bluuebunny/crossref_metadata_2025_split huggingface dataset:
Heart Failure Improvement after Autologous Bone Marrow Mononuclear Cells (ABMMC) Transplantation.
Authors: Carrasco-Yalan Antonio A. , Fernandez-Viña Roberto , Castillo-Aguirre Jorge , Rios Hugo , Alvarez Marco , de Moura R. , Cruz C. , Carrillo A. , Chirinos S. , Ercilla E. , Patel A. , Tumak-Mubarak Jorge 
Published: April 2020

Abstract
Background: Pilot studies suggest that intracoronary transplantation of unselected ABMMC cells may improve Left Ventricular Ejection Fraction (LVEF) in heart failure (HF)
Methods: 18 patients were enrolled and completed 1 year follow up. Patients underwent SPECT evaluation, all had ejection fraction &lt; 35%, 6 patients were randomly allocated to the control group and 12 in the Bone Marrow Cells (BMC) group, median age 65 years old, male/female ratio 17/1; all with ischemic cardiomyopathy. All of cohort had NYHA class of III with maximal medical therapy and median basal LEVF was 28.38% (SD=6.5%). Median number of mononuclear and CD34+ cells infused were 8.1*108 and 1.2*107 respectively in a 50 cc delivered retrograde via coronary sinus approach using balloon occlusion "over wire" for 8 to 10 minutes. No study related adverse events were observed.
Results: After a median time of 21 days, patients in the BMC group had relief of dyspnea symptoms and improvement in functional class. At 1 year, NYHA class improved in 92% of the patients in the BMC group by at least 1 class and no improvement in the control group. Mean improvements of LVEF post BMC transplantation were 6.18% (SD=4.94%) and 6.7% (SD=4.07%) at rest and stress SPECT respectively. Rest LVEF at baseline and after one-year follow up between the BMC and control groups demonstrates significant difference (4.9% vs 1.6%, p=0.006), as well as the comparison of change in stress LVEF in both groups (6.7% vs 0.11%, p&lt;0.001).
Conclusions: Infusion of progenitor cells into the coronary sinus is safe and feasible in the ischemic HF. It is associated with significant improvement in symptoms, functional capacity and LVEF. Larger randomized studies are in progress to build upon this pilot study.
Figure Figure
Autologous Bone-Marrow Stem Cell Support in Acute Anterior Myocardial Infarction.
Authors: Vallisa Daniele , Piepoli Massimo , Cavanna Luigi , Arbasi Cristina , Moroni Carlo , Lazzaro Antonio , Capucci Alessandro , Bernuzzi Patrizia , Arcari AnnaLisa , Bosi Costanza 
Published: April 2020

Abstract
Purpose- Emerging evidence suggests that stem and progenitor cells derived from bone marrow can improve cardiac function and perhaps outcome in patients after acute myocardial infarction (AMI). In this single-centre randomised trial, we evaluated whether intracoronary transfer of autologous bone-marrow stem cells can affect not only global left ventricular (LV) function (nuclear scintigraphy) but also autonomic control (heart rate variability, HRV) and baroreflex sensitivity (BRS) at 12 months’ follow-up.
Methods- After successful percutaneous coronary intervention (PCI) for acute ST-segment elevation myocardial infarction and residual LV dysfunction, 33 patients were randomly assigned to either a control group (n=17) that received optimum postinfarction treatment, or a bone-marrow-stem-cell group (n=16) that received optimum treatment plus intracoronary infusion of autologous bone-marrow stem cells 4±1 days after PCI. Test analyses were done by two investigators blinded for treatment assignment.
Results- Global LVEF at baseline (determined 3±1 days after PCI) was 36.2±9.4 in controls and 35.1±9.8 in the bone-marrow cell group (P=NS). After 12 months, mean global LVEF had increased by 25% in the control group but 41% in the treated group with an anti-remodeling effect in the latter: LVEDV increased by 12% in the control vs 2% in the treated group (P&lt;0.05). This was associated with improved HRV (SD −20ms vs +644 ms in the treated group P&lt;0.01), particularly in the parasympathetic-related oscillation (HF +85.7±216 ms vs. +289.3±366 ms) and higher BRS (+3.4±11.7 vs +8.7±6.3 ms/mmHg) (control vs treated, P&lt;0.05 in all comparisons). Cell transfer did not increase the risk of adverse clinical events, in-stent restenosis, or proarrhythmic effects. In 12 months follow-up 4 exitus (24%) were observed in the control group vs. only 1 (6%) in the treated group
Conclusions- The improvement of left-ventricular systolic function and prognosis in AMI patients by intracoronary transfer of autologous bone-marrow-cells may be mediated by higher vagal tone. (ClinicalTrials.gov number, NCT00437710).
Mesenchymal Stem Cells Improve Wound Healing Via Early Activation of the Matrix Metalloproteinase-9 and Vascular Endothelial Growth Factor: Using a 3-Dimensional Collagen Gel Model.
Authors: Won Jong-Ho , Kim Cheol-Hann , Cho Moon-Kyun , Lee Sang-Cheol , Kim Chan-Kyu , Lee Nam-Su , Hong Dae-Sik , Park Hee-Sook 
Published: April 2020

