bluuebunny/crossref_metadata_2025_split huggingface dataset:
Procedural Apheresis Instrumentation Algorithm Minimizes Platelets Loss during Peripheral Blood Stem Cells Collection.
Authors: Richa Elie M. , Burgstaler Edwin A. , Greiner Carl W. , Bryant Sandra C. , Ege Hilmi , Gastineau Dennis A. , Winters Jeffrey L. 
Published: April 2020

Abstract
Introduction:
Peripheral blood hematopoietic progenitor cell collection is a more popular source of stem cells for transplantation than bone marrow harvest. A number of automated cell separators exist which can be used to perform mononuclear cell collections. These separators collect varying amounts of other cells during the collection process, including them in the mononuclear cell product. Because of this, collection of mononuclear cells can result in significant reductions in platelet count and loss of red blood cells during the procedure. In order to minimize platelet transfusion support, we developed an algorithm to optimize mononuclear cell collections while minimizing platelet losses using the Gambro COBE Spectra and the Baxter Amicus
Methods:
Previously, patients were assigned randomly to any available apheresis machine. Starting August 2nd 2002, patients with a WBC count higher than 35×10 (9)/L were preferably assigned to Amicus for 5 hour collection with processing 1000 ml/cycle or as second choice Spectra for 5 hours run with maximum draw rate 65 ml/Min., for both instruments. If the WBC was lower than 35×10 (9)/L, the preferred machine would be Amicus for 5 hours run at 1400 ml/cycle with maximum draw rate 90ml/Min. or Spectra for 5 hours run with maximum draw 100 ml/Min. We reviewed the difference of pre and post procedure platelet counts of non Hodgkin’s Lymphoma patients undergoing stem cells collection for 6 months before and after the implementation of the above protocol from February 2002 till February 2003. All CBC were performed on the same Coulter analyzer.
Results:
14 patients (2 on Amicus and 12 on Spectra) underwent stem cells collection before August 2nd, 2002. 13 patients (8 on Amicus and 5 on Spectra) were collected after this date. The Fisher’s exact test was used for this study. The median and mean differences in the first group were 141.5 ×10(9)/L and 135 ×10 (9)/L respectively (95% CI, p 0.0183). The second group had a median and mean of 45 ×10 (9)/L and 51 × 10(9)/L (95% I, p 0.0183). The minimum and maximum differences were 36 × 10(9)/L and 200 × 10(9)/L respectively in the first group and −19 ×10(9)/L and 121×10(9)/L for the second group. There was no difference in the numbers of the collections days in both groups in regard to the type of automated cell collector.
Discussion:
The implemented algorithm showed a net advantage. Even though none of the patients required blood transfusions, there was lesser drop of platelets counts with the new settings.
Hemolysis in an Infant Post Small Bowel Transplant: A Case Report.
Authors: Rhamy Jennifer F. , Parks Jodi A. , Houseworth Julie L. , Seidel Judy , Slayten Jay A. , Waxman Dan A. , Smith Daniel S. 
Published: April 2020

Abstract
Introduction: A 4-month-old male infant received a small bowel transplant. He was readmitted at 9 months of age (m.o.a.) with marrow suppression and severe hemolytic anemia. Serologic evaluation demonstrated both cold and warm autoantibodies as well as two alloantibodies (anti-K and -E). T-activation was also present. The patient was unresponsive to Rituxan and IVIG, so therapeutic plasma exchange (TPE) was initiated at 12 m.o.a. A regimen of immunosuppression, IVIG and TPE was mildly successful in abating hemolysis. A third alloantibody was then identified (anti-Jka) at 13 m.o.a., and the patient continued to hemolyze. This case report describes the investigation of severe hemolytic anemia associated with small bowel transplantation.
Methods: Spreadsheets were generated with hematocrit plotted against these variables: Rituxan, IVIG, TPE, RBC transfusions, and serologic findings. Both venous and buccal samples were DNA-typed for RBC antigens. A venous sample was evaluated for chimerism at two different time points after a discrepancy was observed between buccal and venous DNA typing.
Results: The patient’s condition declined precipitously at 9 m.o.a., and stabilized at 10 and 11 m.o.a., then worsened again. Graphs of the hematocrit did not correlate with any one variable. Slight improvement was observed post-TPE, but hemolysis was observed during the procedures. Blood priming was required for all TPE. A heated circuit was devised to avoid extracorporeal cold agglutination. Samples sent for DNA analysis of the RBC antigens showed donor/recipient chimerism in the venous blood. Two of three “alloantibodies” (anti-E and Jka) were probably of donor graft origin against pt RBC. The third antigen was absent in both patient and donor DNA, and the antibody (anti-K) was likely formed in response to transfusion by either pt or donor lymphocytes. There is also a broadly reactive autoantibody. Genetic analysis showed 10% chimerism at 10 m.o.a., followed by 2% chimerism at 13 m.o.a. A decrease in chimerism was qualitatively associated with some decrease in hemolysis.
Conclusion: No one scenario explains all of this patient’s symptoms. The presence of chimerism suggests generation of autoantibodies and production of two alloantibodies by passenger lymphocytes. Alternately, the patient’s own autoimmune response, or a combination of both processes may play a causative role. Reduction of circulating cold and warm autoantibodies and alloantibodies formed by passenger lymphocytes plus priming with alloantigen-negative, least incompatible RBCs may account for the slight improvement seen with TPE.
Ex Vivo Culture and Amifostine Effects in Human Cord Blood.
Authors: Ryoo Hun-Mo , Bae Sung Hwa , Lee Kyung Hee , Hyun Myung Soo 
Published: April 2020

