bluuebunny/crossref_metadata_2025_split huggingface dataset:
Microvesicles Derived from Activated Platelets Enhance the Invasive Potential of Breast Cancer Cells.
Authors: Marquez-Curtis Leah A. , Wysoczynski Marcin , Ratajczak Mariusz Z. , Janowska-Wieczorek Anna 
Published: October 2019

Abstract
There is increasing evidence that platelets contribute to cancer metastasis, and yet platelet concentrates are frequently transfused to cancer patients to treat thrombocytopenia after chemotherapy. Recently we reported that microvesicles derived from activated platelets (PMV) transfer various surface receptors/adhesion molecules to normal and malignant target cells and modulate their biological responses (Blood2001; 98:3143; Exp Hematol2002; 30:450). In this work, we hypothesized that the interaction of PMV with cancer cells increases their invasive and metastatic potential. PMV were isolated from outdated platelet concentrates and pre-incubated with human breast cancer cell lines (MDA-MB-231, BT-549 and T47D), and the effect of PMV on the invasive/metastatic potential of these cancer cells was evaluated. We determined (i) the transfer of the platelet-derived antigen CD41 to cancer cells and the adhesion of these cells to human umbilical vein endothelial cells (HUVEC), (ii) the expression of matrix metalloproteinases (MMPs) by breast cancer cells and their ability to cross the reconstituted basement membrane Matrigel, (iii) the expression of CXCR4, the cognate receptor of the a-chemokine SDF-1, produced in bone marrow, in these cell lines after incubation with PMV, and (iv) the effects of PMV on the interactions of the tumor cells with stroma. We found that PMV transfer platelet-derived CD41 integrin to the surface of breast cancer cells and promote their adhesion to HUVEC. Preincubation with PMV upregulates the mRNA for MMP-9 and protein secretion in invasive breast cancer cells (MDA-MB-231 and BT-549) and enhances their trans-Matrigel chemoinvasion. PMV also transfer CXCR4 to the surface of the breast cancer cells and stimulate the trans-Matrigel migration of MDA-MB-231 cells towards SDF-1, which was abrogated by AMD3100, a CXCR4 antagonist. Finally we found that PMV increase activation of the latent form of MMP-2 constitutively secreted by fibroblastic cells in co-cultures of tumor cells with bone marrow stroma. Thus, we conclude that PMV may enhance the invasive and metastatic potential of breast cancer cells. Because concentrations of PMV are known to be higher in old platelet concentrates than in fresh ones, we recommend that cancer patients should preferably be transfused with fresh platelet concentrates only.
Unique Expression of Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1/CD31) on Embryonic Stem Cells.
Authors: Han Zhong Chao , Li Zong Jin , Wang Zack Z. , Zheng Yi Zhou , Xu Bin , Yang Ren Chi , Scadden David T. 
Published: October 2019

Abstract
Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) is widely used as endothelial marker during vasculogenesis and angiogenesis from embryonic stem (ES) cells. Here, we demonstrate that undifferentiated ES cells expressed high level PECAM-1, but low levels of other endothelium markers, including VE-cadherin, Flk-1 and CD34. Kinetic analysis revealed that the expression of PECAM-1 was sharply down-regulated during early ES cell differentiation. PECAM-1 was expressed in the vascular-like area along with other endothelium makers during differentiation of ES cells. In the absence of angiogenic growth factors, embryonic bodies (EBs) were lack of vascular-like structures, and PECAM-1 was detected in the cells without organized structure. Furthermore, undifferentiated ES cells were found to express all eight known alternatively spliced isoforms of PECAM-1, among them, the expression of PECAM-1 isoforms lacking exon 15, (D15), or 14&15 (D14&15) was predominant. Only the expression of PECAM-1 isoform lacking exon 12&14&15 (D12&14&15) was significantly increased as vascular development of ES cells. These results indicate that PECAM-1 is a constitutive marker of undifferentiated murine ES cells and suggest that the diversity in the expression of cytoplasmic PECAM-1 isoforms may affect its function during vasculogenesis and angiogenesis.
Effect of Native and Cleaved Forms of Antithrombin on Nuclear Factor κB Activation in Endothelial Cells.
Authors: Ni Hongyu , Chauhan Balwant , Olson Steven 
Published: October 2019

