bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analyzing Variations in Blood Utilization Improves Transfusion Practice.
Authors: Arthur Kathleen R. , Menitove Jay E. 
Published: April 2020

Abstract
During the past five years, randomized, controlled trials and clinical guidelines provided data for making evidence-based decisions about blood transfusion. In light of this new information, we monitored transfusion practices/variations among hospitals in our region. We sought to determine whether hospitals actively involved in addressing transfusion utilization and best practices in 1999 (hospitals 1–3) experienced changes when re-surveyed in 2003. As a “control” group, we determined transfusion patterns at 9 hospitals in 2003 (hospitals 4–12), that did not participate in the 1999 survey. We compared autologous transfusion practices involving orthopedic surgery patients at 3 hospitals participating in both surveys and at 8 hospitals contrasting 2003 survey data and 1999 hospital data (no data was available for comparison from one hospital). Data comparisons involved transfusions between Oct.–Dec.1999 vs. Oct–Dec. 2003. In addition we compared transfusion practices involving cardiovascular surgery patients (DRG’s 104–107, 109–111) at 2 hospitals participating in the 1999 and 2003 surveys and 5 hospitals joining the survey in 2003. Among the hospitals participating in 1999, the number of patients donating autologous units for orthopedic surgery declined 68%(88 to 28 patients), the number of patients transfused decreased by 58% (55 to 23patients), and the number of units collected was 68% lower (151 to 49 units). In contrast, at hospitals participating in the survey for the first time in 2003, there was only a 7% decline in the number of autologous units collected (274 to 255); the number of patients donating was unchanged (156 and 157). Of note, hospitals 10 and 12 were affiliated with hospital 3 and followed the same transfusion protocols. The average number of units transfused per patient in 1999 and 2003 in both groups (~1.5 and 1.8) was essentially unchanged. The relatively low number of units infused further questions the need for autologous donations in these patients.
At 2 hospitals participating in both the 1999 and 2003 surveys, the percentage of cardiovascular surgery patients transfused with red blood cells decreased (40% to 29% and 60% to 44%). This compares to a higher percentage of patients receiving transfusions (range 46–80%) at hospitals participating only in 2003. A similar difference appears in the average number of units infused per transfused patient (2.2–3.6 vs. 2.9–4.7). No differences in patient outcomes were attributed to changes in transfusion practices. Blood utilization patterns improved at hospitals and their affiliates committed to transfusion practice benchmarking compared to hospitals not utilizing this monitoring modality. We conclude that utilization variation analysis remains an effective methods for influencing transfusion practice.
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Preparing Citrated Non-Activated Gamma Globulin for Rapid Infusion.
Authors: Shanbrom Edward , Owens William J. 
Published: April 2020

Abstract
Human plasma is our most valuable source of therapeutic proteins, and intravenous gamma globulin (IVIg) is the most widely used plasma derivative. Traditional yields of the product from alcohol fractionation are not optimal; added to this, viral contamination and other well known side effects, including fever and aseptic meningitis, continue to be of concern. A method of precipitating very high yields of cryoprecipitable proteins with sodium citrate (“Supercryoprecipitation”) has been previously described. A by-product of “Supercryoprecipitation” is a very clean cryoprecipitate-free plasma supernatant, with a citrate concentration of 5% to 6% (w/v). When up to 10% (w/v) additional citrate is introduced into the supernatant, there is a preferential precipitation of nearly 100% of the remaining gamma globulin. Immunoglobulin G (IgG) yield was measured by radial immunodiffusion (86.3% ± 12.5%). IgG, being the largest fraction of gamma globulin, is most often used as a measure of product quality. Protein electrophoresis of the citrate precipitation product showed very little contamination by other plasma fractions (albumin: 9.8% ± 3.3%, alpha globulin: 1.6% ± 0.6%, beta globulin: 3.9% ± 1.2%, gamma globulin 84.3% ± 13.8%). Unlike present protein precipitation methods, citrate precipitation does not rely on denaturation, thereby better preserving the native structure of immunoglobulin. This may lead to improved tolerance during infusion, and may allow for more rapid infusion rates, or improved subcutaneous infusion. Stability and safety studies are currently underway. Considering the current unsettled climate in the world, the potential for improved gamma globulin therapies and supply should not be overlooked.
Further Studies of Galectin-1 (Gal-1) on CD34+ Umbilical Cord Blood Cells.
Authors: Etchegoyen Orlando J. , Morales Victor H. , Milone Jorge H. , Fink Nilda E. 
Published: April 2020

