bluuebunny/crossref_metadata_2025_split huggingface dataset:
TLR4-Agonist Stimulated Human Multipotent Mesenchymal Stromal Cells Enhance Both Hematopoietic Progenitors and Myeloid Differentiation
Authors: Ziegler Patrick , Boettcher Steffen , Garavaglia Guido , Manz Markus G. 
Published: April 2020

Abstract
Human mesenchymal stromal cells (MSCs) can replicate as undifferentiated cells in vitro and are capable of differentiating into multiple mesenchymal tissues. In the bone marrow, MSCs are thought to contribute to nurturing stroma involved in maintaining and modulating hematopoiesis. Toll-like receptors (TLRs) function as receptors for different conserved pathogen associated products as well as certain host derived molecules. TLRs are expressed in several hematopoietic and non-hematopoietic cells, and their activation plays a key role in innate and adaptive immune responses to infectious agents, as well as in the development of pathologic conditions like autoimmune disease. In inflammatory conditions like sepsis, cytokines like G-CSF are elevated in serum, CD34+ cells increase in circulation, and bone marrow myelopoiesis is enhanced. We hypothesized that MSCs express TLRs and are capable to respond to TLR agonists by changing their cytokine expression pattern in order to more efficiently support hematopoiesis according to respective needs. In vitro cultured bone marrow MSCs were analyzed for mRNA expression of TLRs. TLR 1, 3, 4, 5, 6, and 9 expression, but not TLR 2, 7, 8, and 10 expression was detectable. Compared to human conventional and plasmacytoid blood dendritic cells, MSCs expressed TLR-3 and TLR-4 at levels up to 2 log higher than did conventional DCs, while TLR 9 expression was low. Upon in vitro stimulation with TLR-4 agonists, mRNA transcripts of flt3-ligand and thrombopoietin increased, and MSCs secreted high amounts of G-CSF, GM-CSF, and M-CSF, whereas stimulation with TLR-9 agonists did not lead to changes in cytokine levels measured in supernatants. TLR-4 stimulated MSC conditioned media enhanced myeloid colony formation from human CD34+ cells about 2.5× fold compared to non-stimulated MSC conditioned media. Furthermore, in co-culture assays of MSCs with CD34+ cells, TLR-4 stimulated MSCs were capable to retain 8 fold more CD34+ cells in divisions 0–3 and maintained overall 2 fold higher numbers of CD34+ cells. In secondary colony formation assays, CD34+ cells from TLR-4 stimulated MSC co-cultures produced about 2.5 fold more myeloid colonies, with some being CFU-GEMM. Moreover, hematopoietic cells taken from TLR-4 stimulated MSC co-cultures were able to engraft conditioned newborn Rag 2−/− gc−/− mice and differentiated into B, T, and myeloid cells, while CD34+ cells from co-cultures of non-stimulated MSCs did not engraft.
These findings thus suggest a regulatory mechanism how, e.g. in gram-negative infection, inflammatory signals are translated via stromal cells into sustained early hematopoiesis and myeloid differentiation to efficiently meet the requests of an ongoing innate immune response.
Histone Deacetylases Are Essential for Chromatin Condensation and Enucleation in Mammalian Erythroblasts
Authors: Ji Peng , Sánchez-Rivera Francisco J , Lodish Harvey F. 
Published: April 2020

Abstract
In the last steps of differentiation mammalian erythroid cells undergo chromatin condensation and enucleation; the latter process does not occur in other vertebrates. Enucleation was critical for the evolution of mammals, as it permits an enhanced hemoglobin concentration – and thus oxygen- carrying capacity – in mammalian red blood cells. We previously reported that Rac GTPases and their downstream forming target mDia2 are required for mouse fetal erythroblast enucleation. We also found that the nucleus undergoes a gradual ~10- fold decrease in volume during erythropoiesis. Since histone deacetylases (HDACs) play important roles in chromatin condensation, we hypothesized that HDACs are involved in mammalian erythroblast enucleation. To test this hypothesis, we purified E13.5 mouse fetal TER119 negative erythroblasts and cultured them in fibronectin-coated plates. Cells were then treated with Trichostatin A (TSA), a pan-HDAC inhibitor, at different times during erythropoiesis. TSA completely blocked enucleation and kinetic studies showed this inhibitory effect occurred earlier than the step catalyzed by the Rac-mDia2 pathway. This indicates that chromatin condensation is required for the final extrusion of the nucleus. We further investigated the activity ofHDAC6 specifically, since HDAC6 is known to interact with mDia2. We found that mDia2is acetylated in vitro and in vivo and that HDAC6 interacts with and deacetylates mDia2. Treatment of purified TER119 negative mouse fetal erythroblasts with a specific HDAC6inhibitor partially blocked enucleation. We conclude that histone deacetylase activities are essential for mammalian erythroblasts to undergo enucleation and that HDACs act both on chromatin condensation and mDia2 deacetylation to promote enucleation.
Dominant Role for SDF-1 in the Vasculo-Angiogenesis Phenotype in Children with Sickle Cell Disease.
Authors: Gee Beatrice E , Manlove-Simmons Jerry M , Huang Yao , Wilson Nana , Stiles Jonathan , Ofori-Acquah Solomon Fiifi 
Published: April 2020

