bluuebunny/crossref_metadata_2025_split huggingface dataset:
In Vitro Characterization of Magnetic Resonance Imaging Contrast Agents for Molecular Imaging.
Authors: Yu Zhi , Grafe Michael , Meyborg Heike , Fleck Eckart , Li Yangqiu 
Published: April 2020

Abstract
The aim of this work was to evaluate the biological properties of one citrate-coated and two different dextran-coated paramagnetic particles with comparable size (iron core 4–10 nm).
Endothelial cells from humans and mice as well as human macrophages were incubated for different time intervals with different particle suspensions. The cellular uptake was semi-quantitatively measured using the Prussian blue staining and, in addition, by cellular iron content. Furthermore the effect of known inhibitors of endocytosis was evaluated. In addition, it was evaluated whether linking of monoclonal antibodies to dextran-coated particles can make them bind specifically to certain cell surface structures.
The results showed that the bEnd.3 cell line, human umbilical vein endothelial cells (HUVECs) and THP-1/macrophage cell lines internalize paramagnetic particles. The ranking of cellular uptake was: VSOP &gt; CMD-coated particles &gt;&gt; CLIO. The carboxydextran-coated SPIO uptake by endothelial cells is reduced by colchicine (50%). Conversely, cytochalasin B down-regulates the endocytosis of citrate-coated particle. Our data imply that the major mechanism of uptake would be pinocytosis for the VSOP and phagocytosis for the carboxydextran-coated particle CMD. The different surface coating can influence not only the quantity of the internalization, but also the pathway of internalization. CLIO linked to CD40 antibodies or to CD62E antibodies bound significantly better than IgG-linked CLIO. This was true especially for the anti-CD40-CLIO constructs where fluorescence increased two fold. Comparable results were observed with anti-CD62E-CLIO constructs; however, increase in fluorescence was higher than with CD40 binding; it increased on 3.9-fold (median) and 4.5-fold (mean).
In conclusions, the binding of antibody-conjugated CLIO to the antigen-expressing cells was specific, with an affinity similar to that of the free antibody. Thus, it seems feasible to use antibody linked SPIOs for molecular imaging.
Prevalence of Anti-HLA and Anti-GPIIB/IIIA Allo-Immunization in Patients with Glanzmann Thromboasthenia: Experience of a Single Center.
Authors: Santoro Cristina , Rago Angela , Antenucci Angela , Chistolini Antonio , Conti Laura , De Vellis Annalisa , Di Renzo Lucia , Laurenti Luca , Perrone Mariapaola , Foa Robert , Mazzucconi Mariagabriella 
Published: April 2020

Abstract
Background. Platelet transfusions, the main therapy of Glanzmann Thromboasthenia (GT), can induce an allo-immunization against HLA antigens and GPIIb/IIIa complexes, with a possible reduction of efficacy of subsequent treatments.
Aims. To investigate the development of anti-HLA antigen and anti-GPIIb/IIIa complexe allo-immunization in GT transfused patients, and to evaluate the efficacy of replacement therapy.
Patients and Methods. From 1975 onwards, we have followed 17 GT patients; 13 type I, 3 type III, 1 not classified; 8 men, 9 women; median age at diagnosis 9.8 years (range 1–44.5); the median age at the time of this study was 35.5 years (range 23.6–68.5). Our patients had shown at least once in their life the following symptoms: 10/17 epistaxis; 5/17 gastrointestinal hemorrhage; 5/17 oropharingeal hemorrhage; 4/17 muscle hematoma; 2/17 bleeding for traumatic injury; 2/17 hemarthrosis; 2/17 hematuria; 1/17 intracranial hemorrhage; 1/17 hematothorax; 1/17 otorrhagia. Five out of 9 women had experienced meno-metrorrhagia. Ten major and 22 minor surgical procedures had been performed. Two spontaneous deliveries and 3 cesarian sections with 5 live births had been reported; moreover, 2 abortions had occurred, 1 spontaneous and 1 voluntary. Globally, 9/17 patients had been transfused with platelets and red blood cells (RBC); 5/17 only with platelets; 2/17 only with RBC. One patient has never been transfused. Platelet transfusions have always been hemostatically effective. All transfused patients have been investigated for anti-HLA and anti-GPIIb/IIIa allo-antibodies,.
Results. Positivity for allo-antibodies could be demonstrated in 4/15 patients (27%): isolated for anti-HLA in 2; isolated for anti-GPIIb/IIIa in 1; combined in 1.
Conclusions. The prevalence of allo-immunization (27%) is inferior to recent literature data (50%). While positivity for anti-HLA (3/15, 20%) agrees with the recently reported data (22%), positivity for anti-GPIIb/IIIa (13%) is inferior (35%). Presence of allo-immunization did not compromise the efficacy of platelet transfusions.
Assessment of Menorrhagia in Females Followed for Bleeding Disorders at the Hemophilia Center of Western New York-Correlation between Clinical Presentation and Laboratory Parameters.
Authors: Bush Eric E. , Kepner James , Smiley Shannon , Belling Linda , Bernstein Zale P. 
Published: April 2020

