raouf , Busque Lambert , Hebert Josee 
Published: April 2020

Abstract
Located at 1p36.3, MEL1 is a member of the MDS1/EVI1 gene family and encodes a zinc finger protein with a N-terminal PR-domain. It has been proposed that the overexpression of a modified version of MEL1 lacking the PR domain is oncogenic, whereas MEL1 retaining the PR domain is anti-tumorigenic. MEL1 is known to be overexpressed in some myeloid malignancies with reciprocal translocation t(1;3)(p36.3;q21) characterized by trilineage dysplasia and poor prognosis. It is suggested that the translocation of RPN1 gene at 3q21 in the vicinity of MEL1 gene might activate MEL1 expression through an enhancer element. Here we characterized a recurrent cryptic translocation t(1;21)(p36.3;q22) that fuses MEL1 to AML1 gene in a blastic transformation of chronic myeloid leukemia (CML). Fluorescence in situ hybridization (FISH) analysis with BAC/PAC clones revealed that the breakpoints are in intron 1 of MEL1 and between intron 1 and exon 8 of AML1. RT-PCR analysis showed that AML1-MEL1, but not the reciprocal MEL1-AML1 was expressed. Many splicing variants are present, and all fusions splice the 5′ end of AML1 that contains the RUNT domain with almost the entire MEL1. Furthermore two fusion transcripts contained open reading frames making possible the translation of two forms of AML1-MEL1 fusion proteins. To investigate if AML1-MEL1 leads to an inappropriate expression of MEL1 we performed a quantitative RT-PCR with primers outside and within the fused MEL1 allowing the detection of the normal and the rearranged allele respectively. Interestingly, our data show that while no normal MEL1 transcript was detected, there was an overexpression of the fused MEL1. These results suggest that similarly to AML1-EVI1 gene, overexpression of MEL1 could be regulated by the AML1 promoter in leukemic cells with the AML1-MEL1 fusion gene and might also play an important role in the progression of CML. Moreover, in contrast to previous studies showing an antioncogenic role for the PR domain, our findings indicate that in some leukemias, the overexpression of MEL1 is not restricted to the MEL1 PR-lacking form. This suggests that the mechanism by which the PR domain has his effect, is more complex than previously thought.
Molecular Characterization of NRG Gene, a Novel Partner, Fused to NUP98 Gene as a Result of the t(3;11)(q29q13;p15) Translocation in an Acute Myeloid/T Lymphocytic Leukemia.
Authors: Chen Sai-Juan , Zhu Yong-Jin , Pan Qin , Gu Bai-Wei , Xie Zhi-Yao , Cai Xun , Pan Jin-Lan , Wu Ya-Fang , Shi Jing-Yi , Li Guo , Xiong Shu-Min , Xue Yong-Quan , Chen Zhu , Chen Sai-Juan 
Published: April 2020

Abstract
The NUP98 gene has been reported to be fused with at least 17 partner genes in leukemia with 11p15 translocation. An adult patient with de novo acute myeloid/T lymphocytic leukemia harboring t(3;11)(q29q13;p15) has been investigated to characterize the genes involved in that translocation. Through molecular cytogenetic analysis, we identified a fusion transcript between NUP98 gene and a novel partner gene named as NUP98 related gene (NRG) at 3q29. Further molecular analysis showed that exon 13 of NUP98 was fused in-frame to exon 10 of NRG. Moreover, the segment from 3q13 to 3q29 translocated at 11p15 had been inverted and accompanied by the deletion of the distal portion of breakpoint at chromosome 3q29. Interestingly, the NUP98-NRG protein showed nuclear and cytoplasmic distribution, a pattern different from that of wild type NUP98 or NRG. When assayed in a GAL 4 reporter system, the fusion gene showed an aberrant trans-regulatory activity. Transfection in HL-60 cells demonstrated that NUP98-NRG could promote cell proliferation, survival and arrest differentiation. Therefore, NUP98-NRG may exert transforming effects by interfering with the cellular mechanism of transcriptional regulation. Our data provide thus new evidence that NUP98-related molecular abnormality is a recurrent genetic event in leukemogenesis.
Derivative (1) t(1;1) (p36; q21) Produces the Fusion Protein with Phosphatase 10 and MEL1 (MDS1/EVI1-Like Gene 1) in a AML (M5a) Case.
Authors: Noguchi Mitsuho , Tashiro Haruko , Shirasaki Ryosuke , Goto Moritaka , Kawasugi Kazuo , Shirafuji Naoki 
Published: April 2020

