bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antiphospholipid Antibodies Impair the Clearance of Apoptotic Platelets.
Authors: Pierangeli Silvia S. , Vega-Ostertag Mariano E. 
Published: April 2020

Abstract
Background: Thrombocytopenia is frequent in patients with the Antiphospholipid Syndrome (APS). The mechanism(s) that lead to abnormal platelet destruction is not understood. Phosphatidylserine (PS) that is exposed in the outer leaflet of the membrane of aged human platelets (AHP) and β2glycoprotein I (β2GPI) mediate their phagocytosis by macrophages without inducing inflammatory or immune responses. We hypothesized that antiphospholipid antibodies (aPL) affect the clearance of AHP and induce immunogenicity of AHP leading to the production of anti-platelet antibodies.
Methods: To examine that question, we studied phagocytosis of AHP by macrophages in the presence of β2GPI, IgG aPL or control IgG (IgG-NHS). AHP labeled with CM-Orange, were incubated with β2GPI and aPL IgG or with IgG-NHS, and added to a monolayer of cultured phagocytes. The fluorescent-positive phagocytes (FPPC) were then tested by fluorescence microscopy. Then the cells were trypsinized and analyzed by flow cytometry. We also examined the immunogenicity (production of anti-platelet antibodies) of AHP treated with IgG aPL and with IgG-NHS by immunizing Balb/c mice with AHP and IgG-aPL or IgG-NHS. The patterns of reactivity of the sera of the immunized mice was examined by immunoblot of human platelet lysates.
Results: aPL IgG produced a significantly lower % FPPC compared to the IgG-NHS-treated cells (15.33 ± 5.31 vs 89.0 ± 7.94, respectively). This was confirmed in the flow cytometric studies: IgG aPL produced significantly lower % of FPPC when compared to IgG-NHS-treated platelets (42.49 ±11.77 vs 78.11± 5.43, respectively).. Mice immunized with AHP and aPL IgG produced significantly higher titers of anti-platelet antibodies (as detected by ELISA) when compared to mice immunized with IgG-NHS (p=0.0033). Furthermore, there was a different pattern of reactivity of the sera with respect to recognition of platelet antigens, when sera of immunized mice were analyzed by immunoblot.
Conclusions: The data indicate that aPL impair the clearance of apoptotic platelets and affect their immunogenicity. This may lead to the production of anti-platelet antibodies and thrombocytopenia in APS.
A Proteomic Approach to the Characterization of the Endothelial Dysfunction in Uremia.
Authors: Carbo Carla , Diaz-Ricart Maribel , Fuste Berta , Cases Aleix , Carrascal Montserrat , Abian Joaquin , Ordinas Antonio , Escolar Gines 
Published: April 2020

Abstract
Deficient hemostasis coexists paradoxically with accelerated atherosclerosis in patients with cronic renal failure (CRF). The high incidence of aterothrombotic events in these patients has been associated with endothelial dysfunction. We have established an in vitro model of cell dysfunction by exposing endothelial cells (EC) to uremic media (Serradell y cols, Thromb Haemost86:1099,2001). However, it is difficult to advance in this research using conventional protein identification techniques. We have applied a proteomic approach to exhaustively characterize the EC dysfunction in uremia. Extracts of confluent EC, grown in the presence of sera from control donors and from patients under hemodialysis, were collected and lysated. Proteins were resolved by molecular weigh (Mr range 10–50kDa) and isoelectric point (pI range 3–10) using two-dimensional gel electrophoresis (2-DE). Spots were visualized by silver staining and further identified by mass spectrometry (MALDI-TOF). A search for significant changes in protein phosphorylation and glycosilation, and overexpression or new presence of proteins was specifically performed in uremic EC. Identification of some of the most prominent proteins revealed molecules related to inflammation (HMGB1, 25148/5.8, Mr/pI; aldose reductase, 35854/6.5; a proteasome component, 29192/5.7) and oxidative stress (superoxide dismutase, 24772/8.3; gluthatione peroxidase, 21899/6.2), both processes associated with CRF. In addition, we observed a significant increase in the protein destrin (18506/8.1), an actin depolymerizing protein. This finding could be related to the previously reported increased detachment of uremic EC when exposed to flowing blood. Therefore, proteomics can be used to compare the proteome of control and uremic EC, helping to reveal information on the molecular basis of the disease. A more exhaustive analysis of 2-DE gels will provide answers and potential therapeutic targets in a short future.
Defibrotide, an Endothelium Protecting and Stabilizing Drug, Has Anti-Angiogenic Potential In Vitro and In Vivo.
Authors: Eissner Guenther , Geissler Edward K. , Iacobelli Massimo , Andreesen Reinhard , Holler Ernst , Koehl Gudrun E. 
Published: April 2020

