bluuebunny/crossref_metadata_2025_split huggingface dataset:
Study of In Vitro Proliferation Potential of CD133 Positive Cells Derived from Umbilical Cord Blood.
Authors: Zhang Ri , Gao Wenjin , Sun Yuanyuan , Miao Jingcheng , Zhang Xueguang 
Published: October 2019

Abstract
Transforming growth factor-beta 1 (TGF-β1) is known to maintain primitive human hematopoietic stem/progenitor cells with polyfunctional role in a quiescent state and CD133 is a new stem cell antigen that may provide an alternative to CD34 for the selection and expansion of hematopoietic cells for transplantation. To investigate the specific effect of TGF-β1 on proliferation and differentiation of CD133 positive cells derived from umbilical cord blood (UCB) during short-term culture in vitro, CD133 positive cells from 20 fresh UCB samples were selected using Miltenyi Biotec’s CliniMACS separation device and were cultured in IMDM medium with 20% FCS in the presence of a cytokine combination of SCF, IL-6, thrombopoietin, IL-3 and Flt3-ligand for up to 2 weeks and TGF-β1 with low concentration was also added to the mediumon day 4. The proliferative response was assessed at day 7, day 10 and day 14 by evaluating the following parameters: nucleated cells (NC), clonogenic progenitors (CFU-GEMM,CFU-GM and BFU-E), and immunophenotypes (CD133 and CD34). The results showed that efficacious expansion of various hematopoietic stem/progenitor cells was constantly observed during the culture. The fold expansion of NC on day7, day10 and day14 expansion were 33.59,224.26 and 613.48, respectively. The fold expansion of CFU-GEMM, CFU-GM and BFU-E on day 10 were 24.89, 41.62 and 49.28, respectively, obviously higher than that without ex vivo expansion (P<0.05). The expansions of CD133+, CD133+CD34+ and CD34+ subpopulation on day 14 were up to 25.83-fold, 16.16-fold and 60.54-fold, respectively. Furthermore the expansion systems with TGF-β1 showed more CD133+ cells than control at every time points. Our datas suggested that the CD133+ cells from human UCB have great expansion potential for ex-vivo expansion. The low concentration of TGF-β1 may delay over-differentiation of hematopoietic stem/progenitor cells.
Gadd45 Genes Participate in Myeloid Cell Fate Determination.
Authors: Gibbs John D. , Mohamed-Hadley Alisha , Liebermann Dan A. , Hoffman Barbara 
Published: October 2019

Abstract
The gadd45 family of genes is rapidly induced by different stressors, including differentiation-inducing cytokines, and there is a large body of evidence that their cognate proteins are key players in cellular stress responses. Gadd45 gene(s) were found to be primary response gene(s) to IL-6 mediated terminal differentiation of M1 monocytic leukemia cells, as well as to G-CSF mediated terminal differentiation of 32Dcl3 cells; furthermore, gadd45 gene(s) were induced by myeloid differentiation inducing cytokines such as IL-3 and GM-CSF in primary murine bone marrow (BM) cells (Zhang et al., Oncogene18:4899–4907, 1999; Abdollahi et al, Oncogene6:165–167, 1990). Induction of gadd45 genes at the onset of myeloid differentiation suggested that Gadd45 protein(s) play a role in hematopoiesis, yet no apparent abnormalities were observed in either the BM or peripheral blood compartments of mice deficient for either gadd45a or gadd45b. Both BM and Hoxb8 immortalized macrophage progenitors induced to differentiate in vitro revealed that deficiency in either gadd45a or gadd45b resulted in loss of macrophages and an increase in neutrophils compared to cells derived from wild type mice. How Gadd45a and Gadd45b influence the expression and activation of transcription factors that regulate myeloid cell determination will be discussed.
Effects of Temperature on the Change of HSP and the Viability of Mobilized Hematopoietic Stem Cells.
Authors: Kim Hawk , Min Young Joo , Baek Jin Ho , Shin Su Jin , Noh Eui-Kyu , Lee Eun-Hee , Park Sang-Kyu , Lee Jung Lim , Park Jae-Hoo 
Published: October 2019

