bluuebunny/crossref_metadata_2025_split huggingface dataset:
Paroxysmal Nocturnal Hemoglobinuria With Budd-Chiari Syndrome Treated With Complement Inhibitor Eculizumab; A Case Report
Authors: Valeri Federica , Borchiellini Alessandra , Schinco Piercarla , Boccadoro Mario 
Published: October 2019

Introduction
Paroxysmal nocturnal hemoglobinuria (PNH) is a rare, acquired haemolytic anaemia caused by somatic mutation in the phosphatidylinositol glycan-complementation class A gene, resulting in absence of a key complement regulatory protein, CD59.
Thrombosis occurs in up to 40% of PNH patients; it usually involves abdominal and cerebral veins and it is the leading cause of death disease related.

Methods
We describe the response to Eculizumab (Soliris, Alexion) in 28 years old male with PNH diagnosed as a consequence of Budd Chiari Syndrome, acute liver dysfunction, mild haemolytic anaemia and thrombocytopenia.

Results
The patient was admitted to the gastroenterology department with acute abdominal pain, fatigue, hemolytic anaemia, thrombocytopenia and transaminitis. Abdominal doppler ultrasonography (US) was immediately performed with detection of Budd Chiari Syndrome, portal vein thrombosis, initial portal hypertension and ascites.
He was started on low dose low molecular weight heparin (platelets < 40x10^9/L), but despite anticoagulation progressive liver damage occurred, with poor pain control and worsening ascites. At the same time, we observed rapid exacerbation of thrombocytopenia and increasing in hemolysis tests with lactate dehydrogenase (LDH) reaching 1766 U/L, unresponsive to steroids administration. Bone marrow biopsy was negative but peripheral blood flow cytometry characterized a large PNH clone (85% total red blood cells). Furthermore, liver biopsy identified advanced stage of idiopathic cirrosis.
Eculizumab therapy was then initiated at a dose of 600 mg weekly for 4 weeks and then 900 mg every 14 days. During the first month, transaminases progressively normalized and platelets settled permanently above 40x10^9/L, allowing therapeutic dose of anticoagulation. LDH dropped from basal value of >1000U/L to 600U/L and progressive reduction in abdominal pain was observed.
Recanalization of portal vein thrombosis was found out at the US doppler after 6 weeks of anticoagulation, but recanalization of sovraepatic veins was not yet detectable.

Conclusions
Currently, after 17 Eculizumab administrations, platelets are 44 x 10^9/L, Hb 11.9 g/dl, AST 26 mg/dl, ALT 55 mg/dl, GGT 123 mg/dl, LDH 518 U/L. No further thrombotic episodes occurred, no ascites was detected as well as portal hypertension signs, performing ultrasonography monitoring.
This case shows that Eculizumab can block intravascular haemolysis and platelet consumption and can improve hepatic failure, allowing full dose of anticoagulant as therapy for current thrombosis or as prophylaxis for future events.

Disclosures:
No relevant conflicts of interest to declare.
Modified Heparin Assay For The Quantitation Of Rivaroxaban
Authors: Sanfelippo Michael , Tillema Veronica 
Published: October 2019

The anticoagulant rivaroxaban is not routinely monitored due to its predictable anticoagulant action. However, drug levels are clinically useful in patients requiring surgery as well as in assessing compliance. We have modified a commercial heparin assay based on inhibition of activated factor X (Xa) that was previously modified for the assay of fondaparinux (Am J Clin Pathol 2009, 132:608-612) by replacing the fondaparinux calibrators with commercial rivaroxaban calibration plasmas. Technoview rivaroxaban calibrator plasmas were purchased from Dia Pharma (West Chester, Ohio). These calibrators were substituted for the fondaparinux calibrators. The assay was run on a Siemens BCS XP, as previously described. The assay was linear in the range of 0 to 650 ng/ml. The limit of detection was 14 ng/ml, and carryover at 285 ng/ml was < 1%. The intra-assay precision at 100 ng/ml was standard deviation of 1.04 ng/ml, coefficient of variation of 1.03%. The inter-assay precision at 105 ng/ml was standard deviation 3.5 ng/ml, coefficient of variation of 3.3%. The assay is unaffected by warfarin, but can be affected by other Xa inhibitors such as heparin, low molecular weight heparin, fondaparinux, and apixaban.