Abstract
DF is a polydisperse mixture of single-stranded polydeoxyribunucleotides which is successfully used in the treatment of hepatic veno-occlusive disease and other endothelial disorders. Recent pre-clinical evidence suggests that DF might also have anti-neoplastic properties. We addressed the question whether this might be due to the prevention of tumor blood vessel formation (angiogenesis). The anti-angiogeneic potential of DF was tested in vitro (Matrigel™ tube formation and aortic ring assay) and in vivo (dorsal skin-fold chamber model). Our results show that DF quantitatively (100%) blocks tube formation of trans-differentiated human endothelial-like cells (ELC) at concentrations corresponding to pharmacologic DF blood levels (100 μg/mL). Similarly, the sprouting of rat aorta endothelial cells in Matrigel™ was prevented by nearly 100%, when DF was applied on a daily basis. In vivo tumor angiogenesis in a human gastric cancer (TMK-1) grown in skin-fold chambers (nude mice) was also attenuated on day 5 by DF, as measured by microvascular density. Although the exact mechanism of DF action remains to be elucidated, initial Western blotting results show that DF reduces phosphorylation-activation of p70S6 kinase, which is a key target in the PI3K/Akt/mTOR signaling pathway linked to endothelial cell and pericyte proliferation and activation. Taken together, our data suggest that while DF is known for its endothelium-protecting function, it also inhibits tumor blood vessel formation, and thus should be considered for further testing as an anticancer agent.
Efficacy and Safety of a New Intravenous Immunoglobulin in Adult Subjects with Chronic Idiopathic Thrombocytopenic Purpura.
Authors: Leibl Heinz , Varga Gyula , Volkova Zuzana , Gasztonyi Zoltan , Hlusi Antonin , Mayer Jiri , Chojnowski Krzysztof , Wolf Hans-Heinrich , Sharkhawy Marlies , Pavlova Borislava G. , Birthistle Karl , Engl Werner , Walter Simone , Ehrlich Hartmut J. 
Published: April 2020

Abstract
Idiopathic thrombocytopenic purpura (ITP) is a common clinical disorder of immune regulation leading to phagocytic destruction of platelets. Intravenous immunoglobulin (IGIV) has been used successfully to increase platelet count in patients with ITP. Baxter has developed a new IGIV (IGIV,10%), a 10% liquid immunoglobulin preparation with 3 dedicated virus reduction steps integrated into the manufacturing process. The efficacy and safety of this product was assessed in a prospective multi-center study in adult chronic ITP patients with platelet counts of 20x109/L or less. A total of 23 subjects were enrolled and treated for 2 to 5 days with a total dose of 2 g/kg. Of 21 subjects included in the per-protocol analysis data set, 15 responded to treatment (71.4%), achieving a platelet count of at least 50x109/L by Day 8. Fourteen of 15 responders reached this level by Day 5. The median duration of platelet response was 25 days and the highest median platelet count in the responders was 182x109/L. A total of 81 infusions were administered to the 23 subjects in the safety analysis data set. Among the 40 non-serious adverse events related to the use of the study drug, 35 were mild, 3 were moderate, and 2 were severe. The most frequent related adverse events were headache and pyrexia. The results of this study demonstrate that IGIV, 10% TVR Solution is effective in the treatment of adult subjects with chronic ITP and has an excellent safety profile.
Radiometric Quantitation of PAIgG Stratified Treatment Response in ITP Patients.
Authors: Estrada Dexter , Schwartz2 Kenneth A. 
Published: April 2020