Abstract
The galectins are a family of carbohydrate-binding proteins that are distributed widely in vertebrate organisms. Each galectin exhibits a specific pattern of expression in various cells and tissues. Although these proteins are found mainly in the cytoplasm, some of them are secreted from cells and interact with other glycosylated molecules at the cell surface or with the extracellular matrix. There is no information about the interaction of Gal-1 and hematopoietic progenitor cells (HPC). We studied the effect of rat recombinant galectin-1 (rGal-1) on the clonogenic capability and the morphologic differentiation of the human umbilical cord blood HPC in methylcellulose cultures. The quantification of total anti-Gal-1 binding capability (total receptors) was also determined by flow citometry. First, the cells of 10 human cord blood units were collected and CD34+ selected using an indirect immunomagnetic method (MidiMACS, Miltenyi, Biotec). Ten thousand events were acquired in a flow cytometer (FACScan, Becton Dickinson), employing 105 CD34+ cells, rabbit anti Gal-1 antiserum and anti-rabbit IgG FITC antibody. Finally the CD34+ cells were cultured in a semisolid methylcellulose medium (Methocult GF+ H4435, StemCell) by triplicate at 37°C, 5% CO2 and 100% humidity during 14 days, employing 24 wells culture plates (Nunc) in presence or not of different concentrations of rGal-1 (2, 4, 6 and 20 μg/ml). We analyzed the number of GEMM-CFU (mix colonies), GM-CFU (granulocyte- macrophage colonies), BFU-E (erythroid colonies) and the presence of signals of abnormal morphologic differentiation. Control of inhibition with thiodigalactoside (TDG) and lactose (Lac) 100 mM was done in every case to determine the assay specificity. Histograms show a total receptor mean expression (mean+/−SEM) in CD34+ cells of 18.2+/−2.44% which was significantly inhibited (p&lt;0.05) with TDG (8.8+/−0.98%) and Lac (13.5+/−3.70%). A statistically significant increase in the number of GEMM-CFU, GM-CFU and BFU-E was seen. These increases were directly proportional to the rGal-1 concentration added (p&lt;0.05) and it was stronger for GM-CFU. This effect was partially inhibited by TDG and Lac. We observed an abnormal increase in the mesenchymal, fibroblastic-like cell growth in methylcellulose cultures. These results showed the expression of Gal-1 in CD34+ human umbilical blood cells membranes. The addition of rGal-1 to cultures would increase the proliferation of HPC and the differentiation to stromal-like cells would partially occur. The mechanism and functional meaning of this activation and differentiation must still been elucidated.
Gene-Chip Analysis of Cord Blood Derived CD34+ Cells Expanded on Bioartificial Materials.
Authors: Oswald Joachim , Steudel Christine , Salchert Katrin , Thiede Christian , Ehninger Gerhard , Werner Carsten , Schuler Ulrich , Bornhaeuser Martin 
Published: April 2020