Abstract
Menorrhagia is a common problem among women. The cause of which, in some individuals, has been attributed to von Willebrand’s disease. The assessment of this condition has been hampered by the lack of easily available and consistent laboratory assessment of levels of this factor that correlate with the patient’s clinical presentation. Therefore we initiated a study to correlate the patient’s self assessment of their menorrhagia with their levels of von Willebrand’s factor (VWF) and platelet function. A peripheral blood assessment chart (PBAC) was utilized for patient self-assessment of blood loss during their menstrual cycle. The PBAC is a validated, well-established method which utilized a point system that quantifies the number and extent of soiling of tampons and pads used during a patient’s menstrual cycle. The PFA-100 has been used as an adjunct for lab assessment of platelet function. This is a high shear-inducing device which simulates primary hemostasis after injury to small vessel. This apparatus consists of a reservoir for whole blood and a small capillary surmounted by a collagen-coated membrane with a central aperture. Platelet agonist which is either epinephrine or ADP is present on the membrane. Closure time is reported as a variable which inversely corresponds with von Willebrand factor levels (or platelet function). Clinical correlation with its results are needed. In addition, blood samples were analyzed for Von Willebrand’s factor antigen (VWFA) and Ristocetin co-factor activity (RCOA) using a CLIA approved laboratory. We explored the relationship between PBAC, PFA-100 and VWFA and RCOA levels. Twenty-six patients were enrolled after obtaining an institutional board approved informed consent. Pearson association estimates and P values to assess the association between one month PBAC scores and PFA-100 showed 0.463 (with a p-value of 0.017). A similar analysis between one month PBAC scores and VWFA and RCOA showed 0.099 (p=0.632) and the association between PFA-100 and VWFA and RCOA showed an inverse relationship of −0.481 (p=0.013). These results confirm a correlation between VWFA and RCOA and assessment of platelet function utilizing a PFA-100. Furthermore, there was a correlation between PBAC scores and the results of the PFA-100. The association between PBAC scores and VWAF screening levels did not show significance. These results suggest further data are necessary to understand the relationship between these variables and further suggests that the PFA-100 may offer a more sensitive assessment of platelet function that correlates with clinical presentation than levels of VWFA or RCOA.
Stability and Sterility of Sucrose-Formulated Recombinant Factor VIII (Kogenate® FS/Bayer) for Use during Continuous Infusion.
Authors: Regan Lisa , Chew Jordan , Vo Khang , Leveque Joyce , Khabbaz Naji , Gorina Eduard 
Published: April 2020

