bluuebunny/crossref_metadata_2025_split huggingface dataset:
Emerging Viruses Are Inactivated by Pasteurization.
Authors: Groener Albrecht , Schäfer Wolfram , Nowak Thomas 
Published: April 2020

Abstract
Emerging zoonotic viruses as West Nile virus (WNV), SARS coronavirus, and avian influenza viruses have raised concern about their potential presence in blood/plasma. However, so far only WNV has been demonstrated to be transmitted via blood transfusion. Pasteurization (heat treatment in aqueous stabilized solution at 60°C for 10 hours) is an integral part of the manufacturing process of ZLB Behring’s coagulation factor concentrates such as Haemate P / Humate-P, Monoclate-P, Beriate P, and Berinin P as well as other plasma-derived medicinal products. Therefore, the virus inactivation capacity of this manufacturing step was evaluated utilizing these emerging viruses or appropriate model viruses in virus spiking studies in order to assess the safety of plasma derivatives. The studies demonstrated that WNV was inactivated very effectively by pasteurization in different plasma derivatives resulting in an inactivation factor of more than 7 log10 and that BVDV (bovine viral diarrhea virus, as WNV a member of the family Flaviviridae) is a suitable model virus for WNV. Furthermore, coronaviruses and influenza viruses, of both avian and human origin, were highly susceptible to inactivation by pasteurization resulting in complete inactivation of virus infectivity within a short period of time. In addition, feline calicivirus, a model virus for hepatitis E virus (HEV), was inactivated very effectively within a short period of time. We demonstrate that pasteurization is an effective method to inactivate the emerging viruses WNV, SARS coronavirus, and avian influenza viruses as well as HEV below detection limit of a sensitive cell culture infectivity assay resulting in a safe plasma derivative.
A Clinical Trial of VVR Rate in Preoperative Autologous Blood Donation by Adults and Pediatrics Patients with or without IV Fluid Infusion.
Authors: Kobayashi Sumiko , Sato Norihiro , Hashino Satoshi , Kondo Takeshi , Koike Takao 
Published: April 2020

Abstract
Background: There are no reports that compared with or without a simultaneous IV infusion in the autologous blood donation of adult patients and pediatric patients in the incidence of vasovagal reactions (VVR) rate. These reactions may decrease return donation and disrupt blood donation activities. Objectives:The purpose of our study is whether we can take autologous blood donation safely from the both groups of adults patients and pediatric patients. To determine whether a IV fluid infusion is required during the autologous blood donation in both of adult and pediatric patients and whether a IV infusion influences the incidence of VVR rate. Study design and Methods: Since 1999, a total of the 922patients (1,654 times donations) were enrolled including 40 pediatric patients: we investigated two groups such as with or without IV infusion. In pediatric patients, their original diseases were almost based on orthopedic disorders. We used exactly same size needle (17.5gage) to the adult and pediatric groups. Results: Ten of 394 times donations in 244 adults patients with the IV infusion group were shown in VVR (2.5%). The other group who had not taken an IV infusion in 375 times donation of 164 adult patients had 1.8% incidence rate of VVR (seven of 375 times donations) during autologous blood donation. There was no significant difference between with or without IV infusion groups of adults. In pediatric patients, one of the ten patients with IV infusion group occurred 10% of VVR and 3 of 30 times donations of without IV infusion occurred same percentage of incident rate (10%). We also investigated what is the risk factor of VVR in pediatric patients. What is the risk factor of pediatric patients? The result was that low hemoglobin data (mean 12.7g/dl vs 14.0g/dl) before donation and below 40kg were high risk groups in VVR incidence (P&lt;0.01). Conclusion: Our study shows that there is no significant change between with or without IV fluid infusion in both groups of patients of adults or pediatrics in autologous blood donation before operations. Therefore it is useful that it can be done safely without IV prevention for VVR, particularly in cases of pediatrics patients.
Evaluation of the Sepacell RS-2000® (LeukoConnect) Sterile Dock Leukocyte Reduction Filter Set with Red Blood Cells Collected using Trima.
Authors: Campbell Sandy C. , Mauldin Jennifer E. , McShan Kristy L. , Elfath M. Dean 
Published: April 2020