Abstract
BACKGROUND: Vasculogenesis is critically important to restoring and maintaining vascular homeostasis in patients who have sickle cell disease. Emerging evidence indicates vasculo-angiogenesis is a complex process involving recruitment of endothelial progenitor cells (EPCs) from the bone marrow and homing of EPCs to sites of vascular injury. Both recruitment and homing of EPCs are intimately regulated by cytokines and growth factors that are released in response to vascular insult. Hitherto, the phenotype of growth factors and cytokines intimately involved in EPC recruitment and homing in patients who have sickle cell disease remains poorly understood.
METHODS: A total of 50 children aged 6–18 years were studied, 30 with Hemoglobin SS, including 11 on chronic red blood cell transfusions, and 20 with normal hemoglobin. We measured plasma concentrations of angiogenesis growth factors/cytokines using a multiplex assay, which permitted analysis of nine factors concurrently, and determined the number and phenotype of EPC colonies derived from peripheral blood samples.
RESULTS: Plasma concentrations of stromal derived growth factor (SDF-1, 1.7-fold), angiopoietin-2 (Ang-2, 2-fold) and erythropoietin (7-fold) were significantly elevated (p&lt;0.001) in children with sickle cell disease compared to control subjects. On the contrary, plasma leptin was significantly lower (2-fold, p&lt;0.001) in the sickle cell disease group, while no significant differences were found for several other angiogenic growth factors including VEGF, HGF, IL-8, PGF and PDGF between children with sickle cell disease and the control group. Peripheral blood-derived EPC colonies were significantly higher (almost 3-fold) in patients who have sickle cell disease compared to the control group. Interestingly, EPC colony number was lower in sickle cell patients on chronic transfusion therapy compared to those who were not on chronic transfusion. There were significant correlations between EPC colony number and plasma concentrations of SDF-1 (r=0.65, p&lt;0.001) and Ang-2 (r=0.36, p&lt;0.05), but no correlation between EPC colony number and plasma erythropoietin and leptin.
CONCLUSION: These data demonstrate for the first time enhanced vasculogenic potential in patients with sickle cell disease at steady-state. SDF-1 enhances the homing of EPCs and is highly expressed in ischemic tissues. The association of elevated SDF-1 and Ang-2 levels with EPC colony number is consistent with the hypothesis that sickle cell disease causes ongoing ischemic vascular injury. SDF-1 may be potentially used as a marker for vascular injury and a therapeutic target that can be developed to augment vascular repair in sickle cell disease.
Differential Effects of TRAFs in the Activation of NF-KappB in Hodgkin’s Lymphoma Cells.
Authors: Guo Feng , Chen Zi-Xing , Sun Aining , Zhou Peng , Wang Wenjuan 
Published: April 2020

