bluuebunny/crossref_metadata_2025_split huggingface dataset:
Production of Monoclonal Antibodies Against ADAMTS13 Determining Its Expression in Different Human Tissues.
Authors: Gao Weiqiang , Su Jiang , Bai Xia , Shen Fei , Ruan Changgeng 
Published: October 2019

Abstract
von Willebrand factor-cleaving protease/ADAMTS13 is a plasma metalloproteinase that degrades unusually large von Willebrand factor multimers (UL-vWF) derived from endothelial cells (ECs) and megakaryocytes (MCs) into small peptides circulating in blood. It is documented that transcript mRNAs of the protease are present in many human tissues; however, the protein expression of ADAMTS13 remains to be elucidated. In the present work, the gene of metalloproteinase domain of human ADAMTS13 was cloned into the multiclone site of pET28a(+). After induced by IPTG, the recombinant protein was purified using a Ni-NTA column and the Bal b/c mice were immunized with the protein. Screened with ELISA, three monoclonal antibodies against the metalloproteinase domain of ADAMTS13 were obtained and two of them, SZ-112 and SZ-113, were further evaluated. Both of them belonged to IgG1 subclass. The quantity of them in ascites were 4 mg/ml, and their titers were as high as 1×10−5. The data of competitive ELISA showed that SZ-112 and SZ-113 recognized different epitopes of the recombinant protein. Western blot results demonstrated that SZ-112 not only reacted against the recombinant protein, but also recognized the full-length recombinant ADAMTS13 protein that expressed in CHO cell line (the vectors containing the ADAMTS13 cDNA sequences were provided by Prof. Sadler JE). The immunoprecipitation results showed that the two antibodies could react to an approximately 200 KDa protein in platelet lysate. Then, the expression panels of ADAMTS13 in human normal tissues were investigated using immunohistochemistry with the monoclonal antibodies. And the protease was found to be present in many kinds of tissues such as liver, spleen, ovary, prostate, bladder, small intestine, thyroid and thymus with significantly positive staining. The protease was also present in lung, uterus, large intestine and heart but stained weakly. We did not found the protease in brain. In most of these organs, the protease was expressed in epithelium of the tissues. While in liver, spleen and thymus, it was mainly presented in a subgroup of the solid tissue cells. Moreover, the preliminary results showed that the expression of ADAMTS13 slightly decreased in liver tissues of patients suffering form hepatitis type B and cirrhosis. In conclusion, our data indicated that two novel monoclonal antibodies against the metalloproteinase domain of human ADAMTS13 were successfully prepared, and the expression of ADAMTS13 in different tissue and specific locality might be associated with the regulation and function of the protease, which would contribute to the further research of the deficiency mechanism of the proteases in some disorders.
Relevance of Thrombophilic Risk Factors on Clinical Phenotype in Patients with Haemophilia A.
Authors: Krause Manuela , Hahn Anika , Peyvandi Ferrial , Scharrer Inge 
Published: October 2019

Abstract
Introduction: By assuming the hypothesis, that a high coinheritance of genetic haemostatic abnormalities influences the clinical phenotype in patients with coagulation disorders, we analysed the bleeding tendency in adult patients with haemophilia A.
Patients: A total of 49 patients with haemophilia A (severe:20 patients, moderate:12 patients, mild:17 patients; median age:44 years, range:22–78 years) and 80 sex-matched healthy subjects (median age:26 years, range:16–58 years) were studied in our haemophilia treatment center. Two of this patients have an inhibitor against factor VIII. 10 of 49 patients receive the factor VIII replacement therapy on demand. In addition to factor VIII activity, FV G1691A mutation and the FII G20210A variant were investigated.
Results: In our study the prevalence of FV G1691A was significantly higher among patients 6/49 (severe:5, moderate:1) than among control subjects 2/80 [12% vs 2.5%; p=0.03 OR 4.9]. We found no differences in the FII G20210A variant in 2/49 patient (moderate:1, mild:1) as compared to 2/80 among controls [4% vs 2.5%; p=0.62 OR 1.63 ]. In none of our patients both defects together were identified.
Within the last year the onset of bleeding was not different in thrombophilic haemophilia patients (n=8) 64 bleeding episodes/year compared with non-carriers (n=41) 284 bleeding episodes/year [8 vs 7 bleeding episodes/year/patient; p=0.72; OR 1.15].
Conclusion: The FV G1691A was found significantly more frequent in the thrombophilic haemophiliacs than in the general population. In our smal study group we could not demonstrate a relevant influence of the FV G1691A and FII G20210A on bleeding tendency. Further studies are needed to confirm whether FV G1691A and FII G20210A plays a role on the influence of bleeding tendency in haemophilia A patients.
von Willebrand Disease in Children with Glycogen Storage Disease Type Ia.
Authors: Yilmaz Aygen , Artan Reha , Akcam Mustafa , Kupesiz Alphan , Yesilipek M. Akif 
Published: October 2019