Disclosures:
No relevant conflicts of interest to declare.
Specific recognition of human leukemic cells by allogeneic T cells: II. Evidence for HLA-D restricted determinants on leukemic cells that are crossreactive with determinants present on unrelated nonleukemic cells
Authors: Sosman JA , Oettel KR , Smith SD , Hank JA , Fisch P , Sondel PM 
Published: October 2019

Abstract
Transplantation of immunocompetent cells present within allogeneic bone marrow has been associated with the elimination of residual host leukemia, both in animal tumor models and in patients receiving marrow transplants for leukemia. This observation has been called the “graft- versus-leukemia effect.” We have attempted to study this phenomenon in vitro by characterizing the cytolytic response of T cells from normal donors after in vitro activation with allogeneic leukemic cells. As expected, most T cells that react against an allogeneic patient's leukemic cells recognize their foreign HLA antigens and lyse the patient's nonleukemic remission lymphoid cells. In addition, we have shown that a small fraction of the T cells recognize and lyse foreign leukemic targets without lysis of nonmalignant remission targets from the same leukemic patient. These T cells have been isolated and characterized as CD3+, CD4+ cells expressing the alpha/beta T cell receptor (TCR). Their lysis appears to reflect specific antigen recognition mediated via the CD3-TCR complex and interactions involving the CD4 receptor. Some of these “leukemic specific” T cell lines, which are restricted by HLA class II molecules, can also lyse occasional nonleukemic cells from certain unrelated donors. This recognition appears to involve crossreactive determinants shared by the leukemic cells and the unrelated allogeneic nonleukemic cells. These specific interactions may represent an in vitro model of the graft-versus- leukemia effect.
Synergistic effect of recombinant interferon-gamma and interleukin-3 on the growth of immature human hematopoietic progenitors
Authors: Kawano Y , Takaue Y , Hirao A , Abe T , Saito S , Matsunaga K , Watanabe T , Hirose M , Ninomiya T , Kuroda Y 
Published: October 2019

Abstract
Purified peripheral blood hematopoietic progenitors from children in early remission from cancer respond to recombinant human interleukin-3 (IL-3), but not to granulocyte colony-stimulating factor (G-CSF). With these purified cells as a target, we studied the effect of recombinant human interferon-gamma (IFN-gamma) on progenitor growth, using both liquid-suspension limiting dilution assay (LDA) and regular methylcellulose culture of progenitors. We found that in LDA with IL-3, IFN-gamma directly stimulated the growth of blood progenitors in a dose- dependent manner with single-hit kinetics, whereas IFN-gamma suppressed the growth of G-CSF-supported progenitors obtained from bone marrow. The stimulatory effect was also observed in methylcellulose culture, but the addition of antibodies for G-CSF, granulocyte-macrophage CSF, IL-1 alpha, IL-1 beta, IL-6, or tumor necrosis factor did not result in a decrease of the colony number, supporting further the possible direct effect of IFN-gamma on progenitor growth. These results suggest that the inhibitory effect of IFN-gamma on hematopoietic progenitors is limited to those in an advanced stage of maturation. IFN-gamma may be one of the essential lymphokines upregulating the growth of human hematopoietic progenitor cells.
Consistent involvement of the bcr gene by 9;22 breakpoints in pediatric acute leukemias
Authors: Suryanarayan K , Hunger SP , Kohler S , Carroll AJ , Crist W , Link MP , Cleary ML 
Published: October 2019

Abstract
To investigate the relationship of bcr-abl fusion mRNAs with childhood acute lymphoblastic leukemias (ALL), we examined 27 pediatric Philadelphia chromosome (Ph1)-positive acute leukemias using a reverse polymerase chain reaction (PCR) procedure. In cells from 24 leukemias, single bcr-abl PCR products were detected that corresponded to breakpoints in the minor breakpoint cluster region (mbcr in intron 1 of the bcr gene) associated with production of the P190 fusion protein. Cells from the three remaining leukemias contained breakpoints in the major breakpoint cluster region (Mbcr) as shown by PCR and Southern blot analyses. These three leukemias also contained low levels of the mbcr PCR product that may have resulted from alternative splicing of the bcr-abl precursor RNA. A screen of 35 additional leukemias from patients who failed therapy before day 180 (induction failures or early relapses) found one case with unsuccessful cytogenetics to express Mbcr- abl RNA. All four children with Mbcr breakpoints had white blood cell levels in excess of 250,000 at presentation (compared with 2 of 24 with mbcr breakpoints) and two had hematologic and clinical features suggestive of chronic myelogenous leukemias (CML) in lymphoid blast crisis. Our results indicate that in Ph1-positive pediatric leukemias, all 9;22 breakpoints occur in one of the two known breakpoint cluster regions in the bcr gene on chromosome 22. The reverse PCR reliably detected all patients with cytogenetic t(9;22) and is capable of detecting additional Ph1-positive leukemias that are missed by standard cytogenetics. Furthermore, the Mbcr-type breakpoint, associated with production of p210, can be seen in childhood leukemias presenting either as clinical ALL or as apparent lymphoid blast crisis of CML, suggesting that t(9;22) breakpoint locations do not exclusively determine the biologic and clinical features of pediatric Ph1-positive ALL.
Use of polymerase chain reactions to monitor minimal residual disease in acute lymphoblastic leukemia patients
Authors: Yokota S , Hansen-Hagge TE , Ludwig WD , Reiter A , Raghavachar A , Kleihauer E , Bartram CR 
Published: October 2019