Abstract
Antithrombin (AT) is a single chain plasma glycoprotein that belongs to a superfamily of serine protease inhibitors (Serpin). AT is the major inhibitor of the serine proteases of the coagulation cascade, notably thrombin and factor Xa. In addition, AT has also been demonstrated to have anti-inflammatory properties. Recently, cleaved and latent forms of AT have been shown to function as antiangiogenic agents. However, the molecular mechanisms by which native AT and cleaved AT exert their anti-inflammatory and antiangiogenic effects remains unknown. In this study, we have investigated the effects of both native and cleaved forms of AT on endothelial cell proliferation and nuclear factor κB (NF-κB) activation in the endothelial cells. Bovine pulmonary artery endothelial cells (BPAE) was used in the study. Human α-antithrombin was purified from plasma by affinity chromatography on heparin Sepharose, followed by anion-exchange chromatography. Cleaved form of AT was prepared by digestion of purified human AT with human neutrophil elastase, followed by chromatography on a heparin Sepharose column. The endothelial cells were stimulated with basic fibroblast growth factor (bFGF) or tumor necrosis factor-α (TNF-α) in culture and incubated with either native AT or cleaved AT. Endothelial cell proliferation was measured by the MTT (methylthiazolyldiphenyl-tetrazolium bromide) cell proliferation assay as well as by counting cell numbers before and after the treatment. The results demonstrate that both native AT and cleaved AT could inhibit bovine pulmonary artery endothelial cell proliferation. These results contrast with those reported with other types of endothelial cells such as human umbilical vein endothelial cells (HUVEC) in which only cleaved form but not native form of AT inhibits cell proliferation. NF-κB activation was detected by the ELISA-based assay using antibodies specific for the activated form of p50 and p65 subunit of the NF-κB. In native AT, the inhibitory activity of endothelial cell proliferation was associated with down-regulation of NF-κB as measured by decreased nuclear p65. However, the cleaved AT showed minimal effect on the NF-κB activation. Our results suggest that although both native and cleaved AT inhibit endothelial cell proliferation, they might use different signal transduction pathways. Moreover, our findings suggest that native and cleaved AT may have differential effects on different types of endothelial cells.
Rituximab in the Treatment of Adults with Chronic Idiopathic Thrombocytopenic Purpura (ITP) and Autoimmune Hemolytic Anemia (AIHA).
Authors: Zalzaleh Ghassan , Jajeh Ahmad , Tamoseviciene Diemante 
Published: October 2019

Abstract
Corticosteroids have been the first line of treatment of ITP since 1950, however some patients do not respond to this treatment (refractory) and some will relapse after its discontinuation. For such patients second line treatments were introduced. Some patients will continue to be refractory to this treatment and need other therapy modality.
Rituximab is a chimeric monoclonal antibody directed against the CD20 antigen exposing B Lymphocytes, causing its depletion. This could alter the production of auto-antibodies in some Auto-Immune diseases and thus could be used in their treatment. Few medical centers had reported using Rituximab in the treatment of refractory (ITP) and (AIHA), yet its definite role could not be determined, and here we share our experience.
Patients with documented diagnosis of ITP or AIHA who were refractory to at least two lines of therapy including steroids were offered to receive Rituximab (375mg/m2 weekly for 4 weeks). 15 patients were enrolled, 10 with ITP, 4 with AIHA, 1 with Coombs negative Hemolytic anemia, and 1 with pure red cell aplasia. One had both ITP and AIHA. 10 were females and 5 males. 5 were >60 years old and 10 were < 60 years old. 2 out of the 10 patients with ITP had also Chronic Lymphocytic Leukemia (CLL). Duration of follow up ranged from 2 months to 17 months (average 7 mos). Of the 10 patients with refractory ITP treated with Rituximab overall response was 60%. 4 were NR (no response), 2 were MR (minimal response: Platelets increased to <50000), 2 were PR (Partial response: Platelets increased to <100000) and 2 were complete response (Platelets became normal). 3 patients of 6 with Hemolytic anemia or PRC aplasia had NR, 1 had MR (Hct <30), and 2 had partial response (Hct 30–35). No complete response was observed in this group. In 3 patients with hemolytic anemia and CLL 1 had MR, 1 had PR and 1 had NR. 2 patients with hemolytic anemia who had NR died as a complication of their disease (one with septic shock and one with severe autoimmune flare up). Only one patient with refractory ITP had mild allergic side effects and did not complete 4 doses. No Rituximab related mortality was observed.
CONCLUSION:
Rituximab therapy had a variable but valuable effect in the treatment of patients with chronic refractory ITP and refractory/ relapsed AIHA. Overall response in our group reached 60%. No clinical or laboratory parameters were found to predict response, although there was a suggestion that males, younger age, and no history of splenectomy have a better chance of response. As we lack an effective alternative treatment in chronic refractory ITP and AIHA, Rituximab use could be a valid option in view of its mild toxicity. Further follow up of our patients and input from other institutions in this regard are needed.
A Ten Year Survey of Chronic Immune Thrombocytopenic Purpura in Canadian Children, 1984–2003: Report of the Canadian Chronic ITP Working Group.
Authors: Ali Syed Kaiser , Belletruti Mark , Barnard Dorothy , Chan Anthony , David Michele , Luke Brian , Price Victoria , Ritchie Bruce , Wu John 
Published: October 2019

