bluuebunny/crossref_metadata_2025_split huggingface dataset:
Circulating Cytokeratin 5+ Progenitor Epithelial Cells Also Express Other Progenitor Cell Markers and Are Necessary for Normal Airway Repair.
Authors: Gomperts Brigitte N. , Burdick Marie D. , Belperio John A. , Strieter Robert M. 
Published: April 2020

Abstract
Background: Human lung transplants have demonstrated recipient epithelial cells in donor organs, supporting the notion of a circulating epithelial progenitor cell. Resident progenitor epithelial cells in the proximal airway are located in the submucosal gland ducts and basal epithelium and express cytokeratin (CK) 5/14. We have identified circulating progenitor CK5/14 positive epithelial cells in the bone marrow and buffy coat of mice. These cells appear to traffic via the CXCR4/CXCL12 biologic axis.
Purpose of study: To further determine additional markers of these circulating progenitor epithelial cells and to disrupt the CXCR4/CXCL12 axis to prevent recruitment of these circulating progenitor cells after airway injury.
Methods used: Flow cytometric analysis of cytokeratin 5 (CK5), CXCR4, CD45, CD34, Sca-1, c-kit expressing cells in the buffy-coat and bone marrow of naive mice and CK5-GFP transgenic mice. Subcutaneous heterotopic implantation of dissected female wild type tracheas into male GFP recipient mice. Immunohistochemical and dual immunofluorescence analysis of GFP and CK5 as well as p63 in tracheal transplants.
Summary of results: FACS analysis of bone marrow and buffy coat from CK5-GFP transgenic mice and wild type mice revealed that these circulating CK5+ progenitor epithelial cells also express Sca-1,CD34 and c-kit. In the heterotopic tracheal transplant model, wild type tracheal transplantation in CK5-GFP recipient mice demonstrated that the recipient circulating CK5-GFP cells contributed to repair of the airway epithelium. Passive immunization of animals bearing tracheal transplants with specific neutralizing anti-CXCL12 F(Ab)2 antibodies resulted in the unexpected phenotype of the airway epithelium demonstrating squamous metaplasia at day 21 post-transplant.. In contrast, animals bearing tracheal transplants passively treated with control antibodies demonstrated normal pseudostratified epithelium at day 21 post-transplantation. Immunofluorescence for p63 in the tracheal transplants demonstrated that all of the cells of the squamous metaplasia were p63 positive in tracheal transplants from animals that had CXCL12 depleted. Analysis of these tracheal transplants with immunohistochemistry and immunofluorescence for GFP and CK5 demonstrated that the squamous metaplasia was solely derived from resident progenitor epithelial cells.
Conclusions: There is a population of CK5+ cells in the bone marrow and circulation of naive mice that also express Sca-1, CD34 and c-kit, which is compatible with a stem cell. These circulating CK5 positive cells contribute to regeneration of the airway epithelium and their recruitment to the progenitor cell niche in the airway appears to be critical for the development of normal pseudostratified columnar epithelium.
Tissue Factor-Exposing Microparticles at the Site of Hemostasis Activation.
Authors: Gartner Verena , Lubscyk Barbara , Kollars Marietta , Kyrle Paul A. , Weltermann Ansgar 
Published: April 2020

Abstract
Introduction: Tissue factor exposing microparticles (TF+ MPs) are capable to activate hemostasis in vitro. The aim of this study was to investigate the impact of TF+ MPs during physiologic activation of hemostasis in vivo in human.
Methods: In a double-blind, cross-over study healthy male volunteers (n=13) were randomized to 7 days treatment with 100 mg aspirin and placebo, respectively. At the end of each treatment period TF+ MPs were determined in venous blood and in blood emerging from a skin incision reflecting hemostasis in an activated state (shed blood). TF+ MPs were analyzed by flow cytometry after staining with annexin V, a TF-antibody and cell-specific antibodies.
Results: Compared to venous blood, the median (interquartile range) number of TF+ MPs was significantly higher in shed blood: 16.2 (11.3 – 20.1) k/ml vs. 8.2 (5.9 – 14.8) k/ml (p = 0.004). TF+ MPs from platelets were higher in shed blood than in venous blood [4.7 (3.7 – 8.6) k/ml vs. 3.3 (2.0 – 5.0) k/ml, p = 0.03], whereas TF+ MPs from endothelial cells were not [1.7 (1.2 – 5.8) k/ml vs. 2.7 (1.7 – 4.5) k/ml, p = 0.9]. Aspirin did not significantly influence the level of TF-MPs both in venous blood and in shed blood.
Conclusion: High levels of TF+ MPs can be detected at the site of physiologic activation of hemostasis in vivo in human.
An Anti-Phosphatidylserine (PS) Antibody, Tarvacin™ That Targets Tumor Blood Vessels Does Not Affect Platelet Function In Vitro.
Authors: Dyszkiewicz-Korpanty Anna M. , Sarode Ravindra , Thorpe Philip E. , Frenkel Eugene P. 
Published: April 2020

