bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Dynamics of Chromatin Modification during RA Induced Promyelocyte Differentiation.
Authors: Harris Robert G. , Krause Diane S. 
Published: April 2020

Abstract
Epigenetic histone modifications have recently been shown to be important in gene regulation and to be correlated with gene expression profiles. In particular, particular modifications at Lysine 4 and Lysine 9 of Histone H3 may be highly indicative of both active and silenced chromatin. Promoters of highly active genes are generally tri-methylated at Lysine 4 and acetylated at Lysine 9 whereas silenced chromatin is unmethylated or mono methylated at Lysine 4 and di or tri-methylated at Lysine 9. NB4 is a human acute promyelocytic leukemia (t15:17) cell line that can be differentiated down the neutrophil lineage by induction with supraphysiological levels of retinoic acid (RA). We have examined post-translational modifications at Lysine 4 and Lysine 9 of Histone H3 at specific promoters using chromatin immunoprecipitation (ChIP) during differentiation of NB4 cells. We have performed quantitative (q) RT-PCR for the Myeloperoxidase (MPO) and Defensin-alpha (Def-a) genes after 0,12, 24, 48 and 72 hours of RA induction of NB4 cells. Concurrently we performed ChIP followed by qPCR for Di- and Tri-methylated Lysine 4 of Histone H3 and acetyl- and di-methylated Lysine 9 of Histone H3 at the promoters of these genes. MPO mRNA is expressed at time 0, and decreases progressively starting at 24 hours such that by 48 hours mRNA levels are negligible. This correlates nicely with a sharp decrease in tri-methyl Lysine 4 and acetyl Lysine 9 levels by 12 hours, with a smaller decrease again at 24 hours on the MPO promoter. By 72 hours a concurrent increase in dimethyl Lysine 9 levels is seen, indicating chromatin silencing. Conversely, def-a is initially not expressed in NB4 cells. After 24 hours of induction with RA, an increase in mRNA levels is seen. This correlates well with a sharp increase of acetyl Lysine 9 in the promoter and a concurrent decrease in dimethyl lysine 9 levels. In conclusion we have shown that during RA induced differentiation of promyelocytic leukemia cells, chromatin modifications slightly precede changes in mRNA expression levels and recapitulate other studies linking Lysine 4 methylation with active transcription and Lysine 9 methylation with gene silencing.
Antagonistic Roles of the ERK and p38 Pathways in Chemically Induced Erythroid Differentiation of Murine Erythroleukemia Cells.
Authors: Blank Volker , Boudia Mansouria Merad , Todorova Margarita , Moore Amy P. 
Published: April 2020

Abstract
Murine erythroleukemia (MEL) cells provide a valuable model to study the molecular events leading to erythroid differentiation. Maturing erythroid cells synthesize large quantitities of hemoglobin, a process requiring the coordinated synthesis of heme and globin. Here, we investigated the role of the ERK and p38 mitogen-activated protein kinase (MAPK) signaling pathways in chemically induced differentiating MEL cells. We showed that treatment of DMSO- or HMBA-induced MEL cells with the MEK1/2 inhibitor UO126, blocking the ERK pathway, results in the increase of beta-globin and 5-aminolevulinate synthase-2 (ALAS-2) transcript levels. We found that addition of the p38 inhibitor SB203580 has the opposite effect, leading to decreased beta-globin and ALAS-2 mRNA levels. The regulation on the transcript level correlated with increased or decreased hemoglobin levels in the presence of MEK1/2 and p38 inhibitors, respectively. Our results suggest that the ERK and p38 pathways play antagonistic roles in DMSO- and HMBA induced erythroid differentiation in MEL cells. This data also provide a novel link between MAPK signaling and the regulation of intracellular heme levels, as ERK and p38 MAPK signaling cascades appear to differentially regulate one of the key enzymes of the heme biosynthesis pathway.
Expression and Functional Differences of CBFA2T (ETO, MTG) Gene Family Members in Hematopoiesis.
Authors: Mortazavi Fariborz , Kadkol ShriHari , Bruno Annette , Baraoidan Kristine , Ackerman Steven , Wickrema Amittha , Saunthararajah Yogenthiran 
Published: April 2020