Abstract
Because treatment response in ITP is variable, it would be useful to be able to assess which patients are likely to respond to conventional treatment. Toward this goal we correlated the number of platelet surface IgG molecules measured with a quantitative radiometric platelet antibody assay (PAIgG) with treatment related changes in platelet count. We tested the hypothesis that patients’ response to conventional treatment could be stratified based on quantitation of the patients’ PAIgG. Data was obtained via a retrospective chart review. Patients with a diagnosis of idiopathic thrombocytopenic purpura (ITP), age 18 or older with an initial platelet count of &lt;150,000, and quantitation of antiplatelet IgG performed at the time of diagnosis were included in the study. Patients were treated using conventional approaches that included steroids, IVIgG, anti-D antibody, splenectomy, and or cytotoxic medications. Patient response to therapy was evaluated as the change in platelet count, computed as the difference in platelet count obtained at diagnosis and the latest stable platelet count available. There were 64 patients with at least 90 days between initial and follow-up platelet counts. Of the 31 patients whose PAIgG were ≤1,000 molecules/platelet (m/plt), only 1 had an initial platelet count of &lt;50,000. Of the 33 patients who’s PAIgG were &gt;1,000 m/plt 16 had platelet counts of &lt;50,000 (p≤0.001. Patients with PAIgG levels &gt;1000 m/plt exhibited a significantly greater increase in platelet count μ=73,000 ±92,000 as compared to those with PAIgG levels ≤1000 m/plt, μ=18,000 ±43,000 (p=0.03). Although a PAIgG level of 500 m/plt is 3 standard deviations above the mean of normals, a greater response to treatment was noted in patients if their PAIgG level was &gt;1000 m/plt. Patients with PAIgG levels between 500 and 1000 m/plt respond in a similar fashion to those with PAIgG levels &lt;499. In addition, more patients with PAIgG levels &gt;1000 m/plt received conventional treatment (19/33 vs. 10/31; p&lt;0.05). In particular, there were more patients who received cytotoxic treatment in this cohort, (11/33 vs. 3/31; p≤0.025). Similar results were observed in 74 patients with at least 30 days between initial and follow-up platelet counts. This study suggests that measuring the amount of antiplatelet IgG stratifies ITP patients into 2 groups: patients with PAIgG ≤1,000 m/plt with platelet counts over 50,000 and patients with PAIgG &gt;1,000 m/plt platelet counts that are less than 50,000. Those patients whose PAIgG is &gt;1,000 m/plt are more likely to receive treatment and have an increase in platelet count with conventional therapy. We conclude that radiometric quantitation of PAIgG is helpful for predicting a patient’s response to therapy.
Hemolytic Uremic Syndrome Is an Immune Mediated Disease: Role of Anti-CD36 Antibodies.
Authors: Rock Gail , Clark William , Sternbach Marion , Kolajova Maria , McLaine Peter 
Published: April 2020

Abstract
Hemolytic uremic syndrome is a disorder in which platelet microthrombi are formed which have a particular propensity to deposit in the kidney microvasculature resulting in impaired renal function and thrombocytopenia. The mechanism of formation of these microthrombi is not known. In this study, we showed that plasma from five adult and six pediatric cases of HUS caused aggregation and release of adenosine triphosphate from normal platelets. The plasma reacted against platelet lysate in a protein blot and all samples showed reactivity against a band at 88kDa corresponding to the membrane antigen CD36. This was confirmed by probing with Mo91, a monoclonal antibody to CD36. CD36 was also identified in the immune complex formed by incubation of patient plasmas with normal platelet lysate. In other studies, bands of 32kDa and 7.7kDa were obtained when purified verotoxin was protein blotted and probed with either patient plasma or with anti-CD36 antibody Mo91 suggesting structural homologies between CD36 and verotoxin. The data support the concept of an immunological pathogenesis for HUS and suggest that molecular mimicry involving one or both of the homologous domains in membrane-bound CD36 and verotoxin lead to the development of antibodies capable of inducing the pathophysiological events characteristic of HUS.
Treatment of Refractory Chronic Immune Thrombocytopaenic Purpura (ITP) with Rituximab: Report of a Cases Series.
Authors: Garcia-Chavez Jaime , Vela-Ojeda Jorge , Gonzalez-Acosta Jose C. , Ovilla-Martinez Roberto 
Published: April 2020