Abstract
Glycogen storage disease, Type Ia (GSD-Ia) is an autosomal recessive disease, characterized by hepatomegaly, hypoglycemia, hyperuricemia, and bleeding diathesis. Easy bruising and epistaxis are common and are associated with a prolonged bleeding time as a result of impaired platelet aggregation and adhesion. Two of four of our GSD-Ia patients were evaluated because of bleeding diathesis and both were diagnosed as von Willebrand’s disease (vWd) with normal platelet count, prothrombin time and activated partial thromboplastin time.
Case 1: Six year old boy diagnosed as GSD-Ia when he was 3 month of age and bleeding diathesis was first recognized 5th year of his life. His bleeding time was prolonged (15 minutes). Measurement of serum factor VIII-related antigen (vWf-Ag) revealed a decrease (% 31) and ristocetin cofactor activity was abnormal (% 37).
Case 2: Nine year old girl diagnosed as GSD-Ia when he was 6 month of age and bleeding diathesis was first recognized 4th year of his life. His bleeding time was prolonged (3.5 minutes). Measurement of serum factor VIII-related antigen (vWf-Ag) revealed a decrease (% 17) and ristocetin cofactor activity was abnormal (% 5.7).
Frequent diagnosis of vWd among our GSD-Ia patients might suggest alterations in membrane glycoprotein synthesis. There is no definitive explanation addresses how these alterations actually cause defective platelet aggregation. Although, the presence of association between these two separate diseases might be coincidental, in literature, diagnosis of vWd among GSD-Ia patients had been reported in only one paper. We conclude that GSD-Ia patients may have a metabolically acquired form of vWd as well as an acquired intrinsic platelet defect. Further molecular studies may be necessary to clarify the etiopathogenesis of this association.
Differential Immunodominance of Human and Porcine Factor VIII in Hemophilia A Mice.
Authors: Healey John F. , Parker Ernest T. , Barrow Rachel T. , Lollar Pete 
Published: October 2019

Abstract
Hemophilia A inhibitor patients and patients with acquired hemophilia A recognize immunodominant epitopes in the A2 and C2 domains of human factor VIII (fVIII). Hemophilia A mice also recognize A2 and C2 domain epitopes when immunized with human fVIII using a dosing schedule that mimics clinical use. We compared the immune responses of hemophilia A mice to human and porcine fVIII using a domain specific ELISA. In this assay, monoclonal antibodies are tested against a panel of six single human fVIII domain hybrid human/porcine fVIII molecules that contain the human A1, A2, ap, A3, C1 or C2 domains. With anti-human antibodies, a positive signal with one of the single human domain proteins identifies domain specificity, whereas loss of signal indicates domain specificity of anti-porcine fVIII antibodies. Exon16 (E16) - disrupted hemophilia A mice (n = 3) received six weekly μ10 g/kg intravenous injections of recombinant B-domain deleted human fVIII and a final 25 μg/kg boost. To obtain comparable inhibitor titers, E16 mice (n = 3) received six weekly injections of μ40 g/kg of recombinant B-domain deleted porcine fVIII. Spleens from high titer mice were fused with NS1 mouse myeloma cells and 485 of the resulting hybridomas were analyzed for fVIII domain specificity (Table). Only two hybridomas secreted antibodies specific for the ap domain. Human fVIII elicited a significantly greater number of antibodies to the A2 domain, whereas porcine fVIII elicited a significantly greater number of antibodies to the A1 and A3 domains (p < 0.01, chi square test). The greater number of anti-C2 antibodies to human fVIII was not significant at the 95% confidence level (p = 0.08). The differential immunodominance of human and porcine fVIII epitopes suggests that it may be possible to design a recombinant hybrid human/porcine fVIII molecule that is less immunogenic than human fVIII in the treatment of patients with hemophilia A.
Domain Specificity of Anti-FVIII MAbs
Mouse ID: Immunogen No. of MAbs A1 A2 A3 C1 C2 CR & MD CR: Cross Reactive MD: Multidomain 1- Human fVIII 95 2 16 2 7 21 23 & 24 2- Human fVIII 126 13 23 1 2 27 39 & 21 3- Human fVIII 54 1 15 2 1 10 9 & 15 4- Porcine fVIII 123 39 7 19 8 16 33 & 0 5- Porcine fVIII 27 13 5 0 0 4 2 & 3 6- Porcine fVIII 60 9 6 12 1 9 13 & 10
Portal Vein Thrombosis a Common Complication Following Splenectomy in Adults Patients with CLL or Myeloproliferative Disease.
Authors: Svensson Magnus , Wiren Mikael , Kimby Eva , Hagglund Hans 
Published: October 2019