Abstract
CD30, a member of the TNF receptor (TNFR) superfamily, is a lymphocyte-specific receptor that is originally recognized as a surface molecule overexpressed on Hodgkin’s lymphoma cells. Engagement of CD30 with its ligand CD30L activates the nuclear factor kB (NF-kB), which are mediated by interactions with TNFR-associated factor (TRAFs), and supports proliferation of Hodgkin’s cells. However, the role of individual TRAFs in the CD30 signaling pathway in Hodgkin’s cells has not been fully addressed yet. In this study, we found that except TRAF3, all other TRAFs were expressed consistently in B cell-derived Hodgkin’s cell lines (L428 and KM-H2) either in mRNA or protein level. In L428 but not KM-H2 cells, the classical (p50-RelA) and alternative (p52-RelB) NF-kB activity were constitutively activated measured by Western Blotting and EMSA. To better understand the CD30 signaling properties in Hodgkin’s lymphoma cells, we silenced the expression of TRAFs individually by means of RNAi. Successful downregulation of TRAF1 protein expression led to the apoptosis of L428 cells, which was companied by the reduction of both classical and alternative NF-kB activity. Furthermore, the expression of targeting genes of NF-kB, such as A20, c-Flip, ICAM-1, and Cyclin D1 was also decreased. Using siRNA targeting TRAF2 expression, the classical NF-kB activity was reduced while the alternative NF-kB activity moderately induced especially upon CD30L treatment, which was companied by the induction of p100 processing and RelB nuclear localization. The survival rate was decreased when TRAF2 was knockdown. TRAF5 knockdown manifested similar results as that of TRAF2. These observations were only took place in L428 cells but not KM-H2 cells. In addition, the phosphorylation of the extracellular signal-regulated kinases (ERK) 1 and 2, and subsequent activation of JunB in Hodgkin’s cells were staying unchanged in individual TRAFs knockdown experiment. Taken together, the study here further investigates the features of CD30-TRAFs-NF-kB signaling pathway in Hodgkin’s lymphoma cells. At least TRAF1, 2, and 5 are involved in CD30 signaling pathway and protecting Hodgkin’s lymphoma cells from undergoing apoptosis. The interaction between TRAFs is necessary to facilitate the signaling and proper activation of classical NF-kB, which is responsible for the antiapoptotic effect of Hodgkin’s cells. Among TRAF1, 2, and 5, the redundant function is unlikely. Furthermore, the sequential activation of ERK and JunB is unlikely dependent on TRAFs-NF-kB pathway in Hodgkin’s lymphoma cells. Thus, these data reveal that the classical NF-kB activity downstream of CD30 signaling is TRAFs-dependent in Hodgkin’s lymphoma cells, which is regulated by the relative level of individual TRAF in a cell-specific manner.
VEBEP Regimen and Highly Active Antiretroviral Therapy (HAART) in Patients (pts) with HD and HIV Infection (HD-HIV): Final Results of the Italian Cooperative Group on AIDS and Tumors (GICAT) Study.
Authors: Spina Michele , Antinori Andrea , Neri Valentina , Rossi Giuseppe , Re Alessandro , Allione Bernardino , Chimienti Emanuela , Talamini Renato , Tirelli Umberto 
Published: April 2020

Abstract
Background: The outcome of pts with HD-HIV is still poor, because the duration of complete remission (CR) is short. To improve the prognosis of HD-HIV, a feasibility study with the VEBEP regimen and HAART was started in previously untreated HD-HIV pts.
Methods: Chemotherapy (CT) included epirubicin 30 mg/m2/day (days 1–3), cyclophosphamide 1000 mg/m2 (day 1), vinorelbine 25 mg/m2 (day 1), bleomycin 10 mg/m2 (day 3) and prednisone 100 mg/m2/day (days 1–3).
Results: Since September 2001, 71 pts have been enrolled. The median age was 41 yrs. The median CD4+ cell count was 248/mm3 and 51% of pts had a detectable HIV viral load. Stage III–IV was present in 50/71 (70%) pts. Histologic subtypes were: MC 70%, NS 20%, LD 4%, LP 2%, unknown 4%. Four toxic deaths were observed (septic shock, PCP, hepatic failure and pneumonia during neutropenia). An absolute neutrophil count &lt;500 was noted in 60% of pts. Grade 3–4 anemia was observed in 38% of pts and severe thrombocytopenia in 22% of pts. Twenty-two per cent of pts had febrile neutropenia with 19 documented infections in 16 pts (4 varicella, 4 bacterial pneumonia, 3 bacterial sepsis, 2 PCP, 1 cerebral toxoplasmosis, 1 esophageal candidiasis, 1 HBV reactivation, 1 HCV reactivation, 1 prostatitis, 1 salmonellosis). CR was obtained in 47/71 pts (66%) and PR in 9/71 pts (13%). With a median follow up of 22 months, only 4 pts have relapsed. OS and TTF at 24 months are 69% and 59%, respectively.
Conclusions: Our preliminary data demonstrate that VEBEP regimen in combination with HAART is feasible and active in pts with HD-HIV. This study was supported by ISS grants.
Increase in the Incidence of Secondary Acute Myeloid Leukemia (2-AML): A Single Institution Experience Over 20 Years.
Authors: Jabbour Elias , Kantarjian Hagop M , Pierce Sherry , Garcia-Manero Guillermo , Cortes Jorge , Faderl Stefan , Ravandi Farhad 
Published: April 2020