Abstract
Purpose: To describe the response rates and time to treatment failure in patients with chronic refractory ITP treated with Rituximab.
Methods: Fourteen consecutive patients with chronic refractory ITP, who didn’t achieve a response to first- and second-line treatments (including splenectomy), were recruited between November 2001 and May 2004. Rituximab was administered at 375 mg/m2 iv weekly’ 4 and all patients were pre-medicated with Diphenhydramine (30 mg single dose) and Hydrocortisone (100 mg IV single dose). The first response assessment was performed at week 4, using the following criteria: complete response, platelet count &gt; 100 x 109/L; partial response, platelet count &gt; 50 x 109/L; minimal response, platelet count persisted below 50 x 109/L but without bleeding or need for platelet transfusion; and no response if the platelet count did not change, or the patients required platelet transfusion or remained symptomatic. The response was considered sustained if it persisted for at least 6 months.
Results: The age range was 17–70 years; the baseline platelet count range was 3–37 x 109/L. Five patients achieved a complete response, four patients a partial response, four a minimal response and one no response. The time to response was from 4–32 weeks and 10 patients showed a sustained response. No patients with a response showed bleeding and no patients required the use of other drugs.
Conclusions: These results suggest that Rituximab should have a significant therapeutic effect and an acceptable safety profile in patients with refractory chronic ITP. However, there is a lack of randomized, prospective studies to demonstrate the therapeutic benefit of this treatment.
Reduced Intensity Allogeneic Transplantation for Non-Hodgkin’s Lymphoma: Extended Follow-Up of an Alemtuzumab-Containing Regimen.
Authors: Thomson Kirsty J. , Morris Emma C. , Milligan Don , Cook Gordon , Cavet Jim , Craddock Charles , Hunter Ann , Schey Steve , Parker Anne , Goldstone Anthony H. , Linch David C. , Peggs Karl S. , Mackinnon Stephen 
Published: April 2020

Abstract
Allogeneic transplantation with reduced intensity conditioning is increasingly being used in patients with non-Hodgkin’s Lymphoma (NHL) who fail standard therapy. We report extended follow-up on 121 patients with NHL, who underwent allogeneic transplantation with reduced intensity conditioning at 8 UK centres. Conditioning was with fludarabine 150mg/m2, melphalan 140mg/m2 and alemtuzumab (60–120mg). Cyclosporin A was administered at 3mg/kg from day-1, and stem cell source was bone marrow or PBSC. Diagnoses were in 3 categories: low grade follicular NHL (n=50), mantle cell lymphoma (n=21), and high-grade NHL (n=50, including transformed low grade disease n=15). Donors were HLA-matched siblings in 75 (62%), and unrelated in 46 (38%), of whom 18 were HLA-mismatched at up to 3/10 loci. 48% of patients had failed previous autologous transplantation. Median follow-up was 35 months (1–78). For the group with low grade follicular NHL (n=50), estimated overall survival (OS) was 76% at 1yr and 67% at 4yrs, and non-relapse mortality (NRM) was 16% at 4yrs. Disease relapse or progression occurred in 12 patients, of whom 8 received donor lymphocyte infusions (DLI), with responses in 6. Current progression-free survival (cPFS) is 68% at 4yrs. For the group with mantle cell lymphoma (n=21), estimated OS was 83% at 4yrs, NRM was 11% at 4yrs and relapse or progression occurred in 6 patients. Three patients received DLI, with non-sustained responses in 2. Current PFS is 43% at 4yrs. For high-grade NHL (n=50), estimated OS was 52% at 1yr and 45% at 4yrs. Prior autologous transplantation was common in this group (72%), and NRM was higher at 34% at 1yr and 40% at 4yrs. Progression/relapse occurred in 15 patients, of whom 10 received donor lymphocytes, with responses in 5. Current PFS is 48% at 1yr and 43% at 4yrs. These results, from patients who were often heavily pre-treated, including having failed autologous transplantation, provide encouraging evidence to support the application of reduced intensity allogeneic transplants in NHL. NRM in low grade follicular or mantle cell lymphoma is low, consistent with the use of T cell depletion, and graft-versus-lymphoma effects can be induced with DLI in a subset of cases. The data in follicular lymphoma, in particular, supports consideration of this therapy earlier in the disease. In high-grade disease, NRM appears to be higher, but durable remissions are attainable in a proportion of patients.
Comparison of Rapid Platelet Function Assay with Whole Blood Platelet Impedance Aggregometry for the Definition of Aspirin Resistance.
Authors: Dyszkiewicz-Korpanty Anna M. , Kim Anne , Burner James D. , Frenkel Eugene P. , Sarode Ravindra 
Published: April 2020