Abstract
It is well known that HSP is influenced by temperature. Here we searched the effects of temperatures on preservation of hematopoietic stem cells (HSC) and the changes of HSP according to the temperatures and cryopreserving agents. The G-CSF-mobilized peripheral blood HSCs were collected from a healthy donor. DMSO, EG and GLY were used for cryopreserving agents. HSP were measured by real-time quantitative PCR, and cell viability were measured by trypan blue stain and MTS assay, respectively. Testing temperatures were 4°C, room temperature (RT, 20°C) and 37°C. Samples were HSCs stored according to testing temperatures without cryopreserving agents (cycle 0), HSCs thawed after first cryopreservation with various cryopreserving agents (cycle 1). The measure was done daily from D0 to D5 at cycle 0, and in 30 minutes interval (0, 30, 60, 90, 120 min) at cycle 1. Whereas more than 98% of cells were viable till 5 days after the harvest of cycle 0 at 4°C, RT and 37°C showed rapid decrease of viable HSCs. By day 3, less than 10% cells were viable at 37°C. At cycle 1, cell viability was rapidly decreased in both DMSO and GLY after 120 min stay at all temperatures. In case of EG, the same patterns were found at RT and 37°C. However, cell viability remained relatively well at 4°C. The expression of HSP 27 were over-expressed by day 2 and suppressed thereafter at all temperatures of cycle 0. At 4°C of cycle 0, the over-expression of HSP 27 was greater than those at RT and 37°C. The expression of HSP 70 was mostly suppressed along times. The suppression at 37°C was greater than those of other temperatures. In case of HSP 90, the overall expression was suppressed except for day 2 at 4°C and RT. At all cases of cycle 0, the expressions at 37°C were lower than those at 4°C and RT. At 4°C of cycle 1, DMSO showed decreased expression of HSP27 and HSP70. In contrast to DMSO, the expression of HSP27 and HSP70 was increased in case of EG and GLY at 4°C of cycle 1. The over-expression of HSP90 at 4°C of cycle 1 in EG and GLY was very low compared with those in DMSO. Overall expressions of HSPs in GLY were lower than in DMSO or EG at RT of cycle 1 except for HSP27. The expression of HSP27, HSP70, HSP90 were all decreased at 37°C of cycle 1 except for HSP70 in EG and HSP90 in DMSO. In conclusion, we should keep cool when storing HSCs without cryopreserving agent. The rapid apoptosis at 37°C could be caused by changed expression of anti-apoptotic HSPs. The expression of HSPs influenced by temperatures and cryopreserving agents may affect the viability of HSCs through the changes of HSPs.
ISWI ATPase Snf2h Is Required for Both Heterochromatin and Euchromatin Structure in ES Cells.
Authors: Podskocova Jarmila , Vargova Karin , Kokavec Juraj , Skoultchi Arthur I. , Stopka Tomas 
Published: October 2019

Abstract
Snf2h is a chromatin remodeling ATPase which along with associated factors has the ability to assemble and slide nucleosomes. Snf2h levels are substantially increased in acute leukemia cells and in aggressive solid tumors, suggesting a role for Snf2h in regulating chromatin structure in malignancy. Snf2h-null mice exhibit early embryonic lethality and Snf2h heterozygous mutant mice are growth-retarded (Stopka 2003, Chong 2007). In this study we have used Snf2h hemizygous (+/−) embryonic stem (ES) cells and mice to examine the consequences of reduced Snf2h stoichiometry on chromatin structure. Using indirect immunofluorescence and confocal microscopy we found that Snf2h in ES cells is localized both in euchromatin and to a lesser extent in heterochromatin. Its expression is markedly reduced in Sn2h +/− cells. Chromatin of Snf2h +/− ES cells exhibited multiple defects including decreased amounts of heterochromatin and a significant decrease of euchromatic marks including histone H3K79 and H4K16 acetylation. In contrast, histone H3K9 acetylation and the level of another chromatin remodeling ATPase, Brg-1, was not altered in Snf2h +/− ES cells as compared to wild-type ES cells. Thus in ES cells Snf2h may be involved in formation of both heterochromatin as well as transcriptionaly active euchromatin. Our data support the role of Snf2h in regulation of chromatin structure in leukemia, currently tested using Snf2h heterozygous animals.
Safety and Antitumor Efficacy of the Proteasome Inhibitor Carfilzomib (PR-171) Dosed for Five Consecutive Days in Hematologic Malignancies: Phase 1 Results.
Authors: Orlowski Robert Z. , Stewart Keith , Vallone Marcy , Molineaux Chris , Kunkel Lori , Gericitano John , O’Connor Owen A. 
Published: October 2019

