bluuebunny/crossref_metadata_2025_split huggingface dataset:
Aspirin Resistance - A Comparison of Platelet Functions and 11-Dehydro Thromboxane B2 and 8-epi Prostaglandin F2α Plasma Concentration.
Authors: Zawilska Krystyna , Zytkiewicz Marcin , Hanszke Ewelina , Gielwanowska Liwia , Psuja Piotr , Turowiecka Zofia , Gaik Irmina , Zozulinska Maria , Duszynska Malgorzata 
Published: April 2020

Abstract
Introduction. Laboratory tests do not reveal any changes due to administration of aspirin (75 – 150 mg/d) in about 25% of patients. The definition of aspirin resistance and parameters of different laboratory tests for its identification have not yet been established.
Aim. The aim of our study was to compare the results of platelet aggregation studies, the closure times in PFA-100, the plasma concentrations of 11-dehydro thromboxane B2 (11-d TxB2) and 8-epi prostaglandin F2α (8-epi PgF2α) in patients receiving aspirin chronically.
Material. The study group consisted of 22 patients taking aspirin for the secondary prevention of ischemic stroke (IS). All patients were at least 6 months after the acute onset, and had a diminished intraplatelet concentration of malonyldialdehyde due to aspirin ingestion.
Methods. Following parameters have been evaluated:
Platelet aggregation induced by either ADP (3,5 and 5,0 μM), collagen (2 μg/ml) or arachidonic acid (AA) (0,6 mM); Closure time in PFA-100 (collagen/epinephrine and collagen/ADP cartridges); Plasma 11-d TxB2 and 8-epi PgF2α concentration measured by ELISA method (Cayman Chemicals).
Aspirin resistance has been determined by the following criteria: the intensity of ADP induced platelet aggregation ≥ 60%, collagen induced aggregation ≥ 70%, AA induced aggregation ≥ 20%, PFA-100 closure time ≤ 165 s, 11-d TxB2 concentration ≥ mean of the control group ± 2 SD.
Results.
The frequency of aspirin resistance in patients after ischaemic stroke Aggregation ADP 3,5 μM ADP 5,0 μM Collagen Arachidonic acid PFA-100 11-d TxB2 63,6% 45,5% 18,2% 13,6% 54,5% 50,0%
Statistically significant correlations have been found between the plasma concentration of 11-d TxB2 and PFA-100 (Col/Epi) closure times and between plasma concentration of 8-epi PgF2α and PFA-100 (Col/Epi) closure times. There was no difference in mean plasma concentration of 8-epi PgF2α between the group of patients and controls.
Conclusions.
The most valid laboratory method of aspirin resistance identification in ischemic stroke patients (besides a mesurement of 11-dehydro thromboxane B2) seems to be PFA-100 closure time. Our PFA-100 studies reveal a much higher percentage of aspirin resistance as compared to former studies. The plasma concentration of 8-epi prostaglandin F2α remains in the normal range in patients taking aspirin for the secondary prevention of ischemic stroke.
Rituximab for the Treatment of Recurrent Thrombotic Thrombocytopenic Purpura.
Authors: Bortolheiro Teresa C. , Castro N. S. , Rodolfo Cançado D. , Carlos Chiattone 
Published: April 2020

Abstract
Background: Thrombotic thrombocytopenic purpura (TTP) is a rare and severe disease, characterized by thrombocytopenia, microangiopathic anaemia, fever, renal failure, and neurological manifestation caused by deposition of von Willebrand factor - platelet rich hyaline thrombi in the arterioles and capillaries. Plasma exchange has decreased the mortality in TTP from almost 100% to 10–30%. However, relapses occur in more than one-third of patients, a subset of whom develop multiple relapses or chronic disease requiring numerous sessions of plasma exchange. This treatment is costly and associated with many adverse reactions. Case
Report: We describe a female patient - diagnosed with TTP in 1996 - who did not achieve a sustained remission with plasma exchange, steroids and vincristine, but who remained in remission after a splenectomy performed in 1997. The patient underwent relapse in 2003 and restarted plasma exchange, with progressive improvement of cytopenias. Plasma exchange was reduced in frequency from daily sessions to every other day. There was a lowering of platelet levels and an increase in LDH levels - steroids and vincristine were re-initiated, but without response. The patient was treated with four, weekly doses (375 mg/m2) of rituximab. Plasma exchange was only required during the first 2 weeks of rituximab treatment, and was discontinued after the second dose of rituximab. The patient remained in complete remission for 9 months, but was then admitted to hospital with femoral thrombosis, and normal LDH and platelet levels. After 40 days of thrombosis therapy, the patient had a further relapse and refused plasma exchange. Treatment with rituximab was restarted as previously, and the patient achieved a complete remission within 2 weeks - now sustained for 11 months.
Conclusion: In comparing this patient with eight previous cases not treated with rituximab, we conclude that rituximab is a very effective treatment for TTP, which is not associated with clinically significant toxicity. Randomized clinical trials are required to confirm these findings.
Expression of Vascular Endothelial Growth Factor (VEGF) Associated with Its Receptor (VEGFR) in Relation to Tissue Factor (TF) in Human Breast Cancer Tissue.
Authors: Wojtukiewicz Marek , Sierko Ewa , Zimnoch Lech , Thorpe Philip , Brekken Ralf A. , Kisiel Walter 
Published: April 2020

