bluuebunny/crossref_metadata_2025 huggingface dataset:
Syndecan-1 is targeted to the uropods of polarized myeloma cells where it promotes adhesion and sequesters heparin-binding proteins
Authors: Børset Magne , Hjertner Øyvind , Yaccoby Shmuel , Epstein Joshua , Sanderson Ralph D.
Published: October 2019

Syndecan-1 (CD138) is a heparan sulfate-bearing proteoglycan present on the surface of myeloma cells where it mediates myeloma cell-cell and cell-extracellular matrix adhesion. In this study, we examined myeloma cell lines for cell membrane localization of syndecan-1. On some cells we note a striking localization of syndecan-1 to a single small membrane protrusion, with the remainder of the cell surface being mostly negative for syndecan-1. Examination of cell morphology reveals that a proportion of cells from myeloma cell lines, as well as primary myeloma cells, are polarized, with a uropod on one end and lamellipodia on the other end. On these polarized cells, syndecan-1 is specifically targeted to the uropod, but in contrast, on nonpolarized cells syndecan-1 is evenly distributed over the entire cell surface. In addition to syndecan-1, several other cell surface molecules localize specifically to the uropod, including CD44 and CD54. Functional assays reveal that myeloma cell lines with a high proportion of polarized cells have a much higher migratory potential than cell lines with few polarized cells. Moreover, the uropod is the cell pole preferentially involved in aggregation of myeloma cells and in adhesion of myeloma cells to osteoblast-like cells. When polarized myeloma cells are incubated with heparin-binding proteins, like hepatocyte growth factor or osteoprotegerin, they concentrate in the uropod. These data indicate that syndecan-1 is targeted to the uropod of polarized myeloma cells and that this targeting plays a role in promoting cell-cell adhesion and may also regulate the biological activity of heparin-binding cytokines.
Identification of the Dombrock blood group glycoprotein as a polymorphic member of the ADP-ribosyltransferase gene family
Authors: Gubin Alexander N. , Njoroge J. Muthoni , Wojda Urszula , Pack Svetlana D. , Rios Maria , Reid Marion E. , Miller Jeffery L.
Published: October 2019

Identification of the 25 known human blood group molecules is of fundamental importance for the fields of erythroid cell biology and transfusion medicine. Here we provide the first molecular description of the “Dombrock” blood group system. A candidate gene was identified by in silico analyses of approximately 5000 expressed sequence tags (ESTs) from terminally differentiating human erythroid cells. Transfection experiments demonstrated specific binding of anti-Dombrock and confirmed glycosylphosphatidylinositol membrane attachment. Dombrock expression is developmentally regulated during erythroid differentiation and occurs at highest levels in the fetal liver. Homology studies suggest that the Dombrock molecule is a member of the adenosine 5′-diphosphate (ADP)–ribosyltransferase ectoenzyme gene family. Genotypic comparisons suggest Doa versus Dob antigenicity results from a single amino acid substitution within an encoded arginine-glycine-aspartic acid (RGD) motif of the molecule.
Comparison of sensitivity to ultraviolet B irradiation between human lymphocytes and hematopoietic stem cells
Authors: Azuma Hiroshi , Ikebuchi Kenji , Yamaguchi Miki , Murahashi Hideaki , Mogi Yuko , Sato Norihiro , Fujihara Mitsuhiro , Hirayama Fumiya , Ikeda Hisami
Published: October 2019

To investigate the clinical applicability of prophylaxis of post-transplant graft-versus-host disease by UV-B irradiation of stem cell preparations, the UV-B sensitivities of human lymphocytes and primitive hematopoietic progenitors were compared. The mononuclear cell fractions (MNC) derived from human cord blood and granulocyte–colony-stimulating factor–mobilized peripheral blood were used. After UV-B irradiation, lymphocyte proliferation ability, hematopoietic colony-forming cells, and apoptotic cells were analyzed. At a dose of 33 J/m2, significant differences were observed in the residual percentages of hematopoietic progenitors and lymphocyte functions [ANOVA, F (5,46) = 19.4; P < .0001], and the difference between CFU-C (85.2% + 24.0%; n = 8) and MLR (12.7% + 12.6%; n = 10) was significant (P < .0001). There were no significant differences in the residual percentages of CFU-C, HPP-CFC, and LTC-IC. Percentages of annexin V-positive cells in the total MNC and the CD34+cell population in MNC after UV-B irradiation were 69.8% and 18.7%, respectively. In conclusion, there was a range of UV-B doses over which T lymphocytes were inactivated but hematopoietic progenitors, including HPP-CFC and LTC-IC, were preserved.
Comparison of sensitivity to ultraviolet B irradiation between human lymphocytes and hematopoietic stem cells
Authors: Azuma Hiroshi , Ikebuchi Kenji , Yamaguchi Miki , Murahashi Hideaki , Mogi Yuko , Sato Norihiro , Fujihara Mitsuhiro , Hirayama Fumiya , Ikeda Hisami
Published: October 2019