Abstract
Continuous infusion of factor VIII (FVIII) is an alternative treatment approach to bolus dosing for providing hemostatic control in patients with hemophilia A undergoing surgery. Continuous infusion maintains consistent circulating FVIII, thus avoiding wastefully high or dangerously low levels. Potential advantages of continuous infusion include improved protection from excessive bleeding, reduced factor consumption, and easier monitoring. Continuous infusion requires FVIII usage parameters that are typically outside of the tested and approved ranges for the product, including extended time in an aqueous state at room temperature and exposure to a large surface area of tubing. Here we present in vitro data on the stability and sterility of a sucrose-formulated recombinant FVIII product (Kogenate® FS/Bayer; rFVIII-FS) under conditions comparable to those that would be experienced during the continuous infusion of patients. Three lots for each of 3 vial sizes (250, 500, and 1000 IU) were tested. Twenty vials of each size were reconstituted in Water for Injection, pooled into a 50 mL volume (100, 200, or 400 IU/mL for the 250, 500, and 1000 IU vial sizes, respectively), and aseptically transferred to pump reservoirs. Two mini-pumps were evaluated (WalkMed 350 Ambulatory Infusion Pump and CADD-Legacy PLUS Ambulatory Infusion Pump Model 6500) using a flow rate of 0.6 mL/hr. Reservoir bags containing reconstituted rFVIII-FS, pumps, and tubing were incubated at 30°C and samples were withdrawn to test coagulation activity via one-stage and chromogenic assays after 0.5, 1, 2, 3, 6, 12, 24, and 48 hours. Sample remaining in the reservoir after 48 hours was tested for sterility. The reservoirs and tubing used in this study were made from polyvinyl chloride (PVC), which has a high FVIII adsorption potential. Results for all 1000 IU lots based on the recoveries obtained at the first time point demonstrated &gt;80% activity at the end of the study using both the one-stage and chromogenic assays on both pumps. Results for all 500 IU lots showed similar recoveries (≥80%) for both pumps at study end using the chromogenic assay. However, the one-stage assay gave higher recovery values for the WalkMed pump (&gt;90%) compared to the CADD pump (&gt;75%). Results for all 250 IU lots demonstrated ≥79% activity at the end of the study using both pumps and both assays. All lots passed the sterility test. These findings indicate that rFVIII-FS is stable in both the WalkMed and CADD pumps and that sterility of rFVIII-FS is maintained for at least 48 hours. This suggests that rFVIII-FS is suitable for use in continuous infusion.
The Effect of the Order of Draw of Evacuated Blood Collection Tubes on Coagulation Parameters in Normal Volunteers.
Authors: Wakeman Lisa J. , Munro Roger C. , Al-Ismail Saad , Benton Ann , Beddall Andrew , Dorward Nick , Morris Keith 
Published: April 2020

Abstract
Introduction: The recommended order of draw for multiple tube collections (NCCLS [CLSI] H3-A5) clearly indicate that citrate tubes for coagulation tests should be taken before any other tubes (except blood cultures) and that a discard tube should be used if specialised coagulation tests are to be performed. This is to avoid the possibility of tissue activation and the theoretical risk of additive carryover on the parameters being measured. Because of the paucity of published evidence, this study was performed to determine the effect of the order of draw and whether the use of a discard tube is really necessary.
Methods: Three consecutive early morning venous samples were collected into siliconised glass B–D Vacutainers containing tri-sodium citrate (Ref: 367691) from 116 healthy laboratory personnel (F= 74; M = 42) aged 20–63 yrs. Age groups were equally represented. Samples were processed on a Sysmex CA1500 analyser within 1 hour of collection. Appropriate CLSI guidelines were followed throughout. All parameters were measured using Dade-Behring reagents: Activated partial thromboplastin time (APTT) (Actin FSL), prothrombin time (PT) and derived fibrinogen (DF) (Innovin), thrombin clotting time (TCT) (Thromboclotin) and Clauss fibrinogen (CF) (Bovine thrombin and Owren’s veronal buffer). For each parameter, the data from each of the three samples were analysed for significant differences by one way analysis of variance (ANOVA).
Results: Data obtained on measurements of basic coagulation parameters are shown in the table below. SDs are shown in parenthesis. (ns = not significant).
Coagulation Parameter Results and Statistical Analysis Parameter First Sample Second Sample Third Sample ANOVA (p) ns=not significant APTT (secs) 28.3 (1.73) 28.3 (1.73) 27.9 (1.64) 0.230 (ns) PT (secs) 10.9 (0.47) 10.9 (0.47) 10.8 (0.45) 0.368 (ns) TCT (secs) 15.8 (1.03) 15.8 (1.02) 15.7 (1.02) 0.740 (ns) DF (g/L −1) 2.44 (0.54) 2.47 (0.55) 2.48 (0.55) 0.866 (ns) CF (g/L −1) 3.03(0.67) 3.04 (0.67) 3.10 (0.67) 0.825 (ns)
No statistically significant differences were found between the first, second or third samples for any of the measured parameters.
Conclusions: The CLSI recommends an order of draw for evacuated blood collection tubes in order to reduce the possibility of tissue activation in coagulation samples and the theoretical risk of additive carryover on the parameters being measured. Until now, this was based largely on theoretical probability. This comprehensive study demonstrates that the use of a discard tube is probably unnecessary since there is no statistical difference in any of the parameters measured between the first, second or third samples. Although this potentially obviates the expensive use of a discard tube in normal subjects, further work is required to determine whether it is necessary when measuring abnormally prolonged parameters in various pathological states.
Unexpected Harmful Side Effects Triggered by Immune Modulation.
Authors: Sternbach Marion , Dragomir Alina , Crowther Mark A. , Holbrook Ann 
Published: April 2020