Abstract
As automated red blood cell (RBC) collections become more prevalent, sterile dock filtration will continue to be an important means of leukoreduction (LR). This study evaluated the performance of the Baxter Sepacell RS-2000 sterile dock leukocyte reduction filter set with RBCs collected using Trima, which were filtered either immediately at ambient temperature, or refrigerated and then filtered 6–8 or 68–72 hours post-collection. 180 RBC units were collected using Trima (Gambro BCT, Lakewood, CO) and filtered with the Sepacell RS-2000 (Baxter Healthcare, Deerfield, IL). Units within Group A (n=60) were filtered at ambient temperature immediately after collection. Units within Group B (n=60) and C (n=60) were refrigerated and filtered at 6–8 or 68–72 hours post-collection respectively. Pre- and Post-filtration hematocrits were performed on the Cell-Dyn 3700 (Abbott Laboratories, Abbott Park, IL) to determine RBC percent recovery. Residual leukocytes in the final product were counted using either the FACSCalibur flow cytometer (BD Biosciences, San Jose, CA). Relevant data is presented in Table 1 below. This study demonstrated that the Sepacell RS-2000 (LeukoConnect) sterile dock filter yielded red cell units that met or exceeded the FDA and AABB standards for leukoreduction when filtering refrigerated red blood cell (RBC) units collected on Trima at 6–8 or 68–72 hours post-collection. 60 out of 60 (100%) LR-RBC units in Groups B and C contained residual white blood cell (WBC) counts at or below the limit of detection of 0.3 x 105. 55 of 60 (92%) LR-RBC units in Group A contained residual WBC &lt; 5x106. 38 of 60 (63 %) LR-RBC units within this group contained residual WBC = 1x106. The hold-time prior to ambient filtration for Group A averaged 1:37 ± 0:18. Filtration was initiated &lt; 3 hours post-collection for all units within this group. There is speculation that a correlation may exist between the temperature of the unit at the time of filtration and leukocyte reduction performance for this filtration condition. This correlation was not assessed by this study and warrants further investigation. It was also noted that 11 out of 180 total units (6%) achieved RBC recovery results slightly less than 85%. Unexpected variability in volume recovery for these units can be attributed to inadequate drainage time of the filter. Allowing additional time for complete drainage of the filter would be expected to result in all units achieving &gt; 85% recovery. Table 1
Parameters Group A Group B Group C Mean ± SD (n=60) (n=60) (n=60) Storage & Filtration Condition Ambient (&lt; 3hrs) 1–6°C (6–8hrs) 1–6°C (68–72hrs) Filtration Time (min) 11 ± 1 24 ± 5 27 ± 6 Residual WBC count (1x106) 1.81 ± 3.33 0.30 ± 0.00 0.30 ± 0.00 % RBC Recovery 88 ± 2 87 ± 2 88 ± 2
Human Endothelial Cells Generated In Vitro from Circulating Progenitors Represent a Safe Tool for Targeting of Myocardial Tissue.
Authors: Koeck Ruth , Wolf Dominik G.F. , Rumpold Holger F. , Ott Harald , Untergasser Gerold , Erdel Martin , Debagge Paul , Gastl Guenther A. , Gunslius Eberhard 
Published: April 2020

Abstract
Various cell sources, culture protocols and characerization steps are being used in the research field of endothelial progenitors and circulating endothelial cells. We generated EPC by plating the mononuclear fraction onto a collagen I-coated well of a six-well-plate in endothelial cell medium. After 6 to 12 days, 1–10 clones with cobblestone morphology appeared and grew rapidly to confluency within a few days. The rapid proliferation of EPC decelerated after 30–40 days, remaining on the same level for additional 30 days and finally stopped after 70–80 days of culture. Immunofluorescence staining and FACS analysis revealed an endothelial phenotype: EC stained positive for vWF, CD31, CD34, KDR, CD105, CD146, bound UEA-1 lectin and took up acetylated LDL. Of note, CD14 and CD45, which are surrogate markers of monocytes and leukocytes, respectively were negative. The expression of ICAM-1 was upregulated by incubation with TNF-a. In addition, EC formed tubes on Matrigel and expressed VEGF-R1 as well as VEGF-R2-mRNA proven by RT-PCR. To gain deeper insight into the regulation of cellular senescence, we performed western blot analyses of cell cycle regulating proteins. In contrast to early passage cells, growth arrested cells from late passages up-regulated p15, p16, p21, whereas p27 and Id1 were significantly dowregulated. Moreover, late-passage cells highly expressed SA-ß-galactosidase. The obvious replicative senescence together with the regular karyogram indicates a relative safety aspect of their application in vivo. To study their homing behaviour to ischemic tissue, we established a model of myocardial infarction, which was induced by ligation of the left anterior descending coronary artery in Rowett nude rats. 5–10x106 PKH red-labeled EPC were injected into the tail vein. Intramyocardial injection served as positive control, while administation of EPC without prior infarction represented the negative control. Rats were sacrificed on day 14 after the operation and cryosections of their hearts were prepared. PKH-positive cells were almost exclusively found in the infarcted left ventricle after i.v.-application, thus showing a high specifity for ischemic myocardium. The results were confirmed by immunohistochemical staining of human CD31.
In summary, EPC generated according to the described protocol represent a possible tool for safe cell-based therapy strategies.
Ex-Vivo Expansion of Multipotent CD133+ Stem Cells with VEGF, FLT3l and SCGF.
Authors: Loges Sonja , Butzal Martin , Fischer Uta , Gehling Ursula M. , Hossfeld Dieter K. , Fiedler Walter 
Published: April 2020