To investigate the clinical applicability of prophylaxis of post-transplant graft-versus-host disease by UV-B irradiation of stem cell preparations, the UV-B sensitivities of human lymphocytes and primitive hematopoietic progenitors were compared. The mononuclear cell fractions (MNC) derived from human cord blood and granulocyte–colony-stimulating factor–mobilized peripheral blood were used. After UV-B irradiation, lymphocyte proliferation ability, hematopoietic colony-forming cells, and apoptotic cells were analyzed. At a dose of 33 J/m2, significant differences were observed in the residual percentages of hematopoietic progenitors and lymphocyte functions [ANOVA, F (5,46) = 19.4; P < .0001], and the difference between CFU-C (85.2% + 24.0%; n = 8) and MLR (12.7% + 12.6%; n = 10) was significant (P < .0001). There were no significant differences in the residual percentages of CFU-C, HPP-CFC, and LTC-IC. Percentages of annexin V-positive cells in the total MNC and the CD34+cell population in MNC after UV-B irradiation were 69.8% and 18.7%, respectively. In conclusion, there was a range of UV-B doses over which T lymphocytes were inactivated but hematopoietic progenitors, including HPP-CFC and LTC-IC, were preserved.
PU.1 inhibits GATA-1 function and erythroid differentiation by blocking GATA-1 DNA binding
Authors: Zhang Pu , Zhang Xiaobo , Iwama Atsushi , Yu Channing , Smith Kent A. , Mueller Beatrice U. , Narravula Salaija , Torbett Bruce E. , Orkin Stuart H. , Tenen Daniel G.
Published: October 2019

The lineage-specific transcription factors GATA-1 and PU.1 can physically interact to inhibit each other's function, but the mechanism of repression of GATA-1 function by PU.1 has not been elucidated. Both the N terminus and the C terminus of PU.1 can physically interact with the C-terminal zinc finger of GATA-1. It is demonstrated that the PU.1 N terminus, but not the C terminus, is required for inhibiting GATA-1 function. Induced overexpression of PU.1 in K562 erythroleukemia cells blocks hemin-induced erythroid differentiation. In this system, PU.1 does not affect the expression of GATA-1 messenger RNA, protein, or nuclear localization. However, GATA-1 DNA binding decreases dramatically. By means of electrophoretic mobility shift assays with purified proteins, it is demonstrated that the N-terminal 70 amino acids of PU.1 can specifically block GATA-1 DNA binding. In addition, PU.1 had a similar effect in the G1ER cell line, in which the GATA-1 null erythroid cell line G1E has been transduced with a GATA-1–estrogen receptor fusion gene, which is directly dependent on induction of the GATA-1 fusion protein to effect erythroid maturation. Consistent with in vitro binding assays, overexpression of PU.1 blocked DNA binding of the GATA-1 fusion protein as well as GATA-1–mediated erythroid differentiation of these G1ER cells. These results demonstrate a novel mechanism by which function of a lineage-specific transcription factor is inhibited by another lineage-restricted factor through direct protein–protein interactions. These findings contribute to understanding how protein–protein interactions participate in hematopoietic differentiation and leukemogenesis.
Antimicrobial proteins and peptides of blood: templates for novel antimicrobial agents
Authors: Levy Ofer
Published: October 2019

The innate immune system provides rapid and effective host defense against microbial invasion in a manner that is independent of prior exposure to a given pathogen.1 It has long been appreciated that the blood contains important elements that mediate rapid responses to infection. Thus, anatomic compartments with ample blood supply are less frequently infected and recover more readily once infected, whereas regions with poor perfusion are prone to severe infection and may require surgical débridement. Blood-borne innate immune mediators are either carried in circulating blood cells (ie, leukocytes and platelets) or in plasma after release from blood cells or on secretion by the liver.
Neutrophil gelatinase B potentiates interleukin-8 tenfold by aminoterminal processing, whereas it degrades CTAP-III, PF-4, and GRO-α and leaves RANTES and MCP-2 intact
Authors: Van den Steen Philippe E. , Proost Paul , Wuyts Anja , Van Damme Jo , Opdenakker Ghislain
Published: October 2019

Chemokines are mediators in inflammatory and autoimmune disorders. Aminoterminal truncation of chemokines results in altered specific activities and receptor recognition patterns. Truncated forms of the CXC chemokine interleukin (IL)-8 are more active than full-length IL-8 (1-77), provided the Glu-Leu-Arg (ELR) motif remains intact. Here, a positive feedback loop is demonstrated between gelatinase B, a major secreted matrix metalloproteinase (MMP-9) from neutrophils, and IL-8, the prototype chemokine active on neutrophils. Natural human neutrophil progelatinase B was purified to homogeneity and activated by stromelysin-1. Gelatinase B truncated IL-8(1-77) into IL-8(7-77), resulting in a 10- to 27-fold higher potency in neutrophil activation, as measured by the increase in intracellular Ca++concentration, secretion of gelatinase B, and neutrophil chemotaxis. This potentiation correlated with enhanced binding to neutrophils and increased signaling through CXC chemokine receptor-1 (CXCR1), but it was significantly less pronounced on a CXCR2-expressing cell line. Three other CXC chemokines—connective tissue-activating peptide-III (CTAP-III), platelet factor-4 (PF-4), and GRO-α—were degraded by gelatinase B. In contrast, the CC chemokines RANTES and monocyte chemotactic protein-2 (MCP-2) were not digested by this enzyme. The observation of differing effects of neutrophil gelatinase B on the proteolysis of IL-8 versus other CXC chemokines and on CXC receptor usage by processed IL-8 yielded insights into the relative activities of chemokines. This led to a better understanding of regulator (IL-8) and effector molecules (gelatinase B) of neutrophils and of mechanisms underlying leukocytosis, shock syndromes, and stem cell mobilization by IL-8.
Cell dose and speed of engraftment in placental/umbilical cord blood transplantation: graft progenitor cell content is a better predictor than nucleated cell quantity
Authors: Migliaccio Anna Rita , Adamson John W. , Stevens Cladd E. , Dobrila N. Ludy , Carrier Carmelita M. , Rubinstein Pablo
Published: October 2019