Abstract
T-cell receptor (TCR) delta gene rearrangements are observed in more than 80% of acute lymphoblastic leukemia (ALL) patients. Moreover, a preferential usage of specific genetic elements has been shown in different ALL subtypes: V delta 1 DJ delta 1 rearrangements predominate in T-ALL, while most B-precursor ALLs show a recombination of V delta 2 to D delta 3. Recently we have proposed a strategy for the detection of minimal residual disease (MRD) based on the isolation of clonospecific probes following the in vitro amplification of V delta 1 DJ delta 1 junctions by polymerase chain reaction (PCR) and now have adapted this method to the preparation of specific V delta 2 D delta 3 fragments. In the present study, clonospecific probes were generated from 11 T-ALL and 16 cALL patients (21 children, 6 adults). The sensitivity of these 27 probes in detecting residual leukemia cells varied between 10(-4) to 10(-6) as determined by semiquantitative evaluation of dilution experiments. PCR analysis of 55 bone marrow (BM) and peripheral blood (PB) samples obtained from the 27 patients during complete clinical remission showed the following results: (1) Evidence for MRD was obtained in the BM of all patients (eight of eight) investigated 2 to 6 months after remission induction and also in 6 of 11 cases on maintenance therapy 7 to 19 months after diagnosis. (2) In contrast, all patients but one (10 of 11) analyzed 6 to 41 months after the termination of treatment lacked apparent evidence for leukemia DNA; the exception was a girl exhibiting 10(-4) to 10(-5) residual cells in her PB 5.5 years after diagnosis. (3) Longitudinal analysis in nine patients disclosed marked individual differences in the intervals between achievement of clinical remission and complete eradication of the leukemia cell clone. (4) Differences in the duration of MRD were not associated with distinct clinical-hematologic features. (5) Detection of residual disease by PCR proceeded clinical relapse in two cases.
Identification and characterization of a low-affinity granulocyte- macrophage colony-stimulating factor receptor on primary and cultured human melanoma cells
Authors: Baldwin GC , Golde DW , Widhopf GF , Economou J , Gasson JC 
Published: October 2019

Abstract
Hematopoietic growth factor receptors are present on cells of normal nonhematopoietic tissues such as endothelium and placenta. We previously demonstrated functional human granulocyte-macrophage colony- stimulating factor (GM-CSF) receptors on small cell carcinoma of the lung cell lines, and others have reported that certain solid tumor cell lines respond to GM-CSF in clonogenic assays. In the current study, we examine human melanoma cell lines and fresh specimens of melanoma to determine whether they have functional GM-CSF receptors. Scatchard analyses of 125I-GM-CSF equilibrium binding to melanoma cell lines showed a mean of 542 +/- 67 sites per cell with a kd of 0.72 +/- 0.14 nmol/L. Cross-linking studies in the melanoma cell line, M14, showed a major GM-CSF receptor species of 84,000 daltons. Under the conditions tested, the M14 cells did not have a proliferative response to GM-CSF in vitro, nor was any induction of primary response genes detected by Northern analysis in response to GM-CSF. Studies to determine internal translocation of the receptor-ligand complex indicated less than 10% of the 125I-GM-CSF internalized was specifically bound to receptors. Primary melanoma cells from five surgical specimens had GM-CSF receptors; Scatchard analysis was performed on one sample, showing 555 sites/cell with a kd of 0.23 nmol/L. These results indicate that human tumor cells may express a low-affinity GM-CSF receptor protein that localizes to the cell surface and binds ligand, but lacks functional components or accessory factors needed to transduce a signal.
Examination of the roles of glycoprotein Ib and glycoprotein IIb/IIIa in platelet deposition on an artificial surface using clinical antiplatelet agents and monoclonal antibody blockade
Authors: Sheppeck RA , Bentz M , Dickson C , Hribar S , White J , Janosky J , Berceli SA , Borovetz HS , Johnson PC 
Published: October 2019