Abstract
The rare CD133+ stem cell population contains both hematopoietic and endothelial progenitors. Successful ex-vivo expansion of this multipotent population would therefore be of great benefit in many clinical settings including stem cell transplantation and gene therapy. We developed a cell culture system containing the recombinant human cytokines vascular endothelial growth factor (VEGF), FLT3 ligand (FLT3L) and stem cell growth factor (SCGF) for ex-vivo expansion of purified human CD133+ stem cells obtained from leukapheresis products from patients pre-treated with G-CSF.
FACS analysis, colony assays and NOD-SCID transplantation studies were performed to monitor stem cell and endothelial phenotype in-vitro and in-vivo. Cultivation with VEGF, FLT3L and SCGF induced a mean 2200-fold increase of total cell counts in 5 weeks. FACS analysis revealed persistence of 6–15% CD133+ stem cells indicating proliferation and survival of primitive hematopoietic stem cells. 5–6% of the proliferating cells expressed the endothelial markers CD144 (VE-Cadherin) and von-Willebrand factor (vWF). Ex-vivo expanded stem cells could be differentiated into adherent endothelial cells after withdrawal of SCGF and FLT3L allowing generation of large numbers of endothelial cells. Colony-assays showed an increase of hematopoietic and endothelial colonies after 5 weeks of ex-vivo expansion indicating simultaneous proliferation of hematopoietic and endothelial precursors under the established culture conditions (CFU-E 60-fold, CFU-GEMM 48-fold, CFU-GM 59-fold, CFU-G 99-fold, CFU-M 1356-fold and CFU-EC 1843-fold).
To assess in-vivo functionality, hematopoietic stem cells expanded ex-vivo for 7, 14, 21 and 32 days were transplanted into sublethally irradiated NOD-SCID mice. For each expansion period, the mean percentage of anti-human CD45 positive bone marrow cells 3 months post-transplantation was 11, 3, 3 and 1%, respectively. Human CD45+ cells for each set of experiments contained a mean of 15, 26, 8 and 32% T-cells (CD3+), 9, 0, 7 and 21% B-cells (CD19+), 24, 2, 2 and 11% monocytes (CD14+), 21, 3, 1 and 12% granulocytes (CD33+) and 19, 37, 44 and 24% stem cells (CD34+) (d7 (n=5), d14 (n=4), d21 (n=7) and d32 (n=6) respectively).
Our experiments showed multilineage engraftment of human stem cells expanded for more than 4 weeks ex-vivo. Therefore our culture system provides a tool to generate large numbers of human stem and endothelial cells for clinical purposes.
HOXB4-Mediated Immortalization of Murine Hematopoietic Stem/Progenitor Cells.
Authors: Zhang Xiao-Bing , Kiem Hans-Peter 
Published: April 2020