There is evidence that the total cellular content of placental cord blood (PCB) grafts is related to the speed of engraftment, though the total nucleated cell (TNC) dose is not a precise predictor of the time of neutrophil or platelet engraftment. It is important to understand the reasons for the quantitative association and to improve the criteria for selecting PCB grafts by using indices more precisely predictive of engraftment. The posttransplant course of 204 patients who received grafts evaluated for hematopoietic colony-forming cell (CFC) content among 562 patients reported previously were analyzed using univariate and multivariate life-table techniques to determine whether CFC doses predicted hematopoietic engraftment speed and risk for transplant-related events more accurately than the TNC dose. Actuarial times to neutrophil and platelet engraftment were shown to correlate with the cell dose, whether estimated as TNC or CFC per kilogram of recipient's weight. CFC association with the day of recovery of 500 neutrophils/μL, measured as the coefficient of correlation, was stronger than that of the TNC (R = −0.46 and −0.413, respectively). In multivariate tests of speed of platelet and neutrophil engraftment and of probability of posttransplantation events, the inclusion of CFC in the model displaced the significance of the high relative risks associated with TNC. The CFC content of PCB units is associated more rigorously with the major covariates of posttransplantation survival than is the TNC and is, therefore, a better index of the hematopoietic content of PCB grafts.
Expression of the collagen receptor glycoprotein VI during megakaryocyte differentiation
Authors: Berlanga Oscar , Bobe Regis , Becker Marion , Murphy George , Leduc Mireille , Bon Cassian , Barry Fiona A. , Gibbins Jonathan M. , Garcia Paloma , Frampton Jonathan , Watson Stephen P.
Published: October 2019

This study examined the expression of the platelet collagen receptor glycoprotein VI (GPVI) in megakaryocyte cell lines and primary megakaryocytes by reverse transcriptase-polymerase chain reaction and by flow cytometry and ligand blotting using the snake venom toxin convulxin. Expression of GPVI is increased in the megakaryoblastic cell lines HEL and CMK on differentiation with the phorbol ester phorbol 12-myristate 13-acetate (PMA), along with the Fc receptor γ-chain (FcR γ-chain). The increase in GPVI expression is associated with marked potentiation of tyrosine phosphorylation and Ca++ elevation in response to convulxin. Syk, linker for activated T cells, and phospholipase Cγ2 (PLCγ2) are among the proteins tyrosine phosphorylated on convulxin stimulation in PMA-differentiated HEL cells. Studies on primary murine megakaryocytes grown in vitro confirmed that GPVI is up-regulated in parallel with functional activation, assessed by measurement of [Ca++]i, during differentiation. The results demonstrate that expression of GPVI is up-regulated along with the FcR γ-chain during differentiation of megakaryocytes.
Tumor-specific recognition of human myeloma cells by idiotype-induced CD8+ T cells
Authors: Li Yiwen , Bendandi Maurizio , Deng Yuping , Dunbar Cynthia , Munshi Nikhil , Jagannath Sundar , Kwak Larry W. , Lyerly H. Kim
Published: October 2019

Immunoglobulin secreted by myeloma cells contains a unique antigenic determinant (idiotype [Id]) that may serve as a tumor-specific antigen. Although Id-protein–specific T-cell responses have been reported in patients with myeloma, it is not known whether primary myeloma tumor cells can present naturally processed Id peptides on their surface as a target. We immunized 2 healthy human stem-cell donors with Id proteins from their recipients. T cells from the immunized donors released high levels of T-helper 1–type cytokines in response to stimulation with myeloma cells from their recipients. The T-cell–mediated cytokine response to tumor cells was blocked by a major histocompatibility complex (MHC) class I monoclonal antibody, whereas the response to soluble Id protein was dependent on MHC class II. To investigate whether Id-specific CD8+ T cells can recognize and kill autologous myeloma cells, we generated T cells from peripheral blood mononuclear cells from a third patient with myeloma by means of in vitro stimulation with autologous dendritic cells pulsed with Id protein. Tumor-specific lysis of myeloma cells was demonstrated by the lack of killing of autologous nonmalignant B cells or natural killer–sensitive K562 cells. Lysis of autologous myeloma targets was restricted by MHC class I molecules. These data represent the first report of class I–restricted T-cell recognition of fresh autologous myeloma targets and formally demonstrate that human myeloma cells can serve as targets of an Id-specific T-cell response.
Treatment with liposome-encapsulated clodronate as a new strategic approach in the management of immune thrombocytopenic purpura in a mouse model
Authors: Alves-Rosa Fernanda , Stanganelli Carmen , Cabrera Juana , van Rooijen Nico , Palermo Marina S. , Isturiz Martı́n A.
Published: October 2019

Immune thrombocytopenic purpura (ITP) is an autoimmune disease related to the presence of elevated levels of platelet-associated immunoglobulin, or autoantibodies. In recent years the importance of macrophage Fcγ receptors in the uptake of platelets in ITP has been confirmed. Although in patients with ITP the platelet destruction occurs in liver and spleen, in this present experimental mouse model the liver was the principal organ of sequestration of sensitized platelets. The uptake in the spleen, bone marrow, lung, and kidneys was negligible and not different from that in control animals. In addition, the trapped platelets did not return to circulation, and new cells derived from the platelet-storage pool or new thrombocytogenesis were necessary to restore the platelet count. The depletion of splenic and hepatic murine macrophages by liposome-encapsulated clodronate (lip-clod) was studied as a new strategy for ITP treatment. Lip-clod inhibits, in a dose-dependent manner, the antibody-induced thrombocytopenia. Moreover, lip-clod treatment rapidly restored (24 hours) the platelet count in thrombocytopenic animals to hematologic safe values, and despite additional antiplatelet antiserum treatment, mice were able to maintain this level of platelets at least up to 48 hours. The bleeding times in lip-clod–treated animals was not different from those in controls, demonstrating that the hemostasis was well controlled in these animals. The results presented in this study demonstrate that lip-clod treatment can be effective in the management of experimental ITP.
Down-regulation of the chemokine receptor CCR5 by activation of chemotactic formyl peptide receptor in human monocytes
Authors: Shen Weiping , Li Baoqun , Wetzel Michele A. , Rogers Thomas J. , Henderson Earl E. , Su Shao Bo , Gong Wanghua , Le Yingying , Sargeant Robert , Dimitrov Dimiter S. , Oppenheim Joost J. , Wang Ji Ming
Published: October 2019