Abstract
Secondary acute myeloid leukemia (2-AML) is a distinctive clinical syndrome occurring after primary malignancy treated or not with chemotherapy (CT), radiotherapy (RT), surgery, or a multimodality strategy. We analyzed the data on 2898 consecutive patients (pts) with AML (WHO criteria) referred to our institution between 1985 and 2005, including 1642 males and 1256 females. The median age was 58 years (range, 13–89 years) for pts with de novo AML (1-AML) (n=2198) and 66 years (range, 18–89 years) for those with 2-AML (n=700). 171 pts (18%) were treated for 2-AML between 1985 and 1994 versus 529 patients (27%) treated between 1995 and 2005 (p&lt;0.001). The predominant primary malignancies included lymphoma (18%), breast (16%), and prostate (13%). Pts had been administered various cytotoxic chemotherapy agents (185 pts, 26%) and RT (102 pts, 15%); 182 (26%) had undergone both modalities, and 231 pts (33%) had undergone surgery alone. At diagnosis, 181 (26%) pts with 2-AML had diploid cytogenetics versus 845 (38%) pts with 1-AML (p&lt;0.001). 252 (32%) pts with 2-AML had clonal abnormalities involving chromosomes 5 and 7 versus 383 (17%) pts with 1-AML (p&lt;0.001), and 49 (7%) pts with 2-AML had 11q abnormalities versus 73 (3%) with 1-AML (p&lt;0.001). Pts undergoing CT/RT had a worse cytogenetic risk profile compared to those undergoing surgery alone with more clonal abnormalities involving chromosomes 5 and 7 (36% versus 24%; p=0.002) and chromosome 11 (10% versus 2%; p&lt;0.001), and less diploid karyotype (19% versus 39%; p&lt;0.001). Median survival time after diagnosis of 2-AML was 6 months with 2- and 5-year survival rates of 18% and 11%, respectively. The median survival for pts treated with surgery alone for the primary cancer was 8 months versus 5 months for those receiving CT/RT (p=0.007); their 2- and 5-year survival rates were 22% and 14% and 16% and 9%, respectively. The median survival for pts treated for their primary cancer between 1985 and 1994 was 4 months versus 6 months for those treated after 1994 (p=0.089); their 2-year and 5-year survival rates were 16% and 9% and 19% and 12%, respectively. In conclusion, the incidence of 2-AML is increasing; patients receiving CT/RT had worse cytogenetic profiles and clinical outcomes than those receiving surgical treatment alone. There remains to be seen whether increased use of targeted therapies as prime modality of therapy in primary cancers will be associated with a decreased risk of 2-AML.
Identification of a Human Natural Regulatory T Cell Micro-RNA Signature and Demonstration of the Major Role Played by miR31 in the Control of Foxp3 Expression
Authors: Rouas Redouane , Kazan Hussein Fayyad , Zein Nabil El , Lewalle Philippe , Rothé Françoise , Akl Haidar , Burny Arsène , Romero Pedro , Martiat Philippe , Badran Bassam 
Published: April 2020

Abstract
Regulatory T cells are the main mediators of dominant tolerance. Their mechanisms of action and applications are subjects of considerable debate at the moment. However, a micro-RNA human Treg signature has not been described yet. We investigated human natural regulatory T-cells derived from cord blood and identified, using TLDA and qPCR, a signature composed of five micro-RNAs (21, 31, 125a, 181c and 374). Among those five, two were considerably under-expressed (miR31 and miR125a). We identified a functional binding site for miR31 in the 3′ UTR of the Foxp3 mRNA. We show that Foxp3 gene expression can be directly regulated by miR31. However, our experiments demonstrate that two distinct mechanisms must cooperate to regulate play the global level of expression of FoxP3, one mediated via miR31, and the second one at the promoter site of the gene. This part of the work is still in progress. In conclusion, not only have we found and validated a miR signature for human natural Treg, but we also unveiled some of the mechanisms by which this signature was related to the control of the Foxp3 protein levels in these cells.
The Src-Like Adaptor Proteins, SLAP and SLAP2, Regulate Flt3-Dependent Dendritic Cell Development.
Authors: Liontos Larissa , McGlade C. Jane 
Published: April 2020