Abstract
HOXB4 overexpression has been shown to be able to expand hematopoietic stem/progenitor cells (HSPCs) in short-term ex vivo culture. Here we found that HOXB4-transduced cells could expand continuously in up-to-eight-month long-term liquid culture. In 3 independent experiments, mouse bone marrow (BM) Sca1+ cells were transduced with HOXB4 or YFP vectors and kept culture in IMDM/10%FCS containing SCF, Flt3-L, and TPO each at 50 ng/ml. After 3–4 weeks of culture, all YFP-expressing cells differentiated and died. However, HOXB4-transduced cells continuously expanded and percentage of HOXB4-expressing cells increased to more than 95% after 4 weeks of culture. Without cytokine support, these cells stopped growing, while a single cytokine, SCF, was able to support the expansion of these cells, albeit at a lower rate. In the presence of SCF, TPO and Flt3-L, these cells expanded robustly with cell doubling time of around 2 days. These cells are morphologically immature by cytospin and Wright-Giemsa staining. CFU assay showed a clonogenicity of 2%. Phenotyping by FACS showed that all the cells expressed c-kit, while some cells expressed Gr-1, Mac-1 and CD3, indicating that HOXB4-immortalized cells spontaneously differentiate into myeloid and lymphoid cells in ex vivo culture. To test the repopulating capability of these cells, 1x107 cells were injected into each sublethally irradiated NOD/SCID mice via tail vein. These cells engrafted in the recipients with 2% in BM and 10% in spleen. Transplantation into lethally irradiated congenic CD45.1 mice is in progress. Immortalization of human CD34+ by HOXB4, however, failed. To determine the mechanisms under which HOXB overexpression immortalizes mouse rather than human HSPCs, we performed Western blot assay with HOXB4-specific antibody. HOXB4 protein levels in human cells decreased continuously from 1–3 weeks posttransduction, whereas expression in mouse cells remained stable. Furthermore, we found that HOXB4 transduced in mouse cells was tyrosine phosphorylated, which is evidenced by treatment of cells with sodium orthovanadate, a protein tyrosine phosphatase inhibitor, treatment of cell extracts with l-phosphatase and immunoprecipitation with phosphorylated tyrosine-specific antibody. Consistent with other reports, no leukemogenesis was observed in our studies as yet. It would be interesting to compare the differences in gene expression profiles between the HOXB4- and HOXA9-immortalized hematopoietic cells, which may unveil the molecular mechanisms governing leukemogenesis. In conclusion, murine HSPCs can be immortalized by HOXB4 overexpression and tyrosine phosphorylation of HOXB4 may be responsible for the self-renewal and immortalization of HSPCs.
The Tetraspanin CD82 Defines Erythroid Committment, Regulates Stem- and Progenitor Cell Survival and Mediates Adhesion.
Authors: Burchert Andreas , Denecke Barbara , Haerle Tobias , Neubauer Andreas 
Published: April 2020

Abstract
CD82 belongs to the tetraspanin superfamily, members of which regulate multiple aspects of cell biology such as T-cell activation, proliferation, differentiation, and adhesion.We have previously shown expression of CD82 on hematopoietic progenitor cells. Here we were seeking to shed light on the functional role of CD82 in hematopoiesis. First, we found that CD82 expression is strongly associated with an erythroid committment as exclusively the CD82bright fraction but not the CD82medium or CD82dim fraction of CD34+ cells gave rise to erythroid colony forming units (CFU). In order to manipulate CD82 activity, we used as surrogate ligand, a previously described, CD82-activating monoclonal antibody (moAb), clone 50F11, to activate CD82 on hematopoietic precursors. We found that 50F11, but not another CD82 specific antibody clone, BL-2, specifically induced tyrosine phosphorylation of CD34+ cells. In Dexter-type long-term cultures (D-LTC) 50F11, and not IgG1 isotype control moAbs significantly inhibited myelopoiesis and the number of CD34+ clonogenic progenitors. Moreover, in long term culture initiating cell (LTC-IC) assays, 50F11as compared to isotype control antibodies substantially inhibited, but not entirely abolished, the number of 5 week-LTC-IC′s, indicating that CD82 activation inhibits progenitor - and stem cell proliferation or self renewal. Finally, plastic immobilized 50F11-antibodies caused a time-, and concentration dependent induction of adhesion of CD34+ cells, which was associated with the formation of F-actin and development of multipolar extensions. Finally, CD82 ligation by 50F11 caused a statistically significant down-regulation of the integrin CD49d (p=0,036) and CD62L (p=0,010).
Together, it is shown that CD34+/CD82high cells characterize an erythroid committment implying a role for this tetraspanin in erythroid hematopoiesis. Activation of CD82 induces adhesion and negatively regulates proliferation of adult stem- and progenitor cells. This implicates a so far unknown role for CD82 in the regulation of early hematopoiesis.
Intracardiac Transplantation of Already Specifically Dedifferentiated Circulating CD34+ Cell Subsets Would Favor Post-Ischemic Myocardial Regeneration.
Authors: Henon Philippe R.G. , Sovalat Hanna , Ojeda-Uribe Mario , Arkam Yazid , Bischoff Nicolas , Monassier Jean-Pierre , Dallemand Robert 
Published: April 2020