Abstract
Elective laparoscopic splenectomy (LS) is performed with increasing frequency in relation to open splenectomy (OS). The advantage with LS is reduced morbidity. Moreover, this method is feasible also in patients with splenomegaly, a patient group with more postoperative complications as bleeding, infections and portal vein thrombosis. Portal vein thrombosis (PVT) is a rare but serious complication of splenectomy. We retrospectively reviewed the medical records of 69 consecutive patients splenectomized for haematological diseases during a five-years period (Jan 1999 to Dec 2003) at the Dep. of Surgery Huddinge University Hospital, with the aim of comparing the results and complications after LS and OS, with focus on thromboembolic events. The follow-up for all patients was performed Jan 31. 2004. Thirty-nine patients underwent LS and 30 OS. There were three conversions (7.7%) from laparoscopic to open surgery due to bleeding and splenomegaly. Accessory spleens were found in 16 of 69 (23%) patients, 6 of 39 (15%) in LS and 10 of 30 (33%) in OS. Thromboembolic complications were seen in seven patients; a) deep vein thromboses in the lower leg in two patients with ITP, both with LS and neither had received thromboprophylaxis, b) PVT in five patients, one after LS and four after OS; three with CLL, (two with a concomitant haemolytic anaemia) and two with a myeloproliferative disease. The five patients with PVT had all splenomegaly and had received thromboprophylaxis with low-molecular-weight heparin. PVT was diagnosed from day 6 to day 111 after splenectomy. Three of the five patients had thrombocytosis, 478, 740 and 1459x 10(9) at the time of PVT-diagnosis. In all the splenectomized patients, two patients had overwhelming post splenectomy sepsis (OPSI). One patient with Evans syndrom died of E. coli sepsis four months after splenectomy and one patient with myelofibrosis had severe pneumococksepsis 44 months after splenectomy. Both had recieved preoperative pneumocock vaccination. Further seven patients died during the follow-up period, five due to infections and bleedings, in all related to progressive malignant lymphoma. One patient died of sudden cardiac arrest 15 months postoperative, and one patient performed suicide. Conclusions: Portal vein thrombosis after splenectomy was seen in 13.5 % of patients with haematological malignancies despite thromboprophylaxis. Patients with splenomegaly and myeloproliferative disease or CLL with haemolytic anaemia have high risk of PVT. We recommend careful observation of postoperative thrombocytosis and extended thromboprophylaxis. Ultrasonography or CT should be considered in all patients with abdominal symtoms after splenectomy. Patients should receive pneumocockvaccination and be informed of the lifelong risk of severe infections.
Lymhoma of the Oral Cavity. A Rare Entity with Favorable Prognosis. Ten-Year Experience of a Single Hematology Unit.
Authors: Siakantaris Marina P. , Tzenou Tatiana , Vassilakopoulos Theodoros P. , Kyrtsonis Marie-Christine , Angelopoulou Maria K. , Dimopoulou Maria N. , Kokoris Styliani I. , Dimitriadou Evangelia N. , Korkolopoulou Penelope , Patsouris Efstratios , Tsaftaridis Panayiotis , Variamis Eleni , Laskaris George , Pangalis Gerassimos A. 
Published: October 2019

