bluuebunny/crossref_metadata_2025_split huggingface dataset:
In Vivo Permanent Marking of Cells with RAG1 Activation History Demonstrates Only a Minor Contribution of the Lymphoid Pathway to Dendritic Cell Development.
Authors: Chong Yong , Mizuno Shin-ichi , Shigematsu Hirokazu , Arinobu Yojiro , Iino Tadafumi , Iwasaki Hiromi , Akashi Koichi 
Published: April 2020

Abstract
Dendritic cells (DCs) have been shown to arise from both myeloid and lymphoid pathways, by evaluating the in vivo reconstituting potential of myeloid- and lymphoid-committed progenitor populations. It has been suggested that these “myeloid” or “lymphoid” DCs may constitute independent DC compartments with different functions. However, evaluation of DC development after conventional adoptive transfer experiments may not correctly represent normal DC development including lineage contribution toward the formation of the DC pool. In order to accurately evaluate the distribution and the function of DCs originated from each lineage, it is ideal to mark their origin in vivo in steady-state hematopoiesis. By crossing RAG1-Cre knockin with yellow fluorescence protein (YFP)-floxed reporter lines, we developed a mouse line in which cells with a history of RAG activation should be permanently marked with YFP as a result of lymphoid commitment. The YFP expression started at the common lymphoid progenitor (CLP) stage (~35%), and the percentages of YFP+ cells progressively increased as they differentiate into mature lymphocytes. More than 99.5% of mature T and B cells were marked by YFP, while &lt;0.5% of granulocyte/monocyte progenitors (GMPs) or granulocytes were YFP+, indicating that this system efficiently and exclusively marks cells of lymphoid origin. We found that only 4.1, 4.6, and 9.3% of DCs were YFP+ in the spleen, the lymph nodes, and the thymus, respectively. This is the formal evidence that the vast majority of DCs in steady-state hematopoiesis originate from stages that have committed to the myeloid lineage, even in the thymus. We then profiled the gene expression of the YFP+ and YFP- DCs at a genome wide level by DNA microarray analysis. Unexpectedly, the gene expression profiles of myeloid and lymphoid-derived DCs are virtually identical. These results collectively suggest that myeloid and lymphoid DCs might use a common developmental program that can be activated even after myeloid or lymphoid commitment. Thus, the DC developmental program is unique, which should be independent of either myeloid or lymphoid commitment program.
Mouse Model for Shwachman-Diamond Syndrome with the R126T Disease Mutation Leads to Severe Growth and Developmental Deficiencies with Impairment of Hematopoiesis.
Authors: Zhang Siyi , Zhong Jian , Gan Olga I. , Dick John E. , Rommens Johanna M. 
Published: April 2020

Abstract
Shwachman-Diamond syndrome (SDS) is a bone marrow failure syndrome characterized by exocrine pancreatic insufficiency and skeletal abnormalities, as well as hematological dysfunction. SDS is caused by mutations in SBDS, a highly conserved gene that has been suggested to be involved in RNA metabolism and/or ribosome biogenesis. It is essential, based on our mouse knock-out model studies and the absence of observed patients with the combination of two null alleles. We have generated an allele with the disease mutation, R126T, in the murine ortholog in order to develop new models of SDS. This mutation has been interpreted to be hypomorphic in nature, as it occurs in combination with the common null disease allele in two patients. Interestingly, these patients also have severe hematological disease phenotypes.
SbdsR126T/+mice develop normally and show no disease phenotypes, in accordance with the recessive inheritance of SDS. However, the SbdsR126T/R126T and SbdsR126T/- mice did not survive birth, and exhibited marked size reduction. The growth difference became apparent in embryos during the mid-fetal period, at E12.5–14.5; there were also noted disturbances of major organs including the skeleton, brain and lung. Comparable deficiencies were noted overall, but the SbdsR126T/- embryos were consistently more severely affected than were SbdsR126T/R126T embryos. Examination of hematopoietic progenitors from the fetal livers of SbdsR126T/R126T and SbdsR126T/- mutant mice also showed marked reductions in BFU-E, CFU-G, CFU-M, CFU-GM and CFU-GEMM numbers, as revealed by standard colony formation assays, when compared to wild type and heterozygote littermates.
In additional studies, primary fibroblast cultures from E16.5 day mutant embryos were found to exhibit slower growth and an extended G1 phase of the cell cycle compared to fibroblasts from wild type embryos. Further, total protein synthesis was measured by incorporation of radio-labeled amino acids and found to be notably reduced in the mutant fibroblasts. The small size, organ deficiencies and early death of the mutant mice together with the cellular deficiencies, which indicate severe autologous growth problems, emphasize the severe consequences of loss of Sbds. These animal disease models provide new avenues for investigation to elucidate basic functions of SBDS and the pathobiology of SDS.
Hematopoietic Specific GATA-1-Improved Cre Mouse for Erythroid-Specific Gene Modification.
Authors: Yoon Donghoon , Kim Bumjun , Kwon Myunghi , Prchal Josef T. 
Published: April 2020