Abstract
New blood vessel formation is an important process in tumor growth and dissemination. The relationship between vessel density and survival of patients was documented in breast cancer patients. Vascular endothelial growth factor (VEGF) is of paramount importance in new blood vessel development. VEGF exerts its biological function through binding to specific receptors (VEGFR-1/Flt-1 and VEGFR-2/KDR/Flk-1). Recent studies revealed that tissue factor (TF), apart from being the main procoagulant in cancer patients, is endowed with biological activities that do not relate to its hemostatic function. There exists a large body of evidence that TF is one of the key regulators of angiogenesis. In vitro experiments demonstrated that TF stimulates the synthesis of VEGF in different cell types. The purpose of the study was to assess the expression of VEGF and VEGF coupled to its receptor in relation to the expression of TF. Studies were performed on breast cancer fragments and metastatic axillary lymph node tissues obtained during surgical treatment of previously untreated patients. The tissues were fixed according to AMeX method and embedded in paraffin. Immunohistochemical staining procedures according to the ABC method and Envision Double Staining System (DAKO) were employed using polyclonal antibodies directed to TF and VEGF and a novel monoclonal antibody that binds to VEGF only when it is associated with its receptor - VEGFR. In breast cancer and lymph node tissues the co-localization of TF and VEGF in cancer cells was observed. Of particular interest was the co-expression of TF and VEGF in association with its receptor - VEGFR in cancer cells and endothelial cells of small blood vessels. Furthermore, VEGF coupled to its receptor was localized in dendritic cells in metastatic lymph nodes, which may indicate its influence on the host immune response system. The observed co-localization of TF and VEGF (including VEGF in association with VEGFR) suggests a functional interrelationship that exists between TF and VEGF in breast cancer in vivo.
Factor VIII A2 Domain Residues 497–510 and 584–593 Comprise the R8B12 Epitope.
Authors: Ansong Charles , Miles Stephen , Fay Philip J. 
Published: April 2020

Abstract
The murine monoclonal antibody R8B12 recognizes the C-terminal region (residues 563–740) of the A2 subunit of factor VIIIa (Fay et al. J. Biol. Chem. 266:20139, 1991), as judged by western blotting. The location of the R8B12 epitope within the A2 subunit is not known. In the present study we used affinity-directed mass spectrometry (Zhao et al. Proc. Natl. Acad. Sci. USA.93:4020, 1996) to map the R8B12 epitope. Purified A2 subunit was subjected to proteolytic digestion with trypsin. The digest was then subjected to immunoprecipitation (IP) using R8B12 IgG, in which peptide(s) are selected that bind the antibody. Subsequently the masses of the affinity-selected peptides were determined directly from the immune complex by MALDI-TOF mass spectrometry. Proteolysis of A2 with trypsin generated a pre-IP peptide fingerprint that covered ~40% of the A2 domain sequence (Figure, left). Analysis of the post-IP peptide fingerprint showed two masses, 1309 and 1654, as representing affinity-selected peptide fragments (Figure, right). A theoretical database search identified the 1309 mass peak as A2 domain residues 584–593 and the 1654 mass peak as A2 domain residues 497–510. Direct sequencing of both mass peaks confirmed the results of the theoretical database search. These residues mapped to regions on the A2 domain of the factor VIII A domain homology model that are surface exposed and proximal to each other. Taken together, the above results suggest that A2 domain residues 497–510 and 584–593 represent a discontinous epitope for R8B12. Furthermore, based upon blotting specificity of R8B12 to the C-terminal portion of A2, we speculate that the latter sequence that forms this epitope makes a substantially greater contribution to the binding energy.
Figure Figure
The Role of Erythropoietin in Vascular Endothelial Cells and Thrombosis.
Authors: LaMontagne Kenneth R. , Bdeir Khalil , Carinato Maria E. , Butler Jeannene , Cines Douglas B. , Farrell Francis X. 
Published: April 2020