Abstract
The CBFA2T (ETO, MTG) family has three similar family members - CBFA2T1-T3. CBFA2T1 (ETO, MTG8) and CBFA2T3 (ETO2, MTG16) are targeted by chromosomal translocations in acute myeloid leukemia. To better understand the usual hematopoietic function of this gene family, we examined the expression of CBFA2T RNA using RQ-PCR in cell-lines and human CD34+ hematopoietic cells during macrophage and erythroid differentiation. RQ-PCR on extracted RNA was performed with an icyclerQ instrument (Bio-Rad) using the Quantitect SYBR Green RT-PCR kit (Qiagen) and in vitro transcribed RNA to construct standard curves. CBFA2T3 was the most highly expressed family member in human CD34+ cells, the erythro-leukemia line K562, the myeloid line MPD, the T cell line Jurkatt and the B-cell line LCL-11. However, CBFA2T3 expression decreased by &gt;50% during both macrophage and erythroid differentiation of human CD34+ cells. In contrast, CBFA2T1 expression was almost undetectable in human CD34+ cells and all cell lines except K562 but increased more than 20 fold during erythroid (but not macrophage) differentiation of human CD34+ cells. Extrinsic over-expression of CBFA2T1, but not CBFA2T2, significantly increased glycophorin-A and hemoglobin A expression in K562 cells, consistent with a regulatory role for CBFA2T1 in erythroid differentiation. CBFA2T2 (MTGR1) was moderately expressed in human CD34+ cells and all the cell lines and demonstrated a 2.5 fold increase in expression with macrophage differentiation but essentially no change with erythroid differentiation of human CD34+ cells. These findings suggest that despite their similarity, the CBFA2T family members have distinctive regulatory roles in hematopoietic differentiation.
In Vivo Induction of Cytokine mRNA and Protein in Mice by the Radioprotectant Steroid 5-Androstenediol.
Authors: Grace Marcy B. , Chang Cheng-Min , Singh Vijay K. , McLeland Christopher , Inal Cynthia E. , Jackson William E. , Whitnall Mark H. 
Published: April 2020

Abstract
We previously showed 5-androstenediol (5-AED) injected sc enhanced survival in mice exposed to whole-body gamma-irradiation, ameliorated radiation-induced neutropenia in mice, dogs, and monkeys, and induced increases in bone marrow GM-CFC in irradiated mice. Radiation-induced decreases in platelets, natural killer cells, red blood cells, and monocytes were also blunted by the steroid. Administration of 5-AED caused functional activation of circulating granulocytes (phagocytic ability), monocytes (oxidative burst), and natural killer cells (surface CD11b expression). To test our hypothesis that 5-AED acts via initiation of a cytokine cascade in hematopoietic tissue, we performed real time quantitative PCR on spleen samples taken from mice and correlated the results with serum ELISA. PCR was performed for mRNAs of GM-CSF, IL-6, IL-10, IL-12, and IFNg. Spleens and serum were taken at 3 time points: 4 h after 5-AED or vehicle (PEG-400) injection, and 4 or 24 h after irradiation (7.5 Gy whole-body gamma) or sham-irradiation. Each group contained 5 mice. Results were analyzed by 2-way ANOVA for PCR results 4 and 24 h after irradiation, and by a t-test 4 h after injection. Radiation alone induced increases in mRNAs for all cytokines at both 4 and 24 h, except for IFNg, which was not elevated at 4 h. 5-AED alone did not induce a significant increase for any cytokine, but the increases bordered on significance for IL-12 at 4 h after irradiation or sham-irradiation (28 h after injection, p = 0.07), and for GM-CSF 24 h after irradiation or sham-irradiation (48 h after injection, p = 0.08). There was a significant positive interaction between radiation and 5-AED for IL-6, 4 h after irradiation (p = 0.006). The interaction effect bordered on significance for IL-12 4 h after irradiation (p = 0.07) and for GM-CSF 24 h after irradiation (p = 0.10). ELISA for serum cytokine protein was measured for G-CSF, IL-6, IL-10, and IL-12. Results were analyzed by the Mann-Whitney test for exact p-values. IL-6 was not detected in serum. G-CSF was detected 4 and 24 h after irradiation or sham-irradiation only in 5-AED-treated mice. IL-10 and IL-12 were detected only 24 h after irradiation, and the values in 5-AED-treated mice were significantly higher than those in vehicle-treated mice (p = 0.02). The results support our hypothesis that the previously observed increases in numbers of circulating innate immune cells and platelets, and functional activation of granulocytes, monocytes, and NK cells, result from a cytokine cascade induced by 5-AED.
Decreased Total Body Irradiation (TBI)-Induced Apoptosis in Murine Hematopoietic Side Population Cells Compared to Non-Side Population Cells.
Authors: Shen Hongmei , Yun Yun , Epperly Michael W. , Greenberger Joel S. 
Published: April 2020