Abstract
A 66 year old man was referred urgently to Hematology for severe, spontaneous bruising, bleeding and acute onset anemia. His Past History revealed long standing, poorly controlled rheumatoid arthritis, ischemic cardiac disease with a prior MI, hypertension and elevated Homocystein. Medications included: Leflunomide, Hydroxychloroquine, Ramipril, Bisoprolol, vitamins B12 and B6 and the recently initiated Adalimumab (Humira), an anti-Tumor Necrosis Factor (TNF) immune modulator. On examination he was covered by extensive ecchymoses, large hematomas and had a knee effusion, feeling tired and light headed. He denied any injury. Hemostatic screening showed a normal PT and a markedly prolonged PTT, first 71, later over 100 seconds. A mix of patient and control plasma did not correct the PTT. An inhibitor was detected. Factors XI and IX were normal, Factor VIII was less than 1%. Factor VIII inhibitor was 60 Units Bethesda. White cell and Platelet counts were normal. Hemoglobin was 91 g/l from a previous 120.
He was treated with Prednisone and Cyclophosphamide as well as Hexacapron. Activated Factor VIIa was on standby and was used on one occasion for a huge lower lip hematoma oozing intractably, that required also topical Aprotinin.
Investigations for an underlying lympho-proliferative disorder or solid tumor neoplasia were negative. His complete response to chemotherapy took several months.
Factor VIII inhibitors in non-hemophiliacs are the commonest autoantibodies against clotting factors. Their incidence is 0.2 to 1.0 case per million per year and mostly associated with hematological neoplasias, solid tumors and post-partum. This appears to be first occurrence of Anti-Factor VIII triggered by this type of immune modulation.
Since Monocyte-Macrophages are a major source of TNF, it is possible, that by the interaction of Adalimumab, as Anti-TNF and the TNF receptor, the antigen presenting cell becomes altered, permitting in a patient with underlying autoimmune disease, a monoclonal B-Lymphocyte expansion with anti-Factor VIII properties.
Factor VII Deficiency - A Restrospective Case Review.
Authors: Katz Randi J. , Steinberg Amir , Klafter Robert 
Published: April 2020

Abstract
Factor VII deficiency is a rare autosomal recessive disorder. Its incidence is thought to be 1 in 500,000. Symptoms vary from mild to severe. Factor VII deficient patients usually do not experience bleeding if their level of factor VII is less than 10%. Manifestations are seen with levels less than 5%, and severe bleeding can occur with levels less than 1%. Surgical hemostasis is obtained with levels greater than 25%. Factor VII deficiency is notable for causing a prolonged PT with a normal aPTT, making the diagnosis easy to determine.
This is a descriptive study based on a population of patients from our community based teaching hospital with severe Factor VII deficiency. We reviewed lab data and found any patients with less than a 5% factor deficiency from March 2004- June 2006. In our experience we have noted varying manifestations in moderate factor deficiency patients. We describe four patients with varying symptoms, all with factor levels less than 3%. Their charts were reviewed, and three of the patients were available for a telephone interview.
Patient A is a 75 yo African American female found to have factor deficiency after an increased Prothrombin time (PT) of 31.2 seconds, INR 5.9, and Partial thromboplastin time (PTT) of 34.9 seconds on routine blood work. This patieny’s factor level was found to be 1.86%. Patient A denied any history of bleeding complications, including an uncomplicated tooth extraction. Patient B is a 46 yo African American female with a long history of bleeding complications. Laboratory data revealed a PT of 26 seconds, INR 4.2, PTT 22 seconds and a Factor level of 1%. She experienced years of heavy menstrual periods and nose bleeds. A hysterectomy was performed secondary to bleeding fibroids. She required multiple doses of novoseven as well as FFP. Patient C was diagnosed at the age of 11. Laboratory results revealed a factor level of 1.87 %, and a PT of 25.1 seconds and an INR of 4.1. She has had minor bleeding events, such as gum bleeding. She did receive FFP prior to a foot surgery, and tooth extraction. Patient D is a 46 year old Hispanic man found to have and INR of 4.3 and, factor VII 1.45. Pt had an uncomplicated cholocystectomy.
This is a descriptive study of four different patients with factor VII levels of less than 2%. These four cases demonstrate the wide range of clinical manifestations that factor VII patients may experience. One patient did require novoseven prior to a hysterectomy. Though, as our cases illustrate, some patients do not experience any bleeding manifestations, and therefore, prophylactic plasma may not be required. Furthermore, treatment of factor VII deficient patients needs to be individualized, and guided by personal history.
Coagulation Studies of our Patients Patient Prothrombin Time INR Factor VII A 31.2 5.9 1.86% B 26 4.2 1% C 25.1 4.1 1.87% D 46 4.3 1.45%
Vascularization of the Synovial Membrane in Hemophilic Synovitis.
Authors: McClain Ramona , Hakobyan Narine , Valentino Leonard A. 
Published: April 2020