Abstract
Introduction: Carfilzomib (CFZ) is a structurally- and mechanistically-novel proteasome inhibitor (PI) of the peptide epoxyketone class that exhibits a high level of selectivity for the unique N-terminal threonine active sites within the proteasome. CFZ is similar to bortezomib (BTZ; VELCADE®), in that it is a potent inhibitor of the chymotrypsin-like activity, but unlike BTZ, CFZ has shown minimal cross-reactivity with the other catalytic sites within the proteasome or across other protease classes. Preclinical human tumor xenograft models indicate that consecutive day (D1, D2) dosing resulting in 48 hours of proteasome suppression was superior to split dosing (D1, D4).
Methods: This phase 1 sequential dose escalation trial tested an intensive proteasome inhibition schedule with CFZ administered as an intravenous push (IVP) daily (QD) for 5 consecutive days on a 14 day cycle. The doses administered ranged from 1.2 mg/m2 to 20 mg/m2. Multiple myeloma (MM), non-Hodgkin’s lymphoma (NHL) that included mantle cell lymphoma (MCL), Hodgkin’s disease (HD), or Waldenström’s macroglobulinemia (WM) were eligible if they had relapsed after 2 or more prior therapies.
Results: A total of 29 subjects have been treated. The minimal effective dose (MED) was 11 mg/m2. A maximum of 80% proteasome inhibition was achieved in peripheral blood and mononuclear cells at ≥15 mg/m2. Five responses have occurred in 14 patients (6 MM, 3 MCL, 1 WM, 4 NHL) enrolled at ≥ the MED: 1 MM partial response (PR), 2 MM minimal responses (MR), 1 WM MR and one macroscopic CR in a patient with gastrointestinal MCL. Responses occurred in patients whose disease had failed therapy with BTZ, immunomodulatory agents, and stem cell transplant. Four NHL patients have had stable disease and remained on treatment for up to 7 months. Dose limiting toxicity reported at 20 mg/m2 included febrile neutropenia (reversible) and grade 4 thrombocytopenia. Grade 3 (25%) or grade 4 events (8%) were primarily hematologic events. Most withdrawals occurred due to the inconvenience of frequent visits, or progressive disease.
Conclusions: This study demonstrates that intensive CFZ dosing leading to sustained proteasome inhibition is well tolerated. The selectivity of the epoxyketone class of PI may promote a favorable safety profile. Furthermore, antitumor responses in heavily pretreated patients suggest non-cross resistant activity. Phase 2 trials in MM and solid tumors are proceeding with an alternate IV schedule. An oral agent suitable for frequent dosing schedules is under development.
Switch to Full-Length of XAF1 mRNA Expression in Myeloma RPMI8226 Cells by DNA Methylation Inhibitor 5-Azacytidine.
Authors: Chen Guanghua , Wu De Pei , Yang Ming Zhen , Tang Xiao Wen , Sun Ai-ning 
Published: October 2019