Abstract
Tarvacin™ is a chimeric anti-PS antibody that is currently in Phase I clinical trials in cancer patients. It acts by targeting PS that becomes exposed on vascular endothelium in tumors in response to oxidative stress in the tumor microenvironment. Tarvacin™ recognizes a complex of PS and the PS-binding protein, β2 glycoprotein I. Host leukocytes are induced to bind to the complex in tumor vessels and destroy tumor vessels by antibody-dependent cellular cytotoxity. However, antibodies directed against PS-associated proteins are also known to elicit anti-phospholipid syndromes (APS). Anti-PS antibodies possibly cause APS by displacing anticoagulant proteins from PS on activated cell or by enhancing the binding of prothrombin; another explanation might be a direct activation of endothelial cells and platelets. The aim of the study was to determine whether Tarvacin ™ induces or interferes with platelet activation caused by ADP, collagen type I or calcimycin in vitro. Blood was drawn from 3 healthy volunteers, aged 31–54, who have not taken any antiplatelet medication for 14 days prior to the study. Dual channel whole blood aggregometer (Chronolog, Havertown, PA, USA) was employed for platelet aggregation studies in whole blood (WB/impedance method) and platelet rich plasma (PRP/optical method). Platelet count in PRP was adjusted to 200 K/μL. Platelet agonists (PS exposure triggers) used in the experiments were as follows: collagen (0.5, 1, 2 μg/mL), ADP (1.25, 2.5, 5, 10 μM), Calcimycin (10, 20, 30 μM) and Calcium ions (1, 2 mmol/L). Tarvacin™ was provided by Peregrine Pharmaceuticals Inc, Tustin, CA. The Anti-CD 20 antibody, Rituxan ™ and physiologic saline were used as controls. Specimens (WB diluted with saline in 1:1 ratio or PRP) with the addition of Tarvacin™ (100 μg/mL) or Rituxan ™ (100 μg/mL) or saline were first incubated on a gentle mixer for 10 minutes; incubation was then continued at 37 ° in the aggregometer well for another 5 minutes. Agonist-induced platelet aggregation was subsequently examined. Platelet aggregation studies in both WB and PRP showed that Tarvacin™ neither induced platelet activation, nor inhibited platelet activation in response to ADP, collagen or calcimycin in vitro. In conclusion, Tarvacin™ does not affect platelet function in the present in vitro assays. Possibly, the epitope on the PS -β2 glycoprotein I complex does not orientate the antibody in a manner that interferes with platelet activation. Alternatively, activated endothelial cells or other factors may be critical to support platelet activation.
2-Oxoglutarate Down-Regulates Expression of Vascular Endothelial Growth Factor and Erythropoietin through Decreasing Hypoxia Inducible Factor-1 α and Inhibits Angiogenesis.
Authors: Imagawa Shigehiko , Matsumoto Ken , Obara Naoshi , Suzuki Norio , Nagasawa Toshiro , Yamamoto Masayuki 
Published: April 2020