Abstract
Animal models of erythropoiesis related genes have been limited by the fact that some of these genes have non-erythroid expression and other functions in addition to erythropoiesis and thus their knock-out may be embryonic lethal. Tissue specific knock-out or knock-in mice models employing GATA-1-Cre and other constructs showed that these promoters are also active in non-hematopoietic tissues, i.e. GATA-1 has activity in early embryonic development and in neuronal tissue. Suzuki et al (Blood, 2002, 100; 2279) isolated the GATA-1 locus hematopoietic regulatory domain (GATA-1-HRD) and demonstrated that the expression of a transgene under its control is limited to the hematopoietic tissue. We generated a transgenic mouse expressing an improved Cre (iCre) under GATA-1-HRD promoter control. This mouse was crossbred with ROSA 26 mouse and the progeny was examined for tissue specificity of iCre expression using beta-galactosidase staining. Brain, spleen, kidney, heart, thymus, liver, lung and ovary were examined for whole organ LacZ staining. All tested organs were negative except kidney and spleen where some positivity was observed. Subsequently, we prepared tissue sections from kidney, spleen and bone marrow and stained with LacZ and anti-beta-galactosidase antibody. Only the bone marrow EpoR expressing cells were positive; the kidney and the spleen cells were negative. Although Suzuki et al previously showed expression of the GATA-1-HRD driven erythropoietin receptor in spleen using RT-PCR, we were not able to find iCre expression in the splenic cells using these approaches. We demonstrate that our transgenic mouse (GATA-1-HRD-iCre) showed a restricted iCre expression in hematopoietic tissue that differs from previous studies of other hematopoiesis specific cre mouse. We conclude that this mouse model should be useful in studies of function of erythroid specific genes.
A Non-Invasive Evaluation of Hemoglobin with a New Optical Sensor.
Authors: Berrebi Alain , Amir Orna , Weinstein Aharon , Monashkin Emmanuel , Holland Paul 
Published: April 2020

Abstract
Background: The performance of the NBM-100, a new non-invasive Hematocrit (Hct) measurement device, was tested in primary and secondary medical care environments. The device utilizes a finger sensor with Occlusion Red/Near-Infrared Spectroscopy (O-RNIRS) technology to non-invasively measure hemoglobin and Hct levels (Figure). A non-invasive Hct measurement has many potential advantages including the prevention of pain and potential transmission of infectious diseases, and the reduced need for trained technicians.
Methods: In order to investigate the use of the device in both primary and secondary medical environments, clinical trials were conducted in a blood donation center (USA), an oncology clinic (USA) and a hematology clinic (Israel). The studies were carried out on a group of 304 subjects (155 Female, 149 Male), ages 18–96 (average age 54.9).
After signing a written consent, all study volunteers were tested non-invasively with the NBM-100 device, and invasively, using a venous sample evaluated on a Cell-Dyn (Abbott) blood analyzer. At the blood donation center the NBM-100 measurements were performed before donation, and the venous blood sample was taken after donation. To neutralize the decrease in Hct occurring during blood donation, as described in published clinical trials, a value of 2.2% was added to the reference venous values obtained from the blood donation center.
Results: The operating staff at all centers found the NBM-100 easy to use and appreciated by subjects. The venous Hct measurements ranged from 15% to 51%. The mean NBM-100 Hct was 36%, the same as the mean venous result. The mean absolute error (MAE) was 2.66% and the relative absolute error (RAE) was 7.9%. The standard deviation of the difference between the venous reference and noninvasive Hct readings was found to be 3.34% and the correlation between the two devices was r=0.9. The bias between the two methods was a negligible 0.40%.
Conclusions: This study supports the use of the NBM-100 device as a useful, safe and accurate non-invasive method for measuring a wide range of Hct in both a hospital (e.g. surgery and blood transfusions) and ambulatory setting (e.g. blood banks). Good agreement was found between the non-invasive NBM-100 and the invasive Cell-Dyn Hct measurements. The non-invasive system was found to be environmentally - and user-friendly by the operating staff. These findings support the high clinical utility of a commercial application of the NBM-100 device.
Altered Immune Response in Severe Malaria Anemia in Children.
Authors: Gordeuk Victor R. , Kasvosve Ishmael , van Dijk Janneke , Weiss Guenter , Debebe Zufan , Nekhai Sergei , Kuddo Thea , Thuma Philip E. 
Published: April 2020