Abstract
Lymphomas of the oral cavity consist <5% of the malignancies of the region. B-cell lymphoma of the diffuse large cell histology is the predominant type (B-DLCL). The purpose of our study was to describe the clinicopathologic characteristics of a group of patients with oral cavity lymphomas and their treatment outcome. We reviewed the records of 46 patients (21 males and 25 females) with oral lymphomas who were diagnosed and treated in our Service, between 1993 and 2003. The mean age was 57 years and the most prevalent site of involvement was the tonsil (41.3%) followed by the tongue (15.2%), while 8.7% had >1 involved sites. Thirty six patients (pts) (78.3%) had clinical stage I or II and 10 (21.7%) advanced stage disease. A monoclonal lymphocytic population was detected in the blood of 7 pts (15.2%) by PCR (IgH or TCR rearrangement), 4 of them with early stage lymphoma. 56.5% of the lesions were diagnosed as B-DLCL and only 10.8% as peripheral T-cell lymphomas.
Nineteen pts with B-DLCL received CHOP or equivalent chemotherapy regimens and 11 CHOP combined with anti-CD20. Chlorambucil was administered in 4 pts with MALT lymphoma and local radiotherapy as the sole treatment in 1. The pts with T-cell lymphoma received CHOP (3) or interferon alfa-2b (2). Thirty eight pts are in complete remission. There were 3 deaths due to the disease (1 MALT, 1 predominantly small cleaved cell and 1 B-DLCL). Three additional pts relapsed and achieved complete response with a second chemotherapy course and 2 pts developed a lymphoma of different histology. In conclusion, oral lymphoma is a rare disease mainly of a diffuse large B-cell histology and a prolonged survival, when treated with chemotherapy (10-year Event Free Survival 82.5%).
Radiolabeled IL-2-Human Serum Albumin Fusion Protein (Albuleukin™) as a Potential New Reagent for Radioimmunoimaging / Therapy of Hodgkin’s Lymphoma.
Authors: Staak Jan O. , Colcher David , Wang Jianyi , Sung Cynthia , Raubitschek Andrew A. 
Published: October 2019

Abstract
Objectives:
CD25 (IL-2Rα) is overexpressed on the cell surface by a variety of malignant diseases including Hodgkin’s lymphoma (HL) and its surrounding lymphoid infitrate. A soluble form of this receptor (sCD25) may limit a targeting agent’s ability to bind to the tumor cells which may result in decreased therapeutic efficacy. Albuleukin is a fusion protein of recombinant native IL-2 and human serum albumin with a molecular mass of 82kD. Albuleukin possesses the immunomodulatory characteristics of IL-2 with a significantly extended circulatory half-life and better localization in lymphocyte rich tissues than IL-2. We investigated the targeting and pharmacokinetic properties of Albuleukin in a human CD25+ HL xenograft model as a potential novel radioimmunodiagnostic/-therapeutic agent.
Materials & Methods:
Albuleukin was conjugated with a DOTA bifunctional chelating agent. DOTA-Albuleukin was radiolabeled with 111In (specific activity 3μCi/μg; 91–100% labeling efficiencies). Human HL (L540) tumor bearing nude mice were injected i.v. with 111In-Albuleukin (5μCi) with and without unlabeled humanized anti-CD25 (daclizumab; 5μg 4h prior) to investigate its potential impact on pharmacokinetics and tumor targeting. Mice were sacrificed at various time points and tumors, blood and major organs were collected and analyzed for radioactivity. Serum samples were analyzed by size exclusion chromatography (SEC-) HPLC. Serial whole body γ-imaging studies (15μCi) were also performed.
Results:
Biodistribution and imaging studies of 111In-Albuleukin exhibited rapid blood clearance (4%ID/g at 24h), specific tumor localization (18%ID/g at 24h) and favorable tumor:blood ratios (22:1 at 48h). Adding anti-CD25 MAb did not alter the biodistribution profile or targeting of Albuleukin. No breakdown products of Albuleukin or formation of complexes between Albuleukin and sCD25 (with or without anti-CD25) were detected by SEC-HPLC in the mouse serum.
Conclusions:
Unlike small MAb fragments of comparable size, Albuleukin did not accumulate in metabolizing organs such as liver and kidneys. The low affinity of Albuleukin to sCD25 may explain the absence of immunocomplexes, as we have observed with the radiolabeled anti-CD25 alone, and unaltered biodistribution when unlabeled anti-CD25 was added. The good tumor targeting of the Albuleukin may be due to the involvement of the cell surface high affinity receptor including beta- and gamma-chains. The rapid blood clearance in concert with low normal organ uptake and good tumor targeting as well as its presumed low immunogenicity in humans makes 111In-Albuleukin a potential new reagent for radioimmunoimaging, and possibly for treatment of CD25+ lymphoma utilizing 90Y.
High Frequency of Aberrant Phenotype in Multiple Myeloma Patients Detected by Multiparametric Flow Cytometry.
Authors: Calheiros Luiz Arthur , Kimura Eliza Y.S. , Almeida Manuella S.S. , Chauffaille Maria de Lourdes L.F. , Bigonha Jandey G. , Sandes Alex F. , Barreto Wolney G. , Escaramuzi Carolina L. , Colleoni Gisele W.B. , Yamamoto Mihoko 
Published: October 2019