Abstract
In oxygenated cells, hypoxia inducible factor-1 (HIF-1)α subunits are rapidly degraded by a mechanism that involves ubiquitination by the von Hippel-Lindau tumor suppressor (pVHL) E3 ligase complex using 2-oxoglutarate as a substrate. This process is suppressed by hypoxia and iron chelation, allowing transcriptional activation. The interaction between human pVHL and a specific domain of the HIF-1α subunit is regulated through hydroxylation of proline residues 402 and 564 by HIF-1α proryl-hydroxylase (PHD). N-oxalyl glycine acts as a competitive inhibitor of HIF-PHDs and this inhibition is in competition with 2-oxoglutarate. We examined the effect of 2-oxoglutarate on the production of vascular endothelial growth factor (VEGF) and erythropoietin (Epo). The expression of VEGF and Epo protein were dose-dependently downregulated in Hep3B cells by the addition of 2-oxoglutarate. The enhancer activity of the HIF-1 binding site of Epo and the promoter activity of VEGF-luciferase were also dose-dependently downregulated by the addition of 2-oxoglutarate. Gel mobility shift assays revealed that the addition of 2-oxoglutarate dose-dependently inhibited HIF-1 binding activity, but did not affect GATA binding activity. Western blot analysis revealed that 2-oxoglutarate dose-dependently inhibited HIF-1α protein in Hep3B, Hela and SW480 cells in hypoxic conditions. However MG132 (the proteasome inhibitor) rescued the inhibition of HIF-1α protein expression by 2-oxoglutarate. Furthermore, under hypoxic conditions, 2-oxoglutarate dose-dependently inhibited tube formation in in vitro angiogenesis assays. These results indicate that 2-oxoglutarate treatment may be useful for the inhibition of tumor angiogenesis through decreasing HIF-1α protein, HIF-1 binding activity, the promoter activity of VEGF and enhancer activity of Epo, and the production of VEGF and Epo. More studies to determine if 2-oxoglutarate inhibits tumor angiogenesis in vivo mouse assay are in progress.
Production of Tissue Factor Pathway Inhibitor in Endothelial Cell Cultures Is Reduced by Estrogens, Selective Estrogen Receptor Modifiers, and a Selective Estrogen Receptor Downregulator.
Authors: Dahm Anders E.A. , Iversen Nina , Birkenes Baard , Sandset Per Morten 
Published: April 2020

Abstract
Low plasma tissue factor pathway inhibitor (TFPI) is a weak risk factor for venous and arterial thrombotic disease. Female hormones strongly influence the level of TFPI in plasma. Oral contraceptives (OC) and hormone replacement therapy (HRT) reduce plasma levels of TFPI by 30–50%, and premenopausal women have lower TFPI than postmenopausal women and men. Moreover, the selective estrogen receptor modifiers (SERM) Tamoxifen and Raloxifen also reduce TFPI in plasma. The effect of these drugs on TFPI may in part explain their prothrombotic effects. The aim of the current study was to study the effects of estrogens and SERMs on the production of TFPI from endothelial cell cultures. Human coronary arterial endothelial cells from Cambrex Bio Science were seeded into 24 well plates and preincubated with 0 (control) or 10 nM of 17β-estradiol (E2), 17α-etinyl-estradiol (EE2), tamoxifen or raloxifene for 48 hours. The cell-medium was then changed and the cells were stimulated with the same concentrations of the same drug for 24 hours before harvesting. TFPI total antigen was measured using a commercial ELISA-kit, and cellular total protein was measured by an improved Lowry assay. The concentration of TFPI total antigen was normalized against the concentration of total protein from the cells. We found that 10 nM of E2, EE2, Tamoxifen and Raloxifene reduced TFPI in the cell medium with 39% (E2), 30% (EE2), 19% (Tamoxifen), and 35% (Raloxifene) compared to untreated cells. Fulvestrant is a selective estrogen receptor downregulater (SERD) known to have no agonist effects, and is also used as a drug against breast cancer. The effect of fulvestrant on TFPI-production was tested in order to later use it as a blocker of the estrogen receptor (ER), to decide whether the effects observed from the other compounds were ER-dependent. Surprisingly, 10 nM fulvestrant reduced TFPI in the cell medium by 12%, and 1000 nM fulvestrant reduced TFPI by 30%. We conclude that the effects of estrogens and SERMs, and interestingly also the SERD fulvestrant, reduce TFPI production in endothelial cells. Based on these results we propose that women using fulvestrant as a drug against breast cancer will have reduced levels of TFPI, and perhaps also increased risk for thrombotic dieseases.
Preparation and Charaterization of MoAb SZ118 and SZ118-scFv Against Platelet Glycoprotein VI.
Authors: Yu Ziqiang , Dong Ningzheng , Bai Xia , Shen Fei , Dai Lan , Ruan Changgeng 
Published: April 2020