Abstract
X-linked inhibitor of apoptosis (XIAP) is the most potent member of the IAP family that exerts antiapoptotic effects by interfering with the activities of caspase-3, -7, -9, while the XIAP-associated factor1 (XAF1), a zinc finger protein, antagonizes XIAP activities, thereby promotes apoptosis.The aberrant silencing of the XAF1 gene has recently been found in various types of cancer cells, which is suggested to be one of the potential mechanisms underlying survival advantages of malignant cells. We investigated the XAF1 expression in myeloma RPMI8226 cells. Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and western blot methods were used for detecting the expressions of XAF1, XIAP, SURVIVIN, β-actin genes in RPMI8226 cells at different time points after treatment with 2.5 μM or 5 μM 5-azacytidine. The apoptosis of RPMI8226 cells was simultaneously detected using flow cytometric assay. Methylation-specific PCR was used to assay methylation status of the CpG sites in the XAF1 promoter. Compared with normal bone marrow mononuclear cells where only a full-length of XAF1 mRNA is detected, RPMI8226 cell expressed a full-length form of XAF1 transcript and a short form of XAF1 transcript. Only a full-length form of XAF1 transcript was detected after RPMI8226 cells treated with 5 μM 5-azacytidine for 120h. More percentages of RPMI8226 cells progressed to early apoptosis after 5 μM 5-azacytidine treatment for 120h. Collectively, our study suggests that epigenetic silencing of XAF1 by aberrant promoter methylation may contribute to the malignant progression of human myeloma. DNA methylation inhibitor 5-azacytidine treatment can increase myeloma RPMI8226 cell XAF1 gene expression and promote apoptosis.
Pegylated Recombinant Human Arginase, Rharg-Peg5,000MW, Targets Cell Cycle Machinery in Multiple Myeloma Cells.
Authors: Liu Delong , Lin Xianghua , Lu Quanyi , Leung Thomas , Cheng Paul N.M. , Chiao Jen-Wei 
Published: October 2019

Abstract
Arginase has been shown to inhibit growth of human hepatocellular carcinoma by depletion of arginine. We have studied the effects of the pegylated human recombinant arginase (BCT-100, rhArg-peg5,000MW) on RPMI8226 cells, a multiple myeloma cell line. This study showed that three day exposure of the myeloma cells to pegylated rhArg at a concentration of 0.08 IU/ml and 0.48 IU/ml resulted in growth suppression of 10% and 70% respectively, as compared to untreated control. Cell cycle analysis revealed significant decreases in the proportion of cells in both S- and G2M-phase and a concomitant increase of cells in G1-phase in a time- and concentration- dependent manner. We further studied the mechanisms of cell cycle arrest induced by the pegylated rhArg. The pegylated rhArg inhibited both cyclin-dependent kinases CDK2 and CDK4, enhanced the expression of the CDK inhibitor p21, and reduced the expression of cyclinD1, D2, and E. The level of phosphorylated Rb protein was also found to be significantly decreased. The regulators of cell cycle have thus been revealed as targets of pegylated rhArg for myeloma growth arrest. The pegylated rhArg may serve as a novel antitumor agent for multiple myeloma.
Globin Genes Induction by Nitric Oxide of Stromal Cell Origin.
Authors: Cokic Vladan P. , Beleslin-Cokic Bojana B. , Noguchi Constance Tom , Schechter Alan N. 
Published: October 2019

Abstract
We have previously shown that nitric oxide (NO) is involved in the hydroxyurea-induced increase of gamma-globin gene expression in cultured human erythroid progenitor cells and that hydroxyurea increases NO production in endothelial cells via endothelial NO synthase (NOS). Here we report that co-culture of human bone marrow endothelial cells with erythroid progenitor cells induced gamma-globin mRNA expression (1.8 fold), and was further elevated (2.4 fold) in the presence of hydroxyurea (40 μM). Based on these results, NOS-dependent stimulation of NO levels by bradykinin and lipopolysaccharide has been observed in endothelial (up to 0.3 μM of NO) and macrophage cells (up to 6 μM of NO), respectively. Bradykinin slightly increased gamma-globin mRNA levels in erythroid progenitor cells, but failed to increase gamma-globin mRNA levels in endothelial/erythroid cell co-cultures indicating that stimulation of endothelial cell production of NO alone is not sufficient to induce gamma-globin expression. In contrast, lipopolysaccharide and interferon-gamma mutually increased gamma-globin gene expression (2 fold) in macrophage/erythroid cell co-cultures. In addition, hydroxyurea (5–100 μM) induced NOS-dependent production of NO in human (up to 0.7 μM) and mouse macrophages (up to 1.2 μM). Co-culture studies of macrophages with erythroid progenitor cells also resulted in induction of gamma-globin mRNA expression (up to 3 fold) in the presence of hydroxyurea (20–100 μM). These results demonstrate a mechanism by which hydroxyurea may induce globin genes and affect changes in the phenotype of hematopoietic cells via the common paracrine effect of bone marrow stromal cells.
3D Grown Human Placental-Derived Mesenchymal Stromal Cells (PLX-I) Support and Potentiate the Engraftment of Human Cord Blood Unfractionated (UFCBC) and Fractionated CD34+ Cells (hCD34+) in Irradiated NOD/SCID Mice.
Authors: Burger Ora , Jacov Osnat M. , Carmi Nirit D. , Ashtamker Galit F. , Pinzur Lena , Mizrachi Einat , Rusanovsky Marina , Bercovich Noam , Moshe Ofer B. , Lior Limor , Meiron Moran , Prather William R. , Meretzki Shai , Treves Avi J. , Kadouri Avinoam , Rowe Jacob M. , Horwitz Edwin M. , Nagler Arnon 
Published: October 2019