Abstract
Background
Human cord blood(CB) is the third most promising source of hematopoietic stem cells to the bone marrow and peripheral blood for replacing depressed bone marrow function in high dose chemotherapy in patient with cancer. The possibility of cord blood transplantation in adults was limited by the amount of cord blood that could be collected. Cord blood transplantation after ex vivo expansion with cytokines have already been tried in adults. Amifostine is a phosphorylated aminothiol that affords broad cytoprotection from the myelosuppressive effects of antineoplastic agents.
Objectives
The purposes of this study were to investigate expansion of progenitor and myeloid cells after ex vivo culture of mononuclear cells(MNCs) in umbilical cord blood with growth factor and characterize hematopoietic activities of amifostine.
Methods
MNCs were cultured and ex vivo expanded into myeloid progenitors by using hematopoietic growth factors(IL-1β, IL-3, IL-6, G-CSF, GM-CSF, SCF, EPO) which are known to stimulate differentiation and proliferation of myeloid progenitors. MNCs exposed to the appropriate amount of amifostine for 15 min were cultured in semisolid media and harvested at 24h intervals, and then apoptosis was assessed by flow cytometry.
Results
Myeloid colonies were successfully produced from MNCs. Maximal expansion was obtained with the combination of IL-3+SCF+G-CSF+GM-CSF. SCF was thought to be the most important growth factor for expansion of myeloid progenitor. Pretreatment with amifostine for 15 min stimulated formation of hematopoietic colonies at clinically relevant concentrations ranging from 1 to 100μM. Increase in colony number compare to control were comparable after pretreatment with amifostine (10μM), and CFU-GEMM and BFU-E were highly responsive. Further enhancement of colony was not observed after prolonging the duration of pre-incubation exposure to 1, 8 and 24 hours. Amifostine enhanced IL-1 and IL-3 induced formation of CFU-GEMM and BFU-E. Incubation of MNCs with amifostine in suspension culture increased recovery of secondary colonies. Treatment with amifostine retarded cell loss and apoptosis, and promoted cell survival at 24, 48 and 72 hours in cytokine-deficient medium.
Conclusions
In conclusion, cord blood MNCs can be successfully expanded into myeloid progenitors by using hematopoietic growth factors. This investigation extend the previously recognized hematologic effects of amifostine, and indicate that in addition to its cytoprotective properties, amifostine is a potent stimulant of hematopoietic progenitor growth.
The Effects of Heavy Ion Particles on Human Megakaryocytopoiesis and Thrombopoiesis.
Authors: Kashiwakura Ikuo , Takahashi Kenji , Monzen Satoru , Eguchi-Kasai Kiyomi , Toki Tsutomu , Abe Yoshinao 
Published: April 2020