Abstract
We prospectively assessed immune markers in children &lt;6 years with severe malarial anemia (hemoglobin &lt;5.0 g/dL; n = 72) and uncomplicated malaria (n = 69) who presented to Macha Mission Hospital in Zambia’s Southern Province. We also studied 70 children &lt;6 years who presented to well child clinics in Harare, Zimbabwe as controls. Compared to controls, children with uncomplicated malaria had significantly higher temperatures and parasite counts, lower hemoglobin and platelet concentrations, higher plasma levels of interferon-gamma, tumor necrosis factor alpha, and interleukin 10 and lower levels of monocyte inhibitory factor (MIF). Compared to uncomplicated malaria, severe malaria anemia was associated with younger age, longer duration of fever and lower temperature on admission. Reticulocyte index and serum concentrations of bilirubin and LDH did not differ between the malaria groups, suggesting that unusually severe extra- or intra-medullary hemolysis did not explain the severe anemia. Higher white blood cell and platelet counts in the severe malaria group suggested that pan-suppression of the marrow was also not the primary cause. Of originally selected measures of inflammation, plasma levels of TNF-alpha and MIF did not differ between the malaria groups, but concentrations of both interferon-gamma and interleukin-10 were significantly lower in the severe anemia group (P &lt;0.006). Additional testing revealed levels of interleukin-1alpha, interleukin-6, and IP-10 to be lower and levels of sFAS to be higher in the children with severe anemia versus uncomplicated malaria (P &lt;0.0005). In a logistic regression model, severe malarial anemia was associated with younger age (P = 0.010), prior treatment with sulfadoxine/pyrimethamine or traditional medicine (P &lt;0.32), lower levels of Interleukin-10 (P = 0.025) and higher levels of sFAS (P = 0.003) and TNFa (P = 0.013). Our results are consistent with a multifactorial cause of severe malarial anemia, possibly including infection with resistant plasmodia, over-expression of TNF-alpha in conjunction with under-expression of IL-10, and increased apoptosis.
SNP Associations with Event Free Survival in Myeloma from Two Phase III Clinical Trials Using the Bank On A Cure Chip.
Authors: Van Ness Brian G. , Crowley John C. , Ramos Christine , Grindle Suzanne M. , Hoering Antje , Haessler Jeff , Jacobus Susanna , Durie Brian G. , Katz Michael , Rajkumar Vincent S. , Barlogie Bart , Johnson David C. , Morgan Gareth J. 
Published: April 2020

Abstract
While there are certain common clinical features in myeloma, the disease shows significant heterogeneity with regard to disease progression, and responses to therapy, affecting both survival and toxicities. Heritable variations in a wide variety of genes and pathways affecting cellular functions and drug responses likely impact patient outcomes. In the Bank On A Cure (BOAC) program we have developed a custom chip that assesses 3,404 SNPs representing variations in cellular functions and pathways that may be involved in myeloma progression and response. The chip has gone through rigorous quality controls checks for high call rates, accuracy, and reproducibility that will be presented. Using the BOAC chip, we have conducted studies to look for SNPs that may identify biologic variations that are associated with good or poor response across a variety of treatments. In this study we looked for SNPs that may distinguish short term and long term survivors in two phase III clinical trials: ECOG E9486 and intergroup trial S9321. E9487 patients were treated with VBMCP followed by randomization to no further treatment, IFN-alpha, or cylcophosphamide; and, although there was variation in survival, no significant differences in survival were noted among the 3 arms of the trial. Patients included in this SNP study from S9321 received VAD induction followed by randomization to VBMCP or high dose melphalan + TBI. SNP profiles were obtained for patients with less than 1 year EFS (n=20 in E9487; n=50 in S9321) and patients showing greater than 3 years EFS (n=32 in E9486; n=41 in S9321). Statistical approaches were performed to identify single and groups of SNPs that best discriminated the survival groups. Previous studies have suggested genetic variations in drug metabolism genes, p-glycoprotein transport, and DNA repair genes may influence survival outcomes. Our results show significant survival associations of genetic variations in genes within these functional categories (eg. GST, XRCC, ABCB, and CYP genes). Although genetic variations were found that were uniquely associated with each clinical trial, several of these genetic variations show survival associations that increase in significance when the two trials were examined as a conglomerate data set. Grouping genetic variations through common pathway approaches using gene set enrichment analysis, as well as clustering or partitioning algorithms, further improve the value of the SNPs as potential prognostic markers of survival outcomes. These results and statistical approaches will be presented, and represent steps toward identifying patient variations in biologic mechanisms important in predicting therapeutic outcomes.
MafG-Null Mice Show Delayed Leukocyte Engraftment Following Whole Bone Marrow Transplant.
Authors: Zafar Abu-Bakr , Li Xiao-Miao , Jorgensen Marda L. , Hu Zhongbo , Bungert Jorg , Engel James , Slayton William 
Published: April 2020