Abstract
The Src-like Adaptor Proteins, SLAP and SLAP2, are hematopoietic adaptor proteins that have been previously shown to act as negative regulators of T- and B-cell signaling. SLAP and SLAP2 work in conjunction with the E3 ubiquitin ligase, c-Cbl, to down regulate the T-cell receptor and other components of the T- and B-cell receptor signaling pathways including ZAP-70 and Syk. SLAP and SLAP2 are expressed in many hematopoietic cell types, including progenitor cells that give rise to cells of both myeloid and lymphoid lineages. Recent evidence indicates a role for SLAP and SLAP2 in regulating hematopoietic receptor tyrosine kinase (RTK) signaling. We have shown that SLAP and SLAP2 interact with CSF-1R/Fms and demonstrated that SLAP2 negatively regulates CSF-1 dependent differentiation of FD-Fms cells. Our recent work demonstrates that both SLAP and SLAP2 interact with the Flt3 receptor in an SH2 domain dependent manner and the interaction was mapped to pY589 and pY591 within the juxtamembrane region of the receptor. To examine the role of SLAP and SLAP2 in Flt3 regulation and signaling in vivo, we utilized Flt3 ligand (Flt3L) to generate dendritic cells from mice lacking both SLAP and SLAP2. Slap1−/− Slap2−/− mice have impaired development of Flt3L-dependent CD11c+ bone marrow-derived dendritic cells (BMDC). In contrast to wild-type mice, Slap1−/− Slap2−/− mice produce 47% ± 9.7% less CD11c+ BMDC after 10 days in culture with Flt3L. Whether the reduction in BMDC numbers is indicative of a defect in the proliferation of Slap1−/−Slap2−/− progenitor cells in response to Flt3L is currently being explored. Although the absolute number of Flt3L-generated BMDC from Slap1−/−Slap2−/− mice is reduced, there are no major differences in the subtype, myeloid (70–90% CD11b+) and lymphoid (4–8% B220+), of DC produced between wild-type and double knock-out mice. To determine whether the reduction in dendritic cell numbers is specific to those generated with Flt3L, we generated BMDC using GM-CSF and IL-4. Both wild-type and double knock-out mice produce similar numbers of CD11c+ BMDC when this combination of cytokines is used. The maturation and activation of Flt3L-generated BMDC from Slap1−/−Slap2−/− mice is being investigated. These data indicate a novel role for SLAP and SLAP2 in the differentiation of Flt3L-dependent dendritic cells.
The Effects of Atorvastatin on Hematological and Inflammatory Parameters.
Authors: Kadikoylu Gurhan , Uysal Hilal Bektas , Yenisey Cigdem , Yavasoglu Irfan , Bolaman Zahit 
Published: April 2020

Abstract
Background: Chronic inflammation may play role in the development of atherosclerosis and its complications. In hypercholesterolemia, the evidences of chronic inflammation such as the stimulation of chemokines and cytokines, increase in endothelial adhesion molecules, and the immune reactions against oxidants on lipoproteins are detected. It was shown that statins had some beneficial effects on lipids, thrombosis, endothelial dysfunction, and smooth muscle proliferation.
Aim: To investigate the effects of atorvastatin on hematological and inflammatory parameters in prospective study.
Material and methods: Forty patients (14 male and 26 female) with primary hypercholesterolemia were treated with 20 mg/day atorvastatin for 12 weeks, according to Adult Treatment Panel for Third Report of National Cholesterol Education Program. All patients received a detailed description of this study and signed consent prior to enrollment. National and local ethical committees approved this study. The exclusion criteria were secondary hypercholesterolemia, acute coronary syndromes, liver and renal dysfunctions, diabetes mellitus, acute/chronic infection and inflammatory diseases, pregnancy, lactation, malignancy, and tendency to bleeding. At baseline, and 12th weeks, lipid levels such as low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), very-low-density lipoprotein cholesterol (VLDL-C), triglycerides (TGs), and hematological parameters such as whole blood cell counts, hemoglobin and fibrinogen levels, CD3, CD4, CD5, CD8, CD14, CD16, CD19, CD40, CD45 using flow-cytometry, inflammatory parameters such as interleukin-1 (IL-1), IL-6, IL-18, tumor necrosis factor-alpha (TNF-a), interferon-gamma, soluble CD-40, intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1, high-sensitive CRP, sedimentation rate, and enzymes including CK, AST, ALT were evaluated. The results were compared with two-paired student’s-t test.
Results: At the end of study, atorvastatin decreased TC (p&lt;0.001), LDL-C (p&lt;0.001), TGs (p=0.006), VLDL-C (p=0.012), and HDL-C (p&lt;0.001). While absolute lymphocyte (p=0.003) and platelet counts (p=0.001) were decreased with atorvastatin treatment, absolute monocyte count increased (p=0.002). On flow-cytometric examination, the expressions of CD14 (p=0.015) and CD19 (p=0.039) on lymphocytes were decreased with atorvastatin. Moreover, atorvastatin decreased the levels of TNF-a (p&lt;0.001), sCD40 (p&lt;0.001), ICAM-1 (p&lt;0.001), and IL-18 (p=0.024). ALT increased at the end of treatment (p=0.041). But important adverse events were not seen in the patients.
Conclusion: The anti-platelet and anti-inflammatory effects of atorvastatin, independent from lipid-lowering effects, may play an important role on the prevention of atherosclerosis, in addition to its beneficial effects on lipid parameters.