Abstract
Chronic Immune Thrombocytopenic Purpura (C-ITP) is a heterogeneous group of disorders manifesting as thrombocytopenia of greater than six months duration. To better understand its natural history, and detect trends of response to therapy and of outcomes, a ten-year retrospective survey of C-ITP diagnosed in Canadian children from age 6 months to under 20 years was conducted in 2004. A standardised form for capture of anonymous data on each patient was completed and returned from 11 institutions, including the three largest Canadian Pediatric Hematology/Oncology centres. Of the total of 243 forms received, 213 cases met the inclusion criteria. Male:female::1:1.3. Mean age at diagnosis of C-ITP was 7.9 years (range 8 months - 17.7 years). Mean platelet count at diagnosis and mean duration of C-ITP were 46 x 109/L (range 1 - 145 x 109/L) and 2.9 years (range 0 - 9.7 years) respectively. Primary ITP was the underlying diagnosis in 85 % of cases. Internal hemorrhage was documented in 1 % of patients. Treatments (either singly or in combination) with drugs, IVIG, anti-D, or monoclonal antibody, were administered in 91 % of cases. Five percent of patients received blood transfusions for treatment of bleeding episodes; 18.1 % underwent splenectomy. Information gained from this data base will be used in developing management guidelines for C-ITP, and will provide the foundation for establishing a Web-based Canadian National C-ITP Registry.
Impedance-Based Whole Blood and Optical-Based Platelet Rich Plasma Aggregation Methods for the Assessment of Celexocib Effects on Platelet Function: A Preliminary Report.
Authors: Dyszkiewicz-Korpanty Anna M. , Frenkel Eugene P. , Sarode Ravindra 
Published: October 2019

AbstractCyclooxygenase-2 (COX-2) is the inducible cyclooxygenase isoform expressed by cells involved in inflammation. Platelets normally lack COX-2; however, the enzyme is expressed in megakaryocytes and newly formed platelets and may play a role in clinical syndromes associated with high platelet turnover (1, 2). Selective COX-2 inhibitors (coxibs), have become the mainstay for treatment of patients with arthritis and also have been widely used in cancer chemoprevention. Coxibs were found to have a safer GI toxicity profile when compared with classic NSAIDS; however, in some patients they may increase the risk of GI bleeding (3). The underlying mechanism has not yet been elucidated. Moreover, the role of coxibs in hemostatic alterations has been a subject of controversy. The aim of the study was to investigate the possible anti-platelet effects of celecoxib (200 mg/d for 7 days) in 7 healthy volunteers (2 females, 5 males; mean age = 35.7 ± 8; mean platelet count = 230 ± 23; mean Ht = 42.3 ± 6). A very sensitive whole blood impedance platelet aggregation assay, previously unused for this purpose, was employed for the assessment of celecoxib influence on platelet function. The test was performed along with a traditional, optical-based platelet rich plasma (PRP) aggregation method. Arachidonic acid (0.5; 0.25 mM), Collagen (2; 1mg/mL) and ADP (20, 10 mM) were used as platelet agonists in both tests.Celexocib did not affect platelet function in neither whole blood nor in PRP. The preliminary results of this study indicate that the increased bleeding tendency in patients treated with celecoxib is probably not related to platelet dysfunction.Assay Arachidonic Acid Collagen ADP 0.5[mM] 0.25[mM] 2[μg/mL] 1[μg/mL] 20[μM] 10[μ M] WB Impedance 17.5±4.7 20.9±3.8 25.4±3.9 25.9±6.1 14.3±2.6 14.1±5 Aggregation: Pre-drug Post-drug 18±4 21±5 27.1±4.7 24±4 18.7±6.2 16.3±7 PRP Optical 118.1±10.3 118.1±6.7 101.4±13.7 110±2.2 104±15 108±8.9 Aggregation: Pre-drug Post-drug 119.6±6.7 112.9±10.2 105.4±11.5 110.4±7.5 102.1±18.3 107.7±9.6
Determination of TAFI in Plasma Using a New Chromogenic Method.
Authors: Wilmer Marianne , Buehler Beatrice , Peter Wikstroem , Hugo Ziegler 
Published: October 2019