Interactions between cell surface receptors are important regulatory elements in the complex host responses to infections. In this study, it is shown that a classic chemotactic factor, the bacterial chemotactic peptide N-formyl-methionyl-leucylphenyl-alanine (fMLF), rapidly induced a protein-kinase-C–mediated serine phosphorylation and down-regulation of the chemokine receptor CCR5, which serves as a major human immunodeficiency virus (HIV)-1 coreceptor. The fMLF binding to its receptor, formyl peptide receptor (FPR), resulted in significant attenuation of cell responses to CCR5 ligands and in inhibition of HIV-1-envelope-glycoprotein–mediated fusion and infection of cells expressing CD4, CCR5, and FPR. The finding that the expression and function of CCR5 can be regulated by peptides that use an unrelated receptor may provide a novel approach to the design of anti-inflamatory and antiretroviral agents.
Mechanisms of HIV-associated lymphocyte apoptosis
Authors: Badley Andrew D. , Pilon André A. , Landay Alan , Lynch David H.
Published: October 2019

Infection with the human immunodeficiency virus (HIV) is associated with a progressive decrease in CD4 T-cell number and a consequent impairment in host immune defenses. Analysis of T cells from patients infected with HIV, or of T cells infected in vitro with HIV, demonstrates a significant fraction of both infected and uninfected cells dying by apoptosis. The many mechanisms that contribute to HIV-associated lymphocyte apoptosis include chronic immunologic activation; gp120/160 ligation of the CD4 receptor; enhanced production of cytotoxic ligands or viral proteins by monocytes, macrophages, B cells, and CD8 T cells from HIV-infected patients that kill uninfected CD4 T cells; and direct infection of target cells by HIV, resulting in apoptosis. Although HIV infection results in T-cell apoptosis, under some circumstances HIV infection of resting T cells or macrophages does not result in apoptosis; this may be a critical step in the development of viral reservoirs. Recent therapies for HIV effectively reduce lymphoid and peripheral T-cell apoptosis, reduce viral replication, and enhance cellular immune competence; however, they do not alter viral reservoirs. Further understanding the regulation of apoptosis in HIV disease is required to develop novel immune-based therapies aimed at modifying HIV-induced apoptosis to the benefit of patients infected with HIV.
Down-regulation of neutrophil functions by the ELR+CXC chemokine platelet basic protein
Authors: Ehlert Jan E. , Ludwig Andreas , Grimm Tobias A. , Lindner Buko , Flad Hans-Dieter , Brandt Ernst
Published: October 2019

The platelet-derived neutrophil-activating peptide 2 (NAP-2, 70 amino acids) belongs to the ELR+ CXC subfamily of chemokines. Similar to other members of this group, such as IL-8, NAP-2 activates chemotaxis and degranulation in neutrophils (polymorphonuclear [PMN]) through chemokine receptors CXCR-1 and CXCR-2. However, platelets do not secrete NAP-2 as an active chemokine but as the C-terminal part of several precursors that lack PMN-stimulating capacity. As we have previously shown, PMN themselves may liberate NAP-2 from the precursor connective tissue-activating peptide III (CTAP-III, 85 amino acids) by proteolysis. Instead of inducing cell activation, continuous accumulation of the chemokine in the surroundings of the processing cells results in the down-regulation of specific surface-expressed NAP-2 binding sites and in the desensitization of chemokine-induced PMN degranulation. Thus, NAP-2 precursors may be regarded as indirect mediators of functional desensitization in neutrophils. In the current study we investigated the biologic impact of another major NAP-2 precursor, the platelet basic protein (PBP, 94 amino acids). We show that PBP is considerably more potent than CTAP-III to desensitize degranulation and chemotaxis in neutrophils. We present data suggesting that the high desensitizing capacity of PBP is based on its enhanced proteolytic cleavage into NAP-2 by neutrophil-expressed cathepsin G and that it involves efficient down-regulation of surface-expressed CXCR-2 while CXCR-1 is hardly affected. Correspondingly, we found PBP and, less potently, CTAP-III to inhibit CXCR-2– but not CXCR-1– dependent chemotaxis of neutrophils toward NAP-2. Altogether our findings demonstrate that the anti-inflammatory capacity of NAP-2 is governed by the species of its precursors.
Induction of B-cell tolerance by retroviral gene therapy
Authors: Bracy Jennifer L. , Iacomini John
Published: October 2019