Abstract
Expansion of hematopoietic stem cells from neonatal cord blood is an important issue for clinical uses since the number of CD34+ cells in individual cord blood samples is limited and often not sufficient for a successful engraftment in adult individuals. In vivo, hematopoietic stem cells reside in the bone marrow in close vicinity to stromal cells and extracellular matrix molecules. We have established a culture system for the ex vivo expansion of CD34+ cord blood cells utilizing fibrillar collagen 1 as a bioartificial matrix to enable cellular adhesion during cell culture. CD34+ hematopoietic stem cells were isolated via immunomagnetic separation from umbilical cord blood after informed consent and cultivated in presence of recombinant cytokines and reconstituted collagen 1 fibrils as matrix. After seven days of cultivation, expansion of cells, expression of surface molecules cells and expansion of colony forming units were assessed. Additionally gene expression profiling was performed with Affymetrix HG U133A chips interrogating 22,253 probe sets. As control, CD34+ cells were expanded in liquid culture without fibrillar collagen.
The overall expansion of CD34+ cells was 4.2 fold + 1.7 compared to 11.1 fold + 2.9 for the control sample. The number of colony forming units (CFU) was increased in the collagen 1 containing samples was elevated (65.1 + 10.3 compared to 26.1 + 7.6 in the control). Gene expression analysis with chip technology showed up regulation of several cytokines (e.g. interleukin 8, interleukin 1a) and also of transcription factors with antiproliferative features like BTG2. The chip data have been verified with quantitative PCR using the Taqman technology.
Our data support the idea that direct contact of CD34+ cells with fibrillar collagen 1 results in a delay in cell cycle progression which prevents a subsequent differentiation into more committed progenitors. Therefore fibrillar collagen 1 may serve as supportive matrix for the ex vivo expansion of cord blood derived CD34+ cells.
Comparative Analysis of Methods for Assessment of Circulating Endothelial Progenitor Cells Indicated Heterogeneity of Subpopulations at Various Stages of Differentiation.
Authors: Deutsch Varda R. , Shmilovich Haim , Abashidze Anastasia , Miller Hylton , Keren Gad , Naparstek Elizabeth , George Jacob 
Published: April 2020

Abstract
The cells of the vasculature are derived from pluripotent stem cells, known as hemangioblasts, which also give rise to blood. Angiogenesis, the formation of new blood vessels is considered to result from the proliferation and migration of mature endothelial cells from existing blood vessels or from the recently described, but not yet well characterized endothelial progenitor cells (EPCs). The number and properties of EPC in disease states is of considerable interest due to the promising therapeutic potential of these cells. EPCs have been shown to be mobilized from the bone marrow and contribute to angiogenesis following vascular injury, organ ischaemia and tumor progression. However, mechanisms that drive the EPC response to injury remain elusive and the lack of definitions of EPC subpopulations and the many methods used by different groups to identify these cells make correlation of results difficult. To begin to understand the potential of these cells, we performed a comparative analysis of several methods used for circulating EPC assessment in 40 healthy individuals (mean age of 33±9 years) and correlated them with humoral factors known to influence their numbers. Peripheral blood mononuclear cells were obtained and evaluated by flow cytometeric analysis with antibodies to CD34, CD45, CD133 and KDR, and the remaining cells grown under endothelial cell conditions for assessment of colony forming unit (CFU) numbers and adhesive properties. Levels of circulating vascular endothelial growth factor (VEGF), erythropoietin and C-reactive protein were determined and correlated with each of the EPC markers. CFU numbers did not correlate with the number of CD34/KDR (VEGFR2) or CD34/CD133/KDR positive cells and negatively correlated with CD34/CD133 (which includes hematopoietic progenitors). CD34/KDR number correlated with CD34/CD133/KDR but not with CD34/CD133. Only the VEGFR2 positive cells populations (CD34/KDR and CD34/CD133/KDR) correlated with VEGF serum levels. The number of EPC adhering to fibronectin and endothelial-cells correlated with CFU numbers but not with EPC membrane markers. Our results indicate for the first time that EPC, like hematopoietic precursors may be a heterogeneous cell population comprised of progenitors at various stages of differentiation, having varied proliferative capacity. This could explain the lack of correlation in results obtained using different methods for quantitatively assessing the numbers of circulating EPC. These data also suggest that co-expression of CD34, CD133and KDR-VEGFR2 is appropriate for defining a population of circulating EPC whereas CFU are more likely to reflect the proliferative capacity of the progenitors.
Acquisition of CD34 Correlates with Increased Hematopoietic and Self Renewal Activity of CD34−CD133+ Cord Blood Cells.
Authors: Götze Katharina S. , Schiemann Matthias , Marz Stefanie , Peschel Christian , Oostendorp Robert A.J. 
Published: April 2020