Abstract
GlycoproteinVI (GPVI ), one of the platelet collagen receptors, plays an important role in platelet activation and thrombosis. GPVI participates in platelet adhesion to exposed collagen under high shear rate condition, and leads to platelet activation and thrombus formation. GPVI is widely recognized as a requisite factor for the formation of platelet aggregates on the collagen surface under blood flow. There were growing evidences showed that impairment of GPVI function could result in a long-term antithrombotic protection and less bleeding side-effect, which potentialized that GPVI might be an interesting target for safe anti-adhesion and anti-thrombotic therapy. Recently monoclonal antibodies directed against GPVI were developed, which impaired the interaction of platelets and collagen. A human soluble recombinant GPVI (rGPVI), consisting of extracellular domain of GPVI, was expressed in E. coli. After immunizing Balb/C mice with the purified rGPVI, a monoclonal antibody, SZ118, was developed. Meanwhile the gene encoding for the light-and heavy-chain variable regions of SZ118 had been cloned by RT-PCR from murine hybridoma cells. After ligated into the vector pET20b(+), expression plasmid pET20b(+)-SZ118-scFv was contructed, and SZ118-scFv fragment was expressed in E.coli. The parallel plate perfusion chamber and platelet aggregometer were used to study the effects of SZ118 and SZ118-scFv on the platelet adhesion and aggregation to collagen. The citrated whole blood with or without SZ118 was perfused over the surface of type I fibrillar collagen (Chrono Log, at 80μg/ml) at high shear rate (1000 s−1) for 5 min. The platelet aggregates were visualized by the phase-contrast microscope, the coverage rates of the platelet aggregates were measured. Monoclonal antibody SZ118 and SZ118-scFv could bind to human platelets exclusively by flow cytometry. SZ118 inhibited collagen-induced platelet aggregation in PRP in dose-dependent manner, nearly full inhibition being attained at the concentration of 110 ug•mL−1. Similarly SZ118-scFv inhibited collagen-induced platelet aggregation with maximum inhitition rate 84.3%±5.6%. And also SZ118 and SZ118-scFv had part effect on convulxin-induced platelet aggregation, but having little effect on ADP. After treating the citrated whole blood with SZ118 or SZ118-scFv at final concentration 50μg/ml at 37° for 10 min, the platelet adhesion on the surface of collagen was markedly inhibited with the coverage rates of 1.03%±0.22%, respectively (22.7%±5.7% for control), which indicated that the platelets adhesion to collagen was inhibited by 95.5%by SZ118 and 68.3% by SZ118-scFv. It was concluded that the MoAb SZ118 inhibited platelet aggregation to collagen and platelet adhesion to immobilized fibrillar collagen under high shear rate condition.
Therapy of Chronic ITP - The Patients’ Perspective in Germany.
Authors: Matzdorff Axel C. , Arnold Gabriele , Pralle Hans , Preiss Joachim 
Published: April 2020

Abstract
Introduction: Adult ITP is a chronic disease. Many patients do not achieve remission despite immunosuppressive therapy. After 1st line therapy with steroids and immunoglobulins splenectomy is usually offered as 2nd line therapy, even to thrombocytopenic patients without bleeding symptoms. However, several non-surgical treatment options have been available for many years and recently anti-CD20 antibodies (rituximab) and anti-D (Cooper, Blood 2002) have been added to this list. This study was intended to asses which treatments are usually offered to patients in Germany.
Methods: In June 2004 and February 2005 we sent out a questionnaire that was returned by 80 patients. 7 questionnaires were excluded because of patient age (&lt;12 y) or minor thrombocytopenia (platelets &gt;100.000/μl).
Results: Age (median 50 years) and gender distribution (38% m, 62% f) are as expected for chronic ITP. Median duration of disease is 5 years (range 1–43 years). 27% of the patients have experienced gastrointestinal/urogenital bleeding, 12% had CNS or eye bleeds. 92% of the patients had been treated with steroids and 24% were splenectomized. Only 31% had been informed by their physicians of rituximab and 12% of anti-D. 35% of the patients had exclusively been informed about steroids, immunoglobulins and splenectomy. 45% of patients had the impression that cost of treatment affected their therapy options.
Conclusions: ITP is a chronic condition and patient histories extend over many years. Standard therapies comprise steroids, i.v. immunoglobulins, and splenectomy. However, splenectomy has a considerable long-term failure rate (33%, Kojouri Blood 2004). Many patients are thrombocytopenic but have little or no bleeding symptoms. Particularly those patients are reluctant to undergo surgery as 2nd line therapy and prefer trying a non-surgical approach first. Anti-D and rituximab are not covered by public health insurance. It is possible that insurance reimbursement policies restrict treatments offered to ITP patients in Germany.
Treatment of Thrombotic Thrombocytopenic Purpura Using Solvent Detergent Treated Plasma.
Authors: Rock Gail , Anderson David , Benny Barrett , Clark William , Leblond Pierre , Sutton David , Sternbach Marion , Wells George ,   Members of the Canadian Apheresis Group,   Canadian Association of Apheresis Nurses
Published: April 2020