The primary immunologic barrier to overcome before clinical xenotransplantation can be successful is rejection mediated by preformed natural antibodies in the host, directed toward a single carbohydrate epitope Galα1-3Galβ1-4GlcNAc-R (αGal) present on porcine tissue, encoded for by the enzyme glucosyltransferase UDP galactose:β-D-galactosyl-1,4-N-acetyl-D-glucosaminide α(1-3)galactosyltransferase (EC 2.4.1.151) or simply αGT. Although we have shown previously that a gene therapy approach could be used to prevent production of natural antibodies specific for αGal, the ability to induce and maintain tolerance after rigorous antigen challenge would be required if similar approaches are to be used clinically. Here, we demonstrate in αGT knockout mice (GT0 mice), which, like humans, contain in their serum antibodies that bind αGal, that the efficient transduction and expression of a retrovirally transduced αGT gene in bone marrow–derived cells induces stable long-term tolerance to the αGal epitope. GT0 mice reconstituted with αGT-transduced bone marrow cells were unable to produce antibodies that bind αGal after extensive immunization with pig cells. Furthermore, using ELISPOT assays, we were unable to detect the presence of B cells that produce αGal reactive antibodies after immunization, suggesting that such B cells were eliminated from the immunologic repertoire after gene therapy. Interestingly, after tolerance to αGal is induced by gene therapy, the antiporcine non-αGal humoral response changes from a predominantly IgM to an IgG response. This suggests that once the natural antibody barrier is eliminated by the induction of tolerance, the antipig response changes to a typical T-cell–dependent response involving isotype switching. Thus, gene therapy approaches may be used to overcome immunologic responses leading to xenograft rejection, and similar gene therapy approaches could be used to overcome autoimmunity.
Imprinting of insulin-like growth factor 2 is modulated during hematopoiesis
Authors: Morison Ian M. , Eccles Michael R. , Reeve Anthony E.
Published: October 2019

The transcription of insulin-like growth factor 2 (IGF-2) is affected by genomic imprinting, a multistep process through which the parental origin of a gene influences its transcription. The maternal copy of IGF-2 is silenced in most human tissues, but in the choroid plexus and the adult liver both alleles of IGF-2 are expressed. This study shows that though in peripheral blood mononuclear cells IGF-2shows paternal allele-specific expression, in total bone marrow both alleles are transcribed. This modulation of imprinting is not attributable to use of the P1 promoter, because transcription from the P3 promoter occurred from both alleles. These results suggest that transcriptional recognition of the IGF-2 imprint can be modulated during hematopoiesis and may facilitate the development of in vitro model systems to study the transcriptional recognition of a genomic imprint.
UTY gene codes for an HLA-B60–restricted human male-specific minor histocompatibility antigen involved in stem cell graft rejection: characterization of the critical polymorphic amino acid residues for T-cell recognition
Authors: Vogt Mario H. J. , Goulmy Els , Kloosterboer Freke M. , Blokland Els , de Paus Roel A. , Willemze Roel , Falkenburg J. H. Frederik
Published: October 2019

Rejection of a graft after human leukocyte antigen (HLA)-identical stem cell transplantation (SCT) can be caused by recipient's immunocompetent T lymphocytes recognizing minor histocompatibility antigens on donor stem cells. During rejection of a male stem cell graft by a female recipient, 2 male (H-Y)–specific cytotoxic T lymphocyte (CTL) clones were isolated from peripheral blood. One CTL clone recognized an HLA-A2–restricted H-Y antigen, encoded by the SMCY gene. Another CTL clone recognized an HLA-B60–restricted H-Y antigen. In this study UTY was identified as the gene coding for the HLA-B60–restricted H-Y antigen. The UTY-derived H-Y antigen was characterized as a 10-amino acid residue peptide, RESEEESVSL. Although the epitope differed by 3 amino acids from its X-homologue, UTX, only 2 polymorphisms were essential for recognition by the CTL clone HLA-B60 HY. These results illustrate that CTLs against several H-Y antigens derived from different proteins can contribute simultaneously to graft rejection after HLA-identical, sex-mismatched SCT. Moreover, RESEEESVSL-specific T cells could be isolated from a female HLA-B60+ patient with myelodysplastic syndrome who has been treated with multiple blood transfusions, but not from control healthy HLA-B60+ female donors. This may indicate that RESEEESVSL-reactive T cells are more common in sensitized patients.
Molecular single-cell analysis of the clonal relationship of small Epstein-Barr virus–infected cells and Epstein-Barr virus–harboring Hodgkin and Reed/Sternberg cells in Hodgkin disease
Authors: Spieker Tilmann , Kurth Julia , Küppers Ralf , Rajewsky Klaus , Bräuninger Andreas , Hansmann Martin-Leo
Published: October 2019

Epstein-Barr virus (EBV) can be detected in the tumor cells of approximately 40% of cases of classical Hodgkin disease (cHD). Clonality studies suggest that infection of the neoplastic Hodgkin and Reed/Sternberg (HRS) cells occurs before tumor clone expansion. In EBV-positive cases, variable numbers of EBER-positive small B cells are sometimes also observed that immunohistologically differ from the neoplastic cells by lack of CD30 and latent membrane protein 1 expression. To analyze the clonal relationship between these EBV+ cells and the HRS cells, single EBV-infected CD30− B cells, as well as HRS cells from 3 cases of EBV-positive cHD were micromanipulated, their immunoglobulin gene rearrangements amplified and then compared with each other. In 2 cases, all small EBV-infected cells were clonally unrelated to the HRS cells. In a third case, 2 of 29 small CD30− cells were found to carry HRS cell-specific rearrangements. Thus, small CD30−EBV-infected B cells in cHD belong to the HRS tumor clone rarely, if at all. In all cases, small clones unrelated to the HRS cell clones were identified among the small EBV+ CD30− cells. The vast majority of small EBV+ CD30− B cells was found to carry somatically mutated V region genes, indicating that in lymph nodes of patients with HD, like in the peripheral blood of healthy individuals, EBV persists in memory B cells.
Heterogeneity in therapeutic response of genetically altered myeloma cell lines to interleukin 6, dexamethasone, doxorubicin, and melphalan
Authors: Rowley Matt , Liu Pocheng , Van Ness Brian
Published: October 2019