Abstract
Erythropoietin (EPO) is an essential glycoprotein hormone that regulates erythrocyte production in hematopoietic tissues by stimulating growth, preventing apoptosis, and inducing the differentiation of red blood cell precursors. Clinically, EPO has been extensively used to increase levels of hemoglobin in the treatment of patients with anemia due to chronic renal failure, cancer chemotherapy, and HIV therapy. Recent studies suggest that the function of EPO and its cognate receptor, EPOR, are not strictly limited to erythroid lineages. EPOR expression has been identified in nonhematopoietic cells and tissues including endothelial, neuronal, and tumor cells. Epoetins have shown benefit in correcting anemia in chronic kidney disease and in chemotherapy-associated anemia in cancer patients. However, the incidence of thrombosis appears to be higher in patients administered EPO versus controls transfused to comparable hemoglobin levels. In particular, incidences seem higher in oncological patients treated with EPO and receiving combined radiation and chemotherapy than in those receiving chemotherapy alone. To address the potential interaction and responsiveness of the vasculature to EPO, we evaluated EPO using vascular endothelial cell assays. Biochemical analysis (eg, receptor expression, ligand-receptor binding, and activation of signaling cascades) as well as cellular analysis (eg, proliferation, migration, and differentiation) will be discussed. To evaluate the effects of EPO on thrombosis we used the in vivo pulmonary micoemboli (ME) model. In this fibrinolysis model, a preformed 125I-fibrin microemboli with a defined particle size is infused into mice intravenously. Microemboli lodge primarily in the lungs and the rate of radioactivity lost is measured over time. Defects in fibrinolysis have been identified in mice lacking either tPA or uPA. Mice were exposed acutely or chronically to EPO and the dissolution of ME was tested. Mice were injected subcutaneously with either a single dose of EPO (2500 IU/kg) and for the chronic studies EPO (1200 IU/kg) or control buffer three times a week for three weeks. The residual radioactivity in the lungs was measured after 10 and 90 minutes. Red blood cell count, hemoglobin, and hematocrit were analyzed weekly to confirm EPO activity. Our results indicate that neither a single (acute) dose of EPO nor chronic treatment using EPO affected the rate of pulmonary fibrinolysis. This suggests that EPO treatment does not significantly impair the function of the pulmonary fibrinolytic system proteins (eg, uPA, tPA, plasminogen, and PAI-1) using the ME model. We cannot exclude EPO’s potential influence on other aspects of coagulation and fibrinolysis.
Biphasic Waveform of the Activated Partial Thromboplastin Time (A2 Flag) Is Associated with Adverse Events in the Coronary Care Unit.
Authors: Bane Christopher , Leya Ferdinand S. , Lewis Bruce E. , Walenga Jeanine M. 
Published: April 2020

Abstract
Objective: An abnormality of the optical transmission waveform obtained during measurement of the activated partial thromboplastin time (aPTT) has been described in association with DIC and sepsis. This biphasic waveform (BWF) is caused by the in vitro formation of Ca2+-induced complexes between VLDL and CRP. A review of the literature revealed no studies on the BWF in cardiac patients. The purpose of this study was to determine whether the BWF is associated with adverse clinical outcomes among patients in a Coronary Care Unit (CCU).
Patients: Consecutive patients admitted to the CCU over a two-month period (n=83) were retrospectively evaluated. Standard of clinical care was not altered.
Methods: Laboratory results including aPTT, aPTT waveform analysis, troponin, beta-natriuretic peptide (BNP), complete blood count (CBC), basic metabolic panel (BMP), and high sensitivity C-reactive protein (hsCRP) were analyzed when available. BWF values beyond a pre-set threshold [5 SD’s lower than the mean value of a normal population (n=40)] were signaled by the A2 Flag. Hospital charts were reviewed to determine coronary artery disease (CAD), hypertension (HTN), diabetes mellitus (DM), hyperlipidemia, and renal dysfunction. Hospital course including invasive and surgical procedures and medical interventions were recorded.
Results: The A2 Flag was detected in 23 of 83 patients (27.7%) during the entire course of CCU treatment. Using a lower Slope_1 threshold value (as had been used in the DIC/sepsis studies) produced an A2 flag in &gt;90% of the patients. Patients in the A2 Flag and no-Flag groups were comparable for age, gender, HTN, DM and hyperlipidemia. Of the patients with an A2 Flag, 73.9% had acute coronary syndrome (encompassing the spectrum of clinical conditions ranging from unstable angina to non-ST-segment elevation MI to ST-segment elevation MI) vs 59.6% in the no-Flag group, 95.6% had CAD vs 83.8%, 52.1% had arrhythmia vs 32.2% (p=0.020), 60.8% had renal dysfunction (creatinine &gt;1.5) vs 32.2% (p=0.002), 47.8% had elevated BNP vs 22.5% (p=0.037), and 73.9% had elevated troponin vs 50.0%. Length of stay was longer in the A2 Flag group (16.2 vs 7.2 days; p&lt;0.001). Six of seven patients that expired had an A2 Flag. One patient with an A2 Flag expired during a subsequent readmission shortly after the course of this study.
Conclusion: The A2 Flag result is reported with the aPTT and thus is a rapid test available at no additional cost. As an adjunct to routine lab testing, aPTT waveform analysis appears to be a novel means to identify a select population of CCU patients that are at higher risk for adverse cardiac events and warrants further investigation. This pilot study is being used to develop a larger prospective study to determine if the A2 Flag can improve patient care through earlier diagnosis and treatment intervention.
Asymptomatic Factor VII Padua in African Americans.
Authors: Tidd Theresa M. , Ptashkin Beverly , Pollak Eleanor S. 
Published: April 2020