Abstract
The use of human umbilical cord blood (hUCB) for transplantation (CBT) in adults is limited due to the low cell number in each unit. Engraftment rates are lower and the time to neutrophils and platelet recovery is longer following CBT compared to bone marrow (BM) transplantation (28 vs 14 days, respectively). Co-transplantation of human hematopoietic stem/progenitor cells (hHSC) with mesenchymal stromal cells in an irradiated NOD/SCID mice model has been demonstrated to promote HSC engraftment. In a typical clinical setting of CBT, unfractionated cord blood cells (UFCBC) are usually used. On the other hand, in the irradiated NOD/SCID mice model, the CD34+ enriched fraction is usually used. Therfore, performing pre-clinical studies using UFCBC would better reflect the outcome in the clinical setting. Using PluriX technology, we expanded human placental derived mesenchymal stromal cells on 3D carriers in a bioreactor (PLX-I) and evaluated their potential to promote the engraftment of UFCBC or hUCB derived fractionated CD34+ cells (hCD34+) in NOD/SCID mice. UFCBC (107) or hCD34+ (5×104) were injected into the tail vein of 7–8 week old NOD/SCID mice along with 0.5×106 or 1×106 PLX-I cells. Following 5–6 weeks, FACS analysis of % human CD45+ cells in mice BM demonstrated that in the PLX-I untreated mice, the 107 UFCBC engrafted in a lower rate compare to 5*104 hCD34+ (6.28% vs. 14.43% respectively, n=10). Furthermore, in both settings, hCD34+ and UFCBC, the addition of PLX-I enhanced the hHSC engraftment 1.7 and 2 fold respectively compared to PLX-I untreated mice (24.13% vs. 14.43% in hCD34+ setting and 12.85% vs. 6.28% in UFCBC setting, n=10). We conclude that UFCBC can be used for hHSC engraftment in NOD/SCID mice model and co-transplantation of these cells with PLX-I may serve as a promising approach for the improvement of the hitherto delayed engraftment following cord blood transplantation.
Molecular Mechanisms of Cell Density-Dependent Growth in Myeloid and Lymphoid Leukemia Cell Lines.
Authors: Murohashi Ikuo , Ihara Noriko 
Published: October 2019