Abstract
Heavy ion particles provide unique properties in radiotherapy. However, they have also been shown to pose high risks for both work at nuclear facilities and astronauts participating in space missions. In a previous study, we demonstrated that in radio-sensitive megakaryocyte progenitor cells, namely colony-forming unit megakaryocytes (CFU-Meg), a degree of X-ray-induced damage was prevented by post-treatment with several cytokines. In this study, we analyzed the effects of heavy ion particles on megakaryocytopoiesis and thrombopoiesis.
The CD34+ CFU-Meg were isolated from human placental and umbilical cord blood using a magnetic isolation kit and then were exposed to a carbon ion beam (LET=50 KeV/mm). They were cultured in a serum free medium supplemented with a thrombopoietin (TPO) alone or a combination of TPO plus other cytokines including stem cell factor, interleukin-3 (IL-3) and Flt3-ligand. The number of CFU-Meg was calculated by a plasma clot technique. The differentiation into megakaryocytes (CD41+) and the release of platelets (CD42a+) in a liquid culture were both analyzed by flow cytometry. The increase of gamma-H2AX, a marker of DNA double-strand breaks (DSBs) was also detected by flow cytometry.
The sensitivity of CFU-Meg to a carbon ion beam was found to be extremely high and could not be lowered by any type of cytokines unlike X-rays. However, treatment with TPO plus IL-3 potentially induced megakaryocytopoiesis and thrombopoiesis at 14 days after the exposure to a carbon ion beam at 2 Gy. The cytokine treatment enhanced the induction of gamma-H2AX in X-ray-irradiated CD34+ CFU-Meg but not in a carbon ion beam-irradiated one.
These results show that not only the downregulation of death signals, but also the repair of DSBs was less strongly promoted by cytokines in CFU-Meg exposed to a carbon ion beam than X-rays. Different treatments therefore are required to protect against megakaryocytopoiesis and thrombopoiesis damage by heavy ion particles.
Growth Factor Independent Succession of Hemangiopoietic Cell Development and Gene Activation Patterns in Murine Embryonic E14 Stem Cells.
Authors: Peters Stefan O. , Seifert Torsten , Middeke Mareike , Fiedler Anja , Vetters Katja , Wagner Thomas , Stoelting Stephanie 
Published: April 2020

Abstract
Introduction:
We use murine embryonic E14 stem cells (ESC) to study cellular and molecular events that occur during early hemangioblastic development. Cells were precultured in standard medium (DMEM, 20% FCS) for 6 days and then kept as embryoid bodies (EBs) for 0, 2, 5, 10, 15, 20, 25, 30 days either in standard or medium supplemented with VEGF or hematotropic cytokines (G-CSF, IL-3, IL-6, SCF, Flt3). Suspended cells were analyzed for CD 31, flk1, CD144, CD133, CD13 and CD 90 positivity by flow cytometry. CD31 and CD13 stained embryoid bodies were examined by fluorescent microscopy. We here focus gene activation studies on pathways implying endothelial and hematopoetic development according to literature including gene array data. cDNA was analyzed for ERG, EGR1, FLI1, HoxD3, PECAM, FLK1, VE-Cadherin, Pu-1, GATA2, SCL, Epo, LMO4, HoxB4, LMO2, and cyclin D1 activation by TaqMan PCR.
Results:
28% of cells show endothelial CD31 positivity in day 6+0 EBs. In contrast, hematopoietic CD90 positivity cannot be detected at day 6+0 and are firstly found at low levels in 2% at day 6+5. Endothelial CD31 positivity increases significantly at day 6+2 to as much as 58% and remains very high at day 6+5. Thereafter, at day 6+10 and later, the proportions of cells showing CD31 positivity drop to a constant level of about 15%. Hematopoietic development remains low throughout all timepoints and amount to maximally 10% CD90 positivity at day 6+10 declining at later time points. Low level of VEGF could be detected in ESC supernatant as early as day 6+0, endothelial cells could not be further augmented by adding VEGF to the culture. ‘Hematotropic’ cytokines occasionally show to lead to minor increases of hematopoietic cell fractions but not to lead to a shift to earlier hematopoiesis.
Quantitative PCR show that genes implying endothelial development (ERG, EGR1, FLI1, HoxD3, PECAM, FLK1, VE-Cadherin) precede characteristic ‘hematopoetic’ gene (Pu-1, GATA2, SCL, Epo, LMO4, HoxB4, LMO2, cyclin D1) activation. Addition of supplemntal VEGF surprisingly downregulate many of the ‘endothelalial’ genes. Main expression patterns remain unaffected by adding cytokines to the culture.
Summary:
We show on cellular and molecular levels that endothelial cell development in EBs precedes hematopoietic development and that the addition of cytokines does not affect this sequence.
Stimulation of Human Hematopoiesis: The Androstene Link.
Authors: Groothuis Jessie R. , Stickney Dwight R. , Malik Ajay , Garsd Armando , Reading Christopher , Dowding Charles , Frincke James 
Published: April 2020