Abstract
TAFI (thrombin-activatable fibrinolysis inhibitor) is a proenzyme similar to carboxypeptidase B and is activated to TAFIa by thrombin and probably plasmin also. Thrombomodulin accelerates TAFI activation by thrombin about 1250-fold. TAFIa is an important inhibitor of fibrinolysis by cleaving C-terminal lysine and arginin residues in the fibrin clot. By this the carboxy-terminal plasminogen binding sites in the fibrin clot are degraded, plasminogen activation by t-PA and in consequence fibrinolysis is most effectively inhibited. Whether elevated TAFI levels correlate with the risk of thrombosis is currently under study. To clarify the role of TAFI in various diseases a simple and rapid assay for determination of TAFI concentration in plasma is desirable. With Pefakit® TAFI we developed a new chromogenic method for determination of TAFI in plasma samples.
Method: TAFI is activated by thrombin/thrombomodulin complex in the presence of calcium ions. The activated TAFI cleaves a 3-thia-arginine peptide substrate whose cleavage products subsequently react with Ellman’s reagent. Color development is followed up by measuring absorption at 405 nm. The method is the result of synthetic work and substrate screening. The assay was optimized to determine TAFI in human plasma. The concentration of TAFI in plasma sample corresponds to the initial reaction rate of substrate cleavage. TAFI level in each sample can be calculated from a linear standard curve.
Results TAFI was measured in plasma of about 50 patients. The plasma levels of TAFI determined with Pefakit® TAFI correlate well with a commercially available chromogenic assay (Actichrome TAFI).
Conclusion: Pefakit® TAFI is a fast and precise method to determine TAFI concentration in plasma. There is a linear correlation between TAFI concentration and measuring signal. Because of its simple practicability the assay can be adapted to most of the common automated analyzers for routine application.
Successful Treatment of Intractable Gastrointestinal Hemorrhage with Low Dose of Recombinant FVIIa (Novoseven®) in Pediatric Patient without Coagulation Deficiencies.
Authors: Kim Ji Yoon , Lee Kun Soo , Kwon Soon Hak , Choe Byung Ho , Sohn Sang Kyun 
Published: October 2019

Abstract
Recombinant FVIIa (Novoseven®) was initially developed for the treatment of bleeding in patient with congenital hemophilia with antibodies against factors VIII and IX and FVII deficiency. The mechanism of action is to induce hemostasis at the site of injury both by forming complexes with exposed tissue factor and by directly activating factor X upon the surface of an activated platelet. More recently it has begun to use gradually in the management of bleeding patient without congenital coagulation defect. We report the successful use of recombinant factor VIIa in a pediatric patient who has intractable gastrointestinal bleeding despite of conventional medical therapy.
Case: The patient is a 13-year-old female and her body weight is only 10 kg, with a prior history of cerebral palsy and epilepsy. Recently she presented a recurrent massive hematemesis and melena probably due to stress ulcer for 8 days. She had no personal or family history of a suggestive bleeding disorder. The patient had a normal platelet count and coagulation screening test. Her hospital course was complicated by recurrent gastrointestinal hemorrhage but an endoscopic intervention was not permitted due to her poor condition. Despite of conventional medication with omeprazole and sucralfate over 20 days, she suffered from recurrent hematemesis and melena. In the gastrointestinal bleeding scan with 99mTc-labeled RBCs, small intestinal hemorrhage was found. The following day, the patient was given three doses of Novoseven® at 100 μg/kg, as bolus doses with 2 hour interval, which resulted in resolution of hematemesis and melena after last dose administration. Although she had mild whole body rash after third dose administration, it was subsided soon after medication of antihistamine and steroid. No evidence of hemorrhage was documented at a follow-up gastrointestinal bleeding scan that was taken after 4 days later of treatment. The patient made a full recovery and no further treatment was required.
Conclusion: We suggest that low dose of recombinant FVIIa (Novoseven®) were effective in treatment of pediatric intractable gastrointestinal hemorrhage.
Germline Polymorphisms Provide Strong Prognostic Information in Intermediate Risk Acute Myeloblastic Leukemia (AML).
Authors: Urbano-Ispizua Alvaro , Brunet Salut , Monzo Mariano , Perea Granada , Navarro Alfons , Esteve Jordi , Berlanga Juan , Ribera Josep-Maria , Bueno Javier , Llorente Andreu , Besalduch Juan , Guardia Ramon , Tormo Mar , Pedro Carmen , Sanchez Juan-Maria , Torres Pio , de Llano Maria-Paz Queipo , Font Llorens , Moraleda Jose-Maria , Marti Josep-Maria , Vivancos Pilar , Peñarubia Maria-Jesus , Nomdedeu Josep , Montserrat Emili , Sierra Jorge 
Published: October 2019