Abstract
Several groups, mainly from Germany and Japan, have recently conducted different phase-I clinical studies in which autologous bone marrow (BM) mononuclear cells (MNCs) were reinjected either directly in the ischemic area or intra-coronary in patients with severe post-infarct cardiac failure, resulting in a significant improvement of myocardium viability and/or reperfusion. However, besides “true” HSCs, BM-MNCs represent a mixture of mesenchymal progenitor cells, angioblasts, and maybe other progenitor cells, which does not allow to identify the type(s) of cells potentially responsible for improvement. Moreover, the obstructed coronary artery was always repermeabilized, which biases the evaluation of posttransplant myocardial reperfusion. We have personally chosen an other original approach using mobilized and purified circulating CD34+ cells. We and others have indeed demonstrated that mobilized CD34+cells were in fact subdivided into various subsets: of course, the most important (~75%) is the truly hematopoietic subset (CD34+/133+), of which a CD38− part is probably close to the very primitive HSC. But other smaller subsets are immunophenotypically characterized either as mature (CD34+/VEGFR-2+) or immature (CD34+/133+/VEGFR-2+) endothelial progenitor cells - thus potentially capable of neoangiogenesis -, or as muscle progenitors (Desmin+) and even more as cardiomyocytes (Troponin-T+). In a phase-I trial benefiting of the approval of the regional ethical committee, patients suffering of post-infarct cardiac failure are selected according to the following criteria: left ventricular ejection-fraction (LVEF) =35%; distinct area of left ventricular-wall akinesis determined by PetScan; candidates for coronary artery by-pass grafting (CABG), but without any repermeabilization of the coronary artery involved in the infarction; age =70 y. After a 6-days mobilization by G-CSF, circulating CD34+ cells are collected, then purified by immunomagnetism and immediately reinjected at d+7 during CABG, all around and within the infarcted area. The first evaluable patients well tolerated cell mobilization - and collection phases, as well as operative and post-operative periods. Three patients have presently a follow-up = 1 y post-transplantation. Two show a striking gain in LVEF (14 and 20% respectively) with an important improvement in myocardium viability, reperfusion and contractility, and finally in exercise capacity (from class IV to class I in New-York Association functional class). Although very encouraging, these results have to be confirmed in further patients.
Cord Blood Mesenchymal Stem Cells Are Enriched for Neural and Glial Progenitors.
Authors: El-Badri (Dajani) Nagwa S. , Saporta Samuel , Liang Xiaomei , Sanberg Cyndy D. , Sanberg Paul R. 
Published: April 2020

Abstract
Human bone marrow stroma is the primary source for mesenchymal stem cells (MSCs), a mixture of multipotent progenitors that differentiate into fibrous, cartilage, adipose, bone and reticular tissue cells. Recent reports suggest that human umbilical cord blood (UCB) is a feasible source for MSCs. In this study, we examined whether MSCs from umbilical cord blood contain progenitors that differentiate into cells of neural and glial lineages. UCB mononuclear cell fraction was cultured in Iscove’s MDM culture medium supplemented with 20% fetal bovine serum and L-glutamine. After 2 weeks in culture, adherent cells formed colony forming unit fibroblast (CFU-F)-like colonies. When adherent cells reached 70% confluency, they were further subcultured in the same medium for three more weeks before replacing IMDM with neurogenic differentiation medium. Cord blood MSCs were devoid of hematopoietic activity since they failed to produce CFU-GM, CFU-E, or CFU-GEMM hematopoietic colonies when cultured in methyl cellulose medium supplemented with the appropriate hematopoietic growth factors. Flow cytometric analysis showed a population of cells devoid of hematopoietic phenotype (CD34−, CD45−, CD38−, CD3−, CD19−). After 7 days incubation in serum free supplemented Neurobasal™ medium, cultured cord blood MSCs expressed neuron cell surface antigen A2B5, neurofilament polypeptide NF200, oligodendrocyte marker 04, and intermediate filament proteins, nestin and vimentin. Strong immunoreactivity was observed with astrocyte marker GFAP. A proportion of cultured MSCs developed into neuron-like cells that were positive for TUJ-1 neural progenitor marker. These results indicate that human UCB-derived MSCs are enriched for neuro-glial progenitors, and could be further developed for the treatment of stroke, spinal cord injury, and neurodegenerative diseases such as like Alzheimer’s, Parkinson’s and Multiple Sclerosis.