Abstract
The reported incidence of aspirin (ASA) resistance ranges from 5 to 30%. Various platelet function assays have been employed to detect aspirin resistance in patients with cardio- and cerebrovascular disease. Such a high proposed incidence of ASA resistance poses a critical need for a rapid point-of -care (POC) platelet function test. Unfortunately, no uniformly accepted definition of ASA resistance exists. Platelet aggregation studies that have been used to define ASA resistance are time consuming and require special technical expertise. The Ultegra Rapid Platelet Function -ASA (RPFA-ASA) has been developed as a POC test that is performed without sample processing. This optical method measures agglutination of fibrinogen-coated beads upon platelet activation with arachidonic acid. In the presence of aspirin effect, however, the agglutination of the beads is inhibited. The described cutoff of ≥ 550 Aspirin Reaction Units (ARU) is termed non-responsiveness to ASA based on a preclinical study and subsequent correlation with epinephrine-induced platelet aggregation in platelet rich plasma. Since RPFA-ASA uses whole blood, we validated its performance characteristics against a classic whole blood platelet aggregation assay (WBA). We studied 50 healthy volunteers, aged 25–75 (24 men, 26 women) with normal CBC, who had not ingested anti-platelet drugs for 14 days prior to the study. Baseline studies included WBA (dual channel aggregometer, Chrono-log Inc., Havertown, PA) using both arachidonic acid (AA -0.5; 0.25 mM) and collagen (1; 2 μg/mL) as well as an RPFA-ASA assay (Accumetrics Inc., San Diego, CA). These studies were repeated after 3 days of ASA (325 mg/d) intake. Based on a review of the literature, we defined an adequate ASA response as a completely inhibited AA-induced platelet aggregation and at least 30% inhibition of collagen-induced aggregation (both concentrations of the agonist). Thus, those with &lt; 30% inhibition of aggregation response to collagen were considered ASA resistant. Eleven subjects were ASA resistant by WBA (20%; 8 females and 3 males (aged 25–63). In contrast, since all 50 subjects achieved ARU values of &lt; 550 ARU, none were recognized as an ASA non-responder by the RPFA-ASA. While the current cutoff of &lt; 550 ARU posed by the Ultegra RPFA-ASA does identify those who have taken ASA, the assay is unable to recognize ASA non-responders. Thus, based on these data, the appropriate cutoff for the recognition of ASA resistance by the RPFA-ASA should be re-adjusted to a significantly lower level to ensure appropriate assay results.
New Approach for Evaluation of Quantitative Risk Factors of Venous Thromboembolism Using Individualized Reference Values.
Authors: Zotz Rainer B. , Gerhardt Andrea , Scharf Rudiger E. 
Published: April 2020

Abstract
The evaluation of quantitative risk factors of venous thromboembolism is characterized by several unsolved problems. The majority of quantitative components of hemostasis are dependent on age, sex, and hormone intake. The cut-off values determined in case-control studies depend on the specific patient/control group and can not be generalized on individuals with other characteristics. Furthermore, the approach does not give reference values dependent on age, sex, and hormone intake which are necessary for risk estimations in clinical practice.
To overcome these disadvantages, we used a multiple regression analysis to create a system of reference values which change continuously depending on age, sex, and hormone intake according to the parameter distribution in healthy controls and calculated the relative risk of hemostatic components (729 patients with first VTE and 675 healthy controls). A significantly increased risk for venous thrombosis was associated with deficiency of protein S activity (odds ratio (OR) 2.8 to 6.1, p=0.0007), free protein S concentration (OR 2.7 to 20.4, p=0.0001), protein C activity (OR 3.2 to 9.4, p=0.0001), antithrombin activity (OR up to 75, p=0.0001), and increased levels of fibrinogen (OR 3.8, p=0.0001), factor VIII:C (OR 3.3, p=0.0001), factor IX (OR 2.3, p=0.0001), factor XI (OR 2.9, p=0.0001), vWF activity (OR 2.4, p=0.0001), and vWF antigen (OR 3.5, p=0.0001).
In contrast to previously published studies, this new approach gives clinically important cut-off values dependent on age, sex, and hormone intake and allows to identify patients at increased risk for venous thrombosis on an individualized basis. Particularly parameters which are highly dependent on age and sex, like protein S activity, can be characterized more precisely using the described procedure. This comprehensive analysis demonstrates that, apart from well-known risk determinants, deficiency of protein S and increased values of FI, FVIII:C, FIX, FXI, vWF, and vWF-Ag are risk determinants predicting venous thrombosis.