Abstract
Background:
Patients with Thrombotic Thrombocytopenic Purpura require immediate therapy with large volumes of plasma. They have been reported to respond well to plasma exchange (PE) with FFP (78%)1 and CSP (95%)2. We initiated a randomized prospective trial with CSP and solvent detergent treated plasma (SDP) intended to show non inferiority.
Methods:
Patients were randomized to receive either of the exchange fluids at 1.5 PV x 3 days then 1 PV x 4–6 days. Laboratory values were followed with ADAMTS-13 levels measured according to Furlan3. The primary end point was survival at 6 months.
Results:
A total of 62 of the required 280 patients entered the study then the supply of SDP was discontinued. 3/35 patients receiving CSP and 1/27 on SDP died before 1 month. These values are not significant but the study is underpowered so statistical analysis is not possible. FVIII and vWF levels were elevated in all patients at entry. 34/50 patients tested also had antibodies to CD36. 9 patients had no ADAMTS-3 at entry, 19 had between 5–50%, 9 patients had 50–100% and 15/62 patients tested had normal levels. Inhibitors were present at between 5–50% levels in 22/50 patients tested whereas 9 patients had complete inhibition of ADAMTS-13 and 18 had normal enzyme activity.
Conclusions:
Patients with TTP appear to respond well to SDP thereby offering a relatively safer therapeutic option for PE. An insufficient number of patients were entered to achieve statistical significance. Both the ADAMTS-13 and the inhibitor levels show great variation and are not prognostic.
Earlier Administration of Rituximab Allows Higher Rate of Long-Lasting Response in Adult Patients with Autoimmune Thrombocytopenia.
Authors: Zaja Francesco F. , Vianelli Nicola N. , Battista Marta M. , Sperotto Alessandra A. , PAtriarca Francesca F. , Tomadini Valentina V. , Fili Carla C. , Tani M. , Baccarani Michele M. , Fanin Renato R. 
Published: April 2020

Abstract
Background
Previous reports highlighted the potential short and mid-term therapeutic activity and safety of rituximab in adult patients with autoimmune thrombocytopenias.
Objectives
Primary objectives of this study were to evaluate the long-term efficacy and toxicity profile. Patients and methods From October 1999 to April 2005, 37 adults patients, median age 54 years, with active and symptomatic autoimmune thrombocytopenias (30 idiopathic thrombocytopenic purpura, 1 idiopathic thrombocytopenia and neutropenia, 4 thrombocytopenia and concomitant undifferentiated connective tissue disease, 2 thrombocytopenia and concomitant B-cell lymphoprolipherative disorders) that had relapsed or were refractory to at least a full course of steroid therapy received weekly infusions of rituximab 375 mg/m2 for 4 weeks. The median interval from diagnosis to rituximab was 34 months (1–264 months) and the platelets median count was 11 x 109/L. The following parameters of efficacy and toxicity were considered: rate of complete and partial response, time to initial and maximum response, relapse rate, relapse free survival, treatment free survival, short and long-term toxicity. The possible prognostic influence of some clinical and laboratory parameters on the patients outcome were also analyzed.
Results
Complete and partial response (platelets count ≥ 100 x 109/L and ≥ 50 x 109/L) were 20/37 (54%) and 7/37 (19%), respectively. In most of the patients the time to initial and maximum response was prompt, already before the second administration of rituximab. The median period of observation from treatment was 22 months (1–48 months). Nine out 27 responding patients relapsed; 18/37 patients (49%) remained relapse free and 20/37 (54%) did not necessitated further therapy. A shorter interval period from the time of diagnosis and rituximab administration (≤ vs &gt; 36 months) was associated with a lower relapse free survival (p= 0.03). During the period of rituximab administration, 2 patients experienced short term toxicity with one case of serum sickness syndrome; no infectious or other significant long term toxic complications were documented..
Conclusion
Rituximab administration may allow to achieve long-lasting remission in nearly 50% of patients suffering from autoimmune thrombocytopenia with good toxic profile. The possibility to achieved long lasting response appeared related with an earlier timing of administration.