Abstract
The side population (SP) cells of the murine bone marrow (BM) contain primitive hematopoietic stem cells (HSCs). To understand the effect of irradiation on these cells, C57BL/6NHsd mice were irradiated to 10 Gy total body dose and sacrificed 24 hours later. The bone marrow was removed and stained with Hoechst dye, and Annexin V as an early marker of apoptosis. The SP cells were analyzed by flow cytometry. Following irradiation, there was no change in the frequency of SP cells in total nucleated BM cells between non-irradiated (0.081 ± 0.033%) and irradiated mice (0.093 ± 0.027%). In contrast, there was a decrease in total nucleated BM and SP cells. SP cells decreased by 60.1% from 44873 ± 22099 SP cells per harvest to 17550 ± 4805 SP cells per harvest 24 hours after 10 Gy. The number of lymphocytes in the irradiated BM decreased from 25.7% to 4.5% after irradiation. Since the majority of the SP cells are located in the lymphocyte sorting region, there was a 6-fold decrease in the percent of SP cells found in the live lymphocyte region following irradiation (0.469 ± 0.185% compared to 2.923 ± 1.913%).
Annexin V was used to stain for the appearance of phosphatidylserine (PS) on the cell surface as an early marker for apoptosis. The appearance of Annexin V positive cells did not increase significantly with irradiation in SP compared to non-irradiated SP cells (26 ± 8% and 25 ± 12%, respectively). In contrast, the percent of apoptotic cells in the non-SP fraction was significantly increased following irradiation compared to the non-irradiated cells (35 ± 8% and 25 ± 6%, respectively) (p=0.02). Our data indicate that irradiation was not inducing apoptosis in SP cells but in non-SP cells. It might suggest that SP cells could be more resistant to radiation damage. Future investigations will be performed to understand the functional activity of SP cells from lethally irradiated mouse BM and the molecular mechanism of those SP cells in resistance to irradiation damage.
Characterization of Non-Expanded Mesenchymal Progenitors Cells from Normal Adult Human Bone Marrow.
Authors: Nathalie Boiret , Chantal Rapatel , Richard Veyrat-Masson , Laurent Guillouard , Jacques Guerin Jean , Pascale Pigeon , Stephane Descamps , Stephane Boisgard , Marc Berger G. 
Published: April 2020

Abstract
Adult Bone Marrow (BM) mesenchymal stem /progenitor cells (MS/PC) are a potentially useful tool for cell therapy and tissue repair. Most of the studies related to their characterization have been carried out on cells selected by adherence and expanded in vitro. We have developed a simple means of directly identifying mesenchymal subsets from fresh BM.
First, we adapted our tools on MS/PC enriched by short time adherence (D1-3) before any cell division. Acquisition and analysis of the CD45-CD14- cells was performed on a EPICS Elite flow cytometer (Coulter, Miami, FL, USA). We have already shown that all CFU-F were included in adherent CD45-CD14- subset. When there was a sufficient number of collected adherent cells, the sub populations, defined by the expression of CD73, CD105, CDw90, or CD49a on CD45-CD14- cells, were sorted and plated in CFU-F assay. Then, we adapted this procedure to un-manipulated fresh BM mononuclear cell (MNC).
Adherent CD45-CD14- cells expressing the CD73 or the CD49a antigen contained all the CFU-F even though the CD105+ and CDw90+ subsets comprised less than half the total. In fresh un-manipulated BM MNC, CD73 and CD49a were also highly discriminative and allowed up to a 3 log enrichment of CFU-F when compared to BM MNC. The expression of CD105 and CDw90 appeared non-specific on fresh mesenchymal progenitors. Usual culture conditions up-regulated most of the tested antigens, in particular CD73 and CD49a with a significant increase in mean of fluorescence.
Antigen expression on CD45-CD14- subset
Conclusion: The CD45-CD14-/CD73+ and CD45-CD14-/CD49a+ phenotypes identified subsets containing all mesenchymal progenitors and sufficiently enriched to detect them in fresh BM, enabling evaluation of mesenchymal content of BM collections for cell therapy. Futhermore, this study argues for the existence of differences in antigen expression between un-manipulated mesenchymal cells (MC) and in vitro expanded MC and for the feasibility of distinguishing cell subsets in MC populations.
CD73+ CD105+ CDw90+ CD49a+ Phenotypic analysis and plating afficiency of adherent (D1-3) and un-manipulated CD45-CD14- cells. The proportion of each subset was expressed as a percentage of CD45-CD14- cells (a). The proportion of CFU-F obtained from the positive subset was reported in percentage of the total CFU-F obtained from the two subsets (positive and negative for the same antigen) (b). % (a) adherent 18.3 21.1 17.4 21.1 un-manipulated 5.2 17.4 4.5 4.4 CFU-F % (b) adherent 100.0 45.4 49.0 95.2 un-manipulated 100.0 50.5 65.6 99.7
Karyotypic Evaluation of Expanded Rat MAPCs.
Authors: Rundlett Stephen E. , Boozer Sherry L. , Suntharalingam Piratheepan , Verfaillie Catherine M. , Deans Robert 
Published: April 2020