Abstract
There is great progress on clinical application of stem cells. Stem cells are self-renewing and are potent in differentiating into multiple tissues. It has been reported that mesenchymal stem cells (MSCs) had successfully improved skin-substitute wound healing. We investigate the effects MSCs on wound healing using a 3D collagen gel system in a full thickness large-sized skin defect rat model. Three full thickness skin and tissue defects of 2 x 2 cm in size were excised on the backs of female SD-rats and replaced with 3D collagen gel contains 2 millions of male rat MSCs (rMSCs) on one site. Two different sites were control. One replaced with 3D collagen gel without rMSCs and the other site was empty. Wound size, histology and protein expression were evaluated at 3, 7 and 14 days after injury. Cultured rMSCs were confirmed to be negative for hematopoietic markers by flow cytometry and to be capable of differentiating into osteocytes, chondrocytes, and adipocytes in vitro. The wound size was significantly smaller in the rMSCs-treated sites than control sites (P &lt; 0.01) and the most effect of wound healing was obtained at day 7. A lot of rat Y chromosome positive cells were found in rMSCs treated sites. The expression of matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF) were increased on the rMSCs-treated sites at day 3 but on the other hand those levels on the control sites were increased at day 7. This data suggest that MSCs accelerate wound healing and 3D collagen gel model can stably introduce MSCs to the defected skin area. Early activation of MMP-9 as well as previously known VEGF may play crucial roles in dermal remodeling and accelerate the processes of dermal and epidermal wound healing after MSCs treatment.
Megakaryocytes and Functional Platelets Obtained from Human Subcutaneous Adipocytes in an In Vitro Differentiation System.
Authors: Saito Emi , Matsubara Yumiko , Suzuki Hidenori , Ikeda Yasuo , Murata Mitsuru 
Published: April 2020

Abstract
Because of the difficulty in obtaining the sufficient amounts of hematopoietic stem cells from bone marrow, peripheral blood, and cord blood cells, studies of an in vitro experimental system that allows the production of abundant number of megakaryocytes (MKs) and platelets are currently the focus of research. Here, we present a novel system where human subcutaneous adipocytes were successfully differentiated into MK lineages in an in vitro liquid culture system. We also show that subcutaneous preadipocytes could be successfully transfected with vectors, to obtain modified MKs. Primary human subcutaneous preadipocytes (Cambrex Bio Science Walkersvile, Inc. Walkersville, MD, USA) were cultured in conditioned media to differentiate into mature adipocytes. Cells were cultured in serum-free media containing thrombopoietin for differentiation into MK lineages. The MKs or platelets were counted by flow cytometric analysis on day 14 using the relative value of CD41(+)/propidium iodide(+) cells or platelet size CD41(+) cells, respectively, versus 107 subcutaneous preadipocytes on day 0. The MK and platelet cell count was approximately 9600/ 107 and 2200/ 107, respectively. Morphological analysis with electron microscopy demonstrated that MKs, which had typical organelles such as granules, demarcation membrane, and nuclei, and platelets, which had typical contents such as granules, mitochondria, and open canalicular system, were successfully obtained from subcutaneous preadipocytes. We then attempted to transfect subcutaneous preadipocytes with vectors to obtain transformed MKs. Two glycoprotein (GP) Ib alpha polymorphisms, 145Thr/Met and 1–4 repeats of variable number tandem repeat of 13 amino-acid sequences, were used as the marker of gene transfer, which were detected by PCR-restricited fragment length polymorphism (RFLP) and Western blot analysis, respectively. Subcutaneous preadipocytes with the 145Thr/Thr and 1 repeat sequences were transfected with the expression vector carrying GPIb alpha with the 145Met and 4 repeats sequences. PCR-RFLP analysis with gel electrophoresis was performed on each RNA sample from the expression vector-transfected and non-transfected cells. Non-transfected cell sample had bands corresponding to the 145Thr sequence position, while the expression vector-transfected cell sample had bands corresponding to both the 145Thr and Met sequences. Western blot analysis with an anti-GPIb alpha monoclonal antibody, LJ-Ib alpha1 (a generous gift from Dr. ZM Ruggeri, The Scripps Research Institute, La Jolla, CA, USA), showed bands of the expected size corresponding to 1R and 1R/4R. In summary, we established an in vitro culture system to produce MKs and platelets from subcutaneous adipocytes. We were also able to obtain transfected MKs from subcutaneous preadipocytes.
Identification and Characterization of Zebrafish Tissue Factor and Its Role in Embryonic Blood Vessel Development.
Authors: Zhang Jing , Handin Robert I. 
Published: April 2020