Abstract
5-Androstene-3β, 17β-diol (AED) is a naturally occurring member of the androstene series of steroids. In models of chemotherapy and radiation-induced mylosuppression, AED expanded hematopoietic stem and progenitor cells (HSPC), decreased need for clinical support and increased survival. We studied hematopoietic activity of AED vs. Placebo in healthy adult (20–64 years, n=9) and elderly (65–72 years, n=9) subjects. AED was safe and well tolerated. Complete blood counts were serially obtained between study days 1–56. Bone marrow (BM) samples were obtained on days -1 and 7. AED produced significant increases in average neutrophil counts (NT) and platelet counts (PLT) as compared to placebo. Adult NT significantly increased between days 2–7 (40–73%, p=0.05) and persisted through day 28 (41.5%); adult PLT increased (19.9%) by day 14, and achieved significance by day 21 (15.6%, p=0.01) and increases persisted through day 28 (19.4%, p=0.02). In elderly subjects, NT significantly increased by 56–96% (days 4–6, p=0.05) which persisted through day 28 (30.7%). Fewer elderly subjects demonstrated robust PLT responses; maximum increase in PLT was seen on day 14 (31.5%) and persisted through day 28. BM samples at day 7 showed increased cellularity in AED but not placebo-treated subjects which was consequently reflected as an increase in peripheral blood NT and PT in AE-treated subjects. BM phenotype changes and maturation potential were estimated from immunophenotyping and CFU assay in vitro. BM precursor populations were elevated in most treated subjects on day 7; this reflected an activity of AED on precursor cells that resulted in a decrease in the CD34+CD38− and CFU-F populations and an increase in erythrocyte (BFU-E, CFU-E), and granulocyte (CFU-GM) precursors and megakaryocytes. Elderly patients (but not adults) had increased levels of two angiogenic factors; vascular endothelial growth factor and plasminogen activator inhibitor-1 in plasma (day 5) and in BM (day 7). These findings suggest that AED has a regenerative role in elderly BM and that the stimulation of angiogenic factors precedes an increase in BM cellularity which in turn leads to an increase in circulating terminally differentiated blood elements.
Fresh and Cryopreserved Umbilical Cord Blood Side Population Cells Have Comparable Immunophenotype and Function but Are Not Characterised by ABCG2 Expression.
Authors: Alt Ruediger , Pelz-Ackermann Oliver , Egger Dietmar , Niederwieser Dietger W. , Cross Michael 
Published: April 2020

Abstract
The side population (SP) phenotype resulting from efflux of the fluorescent dye Hoechst 33342 has been associated with high quality stem cells from both hematopoietic and non-hematopoietic organs. While expression of the transporter proteins MDR1 and ABCG2 has been strongly implicated in the SP phenotype of murine bone marrow stem cells, relatively little is known about the SP in human umbilical cord blood. The aim of this study was to investigate the relationship between the expression of common stem cell markers, the SP phenotype and the expression of the MDR1 and ABCG2 transporters in both fresh and cryopreserved umbilical cord blood. Assays of long-term Cobblestone Area Forming Cell (CAFC) activity at week 13 confirmed a 22 fold enrichment (p=0,002) of CAFC potential in the sorted SP (0,11% +/−0,06%, n=7) compared to the control living lymphocyte population subjected to Hoechst-free preparation and sorting (0,005% +/−0,0023%, n=6). SP cells from fresh (n=3) and cryopreserved (n=4) blood samples performed equally well in CAFC assays (p=0,28). A multiparameter FACS analysis involving a combination of four colors plus SP controlled with verapimil revealed the CD133+ and CD34+ cells to be distributed towards the upper part of the SP plot and to extend into the verapimil-sensitive SP. However, only 20,5% +/−15,6% (n=9) of the SP cells expressed either CD34 or CD133. ABCG2+ cells were found to be distributed throughout the main population but to be specifically absent from the side population. Fresh and cryopreserved blood samples were similar in all of these respects. In contrast to the SP cells, the MACS-purified ABCG2+ population failed to grow in cytokine-supplemented media and yielded no colonies in the CAFC assay. Semi-quantitative RT-PCR assays comparing cDNA prepared from the SP to that from the entire mononuclear population revealed a moderate but consistent enrichment of MDR1 mRNA in the SP, but no enrichment of ABCG2 mRNA. A similar comparison of the MACS-separated CD133+ (n=4) and CD133− (n=3) cells revealed the CD133+ population to be enriched (4 fold) for MDR1 but depleted (19-fold) for ABCG2 mRNA. We conclude that ABC transporters other than ABCG2 are responsible for the SP phenotype in human UCB; that MDR1 remains a potential candidate in this respect and that the frequency, immunophenotype and CAFC function of SP cells from fresh and cryopreserved blood are indistinguishable.
Histone Deacetylation Makes an Important Contribution to Chromatin Condensation and Enucleation during Murine Erythroblast Terminal Differentiation.
Authors: Krauss Sharon Wald , Popova Evgenya , Short Sarah A. , Lee Gloria , Villalobos Jonathan , Koury Mark J. , Grigoryev Sergei , Chasis Joel Anne 
Published: April 2020