Abstract
Objective: A 25 years-old Japanese male was diagnosed as AML (M5a) with 46, XY, der (1) t(1; 1) (p36; q21). To determine the leukemogenesis in this case, we analyzed the fusion product of this chromosomal translocation.
Method: After obtaining informed consent, RNA was extracted from his bone marrow cells (blastic cells were occupied in almost 100%) with GTC method, and poly-A rich RNA was separated with oligo-dT latex particles. 1st strand cDNA was synthesized with oligo-dT primer. Oligo-dA stretch was added with TdT to make 5′ RACE. 1st PCR was performed with the synthetic antisense-1 primers from several candidates including p73, and MEL1, and with QT primer (5′-TGAGCCAGAGTGACGAGGACTCGAGCTCAAGCT17-3′) as a sense primer. 2nd PCR was performed with internal antisense-2 primers from several candidates, and Q0 primer (5′-CCAGTGAGCAGAGTGACG-3′) as the sense primer. The reaction of DNA sequencing was performed with BigDye Terminator Cycle Sequencing kit (Applied Biosystems), and analyzed with ABI Prism 3700 DNA analyzer.
Result and Discussion: The fusion cDNA product was obtained, which was consisted of MEL1 and Phosphatase 10. The breakpoint of MEL1 was exon 4, and PR domain of MEL1 was disrupted. This domain is reported to be the key function of MEL1, and when MEL1 without PR domain is expressed, myeloid differentiation is blocked. In this case the fusion of Phosphatase 10 and MEL1 is seemed to be the main cause of leukemogenesis.
Activation of the Unfolded Protein Response in Acute Leukemia and Myeloma.
Authors: Yujiri Toshiaki , Tanimura Atsuko , Tagami Kozo , Nakamura Yukinori , Tanizawa Yukio 
Published: April 2020

Abstract
Accumulation of unfolded or misfolded proteins within the endoplasmic reticulum triggers the unfolded protein response (UPR). Evidence from several studies suggests that the UPR is activated in various tumors and might play a crucial role in tumor growth. For example, in vitro activation of the UPR alters the sensitivity of tumor cells to chemotherapeutic agents. The role of the UPR in hematological malignancies remains unclear, however. We therefore used real-time RT-PCR to quantitatively assess expression of UPR-related genes in clinical samples [peripheral blood (PB), bone marrow (BM) and lymph node (LN)] from 9 newly diagnosed patients with acute myeloid leukemia (AML), 5 with acute lymphoid leukemia (ALL), 4 with multiple myeloma (MM), 12 with Non-Hodgkin’s lymphoma, 1 with Hodgkin’s lymphoma and 6 with infectious lymphadenopathy. Thirteen healthy controls were also examined. Evaluated was the mRNA expression of a spliced form of X-box DNA-binding protein (XBP1), ER degradation enhancing a-mannosidase-like protein (EDEM), C/EBP-homologous protein (CHOP) and glucose regulated protein 78 (GRP78). We found the transcription levels of all four of these genes to be significantly higher in leukemic blast cells and myeloma cells than in control BM cells or PB mononuclear cells. In particular, leukemic cells from Philadelphia chromosome-positive ALL and aggressive MM cells showed higher levels of UPR-related gene expression. Thus, activation of the UPR might be associated with the malignant characteristics of leukemia and myeloma. By contrast, expression of UPR-related genes was not increased in lymphoma cells compared to LN cells from the patients with infectious lymphadenopathy.
The present study provides the first evidence that UPR-related genes are activated in acute leukemia and multiple myeloma cells. Thus, activation of the UPR might play an important role in the development and progression of some hematological malignancies. More comprehensive in vitro studies will be necessary to determine the magnitude of the role played by the UPR, to identify the most important components of the pathway, and to determine which stages of tumor development are regulated by the UPR.
Analysis of IGFBP7 Expression Level and Its Correlation with Associated Genes in AML Patients.
Authors: Hu Shao-yan , Chen Zi-xing , Cen Jian-nong , Gu Wei-ying , Zhao Ye , Gu Min 
Published: April 2020