Abstract
Tissue factor (TF) is a trans-membrane glycoprotein that is the cellular receptor for coagulation factor VII/VIIa. Activation of factor VII by TF is well established as the primary regulator of blood coagulation. Although TF has been implicated in such diverse processes as angiogenesis and tumor metastasis, these other functions have been difficult to study, as the knockout of murine TF is lethal at day 8.4 of embryonic life. Because of their hardiness, their rapid maturation sequence and their optical clarity, the zebrafish is an increasingly popular model for studying embryonic development. In order to study the role of TF in embryonic development we have cloned zebrafish TF cDNA. The 1.36kb orthologous cDNA predicts an ORF encoding a 293 amino acid (aa) protein with a 26 amino acid (aa) signal peptide. The predicted mature protein contains a 230 aa extra-cellular domain, a 20 aa trans-membrane segment, and a 17 aa cytoplasmic tail. The predicted amino acid sequence of zebrafish TF has 31% and 32% overall identity with human and mouse TF respectively. The zebrafish TF gene, like the human and mouse genes, has 6 exons spanning 14kb of genomic DNA. Whole-mount in situ hybridization with a TF anti-sense Riboprobe revealed that TF is detected as early as 18 hours post-fertilization (hpf) in developing head structures and brain. At 20 hpf, TF is expressed in the vascular system and highly expressed in the hindbrain. By 48 hpf, TF is seen in the brain, heart, liver, lens and gut. To investigate the expression of zebrafish TF in vivo, we generated transgenic zebrafish lines that express enhanced green fluorescent protein (EGFP) under the control of the TF promoter. A ∼ 2.0 kb promoter fragment upstream of the first exon of the TF gene was sufficient to drive expression of EGFP in a manner that recapitulated the expression of native TF. We inhibited TF expression by injecting anti-sense oligonucleotide morpholinos into the Tg(fli1:EGFP) line, in which there is robust expression of EGFP in vascular endothelial cells and their angioblast precursors. At 24 hpf the development of trunk capillary beds was delayed and, at 48 hpf, most of the morphants developed pericardial edema and impaired formation of inter-somitic vessels. These studies demonstrate that TF is expressed in multiple organs during zebrafish embryogenesis and that TF expression is necessary for the proper migration of endothelial cells into developing blood vessels during the process of embryonic vasculogenesis.
An In Vitro and In Vivo Evidence That Interleukin-8 Is a Pivotal Hypoxia-Regulated Pro-Angiopoietic Factor Secreted by Human Rhabdomyosarcomas (RMS) - Therapeutic Implications.
Authors: Wysoczynski Marcin , Wan Wu , Kucia Magda , Ratajczak Janina , Ratajczak Mariusz Z. 
Published: April 2020

Abstract
Rhabdomyosarcoma (RMS) is the most common soft-tissue sarcoma of adolescence and childhood that frequently infiltrates bone marrow (BM). We have demonstrated that the stromal-derived factor (SDF)-1 (Blood2002;100:2597), hepatocyte growth factor (HGF) (Cancer Res. 2003;63:7926) and leukemia inhibitory factor (LIF) (Cancer Res. 2007;67:2131) play an important role in RMS metastasis. Since RMS tumors are very well vascularized we asked if these factors are secreted by RMS cells and if they stimulate angiogenesis. To address this issue, first we evaluated expression of SDF-1, HGF and LIF as well as several other proangiopoietic factors (IL-8, VEGF, FGF-2) in 9 human RMS cell lines. Expression of these factors was evaluated in both normal steady state conditions and in conditions that lead to overexpression of hypoxia inducible factor-1 (HIF-1a) due to hypoxia or exposure to CoCl2. We found by RQ-PCR and confirmed by ELISA assays that from all the factors evaluated, IL-8, which expression was very low in steady state conditions was very highly upregulated in all RMS cells lines during hypoxic conditions (∼30–135 times). This selective upregulation of IL-8 was somehow unique for RMS cells, and we did not observe similar phenomenon in several other solid tumor cell lines, where IL-8 was usually already very highly expressed in steady state conditions. To evaluate a biological significance of IL-8 in RMS angiogenesis, we collected conditioned media (CM) from RMS cells and found that they stimulate in IL-8 dependent manner in HUVEC i) phosphorylation of MAPKp42/44 and AKT, and ii) chemotactic responses. Pre-clearing of RMS-derived CM with antibodies against IL-8 significantly inhibited chemotaxis of HUVEC. Finally, by employing shRNA we inhibited expression of IL-8 in human RH-30 cells and noticed that such cells in vivo, if injected into skeletal muscles of immunodeficient mice, have the reduced ability to grow tumors and chemoattract co-transplanted human endothelial cells. We conclude that IL-8 is a pivotal pro-angiopoietic factor that is released by human RMS cells in hypoxic conditions, and that the targeting of IL-8 may prove to become a novel efficient strategy to inhibit RMS growth.
Reduced Genotoxic Risk Using Foamy Viral Vectors for the Treatment of Canine Leukocyte Adhesion Deficiency
Authors: Bauer Thomas R. , Hai Mehreen , Adler Rima L. , Allen James M. , Tuschong Laura M. , Olson Erik M. , Hickstein Dennis D. , Russell David W. 
Published: April 2020