Abstract
Megakaryocytes, which give rise to platelets, constitute a small population of cells. However, molecular aspects of megakaryocyte development are poorly understood. MafG is the small subunit of the transcription factor, NF-E2, which plays a major role in terminal megakaryocyte differentiation and platelet release. We sought to determine the effect of impaired platelet release due to targeted ablation of the mafG gene following a hematopoietic stem cell transplant on engraftment kinetics. We performed three types transplants:C57Bl/6 Ly5.1 murine bone marrow (mafGWT/WT bm) into lethally irradiated mafG null hosts expressing green fluorescent protein, (GFP),mafG-null/GFP bm into lethally irradiated C57BL/6 mafGWT/WThosts, andmafGWT/WT GFP bm into lethally irradiated C57BL/6 mafGWT/WT hosts.
We measured peripheral cell counts, the presence of circulating donor-derived cells by flow cytometry, changes in the cellularity of the bone marrow and splenic weight, and megakaryocyte size and concentration on day 5, 7, 14, and 28 post transplant. We found:Platelet engraftment kinetics differed when mafG null animals served as donors or recipients. Specifically, platelets reached their lowest point at day 7 when mafGWT/WT or mafG null bm was transplanted into wildtype hosts. When mafGWT/WT bm was transplanted into mafG null hosts, platelets reached a low point on day 5.MafG null hosts transplant with mafGWT/WT bm had 5-fold lower circulating platelet levels than mafGWT/WT hosts at day 5.When mafGWT/WT served as donors, circulating platelets began to increase after day 7, while animals receiving mafG null donor cells had platelet counts that continued to diminish.Splenic hematopoiesis was markedly decreased when mafGWT/WT bm was transplanted into mafG null hosts. In contrast, when mafG null bm was transplanted into mafGWT/WT hosts, splenic hematopoiesis was increased 2.5 fold on day 28 compared to animals receiving mafGWT/WTdonor cells.Mortality rate was much higher in mafG null hosts.Leukocyte engraftment was delayed when mafGWT/WT bm was transplanted into mafG null hosts.
These results suggest that host factors determine the timing of the platelet nadir following bone marrow transplant. Furthermore, the mafG protein, contributes in previously unrecognized ways to splenic hematopoiesis and successful engraftment, possibly through its function in promoting megakaryocyte development and platelet release.
MicroRNA hsa-mir-155 Blocks Myeloid and Erythroid Differentiation of Human CD34+ Cells.
Authors: Georgantas Robert W. , Hildreth Richard , Morisot Sebastien , Alder Jonathan , Civin Curt I. 
Published: April 2020