Abstract
IGFBP7 (insulin-like growth factor binding protein 7) is a newly-identified protein which can bind with insulin-like growth factors(IGFs), with 45%~65% homology to other IGF binding proteins. Several studies reported that IGFBP7 may be involved in the development of meningioma, breast cancer and prostate cancer, while its correlation with human leukemia had not been reported yet. We have observed an induced expression of IGFBP7 in human leukemia K562 cells which were transfected with and overexpressed RbAp46 gene. Besides, the expression level of WT1, RbAp46 and IGFBP7 was coordinately regulated during the induced differentiation of NB4 acute promyelocytic leukemia cells (data not shown), suggesting a role of IGFBP7 gene in human leukemia.. To investigate the expression level of IGFBP7(previous known as Insulin-like growth factor binding protein related protein 1)in bone marrow (BM) of leukemia patients and its correlation with other leukemia biomarkers, Real-time quantitative reverse transcription polymerase chain reaction (QRT-PCR) method was established for detecting IGFBP7 expression levels in BM of 127 patients with acute leukemia (AL), 24 with chronic myelogenous leukemia in chronic phase (CML-CP), 9 with CML in blast crisis (CML-BC) and 27 with non-leukemias. The M-Estimators of RbAp46N in 63 patients with newly diagnosed AMLs were higher than those of 27 with ALLs and 27 non-leukemic controls and 10 with AMLs in complete remission(CR)(193.80 v.s 88.84, 81.30 and 67.82, respectively). Although IGFBP7N was 101.13 in CML-BC and 168.77 in CML-CP, difference between them was not statistically significant. Difference in IGFBP7N values among CML-BC, AML, ALL and control groups was also not significant. The M-Estimators of IGFBP7N were lowest in M4 and highest in M5 among initial acute leukemia subtypes, and there was statistical differences between M4 and M5, and between M4 and M2. IGFBP7N expression level was not correlated to the expression of fusion genes BCR/ABL and AML1/ETO, but it was positively correlated with the expression level of PML/RARαfusion gene, the WT1 and RbAp46 gene. The correlation coefficiency was 0.71, 0.31 and 0.52, respectively. Our results suggested that IGFBP7 might act in WT1 mediating pathway to participate in acute myelocytic leukemias.
Tax Ubiquitylation and Sumoylation Control the Two Distinct Steps of NF-κB Activation.
Authors: Nasr Rihab , Chiari Estelle , El-Sabban Marwan , Mahieux Renaud , Kfoury Youmna , Abdulhay Mohammad M. , Hermine Olivier , de Thé Hugues , Pique Claudine , Bazarbachi Ali 
Published: April 2020

Abstract
Activation of the NF-κB pathway by the HTLV-I oncoprotein Tax plays a mandatory role in the proliferation and transformation of infected T cells. We have previously demonstrated that Tax is ubiquitylated, mostly on lysine residues located in its carboxy-terminal half. In this study, we investigated the contribution of Tax ubiquitylation on its ability to transactivate the NF-κB pathway. We show that the integrity of C-terminal lysines, and Tax ubiquitylation, are critical for Tax binding to IKK, IKK activation and nuclear translocation of NF-κB. We also report that Tax is post-translationally modified by SUMO on lysine residues also targeted by ubiquitin and that Tax sumoylation occurs in the nucleus and is required for the terminal steps of NF-κB activation. Conversely, Tax ubiquitylation and sumoylation are not involved in CREB activation. Thus, the differential ubiquitylation or sumoylation of the same lysines in Tax regulates essential events controlling the NF-κB pathway, revealing how distinct cellular modifications enrich the functions of this versatile oncoprotein.