Abstract
Summary
The clinical utility of an ex-vivo expanded stem cell population is dependent on both safety and efficacy. As an initial step toward ensuring that expanded adult stem cell populations are safe, we harvested bone marrows from four different strains of rat and cultured mononuclear cells under low serum media shown previously to allow for propagation of Multipotent Adult Progenitor Cells (MAPCs). We then determined the karyotype and tri-lineage potential for these expanding populations at numerous population doublings. We observe that an average of 6.7% of cells within these cultures are tetraploid and 2.7% of cells show other karyotypic alterations. In spite of this low level of karyotypic instability, no persistent chromosomal abnormalities are present in these cultures after more than 100 population doublings. As measured by qPCR and indirect immunofluoroescence for lineage specific marker genes, these cells have tri-lineage potential and are therefore considered to be MAPCs. These data support and extend previous conclusions that MAPC cultures with a nearly normal karyotype can be routinely established and expanded from rat bone marrow providing important standards for performance and safety in pre-clinical disease models.
Recurrent Chromosomal Rearrangements and a Homozygous Deletion Spanning 1-Mb Region at 10p11.2 in Adult T-Cell Leukemia/Lymphoma.
Authors: Hidaka Tomonori , Nishida Kazuhiro , Shimizu Daisuke , Sasaki Hidenori , Yamashita Kiyoshi , Kubuki Yoko , Nomura Kenichi , Taki Tomohiko , Tsubouchi Hirohito , Morishita Kazuhiro , Taniwaki Masafumi 
Published: April 2020

Abstract
Cytogenetic analyses and their correlation with clinical features are still limited in mature T-cell malignancies. The cytogenetics of adult T-cell leukemia/lymphoma (ATLL) is complicated by many structural and numerical abnormalities of chromosomes. Thus, it is occasionally difficult to define karyotype of ATLL only by chromosomal banding method. Using multicolor spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH), we performed molecular-cytogenetic studies on 38 patients with adult T-cell leukemia/lymphoma (ATLL) and 7 established cell lines. In SKY analysis, chromosomal bands involved in rearrangements were identified based on the inverted images of DAPI staining. Chromosome no.7 (25 cases) and no.14 (30) were frequently involved in structural rearrangements including translocations, inversions, insertions, isochromosomes, and deletions. Breakages frequently occurred at 10p11-13 (21 cases, 46.6%), and 14q11.2 (16, 35.6%), 14q32 (12, 26.7%), 7q22 (10, 22.2%), 9q22 (10, 22.2%), 13q14 (9, 20.0%), 22p11 (8, 17.7%), 8p11 (7, 15.5%), 18p11 (7, 15.5%), 21q22 (7, 15.5%) and 19q13 (6, 13.3%). Recurrent partners associated with 10p11.2-13 rearrangements were 21q22 (3 cases), 13q14 (2) and 14q32.1 (2). Partner chromosomal breakpoints involving 14q11.2 rearrangements were 14q32 (3 cases), 8p11.2-21 (3), and 11q13.3 (2), while those involved in 14q32 rearrangements were 14q11 (3) and 10p11-13 (2). Using FISH with 37 bacterial artificial chromosome clones, we analyzed breakpoints at 10p11.2-13 in two cell lines KOB4 and KK1 and one patient (HH) by walking a segment ranging from 10p13 to 10p11.2. Among 37 BAC clones used as FISH probes, 9 were assigned to 10p15, 3 to 10p14-13, and the remaining 25 to 10p11.2-12. Large genomic deletions were detected at 10p11.2 on der(10) in two cell lines. In patient HH, a homozygous deletion was identified at 10p11.2, spanning the region of 1 Mb in size. Chromosome 14q32 translocation was associated with shorter survival (median survival time; 4.2 months vs. 8.0 months), although statistically not significant. As for 10p11.2-13 and 14q11.2 rearrangements, there was no significant difference of survival between patients with and without respective abnormalities. In conclusion, SKY and FISH analysis identified recurrent chromosomal rearrangements and a homozygous deletion at 10p11.2 in ATLL. Our molecular analysis is now focusing on this genomic region to clone genes associated with development of ATLL.
Expression of the Chk2 Gene Is Downregulated in Hodgkin’s Lymphoma Cell Lines Via Epigenetic Mechanisms.
Authors: Kato Naoko , Kondo Takeshi , Tsukada Junichi , Tanaka Yoshiya , Minami Yasuhiro , Tanaka Junji , Imamura Masahiro 
Published: April 2020