Abstract
In this study we tested whether distinct patterns of functional genomic polymorphisms in genes involved in drug metabolic pathways (GSTT1, SULT1C2, CD-EX, TOP2A, SXR-1, SXR-2), DNA repair (XPD, XPA, XPG, ERCC1), angiogenesis (VEGF-1, VEGF-2), and multidrug resistance (MDR1-1, MDR1-2) predict clinical outcome in patients with AML. Two hundred adult patients ≤60 year-old with primary AML (M3 excluded), enrolled in the AML-99 protocol of the CETLAM group between November-98 and July-03, have been studied. Induction therapy consisted of idarubicin, intermediate dose ara-C (IDAC) and VP16. Intensification included mitoxantrone and IDAC. Patients were following treated with high-dose ara-C or transplantation depending on the risk category. According to the MRC cytogenetic classification, 32 patients had a favourable karyotype, 23 had abnormalities indicating poor prognosis, and 110 patients were included in the intermediate prognosis (IP) group. We focused on IP group patients with the aim of improving its prognostic stratification. The characteristics considered were: age (<50 years vs ≥50), WBC (≤20x109/l vs >20x109/l), FAB classification, MLL rearrangement, internal tandem duplication of FLT3 (ITD-FLT3), induction courses to achieve complete remission (CR) (1 vs 2), and germline polymorphisms of the above-mentioned genes. Prognostic variables in the univariate analysis (Kaplan-Meier method) with a p value ≤0.2 (log-rank test) were included for the multivariate analysis (proportional hazard method). Of the 110 patients included in the IP group, 86 (78%) achieved a CR, 11 (10%) were chemoresistant and 13 (12%) died during induction. After a median follow-up of alive patients of 24 months (range 5–64), overall survival was of 31% at 5 years. In the multivariate analysis, adverse prognostic variables for survival were polymorphism of XPA (RR=3.4; p=0.02) and of MDR1-1 (RR=2.1; p=0.02), and WBC >20x109/l (RR=2.1; p=0.02). Increased risk of relapse was associated with polymorphism of SULT1C2 (RR 4.1; p=0.004), ITD-FLT3 (RR 3.3; p=0.003), VEGF2 (RR 2.8; p=0.04) and MDR1-1 (RR 2.4; p=0.02). Finally, in the multivariate analysis for refractoriness to chemotherapy, XPA polymorphism was the only independent factor increasing the risk (RR=14; p=0.02). In conclusion, germline polymorphisms, which can easily be analyzed from DNA of peripheral blood, have independent prognostic value in intermediate risk AML.
Generation of Prothrombinase According to Hicks and Pitney Test. Comparison of the Levels of F1+2 Simultaneously in Hemophiliacs with and without Thrombophilic Mutations and Normal Subjects.
Authors: Makris Pantelis P.E.M. , Girtovitis Fotios F.I.G. , Tsaousidou Maria M.K.T. , Pithara Eleftheria E.P. , Foka Zoi Z.I.F. , Samama Michel Meyer M.M.S. 
Published: October 2019

Abstract
It has been seen that a significant number of hemophiliacs show a co-existence of mutations related with an increased risk of thrombosis (Foka et al 2003). It has also been suggested that the existence of these genetic defects that manifestate with either hemorrhage or thrombosis, may actually protect these patients from recurrent bleeding episodes. In order to study the coagulation mechanism in these patients the Hicks & Pitney test was selected, that examines the generation of prothrombinase. Materials and Methods. 64 subjects were studied (56 hemophiliacs 42±15 year old and 8 normal subjects 45±10 year old). 37 without mutations, and 19 others with mutations (12 with FVLeiden and FII G20210A and 7 homozygotes in MTHFR). The test was applied to 2 groups (one with hemophiliacs and one with normal subjects). The levels of F1+2 and TAT were also measured in every incubation phase, as markers of the generation of prothrombinase and thrombin.
It can be seen in the charts that follow that hemophiliacs with no mutations show a gradual prothrombinase generation. On the contrary, hemophiliacs with mutations show a rapid generation that is also expressed by the concomitant increase in the levels of F1+2 and TAT. A similar response was observed in the group of normal subjects.
This can be explained as an adjustion of the coagulation mechanism of hemophiliacs with coexistence of thrombophilic mutations, possibly capable of preventing the high frequency and severity of bleeding episodes.
Figure Figure