Abstract
CD34 is a sialomucin expressed on hematopoietic cells, endothelial cells and muscle satellite cells. Within the hematopoietic system, CD34 expression has been associated with very immature progenitor cells as well as hematopoietic stem cells (HSC), and it is widely used to assess stem cell activity in clinical protocols. In the past, HSC activity was thought to be retained exclusively in the subset of cells expressing CD34. This view has been challenged by recent observations in mice in which HSC activity was also found in the CD34-negative fraction. These findings have since been reproduced using human marrow and cord blood cells. However, the exact relationship between CD34+ and CD34− stem cells remains unclear. We investigated the regulation of CD34 expression as dependent on cell division history. To follow cell division, human cord blood cells were labeled with the fluorescent dye CFSE. Lin-CD34−CD133+CFSE+ (CD34−) and CD34+ populations were almost indistinguishable in their ability to produce CAFCweek6 content. After three days of serum-free culture with stem cell factor, Flt3 ligand and thrombopoietin, almost all initially CD34− cells had acquired expression of CD34, including all undivided cells. We found that, in cultures initiated from CD34− cells, virtually all CAFCweek6 were produced from the divided, now CD34+ cells, indicating these cells had self-renewed. In contrast, similar cultures from initially CD34+ cells demonstrated that hematopoietic activity associated with the undivided cell fraction. We did not find any hematopoietic activity in the cell fraction that remained CD34− or the fraction that lost CD34 after division. Analysis of mRNA expression showed that CD34− and CD34+ cells expressed almost equal levels of CD34, AC133, Flt1, Flk1 and Flt4, while CD34− cells expressed significantly lower levels of Tie1 and Tie2 than CD34+ cells. The expression of CD34 message in CD34− cells was explained by our observation that these cells contained intracellular CD34, indicating that they are “primed” to express the antigen on their cell surface. In conclusion, Lin−CD34−CD133+ cells acquire expression of CD34, even in the absence of cell divisions. These CD34− cells self-renew more rapidly in vitro than cells initially expressing CD34, and self-renewal is preceded by acquisition of CD34 antigen.
Identification of Quantitative Trait Loci Regulating Steady State Hematopoiesis and Progenitor Mobilization in AKRB6F2 Mice.
Authors: van Os Ronald , Ausema Albertina , Vellenga Edo , de Haan Gerald 
Published: April 2020

Abstract
The hematopoietic system is a complex organized tissue with a hierarchical structure. Identification of organizational pathways within the hematopoietic system would be relevant for a better understanding of hematopoiesis in health and disease. We have analyzed numerous hematopoietic parameters in AKRB6F2 mice, a cross between AKR and C57Bl/6 mice that differ in various stem cell traits. Specifically, we have measured stem cell numbers, progenitor cell cycling, G-CSF induced stem cell mobilization, as well as neutrophil numbers in blood and bone marrow. We used these data to map quantitative trait loci for these parameters in order to assess whether distinct traits are associated with each other and by which genetic loci these parameters are regulated. When the different hematopoietic parameters were compared, surprisingly few significant correlations were found and only two quantitative trait loci (QTL) could be identified for all hematopoietic parameters. We confirm that a region on chromosome 11 is involved in progenitor cell cycling and we found an additional QTL for stem cell numbers, measured as CAFC day 35 per femur, on chromosome 2. A suggestive QTL was found for the number of progenitors in the bone marrow and the number of neutrophils in the blood. In the first of two series of offspring mice we found a strong association between stem cell mobilization and B6 alleles at the chromosome 3 locus D3Mit88 or AKR alleles on chromosome 15 (D15Mit241). However, this could not be confirmed in the second series. In addition, breeding of an F3 generation from the best mobilizers (6479 ± 1544 CFU-GM/ml blood vs. 3481 ± 3076 overall) of each series did not result in high mobilization capacity of the offspring (1333 ± 844 CFU-GM/ml). Taken together, our data illustrates that different hematopoietic parameters are independently regulated. This suggests that hematopoiesis is highly structured and regulation at all levels of stem cell development can lead to distinct mechanisms of stem cell maintenance resulting in normal levels of mature blood cells. Additionally, variation based on epigenetic determinants that do not transmit to the next generation may play an important role in the regulation of steady state hematopoiesis and stem cell mobilization.
Recurrent Abnormalities in TGIF RNA Sequence in Blasts from Patients with Acute Myeloid Leukemia.
Authors: Hamid Rizwan , Vnencak-Jones Cindy L. , Martincic Danko , Means-Powell Julie A. , Cogan Joy D. , Brandt Stephen J. 
Published: April 2020