Because there is no known genetic abnormality common to all patients with myeloma, it is important to understand how genetic heterogeneity may lead to differences in signal transduction, cell cycle, and response to therapy. Model cell lines have been used to study the effect that mutations in p53 and rascan have on growth properties and responses of myeloma cells. The U266 cell line has a single mutant p53 allele. Stable expression of wild-type (wt) p53 in U266 cells results in a significant suppression of interleukin (IL)-6 gene expression and in the concomitant suppression of cell growth that could be restored by the addition of exogenous IL-6. Expression of wt p53 also leads to cell cycle arrest and protection from doxorubicin (Dox)- and melphalan (Mel)-induced apoptosis. The addition of IL-6 resulted in cell cycle progression and blocked p53-mediated protection from apoptosis. ANBL6 is an IL-6–dependent cell line that is sensitive to dexamethasone (Dex), Dox, and Mel. IL-6 is able to protect ANBL6 cells from Dex- and Mel- but not Dox-induced apoptosis. To study the effect of an activating mutation in ras, the ANBL6 cell line transfected with either a constitutively activated N- orK-ras gene was used. Both N-ras12 andK-ras12 genes were able to protect ANBL6 cells from apoptosis induced by Dex, Dox, and Mel. These data show that changes inras or p53 can alter the myeloma cell response to IL-6 and demonstrate that the genetic background can alter therapeutic responses.
Identification and characterization of CKLiK, a novel granulocyte Ca++/calmodulin-dependent kinase
Authors: Verploegen Sandra , Lammers Jan-Willem J. , Koenderman Leo , Coffer Paul J.
Published: October 2019

Human granulocytes are characterized by a variety of specific effector functions involved in host defense. Several widely expressed protein kinases have been implicated in the regulation of these effector functions. A polymerase chain reaction–based strategy was used to identify novel granulocyte-specific kinases. A novel protein kinase complementary DNA with an open reading frame of 357 amino acids was identified with homology to calcium-calmodulin–dependent kinase I (CaMKI). This has been termed CaMKI-like kinase (CKLiK). Analysis of CKLiK messenger RNA (mRNA) expression in hematopoietic cells demonstrated an almost exclusive expression in human polymorphonuclear leukocytes (PMN). Up-regulation of CKLiK mRNA occurs during neutrophilic differentiation of CD34+ stem cells. CKLiK kinase activity was dependent on Ca++ and calmodulin as analyzed by in vitro phosphorylation of cyclic adenosine monophosphate responsive element modulator (CREM). Furthermore, CKLiK- transfected cells treated with ionomycin demonstrated an induction of CRE- binding protein (CREB) transcriptional activity compared to control cells. Additionally, CaMK-kinaseα enhanced CKLiK activity. In vivo activation of CKLiK was shown by addition of interleukin (IL)-8 to a myeloid cell line stably expressing CKLiK. Furthermore inducible activation of CKLiK was sufficient to induce extracellular signal-related kinase (ERK) mitogen-activated protein (MAP) kinase activity. These data identify a novel Ca++/calmodulin-dependent PMN- specific kinase that may play a role in Ca++-mediated regulation of human granulocyte functions.
Low B-cell and monocyte counts on day 80 are associated with high infection rates between days 100 and 365 after allogeneic marrow transplantation
Authors: Storek Jan , Espino German , Dawson Monja A. , Storer Barry , Flowers Mary E. D. , Maloney David G.
Published: October 2019

To ascertain which mononuclear cell subset deficiency plays a role in the marrow transplant recipient's susceptibility to infections, mononuclear cell subset counts were prospectively determined in 108 patients on day 80. Infections occurring between day 100 and 365 were recorded by an investigator blinded to the subset counts. In univariate analyses, the counts of the following subsets showed a significant inverse correlation with infection rates: total B cells, IgD+ B cells, IgD− B cells, total CD4 T cells, CD28+ CD4 T cells, CD28− CD4 T cells, CD45RAlow/− CD4 T cells and monocytes. In multivariate analyses, the counts of the following subsets remained significantly inversely correlated with the infection rates: total B cells (P = .0004) and monocytes (P = .009). CD28− CD8 T-cell counts showed no correlation with infection rates. In conclusion, the susceptibility of patients to infections late posttransplant may be due in part to the slow reconstitution of B cells and monocytes.
Dermal and pulmonary inflammatory disease in E-selectin and P-selectin double-null mice is reduced in triple-selectin–null mice
Authors: Collins Robert G. , Jung Unsu , Ramirez Maricela , Bullard Daniel C. , Hicks M. John , Smith C. Wayne , Ley Klaus , Beaudet Arthur L.
Published: October 2019

In the initial phase of an inflammatory response, leukocytes marginate and roll along the endothelial surface as a result of adhesive interactions between molecules on the endothelial cells and leukocytes. To evaluate the role of the 3 selectins (E, L, and P) in leukocyte rolling and emigration, a null mutation for L-selectin was introduced into previously described embryonic stem cells with null mutations in the genes for both E-selectin and P-selectin (E/P double mutants) to produce triple-selectin–null mice (E-selectin, L-selectin, and P-selectin [E/L/P] triple mutants). Triple-selectin homozygous mutant mice are viable and fertile and only rarely develop the severe mucocutaneous infections or pulmonary inflammation characteristic of E/P double-mutant mice. Surface expression of L-selectin was undetectable in triple-mutant mice on fluorescence-activated cell-sorter analysis of peripheral neutrophils. Pathological studies revealed moderate cervical lymphadenopathy and lymphoplasmacytic infiltrate, but these were less extensive than in E/P double-mutant mice. Neutrophil emigration during thioglycolate-induced peritonitis was significantly reduced at 4, 8, and 24 hours (35%, 65%, and 46% of wild-type values, respectively). Intravital microscopy of the cremaster muscle revealed almost no rolling at times up to 6 hours after exteriorization, with or without addition of tumor necrosis factor α. The small amount of residual rolling was dependent on α4-integrin. The occurrence of skin and pulmonary disease in E/P double-mutant mice but not E/L/P triple-mutant mice suggests that deficiency of L-selectin alters the inflammatory response in E/P mutants.
Allogeneic blood cell transplantation following reduced-intensity conditioning is effective therapy for older patients with myelofibrosis with myeloid metaplasia
Authors: Devine Steven M. , Hoffman Ron , Verma Amit , Shah Rajul , Bradlow Basil A. , Stock Wendy , Maynard Vera , Jessop Ellen , Peace David , Huml Marianne , Thomason Deana , Chen Yi-Hsiang , van Besien Koen
Published: October 2019