Abstract
Background: Symptomatology in congenital human FVII deficiency with FVII:C (Factor VII Coagulant Activity) levels &lt; 1% ranges from asymptomatic to severe hemorrhagic problems. Additionally, FVII:C differs markedly depending upon the source of tissue factor (TF) in the thromboplastin utilized for measuring FVII:C. FVII deficiency reported in 1978 in patients from Padua, Italy was later shown to be associated with an arginine (R) to glutamine (Q) mutation at FVII amino acid 304. A study by Triplett et al. from 1985 reported FVII deficiency in 26 patients including 16 Caucasians, 1 Hispanic and 9 blacks. Of these patients, all 9 black patients were asymptomatic whereas the other 17 patients had a clinically relevant bleeding disorder. Since that time there have been no reports describing the genetic cause of FVII deficiency in asymptomatic African American individuals.
Methods: Plasma was obtained from patient blood samples obtained in 3.2% sodium citrate. FVII:C levels were performed using one-stage clotting assays with either rabbit brain thromboplastin (Simplastin Excel, Biomerieux, Durham, NC) or lyophilized recombinant human TF (Innovin, Dade Behring, Inc., Newark, DE). Factor VII antigen levels (VII:Ag) were measured using an enzyme-linked immunoabsorbent assay (American Bioproducts Co, Parsippany, NJ). A normal pool was constructed by mixing equal volumes of plasma from greater than 20 healthy control subjects.
Results Patient # Prothrombin Time in seconds FVII:C Simplastin Excel FVII:C Innovin FVII:Ag Genotype Phenotype 1-African American 15.6 16% 33% 61% Heterozygote R304Q Minor Nose Bleeds 2- African American Prolonged 26% 54% 67% Heterozygote R304Q Asymptomatic for bleeding 3- African American 14.5 21% 64% NA Heterozygote R304Q Asymptomatic for bleeding
Conclusions: We show that FVII deficiency in three asymptomatic African American patients at our specialty laboratory was due to FVII-Padua (R304Q). An additional ten African American patients showed prolonged prothrombin times and FVII:C values consistent with the laboratory phenotype of FVII-Padua. This diagnosis prevented inappropriate blood transfusions prior to surgery in several of these patients.
A Novel Congenital Bleeding Disorder Related to High Plasma Heparin-Like Anticoagulant Activity.
Authors: Wang Zhaoyue , Yang Haiyen , Bai Xia , Zhang Wei , Ruan Changgeng 
Published: April 2020