Abstract
To identify genes with prognostic potential in acute myeloid leukemia (AML), we applied microarray analysis and real-time PCR to RNA from cryopreserved bone marrow or venous blood samples from 61 patients with newly diagnosed or relapsed AML, and the expression of one gene, TG-interacting factor (TGIF) was found to be highly predictive of relapse and survival. TGIF is a transcriptional repressor belonging to the TALE (three amino acid loop extension) class of homeobox proteins, and deletion or mutation of a single allele of TGIF is associated with the craniofacial genetic disorder holoprosencephaly (HPE). TGIF transcripts in blasts from patients with AML have now been analyzed for mutations using reverse transcriptase-based PCR followed by dideoxy fingerprinting and automated sequencing. Using this approach, we identified the identical, aberrantly spliced TGIF transcript in independent RNA samples from a subset of patients (10/48) with low TGIF expression and poor long-term survival. Sequencing of this novel product showed that it lacked part of exon 1, all of exon 2, and most of exon 3 and eliminated all of the known interacting domains of TGIF (DNA binding, SMAD2, and mSin3A interacting). In contrast, Southern blot analysis of DNA from these patients did not identify any gross abnormalities in TGIF gene structure. This in combination with the RNA results suggests the likelihood of aberrant splicing. In sum, these results identify a novel, recurrent genetic abnormality in AML. The presence of aberrant splicing in AML contrasts with HPE, which is associated with mono-allelic mutations or deletions. As the protein product of this transcript would not be expected to have any of the known functions of TGIF in the cell, reduced expression of functional protein product may have a role in relapse or progression of AML.
Dissecting the Mechanism of Endogenous GATA-2 Transcription: GATA Factor Interplay at Multiple Upstream Regulatory Regions Establishes Long-Range Transcriptional Control.
Authors: Bresnick Emery H. , Martowicz Melissa M. , Grass Jeffrey A. , Boyer Meghan E. 
Published: April 2020

Abstract
Given the simplicity of the DNA sequence that mediates binding of GATA transcription factors, GATA motifs reside throughout chromosomal DNA. However, analyses of GATA-1 occupancy by chromatin immunoprecipitation (ChIP) revealed that GATA-1 discriminates exquisitely among these sites. GATA-2 selectively occupies the −2.8 kb region of the GATA-2 locus in the active state, despite there being numerous GATA motifs throughout the locus. The GATA-1-mediated displacement of GATA-2 is tightly coupled to repression of GATA-2 transcription. We have used high-resolution ChIP to show that GATA-1 and GATA-2 occupy two additional regions, −3.9 kb and −1.8 kb of the GATA-2 locus. GATA-1 and GATA-2 had distinct preferences for occupancy at these regions, with GATA-1 and GATA-2 occupancy highest at the −3.9 kb and −1.8 kb regions, respectively. Activation of an estrogen receptor fusion to GATA-1 (ER-GATA-1) induced similar kinetics of ER-GATA-1 occupancy and loss of GATA-2 occupancy at the sites. As the distinct preferences for GATA factor occupancy were not evident from electrophoretic mobility shift assay analysis in vitro, establishment of the preferences requires the native nucleoprotein structure of the GATA-2 locus. In the transcriptionally active state, the −3.9 kb and −1.8 kb regions formed strong DNaseI hypersensitive sites (HSs), and the −2.8 kb region formed a weak HS. Whereas ER-GATA-1-instigated repression abolished the −1.8 kb HS, the −3.9 kb HS persisted in the inactive state. Transient transfection analysis provided evidence that the −3.9 kb region functions distinctly from the −2.8 kb and −1.8 kb regions. Molecular studies, including capturing chromosome conformation (3C) analysis, are underway to dissect the structure/function of the upstream regulatory regions. Based on the high sequence conservation and occupancy by GATA factors in vivo, targeted deletions of the −2.8 kb and −1.8 kb regions were generated. Mutant mice are being developed, and functional analysis is being conducted via in vitro differentiation of homozygous mutant ES cells. Our results support a model in which dynamic changes in GATA factor occupancy and chromatin structure at the −3.9 kb, −2.8 kb, and −1.8 kb regions are intimately controlled by Friend of GATA-1 and additional coregulators. This GATA factor interplay is essential for GATA-2 transcriptional regulation and therefore the control of hematopoiesis.