Abstract
The mechanism of platelet thrombus growth on an artificial surface is incompletely understood. While glycoprotein (GP)Ib and GPIIb/IIIa are required for normal attachment and thrombus formation on subendothelium, their roles in platelet deposition to artificial surfaces remain unclear. Using selected platelet inhibitors (aspirin [ASA], low molecular weight dextran, monoclonal antibodies 10E5 [v GPIIb/IIIa], and 6D1 [GPIb]) we examined the mechanism of platelet deposition to polyethylene (PE) surfaces under steady laminar and oscillatory flow conditions. Polyethylene-100 (PE-100) tubes (0.86 mm internal diameter) were perfused under steady laminar flow with citrated human whole blood reconstituted with 111indium-labeled platelets at 312 seconds-1 shear rate in the presence and absence of platelet inhibitors. The effect of oscillatory flow on platelet deposition was examined in a microwell system using 3/16-inch diameter discs of National Heart, Lung, and Blood Institute primary reference PE as the test surface. ASA and dextran did not significantly (P greater than .05) inhibit platelet deposition in laminar flow (not tested in oscillatory). Antibody 10E5 was a potent inhibitor (laminar less than 1%, P less than .0001, oscillatory less than 1.6%, P less than .01) of platelet deposition in both systems, and in this case, true adhesion (first attached layer) was blocked. Antibody 6D1 unexpectedly inhibited 70% of platelet deposition (P less than .01) in steady laminar flow and 56.5% in oscillatory flow (P less than .01). Scanning electron microscopy demonstrated platelets atop platelets in the controls, rare platelets in the 10E5 group, and a patchy monolayer of platelets in the 6D1 group. Transmission electron microscopy of cross-sections confirmed these observations. We conclude that the adhesion of the first platelet layer to an artificial surface requires GPIIb/IIIa. The data also suggest that GPIb is required for the development of the second layer in vertical platelet thrombus growth.
Expression of interleukin-4 receptors on early human B-lineage cells
Authors: Law CL , Armitage RJ , Villablanca JG , LeBien TW 
Published: October 2019

Abstract
Interleukin-4 (IL-4) regulates multiple stages of the antigen-dependent phase of B-cell development. However, its precise role in regulating B lymphopoiesis in bone marrow is not as well defined. We examined whether surface IgM- normal and leukemic human B-cell precursors (BCP) expressed IL-4 receptors using biotinylated IL-4. Constitutive expression of IL-4 receptors was detected on both normal and leukemic BCP. A higher percentage of normal BCP (82% +/- 15%) expressed IL-4 receptors compared with leukemic BCP (44% +/- 8%). Using mean fluorescent intensity as an indicator of receptor level on the IL-4 receptor positive cells, normal (91 +/- 41) and leukemic (44 +/- 37) BCP expressed comparable numbers of receptors. IL-4 induced the expression of CD23 on 30% of the leukemic BCP cases examined. IL-4 induced CD23 on surface IgM+ fetal bone marrow lymphoid cells but not on the surface IgM- normal BCP, despite the presence of detectable receptors on the surface IgM- cells. IL-4 did not stimulate proliferation of normal BCP, nor could it enhance the effect of recombinant IL-7 or low molecular weight B-cell growth factor. However, IL-4 increased the expression of surface IgM and surface Ig kappa on in vitro differentiated pre-B cells. Our collective results identify no role for IL-4 in the proliferation of normal or leukemic BCP, but identify a role in the enhancement of surface Ig expression during pre- B to B-cell differentiation.
Emergence of a B-cell lymphoblastic lymphoma in a patient with B-cell chronic lymphocytic leukemia: evidence for the single-cell origin of the two tumors
Authors: Pistoia V , Roncella S , Di Celle PF , Sessarego M , Cutrona G , Cerruti G , Boccaccio GP , Grossi CE , Foa R , Ferrarini M 
Published: October 2019