Abstract
Gammaretroviral vectors used in recent gene therapy clinical trials have lead to several successes, such as in the treatment of X-linked severe combined immunodeficiency disease, but have also resulted in insertional activation of nearby oncogenes, leading to leukemia in four patients. We previously reported the successful treatment of four dogs with canine leukocyte adhesion deficiency (CLAD), a lethal genetic immunodeficiency disease caused by defects in the leukocyte integrin CD18, by transplanting foamy viral (FV) vector (deltaphiMscv-cCD18) - transduced, autologous CD34+ hematopoietic stem cells. To date, more than 2 years post transplant, all four dogs have maintained CD18+ leukocyte levels ranging between 5–10%, completely reversing of the CLAD phenotype, and have no clinical or laboratory evidence of hematological malignancy. To assess the potential genotoxicity of the FV gene therapy in the treatment of CLAD, we compared the insertion sites (ISs) found in the FV vector treated CLAD dogs with ISs found in CLAD dogs treated by gammaretroviral (RV) vectors (PG13/Mscv-cCD18). Insertion sites were identified by DNA sequence analysis of ligation-mediated PCR (LM-PCR) or linear amplification-mediated PCR (LAM-PCR) amplicons and subsequent comparison to the dog genome (canFam 2.0, May 2005). Insertion site analysis was performed for integrations that were in or within 50 kb of Refseq genes (using mouse/human orthologs). Analysis of the ISs revealed a reduced preference for FV vector integrations near transcription start sites compared to RV vector integrations (41% vs. 48%), fewer integrations near potential oncogenes (11% vs. 16%), and fewer integrations within genes in general (41% vs. 52%), in the FV vector treated animals compared to the RV vector treated animals. These clinically relevant data suggest that a reduced insertional mutagenesis potential exists when using FV vectors compared to RV vectors, and support the use of FV vectors in the treatment of human hematopoietic stem cell diseases such as LAD.
The Ratio of Serum Transferrin Receptor and Serum Ferritin in the Diagnosis of Iron Deficiency Anaemia in Young Children
Authors: Park EunSil , Kim In-Suk , Seo JiHyun , Lim JaeYoung , Park ChanHoo , Woo HyangOk , Youn HeeShang 
Published: April 2020