Abstract
Normal hematopoietic stem cells have been shown to be maintained through interaction with their environmental niches, such as osteoblastic and endothelial ones. The growth of leukemia cells has been shown to be stimulated by environmental niches (paracrine growth) or by cell-to-cell interaction and/or excreted factors of leukemia cells (autocrine growth). The growth of myeloid (MO7-E and HL-60) and lymphoid (Raji, U-266, Daudi and RPMI-1788) leukemia cell lines cultured at various cell densities in serum free medium (Sigma H 4281) with 1% BSA was evaluated. The cells cultured at higher cell densities (cultured cell densities of more than 105/ml) showed logarithmic linear increases in cell number, whereas those at lower cell densities (cultured cell densities of less than 104/ml) ceased increasing cell number. Supernatants of myeloid leukemia cells stimulated the growth of autologous clonogenic cells, but not those of lymphoid leukemia cells. Neutralizing antibodies (Abs) against various hematopoietic growth factors failed to inhibit cell growth except for anti-VEGF Ab, which significantly decreased HL-60 leukemia cell growth. In contrast, anti-TNF-α Ab significantly stimulated the growth of the HL-60 cells. To clarify the nature of the cultured cell density on the growth of leukemia cells, leukemia cells were cultured at higher cell densities (group H, cultured cell densities of 106/ml) or at lower cell densities (group L, cultured cell densities of 104/ml). After culture of 3-, 6-, 10-, and 24-hr, cells were serially harvested and total cellular RNA was extracted. Gene transcript levels were determined by using Real-Time PCR. Gene transcripts examined in the present study were as follows: Jagged-1, -1, Notch-1, -2, -3, Ang-1, -2, Tie-1, -2, Wnd3a, Wnd5a, β-Catenin, γ-Catenin, N-Cadherin, Cyclin D1, p16, p21, HOXA6, HOXA7, HOXA10, HOXB4, and Mef2c. At 24-hr cultures, transcripts of myeloid leukemia cell lines for Bmi-1, Wnt-3a, β-Catenin and γ-Catenin were higher, and those of lymphoid leukemia cell lines for Notch 1, 2, and 3 were higher in group H compared with group L. Transcript levels for Wnt5a were higher at 10-hr culture (HL-60 and Raji), those for HOXA7 at 30–10-hr (MO-7E, U-266 and Raji), and those for Mef2c at 3-hr (MO-7E, U-266 and Raji) in group H compared with group L. Taken together, our present results favor the conclusions that genes related to transcription factors and growth factors are sequentially and differentially expressed through cell-to-cell interaction of leukemia cells. The nature of the leukemia cell-to-cell interacrtion, which is related to the growth advantages of leukemia stem cells over normal hematopoietic stem cells, remains to be further clarified.
FISH Analysis of IgH Rearrangement by 4 Kinds of Probes Involving IgH Genes in Multiple Myeloma: Improved Sensitivity for the Detection by Use of IGH Break-Apart Probe.
Authors: Chung Hye Yoon , Kim Miyoung , See Cha Ja , Min Hyun Jung , Yoon Sung-Soo , Cho Han Ik , Lee Dong Soon 
Published: October 2019

Abstract
Background: IgH (immunoglobulin heavy chain) gene rearrangement is known to be the most frequent chromosome change in multiple myeloma. The detection of this genetic change is conveniently done by using fluorescence in situ hybridization (FISH) method recently. The aim of this study is to determine the utility of most commonly used probes, IGH/CCND1 dual color, dual fusion probe (Downers Grove, IL, USA), IGH/BCL2 dual color, dual fusion probe (Downers Grove, IL, USA), IGH/FGFR3 dual color, dual fusion probe (Downers Grove, IL, USA) and IGH dual color break apart rearrangement probe from Vysis Products (Downers Grove, IL, USA).
Methods: We applied four different probes of IgH FISH on 202 Korean patients with multiple myeloma for comparing the utility of four probes.
Results: 84 of 202 patients (41.6%) had the IgH gene rearrangement. 44 of 84 patients (52.4%) showed positive to all four probes, but 40 of 84 patients (47.6%) showed discrepancy. IGH dual color break apart rearrangement probe showed highly detectable rate (41.6%) compare to IGH/CCND1 (31.2%), IGH/BCL2 (27.7%) and IGH/FGFR3 (24.3%). Among patients who showed discrepancy between four probes, 20 of 40 patients (50.0%) were only positive to IGH dual color break apart rearrangement probe.
Conclusions: The IGH break-apart probe was qualitatively and quantitatively better than three other probes at initial diagnosis and during follow-up of myeloma. In conclusion, it would be most efficient and advisable to use the IGH break-apart probe at first, and then to identify the translocation partner in case of positive rearrangement of IgH.