Abstract
A patient is described who presented with a chronic lymphocytic leukemia (CLL) and later developed a lymphoblastic lymphoma. The cells from the CLL were typical mature B lymphocytes as could be assessed by morphologic, cytochemical, and surface marker analyses. The cells from the lymphoblastic lymphoma were immature B cells that expressed CD10, CD20, and HLA-DR markers, but not surface Ig or cytoplasmic mu chains, and were negative for terminal deoxynucleotidyl transferase (TdT). The cells of two continuous cell lines, obtained from the bone marrow and the peripheral blood of the patient, had the same phenotype as the lymphoblastic lymphoma cells, did not contain the Epstein-Barr virus genome, and displayed malignant features in vitro, including the capacity to form colonies in agar. The two cell lines also shared identical chromosomal abnormalities, a finding which suggests that they derived from the same malignant cell already present in vivo. Such chromosomal abnormalities were not seen in the karyotype of the peripheral blood cells at the onset of the disease. Analysis of the Ig heavy chain genes using a DJ-specific probe showed the very same monoclonal rearrangement in the cells from the B-CLL, the lymphoblastic lymphoma and the two cell lines, thus demonstrating their common clonal origin. By contrast, a monoclonal rearrangement of the lambda chain gene locus was found in the B-CLL cells only, a finding consistent with their exclusive capacity to express surface IgM lambda. This patient represents a rare case in whom a chronic lymphoproliferative disorder with mature malignant cells transforms into a lymphoblastic lymphoma characterized by cells frozen at a very early maturational stage. The possible mechanisms leading to such transformation within the same cell clone are discussed.
Molecular basis of the enhanced susceptibility of the erythrocytes of paroxysmal nocturnal hemoglobinuria to hemolysis in acidified serum
Authors: Wilcox LA , Ezzell JL , Bernshaw NJ , Parker CJ 
Published: October 2019

Abstract
When incubated in acidified serum, the erythrocytes of paroxysmal nocturnal hemoglobinuria (PNH) are hemolyzed through activation of the alternative pathway of complement (APC), but normal erythrocytes are resistant to this process. PNH cells are deficient in decay- accelerating factor (DAF), a complement regulatory protein that inhibits the activity of both the classical and the alternative pathways. However, deficiency of DAF alone does not account entirely for the aberrant effects of acidified serum on PNH cells. Recently, we have shown that PNH erythrocytes are also deficient in another complement control protein called membrane inhibitor of reactive lysis (MIRL) that restricts complement-mediated lysis by blocking formation of the membrane attack complex (MAC). To determine the effects of the DAF and MIRL on susceptibility to acidified serum lysis, PNH cells were repleted with the purified proteins. DAF partially inhibited acidified serum lysis by blocking the activity of the amplification C3 convertase. MIRL inhibited acidified serum lysis both by blocking the activity of the MAC and by inhibiting the activity the C3 convertase. When DAF function was blocked with antibody, normal erythrocytes became partially susceptible to acidified serum lysis. By blocking MIRL, cells were made completely susceptible to lysis, and control of C3 convertase activity was partially lost. When both DAF and MIRL were blocked, the capacity of normal erythrocytes to control the activity of the APC and the MAC was destroyed, and the cells hemolyzed even in unacidified serum. These studies demonstrate that DAF and MIRL act in concert to control susceptibility to acidified serum lysis; of the two proteins, MIRL is the more important. In addition to its regulatory effects on the MAC, MIRL also influences the activity of the C3 convertase of the APC. Further, in the absence of DAF and MIRL, the plasma regulators (factor H and factor I) lack the capacity to control membrane- associated activation of the APC.
Fetal liver hematopoietic stem cells as a target for in utero retroviral gene transfer
Authors: Clapp DW , Dumenco LL , Hatzoglou M , Gerson SL 
Published: October 2019

Abstract
Retroviral-mediated gene transfer into hematopoietic precursors often results in only short-term gene transduction in vivo. Loss of the transduced genetic material over time may be caused by the limited ability of retroviral infection to transduce genes into early, pluripotent hematopoietic stem cells. Because fetal liver contains actively proliferating multipotential stem cells that should be more susceptible to retroviral-mediated gene transfer than quiescent cells derived from adult bone marrow, these cells may be an ideal target for gene transduction. Furthermore, physiologic expansion of these cells during development obviates the need for marrow ablation during gene therapy in vivo. We performed in utero gene transfer by injecting high titer replication-defective retrovirus in vivo into the livers of 11, 14, 16, and 18 day gestation rats. After birth, the rats were analyzed for the presence of proviral integration and gene expression. The efficiency of gene transfer into bone marrow cells was greatest in rats infected at day 14 to 16 of gestation. In rats killed at 1 to 26 weeks of age, gene transfer was detected by Southern analysis in 48% and by polymerase chain reaction in 86% of bone marrow samples. The provirus was also detected in white blood cells, the granulocyte-macrophage colony-forming unit, thymus, spleen, liver, and lung. The presence of the transgene in bone marrow and other hematopoietic tissues at 26 weeks of age suggests that early hematopoietic precursors present in the fetal liver are susceptible targets for gene transfer and that these cells become resident in the bone marrow of the adult animal. This model is a new technique for gene transduction into proliferating hematopoietic cells in vivo that avoids bone marrow transplantation and has potential application in the correction of genetic defects in utero.