Abstract
Multiple Myeloma (MM) is a B cell lymphoproliferative disease with clonal plasma cell accumulation in bone marrow. Multiparametric flow cytometry (MFC) is an usefull tool to distinguish MM cells from normal plasma cells. Normal plasma cells are characterized by the expression of CD19+, CD45++, CD38++, CD138++, cytoplasmic immunoglobulin light chains (κ and λ) and CD56- while most MM plasma cells lose CD19, CD45 and gain CD56. In addition, many other antigens may be expressed by myeloma cells such as myeloid or lymphoid lineage associated antigens and these abnormal antigen expression is known as aberrant phenotype (AP). We studied 29 MM patients at diagnosis, in attempt to evaluate AP, it’s frequency and relation to prognostic parameters. The following monoclonal antibodies were used: CD45, CD38, CD138, CD56, CD19, CD20, CD22, CD10, CD13, CD14, CD33, CD117, CD28 and CD40, conjugated to FITC, PE, PerCP and APC) and acquisition / analysis were done through flow cytometer (FACS calibur, BD, San Jose) using CELL QUEST software (BD). Plasma cells were identified by the expression of CD38, CD138 and CD45 and the monoclonality confirmed by immunoglobulin light chain restriction. Our results showed presence of at least 2 AP in all cases : 2 AP (7 patients), 3 AP (12 patients), 4 AP( 5 cases), 5 AP (4 cases) and 8 AP in one case. The most frequent APs were CD45−, CD56+, CD117+, CD13+, CD33+, and were observed in 88% of patients. The most frequent AP association was CD56+/CD45− (40%), followed by myeloid antigen associated phenotypes (CD117, CD33, CD13). The lymphoid antigens expression was more observed in patients with large number of AP (>4 AP). CD56- patients presented serum β2-microglobulin and ionic calcium labeling levels higher than CD56+ patients (p=0,02) showing the usefulness of this antigen as prognostic marker. Morphological analysis showed that the majority (55%) of plasmablastic cases expressed >2 myeloid antigens against 18% of mature plasma cell morphology cases. These results allow us to conclude that MM express high frequency of AP, highlighting the importance of CD56 as a prognostic factor. MFC may be useful to the immunological detection of minimal residual disease in a great majority of MM patients and we suggest the panel CD45, CD56, CD117, CD33 and CD13 for this purpose, in addition to CD38 and CD138.
Selective Inhibition of JAK2 Driven Erythroid Differentiation of Polycythemia Vera Progenitors
Authors: Geron Ifat , Abrahamsson Annelie , Barroga Charlene , Kavelerchik Edward , Gotlib Jason R. , Hood John D. , Durocher Jeffrey , Mak Chi Ching , Noronha Glen , Soll Richard , Tefferi Ayalew , Kaushansky Kenneth , Jamieson Catriona H.M. 
Published: October 2019

Abstract
Myeloproliferative disorders (MPDs), diagnosed in 15,000 individuals annually, are characterized by overproduction of lineage-committed blood cells, thrombotic events, occasionally marrow fibrosis or progression to acute myelogenous leukemia and frequently a point mutation (V617F) in the JAK2 kinase. Mutant JAK2 expression results in erythrocyte overproduction in almost all cases of polycythemia Vera (PV). We investigated the molecular mechanisms driving erythroid skewed differentiation and the capacity of a selective JAK2 inhibitor (TG101348) to normalize PV progenitor differentiation. Both JAK2 V617F+ transduced cord blood and PV progenitor erythroid colony formation was potently inhibited by TG101348 (300 nM). Bioluminescent immunocompromised mouse transplantation studies revealed that TG101348 (150 mg/kg) treatment significantly reduced human erythroid engraftment by both PV and JAK2 V617F+ cord blood progenitors. Moreover, the imbalance between GATA-1 and PU.1 transcripts typical of JAK2 V617F+ progenitors normalized after treatment with TG101348. In addition, TG101348 inhibited both JAK2-mediated STAT5 phosphorylation and AKT-regulated GATA-1 phosphorylation in an erythropoietin responsive cell line suggesting a dual mechanism for the erythroid inhibitory effects of TG101348. Because of selective inhibition of JAK2 V617F+ progenitors both in vitro and in vivo, TG101348 may be an effective molecularly targeted inhibitor of JAK2 V617F+ driven MPDs in clinical trials.