Abstract
Chk2, the mammalian homologue of the yeast Rad53 and Cds1 genes, encodes a nuclear serine/threonine kinase that plays a crucial role in the DNA damage response and helps guard the integrity of the genome by regulating cell-cycle checkpoints, DNA repair and apoptosis. Furthermore Chk2 is regarded as a tumor suppressor gene. Alterations in tumor suppressors that are involved in the DNA damage response have been reported to be frequently involved in the pathogenesis of lymphoid malignancies. We investigated the expression levels of the genes encoding Chk2 in nine cell lines from lymphoid malignancies, including three Hodgkin’s lymphoma (HL) cell lines. We found that all three HL cell lines exhibited a drastic reduction in Chk2 mRNA and protein expression compared to the other cell lines without any apparent mutation of the Chk2 gene. Accumulating evidence demonstrates the importance of posttranslational modification of histone proteins in addition to DNA methylation, as epigenetic mechanisms involved in the organization of chromosomal domains and gene regulation. It is now generally appreciated that hyperacetylated histones H3 and H4 are associated with activated genomic regions, while hypoacetylation of histones H3, H4 and methylation on H3-lysine 9 results in gene repression and silencing. Therefore we tested the possibility that epigenetic mechanisms are involved in this aberrant expression of the Chk2 gene in these cells using the histone deacetylase (HDAC) inhibitors trichostatin A (TsA) and sodium butyrate (SB). Expression of Chk2 in HL cells was restored following treatment with the TsA and SB, or 5Aza-dC. Chromatin-immunoprecipitation (Chip) assays revealed that treatment of HL cells with TsA, SB or 5Aza-dC resulted in increased levels of acetylated histones H3 and H4, and decreased levels of dimethylated H3 lysine 9 at the Chk2 promoter. These results indicate that expression of the Chk2 gene is down-regulated in HL cells via epigenetic mechanisms. Furthermore we examined whether the down regulation of Chk2 gene in HL cells may be involved in apoptosis resistance to irradiation (IR). We found that upregulation of Chk2 in HL cells following treatment by SB resulted in increased susceptibility of the cells to IR. Therefore, our findings suggested that altered epigenetic regulation of Chk2 gene in HL cells results in the down-regulation of Chk2 kinase and abrogates DNA damage response signaling in the cells
Different Subtypes of Difffuse Large B-Cell Lymphomas Identified Using Microarrays and Phenotyping.
Authors: Poulsen Christian Bjorn , Borup Rehannah , Nielsen Finn Cilius , Borregaard Niels , Hansen Mads , Gronbaek Kirsten , Ralfkiaer Elisabeth 
Published: April 2020

Abstract
The mRNA expression profile in diagnostic, pretreatment samples from 52 primary nodal diffuse large B-cell lymphomas (DLBCL) were investigated using a commercially available oligonucleotide array (the Affymetrix Genechip HU133A) that contains &gt;22.000 probe sets representing 18.400 transcripts. Hierarchical clustering indicated that 3 subtypes could be distinguishes. These included a GCB type (n=20) that showed an expression profile as similar to germinal center B-cells, an ABC type (n=25) that showed a profile as in vitro activated B-lymphocytes, and an intermediate group, type-3 (n=5) that did not fit into either of these categories. Two samples could not be classified due to inconsistency in clustering. Correlation with the genotype and phenotype showed that markers of germinal center B-cells (CD10, Bcl-6, t(14;18)) were prevalent but not exclusive to the GCB type. By contrast, MUM1 (a marker of terminal B-cell differentiation) was only expressed in the ABC type and in type-3. These results are similar to results obtained using a custom-made, spotted array - the Lymphochip (Alizadeh et al. Nature 2000; Wright et al, PNAS 2003). Comparison with the clinical features indicated that patients with GCB types of lesions showed a better initial response to CHOP or CHOP-like regimens than the remaining patients. However there was no significant difference in over-all survival. It is concluded that gene profiling with Affymetrix Genechips can be used to distinguish between GCB and ABC types of DLBCL and that these subtypes are likely to represent different, biological entities. Affymetrix Genechips should constitute a useful platform for prospective investigations, which are needed to elucidate whether gene profiling in DLBCL can be of use in choosing which treatment should be offered to patients.