Abstract
In a large microRNA-array and bioinformatics study, we determined all of the microRNAs (miRs) expressed by human CD34+ hematopoietic stem-progenitor cells (HSPCs) from bone marrow and G-CSF mobilized blood. When we combined miR expression data, mRNA expression data from a previous study (Georgantas et al, Cancer Research 64:4434), and data from various published miR-target prediction algorithms, we were able to bioinformaticly predict the actions of miRs within the hematopoietic system. MircoRNA hsa-mir-155 was highly expressed in CD34+ HSPCs, and was predicted by our bioinformatics database to target several HSPC-expressed mRNAs (CREBBP, CXCR4, Jun, Meis-1, PU.1, AGTRI, AGTRII, Fos, and GATA3) that encode proteins known to be involved in myeloid and/or erythroid differentiation. We used luciferase-3′UTR reporter constructs to confirm that protein expression from these mRNAs were in fact down regulated by microRNA. As an initial test of mir-155′s effect on hematopoietic differentiation, K562 cells were transduced with hsa-mir-155 lentivirus and then exposed to TPA to induce megakaryocyte differentiation, or to hemin to induce erythroid differentiation. Compared to controls, miR-155 reduced K562 megakaryocyte differentiation by ~70%, and erythroid differentiation by &gt;90%. Thus, mir-155 appears to be sufficient to inhibit both megakayrocyte and erythroid differentiation. K562 proliferation was not affected by mir-155, showing that the differentiation block was not due to cell cycle arrest. MicroRNA hsa-mir-155-transduced human mobilized blood CD34+ cells generated &gt;70% fewer myeloid and erythroid colonies than controls in colony forming (CFC) assays, further indicating that mir-155 blocks both myeloid and erythroid differentiation. We are currently further testing the effects of mir-155 on differentiation of CD34+ cells in vitro, and also in vivo on their ability to engraft immunodeficient mice.
The Pim-1 Kinase Is a Direct Target of the Hoxa-9 Homeodomain Protein and a Mediator of Hoxa-9’s Effects on Hematopoiesis.
Authors: Lawrence H. Jeffrey , Hu Yu-Long , Passegue Emmanuelle A. , Fong Stephen , Largman Corey 
Published: April 2020

Abstract
The Hoxa-9 homeodomain protein exerts dramatic effects in hematopoiesis. Loss of Hoxa-9 function in blood cells severely inhibits proliferation of primitive hematopoietic cells and impairs in vivo hematopoietic stem cell (HSC) function. The defects in Hoxa-9−/− animals are more striking than any other single or compound Hox mutation reported to date, and more dramatic than that seen in animals lacking the entire Hoxb cluster of homeobox genes, presumably because Hoxa-9 is one of the most highly expressed members of the 39-member Hox gene family in HSC. Conversely, enforced expression of Hoxa-9 markedly enhances the proliferation of primitive blood cells, expands HSC and leads inevitably to a fatal acute myeloid leukemia in mice. Despite these potent biological effects, the pathways by which Hoxa-9 acts in blood cells are unknown, and although Hoxa-9 is a DNA-binding transcription factor, no direct physiologic target genes have been identified in hematopoietic cells. Our previous gene expression profiling studies suggested that Hoxa-9 positively regulates the Pim-1 serine-threonine kinase, an oncogene with important antiapoptotic and pro-proliferative effects in blood cells. In addition, the hematological phenotypes of Hoxa-9 and Pim-1 deficient animals are strikingly similar, and include a blunted response to myeloid cytokines, T- and B-cell defects, and microcytosis of red blood cells. Using chromatin immunoprecipitation assays, we show that Hoxa-9 protein binds directly to the Pim-1 promoter/enhancer region in hematopoietic cells. Enforced expression of Hoxa-9 significantly induces Pim-1 mRNA and protein in blood cells in a DNA-binding dependent manner. Furthermore, Pim-1 protein levels are diminished in lin- blood cells from Hoxa-9−/ − fetal liver, showing that Hoxa-9 is required to maintain normal levels of Pim-1 in primitive hematopoietic cells. By real-time PCR, we find that Hoxa-9 and Pim-1 mRNA levels track together in highly purified subpopulations of long-term and short-term HSC and in common myeloid and granulocyte-monocytes progenitors (CMP/GMP). Induction of Pim-1 protein by Hoxa-9 significantly increases the phosphorylation and inactivation of the pro-apoptotic BAD protein, a known target of the Pim-1 kinase. Hoxa-9−/ − marrow cells show increased levels of apoptosis and decreased cell cycle progression and proliferation compared to wild-type cells, and these defects are significantly improved by the re-introduction of Pim-1. These data provide strong evidence that Pim-1 is a direct transcriptional target of Hoxa-9 and a key mediator of its antiapoptotic and pro-proliferative effects on primitive hematopoietic cells. Since Hoxa-9 is upregulated in many cases of human acute myeloid leukemia, Pim-1 may serve as a useful therapeutic target in Hoxa-9 driven leukemias.