Abstract
Mammalian erythroid differentiation is characterized by progressive chromatin condensation and culminates in nuclear extrusion. Core histone modifications are known to play a significant role in formation of heterochromatin in several other differentiated cell types. To elucidate molecular mechanisms governing changes in mammalian erythroblast chromatin preceding enucleation, we used a model system in which Friend virus-infected murine spleen erythroblasts undergo terminal differentiation and enucleation in vitro during 0–44 hours. At the conclusion of terminal differentiation of murine erythroblasts, we found that histone H3(K9) dimethylation markedly increased while acetylation of histone H3(K9) and histone H4(K12) dramatically decreased, strongly correlating with nuclear chromatin condensation. To test the effects of altering histone deacetylation on chromatin condensation and enucleation, we added either 1 or 2 mM sodium butyrate, an inhibitor of histone deacetylase, to cultures of mouse erythroblasts and compared specific parameters of differentiation with untreated control cells, which normally terminally differentiate at 44 hrs. Western blotting revealed that control cells had the predicted dramatic decrease in histone H4(K12) acetylation while 1 mM or 2mM butyrate-treated erythroblasts did not show diminished levels of acetylation. Numbers of erythroblasts were 5- and 8-fold higher at 44 hrs in cultures treated with 1 and 2mM butyrate, respectively, compared to untreated control cells and there was a correlated marked reduction in expelled nuclei and young reticulocytes. We also found that nuclei in the butyrate-treated erythroblast populations were 1.5 fold larger relative to nuclei with condensed chromatin in terminally differentiated erythroblasts in control cultures. Evaluation of hemoglobinization of erythroblasts by benzidine/hemotoxin staining showed that hemoglobin synthesis continued after butyrate treatment, indicating cell viability and the absence of significant butyrate-induced apoptosis. In sum, these data show that butyrate exposure maintains erythroblasts in a nucleated state with relatively uncondensed chromatin and inhibits them from normally differentiating into young reticulocytes. Thus widespread deacetylation of histones appears to be necessary to achieve extensive chromatin condensation during mammalian erythroid maturation. Our data add support to the attractive idea that during normal mammalian erythropoiesis chromatin condensation may be essential for the transition from nucleated erythroblasts to enucleated reticulocytes.
Inhibition of p38 MAPK Reduces Ionizing Radiation-Induced Hematopoietic Cell Injury.
Authors: Liu Lingbo , Zhou Daohong , Wang Yong 
Published: April 2020

Abstract
Our previous studies demonstrated that exposure to ionizing radiation (IR) causes long-term bone marrow (BM) damage by induction cellular senescence in hematopoietic cells. The senescent hematopoietic cells exhibited an elevated activity of the senescence associated β-galactosidase (SA-β-gal), a biomaker for senescent cells, and expressed increased levels of p16Ink4a (p16), whose expression has been implicated in the establishment and maintenance of cellular senescence. Activation of the p38 MAPK pathway has been implicated in the induction of cell cycle arrest and senescence in response to a variety of stressors in part by up-regulating the expression of p16. Therefore, in the present study we examined the role of the p38 MAPK pathway in IR-induced hematopoietic cell senescence. The results showed that exposure of male C57BL/6 mice to a sublethal dose (6.5 Gy) of total body irradiation (TBI) caused a sustained activation (&gt;4 weeks) of p38 in Lin− c-kit+ Sca-1+ (LKS+) hematopoietic stem cells (HSCs). This activation was associated with a persistent up-regulation of p16 and quantitative and qualitative reduction of LKS+ HSCs. Inhibition of p38 activity with SB202190 (a specific p38 inhibitor) attenuated IR-induced inhibition of the hematopoietic function of BM hematopoietic cells in an in vitro colony forming cell (CFC) assay. Moreover, the production of hematopoietic progenitor cells (HPCs) in SB202190-treated BM cells was 7.5 fold higher than that of irradiated cells without SB202190 treatment after five weeks of long-term BM cell culture (LTBMC); and the cells treated with SB202190 after IR retained their clonogenic function while the control irradiated cells lose their ability to form colonies. These findings suggest that p38 may play an important role in mediating IR-induced suppression of hematopoietic cell function and pharmacological inhibition of the p38 pathway may potentially be developed as novel therapeutic strategy to ameliorate IR- and chemotherapy-induced BM toxicity.