Abstract
Hemophilia is due to the deficiency of the activity of coagulation factor VIII or IX and results in bleeding. Hemarthrosis frequently leads to synovitis and arthropathy in adolescents and adults with severe disease. The molecular basis for this disorder is only recently being interrogated in detail (Wen, Jabbar et al. 2002; Hakobyan, Kazarian et al. 2004; Valentino, Hakobyan et al. 2004; Hakobyan, Kazarian et al. 2005). In addition to proliferation of synovial fibroblasts, vessels are prominent and likely play a fundamental role. We hypothesized that synovial hyperplasia was limited by the degree of tissue vascularization and a direct relationship between the expansions of these two tissues existed. To test this hypothesis, mouse knee joints were injured by blunt or surgical trauma to induce hemarthrosis or blood was directly injected into the joint capsule to induce synovitis then the knees isolated and the tissues examined by seven reviewers blinded to the experimental conditions for the extent of synovial and vessel hyperplasia (Valentino and Hakobyan 2006, in press) and their individual scores averaged. The distribution of the scores for the 271 sections is given in the table. A direct relationship (r2=0.90, p&lt;0.001) between synovial and vessel hyperplasia was present (figure). It is likely that the synovial membrane can expand up to a point (grade 1) but beyond this, vessel hyperplasia is necessary and that progressive hypertrophy of the synovial membrane is dependent upon the development of vessels to provide oxygen and nutrients. The production of vascular trophic factors by synovial fibroblast cells is under investigation.
Types and numbers of hyperplasia Hyperplasia 0 1 2 3 Synovial 149 67 18 37 Vascular 180 58 19 14 Figure Figure
Characterization of PEGylated Factor VIII Molecules.
Authors: Severs Joanne , Samuels Nicole , Teare John , Tollner Amy , Regan Lisa 
Published: April 2020

Abstract
Patients with hemophilia A typically require frequent treatment with replacement Factor VIII (FVIII) products due to the relatively short circulating half-life of FVIII. Increasing the half-life of FVIII would profound impact the lives of patients with hemophilia A, especially children, by reducing the frequency of infusions. The addition of polyethylene glutamate (PEG) to therapeutic molecules or their carriers has been demonstrated to increase the survival of such molecules in the circulation. To create a longer circulating product, we have developed novel FVIII molecules using site specific PEGylation techniques. We present a characterization of these PEGylated FVIII molecules in terms of the site of PEGylation, the binding of von Willebrand Factor (vWF), and the effect of PEGylation on thrombin digestion, and we compare the one-stage and chromogenic potency test results. The site of PEGylation was determined using MALDI-mass spectrometry. A vWF binding assay was developed using surface plasmon resonance technology (Biacore). The effect of PEGylation on thrombin digestion of the novel constructs was evaluated using MALDI-mass spectrometry, thrombin activation, and potency decay curves following thrombin activation. All novel constructs were PEGylated on the expected target site based on the site of introduced mutation. PEGylation of FVIII appeared to have a slight reduction on vWF-binding capability. The site of PEGylation reduced the ability of thrombin to digest FVIII, as assayed by MALDI-mass spectrometry analysis and by thrombin digestion and decay curves. The size and position of the PEG molecule influenced the extent of FVIII thrombin sensitivity. The one-stage potency assay was more sensitive to the size and position of the attached PEG molecule than the chromogenic potency assay. We report that some PEGylated FVIII molecules are less sensitive to thrombin activation than unmodified FVIII, and that the size and position of the PEG molecule can significantly affect the potency of FVIII. The findings of this study suggest that site-directed PEGylation of FVIII may be a useful technique for the development of FVIII molecules with extended activity in circulation and that careful choice of size and position of the PEG molecule is critical.