Abstract
Heparin or heparin-like compounds present in human plasma in minute amounts. It has been reported that a very few patients with such diseases as plasma cell neoplasms, acute monoblastic leukemia and acquired immune deficincy syndrome have an increased plasma heparin-like anticoagulant activity. Recently, we found a 10-year-old girl who was physically and developmentally normal, but had recurrent episodes of prolonged bleeding and hematoma starting in her early childhood, which could be stopped by transfusion of fresh frozen plasma or prothrombin complex concentrate. The coagulation tests of her plasma were regularly repeated since she was 2 years old, and always revealed a markedly prolonged APTT (61.8–104 seconds, normal 28–40 seconds) and TT (36–50.1 seconds, normal 14–21 seconds), and a slightly prolonged PT (15.9–25 seconds, normal 11–14.5 seconds). Fibrinogen, prothrombin and other coagulation factors as well as anticoagulant and fibrinolytic systems were all normal. The results of immunologic measurements were either negative or within normal ranges. Treatment of the patient’s plasma in vitro with either protamine or heparinase could completely normalize the coagulation abnormalities, but not with normal plasma. The anticoagulant activity of her plasma corresponded to 0.2 heparin U/mL when measured by a TT assay using normal plasma as substrate and standardized with porcine heparin. Her plasma heparin concentration was 0.22 heparin U/mL when measured using a colometric assay. In ex vivo study, the abnormal coagulation tests could effectively be corrected when the patient was intravenously administed with protamine. Considering these characteristic laboratory features of the patient, we suppose it would probably represent a novel congenital bleeding disorder related to high plasma heparin-like anticoagulant activity which, to our knowledge, had not been described before.
Increased Levels of Platelet-Derived Microparticles in Patients with DIC.
Authors: Kawasugi Kazuo , Noguchi Mizuho , Tashiro Haruko , Gotoh Moritaka , Shirafuji Naoki , Matsuda Juzo , Miyazawa Yukihisa , Kinoshita Tadatoshi 
Published: April 2020

Abstract
Circulating microparticles of various cell types are present in healthy individuals and individuals in various disease states. In these microparticles, platelet-dreived microparticles (PMPs) may be implicated in thrombosis and haemostasis. However, little is known about the role of PMPs in patients with DIC. To investigate the role of PMPs, we measured plasma levels of PMPs in patients with sepsis-associated DIC (n=20) and acute promyelocytic leukemia (APL)-induced DIC (n=5). Plasma samples from those patients groups were assayed for PMPs levels by enzyme-linked immunosorbent assay (ELISA, Japan Immunoresearch Laboratory, Japan). Also, sP-selectin levels were determined by ELISA.
The thrombin antithrombin complexes (TAT) levels were higher in both types of DIC patients as reported by others. In the septic patients with DIC, we found significant elevation in PMPs levels as compared with normal controls. However, we did not find any difference in plasma levels of PMPs in the APL patients with DIC. The sP-selectin levels were elevated in the septic patients with DIC. In early phase of septic patients with DIC, plasma levels of PMPs tend to be higher than that in late phase of septic patients with DIC.These results suggest that plasma levels of PMPs may be candidates for a marker of septic DIC. It appears that PMPs may contribute to the processes of septic DIC.
Sepsis associated DIC APL with DIC Controls (n=20) *(P&lt;0.05) significantly different controls PMPs 34.1 ± 50.1* 9.6 ± 3.6 6.5 ± 0.9 U/ml TAT 25.4 ± 15.5* 22.5 ± 15.3* &lt;;3 μ g/ml
Autoimmunity and Hypercoagulability in Anderson Fabry Patients from Argentina.
Authors: Aggio Mario C. , Martinez Pablo A. 
Published: April 2020

Abstract
Autoimmunity secondary to chronic immunostimulation by glucocerebrosides has been reported in Gaucher disease, as well as in Anderson Fabry disease (AFD) by others and us.
AIMS: To get further insight on this association, we looked for antinuclear and antiphospholipid-related autoantibodies in a series of AFD Argentine patients.
METHODS/PATIENTS: 35 patients, 16 hemizygous and 19 heterozygous from 4 families, were studied for: 1) autoantibodies to extractable nuclear antigens (ENAs): (SSA/RO, SSB/La, Sm/RNP, and Jo-1); 2) anti-double stranded DNA (dsDNA), and 3) antiphospholipid antibodies: lupus anticoagulant (LA), anticardiolipin antibodies (ACA), and antiphosphatidylserine antibodies, (APA).
RESULTS: At least one autoantibody was detected in 18 patients (51.4%). They were more frequent found in hemizygous (68.8%) than in heterozygous (42.1%). Elevated titers of ENAs were seen in 11.4% (n=4). Positive for LA were 34.3% (n=12) and for ACA 14.3% (n=5). Three patients shared two autoimmune markers. Anti dsDNA and APA were never present.
CONCLUSIONS:
There was an unexpected high incidence of autoimmune responses: one every two AFD patients showed at least one autoantibody; They were more frequent in hemizygous than in heterozygous cases. The pathologic implications of their presence is still not known, although it is probable that antiphospholipid antibodies contribute to hypercoagulability in AFD.
Autoimmunity seems to play an important role in some pathogenic disturbances seen in other lysosomal storage diseases. So, humoral/clinical correlations should be followed closely to detect the eventual outcome of collagen diseases or other clinical manifestations of autoimmunity in AFD.