Abstract
The incidence of iron deficiency anaemia in 6∼24 month old infants due to increase in iron demand for growth spurt is reported ranged 10 to 40%. However this age group has a common acute illness such as urinary tract infection, pneumonia, and other viral infections. The aim of this study is to evaluate that iron parameter and acute phase reactant are useful parameters in differentiating anaemia by infection from anemia by iron deficiency and the mixed anaemia of these. Among 6–24 months of the infants who visited Gyeongsang Univeristy Hospital for 7 years from 2000 to 2006, 131 infants were enrolled. Hemoglobin(Hb), serum ferritin(SF), serum transferrin receptor(STfR), C reactive protein(CRP), interleukin-6(IL-6), prohepcidine were checked. The subgroup of anaemia of inflammation(AI) was defined as Hb &lt;11 g/dL and SF &gt;50 μg/L, the subgroup of iron deficiency anaemia(IDA) as Hb &lt;11 g/dL and SF &lt;12 μg/L and the normal group as Hb ≥11 and SF ≥12 μg/L. The mean STfR in the subgroup of AI, IDA and normal was 3.89(±2.64), 10.6(±4.95) and 3.96(±1.24), respectively. The mean STfR/Log SF of subgroup was 1.87(±1.55), 36.11(±71.5), 2.31(±0.97), respectively. The mean Log(STfR/SF) was statistically significant between 3 subgroup. All IDA group had Log(STfR/SF) &gt;2.55 whereas in all subjects classified as AI it was &lt;2.55, thus clearly separating two. The mean IL-6 of AI was significantly higher than IDA subgroup and the mean prohepcidine of AI was significantly lower than the normal group. Calculating Log(STfR/SF) is a useful criteria in classification of the iron status. Prohepcine has nothing to do with AI. Iron signal predominant over inflammatory signal in AI.
The Mean(±SD) of STfR, STfR/LogSF, Log (TfR/SF), CRP, IL-6 and Prohepcidine in Subgroups. AI IDA Normal Same letters mean that are not significantly different (P &lt;0.05) AI, anaemia of inflammation; IDA, iron deficiency anaemia Subgroup(%) 33(25) 29(22) 69(53) Hg(g/dL) &lt;11 &lt;11 ≥11 SF(μg/L) &gt;50 &lt;12 ≥12 STfR mean(±SD) 3.89(2.64)a 10.6(4.95) b 3.96(1.24) a STfR/LogSF mean (±SD) 1.87(1.55) a 36.11(71.5) b 2.31(0.97) a Log (TfR/SF) mean(±SD) 1.30(0.56) a 3.29(0.43) b 1.76(0.43) c CRP mean(±SD) 28(39.2) a 7.6(9.6) b 17(28.0) a IL-6 mean(±SD) 6.1(10.5) a 2.0(6.3) b 4.78(11.3) c Prohepcidine mean(±SD) 204(70.5) a 234(144) a 301(120.6) b Fig.1. Log(TfR/SF) in the subgroup. AI, anaemia of inflammation; IDA, Iron deficiency anaemia. Dotted lines indicate the cut-off value at Log(TfR/SF)=2.55 Fig.1. Log(TfR/SF) in the subgroup. AI, anaemia of inflammation; IDA, Iron deficiency anaemia. Dotted lines indicate the cut-off value at Log(TfR/SF)=2.55
The Immunosuppressor Sirolimus Reduces the Number and Maturation of Colony-Forming Units of Erythroid Progenitor Cells.
Authors: Diaz-Ricart Maribel , Arellano Edgar Marcelo , Rovira Jordi , Jou Josep M. , Campistol Josep M. , Vives-Corrons Joan L. , Escolar Gines , Diekmann Fritz 
Published: April 2020

Abstract
Introduction: Kidney transplantation is the treatment of choice in patients with end-stage renal failure. However, the immunosuppression has severe side effects, such as chronic allograft nephropathy, cardiovascular morbidity and neoplasia. An additional effect of inhibiting mTOR is microcytic anemia. We have explored the pathophysiology of this effect.
Methods: Erythroid progenitor cells were isolated from peripheral blood of healthy control persons (HC; n=8), or kidney transplant recipients with chronic sirolimus treatment with (SRL+MC; n=8) or without (SRL-MC; n=8) microcytosis. The isolated progenitor cells were then cultured in a semi-solid medium, containing 3 U/ml erythropoietin, in the absence or presence of SRL (5ng/mL) for 14 days. Burst forming unit erythroid (BFU-E) derived colonies were then counted through an inverted microscope considering that each colony consists of more than 40 cells. Cultures were performed in duplicate and colonies were counted in the entire culture dish.
Results: Hemoglobin was 13.17 (SRL+MC) and 13.23 (SRL-MC) g/dL. RBC count was 5.22 and 4.55x106 /μL in SRL+MC and SRL-MC respectively. MCV was 76 in SRL+MC and 87 fL in SRL-MC. Presence of SRL in the culture medium led to a decrease in the number of colonies in healthy controls and kidney transplant patients (43.2±4.5 vs. 28.5±3.6 BFU-E derived colonies, Mean±SEM, p&lt;0.05). Similar differences were seen if the three groups were analyzed separately (HC, SRL-MC, SRL+MC). This effect was not observed in the presence of tacrolimus, an immunosuppressor with different mechanism of action. Interestingly, culture dishes corresponding to SRL+MC patients contained a higher number of colonies when compared to SRL-MC and HC.
Conclusion: mTOR inhibition leads to a reduced number of erythroid colonies in culture. There is a higher number of circulating progenitor cells in the microcytic patients. Both these observations indicate that microcytosis may be compensated by a higher number of erythrocytes and circulating progenitor cells.