Standard myeloablative conditioning prior to allogeneic hematopoietic stem cell (HSC) transplantation has been associated with significant toxicity in patients older than 45 years of age with myelofibrosis with myeloid metaplasia (MMM). We sought to evaluate the efficacy of a reduced-intensity conditioning regimen for allogeneic HSC transplantation in this setting. A regimen consisting of fludarabine (30 mg/m2 intravenously daily for 5 days) and melphalan (70 mg/m2 intravenously daily for 2 days) followed by transplantation of filgrastim-mobilized peripheral blood cells from HLA-identical siblings was administered to 4 older patients (median age, 56 years; range, 48-58 years) with advanced MMM. All patients achieved prompt neutrophil and platelet engraftment and have experienced a significant regression of splenomegaly and bone marrow fibrosis. All now have normal bone marrow cellularity. With a median follow-up of 13 months (range, 11-19 months), all 4 patients are alive with stable full-donor hematopoietic chimerism. These results support the feasibility and effectiveness of reduced-intensity conditioning prior to allogeneic HSC transplantation for older patients with advanced MMM.
A child with Gradenigo syndrome presenting with meningism: a case report
Authors: Athapathu A. S. , Bandara E. R. S. , Aruppala A. A. H. S. , Chandrapala K. M. A. U. , Mettananda Sachith
Published: October 2019

Background

The symptoms of meningitis which include fever, headache, photophobia and irritability along with abducens nerve palsy pose a diagnostic dilemma requiring urgent attention. Here we report how such a dilemma was methodically and sequentially resolved using anatomical knowledge supported by neuroimaging and the eventual diagnosis of Gradenigo syndrome was made.

Case presentation

A 6-year-old previously healthy boy from Sri Lanka presented with high grade fever, headache, photophobia and left eye pain for 10 days and diplopia for 2 days duration. Neurological examination was unremarkable except for left sided abducens nerve palsy. He had high inflammatory markers and white blood cell count. A tentative differential diagnosis of acute bacterial meningitis complicated by cerebral oedema, acute hydrocephalus or cerebral abscess was made. However, non-contrast CT brain, cerebrospinal fluid analysis and electroencephalogram were normal leading to a diagnostic dilemma. MRI brain with contrast performed 3 days later due to limited resources revealed left mastoiditis extending to petrous temporal bone confirming Gradenigo syndrome.

Conclusion

This case report highlights the importance of a thorough physical examination in children presenting with unrelated neurological symptoms and signs. Unilateral abducens nerve palsy raises the suspicion of increased intracranial pressure and neuroimaging is vital in diagnostic uncertainties. Gradenigo syndrome emphasises the importance of incorporating anatomical knowledge into clinical practice.
Vaccinating outside Section 7a immunisation programmes: Who, how and why?
Authors: Bosanquet Nick , Davies Tim , Donovan Helen , Heffernan Catherine , Kassianos George , Kirkham Karen , Lonergan Kerry , Nicholson Ellen
Published: October 2019

On 23 April 2019, the authors of this roundtable discussion met to deliberate the vaccinations that fall outside the Section 7A immunisation programme. The three-hour meeting resulted in the production of this article
Brexit risks for the NHS remain
Authors: Moffat Polly
Published: October 2019

As the deadline for leaving the EU looms closer, the NHS still faces uncertainty about the availability of medicines and what that will mean for patients in the event of no deal
Supporting advanced bilingual learners
Authors: Wilson Dr Ruth
Published: October 2019

Drawing on the research evidence and best practice, Dr Ruth Wilson advises on supporting advanced bilingual learners to develop the necessary academic writing skills
A huge impact on reading
Authors: Lee-Potter Emma
Published: October 2019

From reading challenges to awards and visits, Ros Harding – the School Librarian of the Year 2019 – has been recognised for the wide range of literacy initiatives she undertakes with students. Emma Lee-Potter finds out more
Martian channels: Imagining interplanetary communication at the dawn of the radio age
Authors: Enns Anthony
Published: October 2019

In 1899 radio pioneer Nikola Tesla claimed to have received a wireless transmission from the planet Mars, which not only confirmed the existence of intelligent life but also invited a response. The public was immediately captivated by the idea that Mars was attempting to communicate with the Earth, and this widespread fascination reflected not only a belief in the existence of extraterrestrial life but also a notion of radio as a transnational medium that could potentially unite the world by making terrestrial borders obsolete. It may seem strange, therefore, that this fascination culminated in Orson Welles’ famous radio adaptation of H. G. Wells’ novel The War of the Worlds, in which radio was represented not as a medium of interplanetary communication but rather as an emergency broadcast system that warned Americans of an extraterrestrial invasion. Through a closer examination of the history of the idea of interplanetary communication, this article explores how radio was initially conceived as a medium that transgressed social, political and linguistic boundaries and how this utopian promise was later displaced by the idea of radio as a medium that served to construct and reinforce national borders and identities.
Contextualizing Australian radio art internationally
Authors: Black Colin
Published: October 2019

Is Australian radio art practice qualitatively different from international practice? Has there been an Australian approach to radio art that was internationally perceived? Drawing on interviews of national and international radio art practitioners and art theory to analyse important contributions to the field, this article contextualizes Australian radio art against international practice to arrive at nuanced answers to these questions.
Book Reviews
Authors: Lewis Peter , MacLennan Anne F. , McHugh Siobhan
Published: October 2019

The Cultural Work of Community Radio, Katie Moylan (2019) Rowman and Littlefield, 180 pp., ISBN: 978-1-78348-932-9, h/bk, £97.05, e-book, £41.52

A Companion to the History of American Broadcasting, Aniko Bodroghkozy (ed.) (2018) Hoboken, NJ: John Wiley & Sons Inc., 512 pp., ISBN: 978-1-11864-615-1, h/bk, $215; e-book $172.99

Podcasting: New Aural Cultures and Digital Media, Dario Llinares, Neil Fox and Richard Berry (eds) (2018) Cham: Palgrave Macmillan, 332 pp., ISBN: 978-3-31990-055-1, p/bk, £22.49, e-book, £21.37

Podcasting: The Audio Media Revolution, Martin Spinelli and Lance Dann (eds) (2019) London and New York: Bloomsbury Academic, 272 pp., ISBN: 978-1-50132-868-8, p/bk, £19.99, e-book, £14.03
Clinical and karyotypical characteristics of the Shereshevsky-Turner syndrome
Authors: Tarskaya L. A. , Prytkov A. N. , Trubnikov V. I. , Patyutko R. S. , Kasatkina E. P. , Kozlova S. I.
Published: October 2019

The incidence of 18 phenotypical signs was analyzed in 82 patients with the Shereshevvky-Turner syndrome. The minimal diagnostic signs occurring with at least 50% incidence have been singled out. Comparative analysis of the distribution of clinical signs in groups with different variants of karyotype disorders showed that the group of patients with the mosaic karyotype (45,X/40,XX) was the least loaded with pathological signs. Groups with karyotype 48,X and structural disorders are similar in the distribution of phenotypical signs. Multidimensional statistical cluster analysis of the matrix of summary phenotypical distances was carried out and a cluster singled out which permitted unification of 4 groups of patients with different karyotype disorders. The results are compared with published data of molecular cytogenetic studies.
Non-invasive method for determining the level of stable intrathyroid iodine in the diagnosis and control of treatment of thyroid diseases (lecture)
Authors: Tomashevsky I. O.
Published: October 2019

The technology of non-invasive determination of the level of intrathyroid stable iodine (ISI), called X-ray fluorescence analysis or X-ray fluorescence scanning, began to be used since 1968 and is currently used in approximately 10 radiological laboratories around the world as a unique research methodology. Only in recent years have the technical difficulties of this diagnostic method been overcome, and it is gradually becoming more accessible. Since 1974, the efforts of the staff of the Radiological Scientific Center of the Russian Academy of Medical Sciences (Obninsk) and the clinical hospital of the Central Health Unit No. 119 of the Federal Administration for Biomedical and Extreme Problems at the Ministry of Health of the Russian Federation (Khimki) carried out the necessary scientific research and created an analyzer, a method for determining the level ISI and interpretations of the results obtained, allowed for use in Russia as serial laboratory technologies. The specified diagnostic method over the past few years has been successfully used in these institutions when examining more than 3,000 patients with various thyroid diseases (thyroid gland) and continues to be used in the comprehensive diagnosis of thyroid pathology. The purpose of this lecture is to familiarize endocrinologists with the diagnostic capabilities of this method.
ISRÂ’ÎLÎYÂT DALAM KITAB ḤÂSHIYAH AL-ṢÂWÎ ‘ALÂ TAFSÎR JALÂLAYN KARYA AḤMAD AL-ṢÂWÎ
Authors: Mahfud Hakim
Published: October 2019

Al-ṣâwî’s attitude toward Isrâ’îlîyât history (attitude in the book of ḥâshiyah) is reflected in his collection of such histories. In many interpretations, Isrâ’îlîyât’s history is widely cited, but the quotation does not include any evidence of the truth of the story, nor the quality of the history. In this article, the author examines al-ṣâwî’s attitude toward the Isrâ’îlîyât story contained in ḥâshiyah al-ṣâwî’s alâ Tafsîr Jalâlayn. This study aimed to find out whether al-Sawi was critical of the Israeli story. The author uses the library reseach method (literature). To obtain information from various sources, the author uses the documentation method then analyzes the data using a discourse analysis method developed by Tuen A.van Djijk namely macro structure, superstructure, and microstructure. The results of this study reveal that al-Sawi does not always provide comments on the quoted Isra'iliyat story. There are several stories that he included in his sanad and gave an explanation of the truth of its history. He also does not display sanad from the history cited. Likewise with the history of the record, sometimes al-Sawi comments, but also the opposite.Keyword: Israiliyyat, attitude of al-ṣâwî, and ḥâshiyah al-ṣâwî
MURATTAL DAN MUJAWWAD AL-QUR’AN DI MEDIA SOSIAL
Authors: Masrurin ‘Ainatu
Published: October 2019

Qur’an as self-referential text within the word Quran and Qul represents the moral dimension involved. It is a dimension when the Quran frequently is applied as a narrative object in daily life by which, according to Frédéric Deny, it is called Performative. In aesthetic reception discourse, Qur’an as a text is placed as an object approached beautifully. For instance, it’s voiced by sound and rhythm called murattal or Mujawwad. The oldest quranic recording is found in 1855 by S Hurgronje in which it was a starting point for the Quran to recept digitally in media matters. Around 2000, Quranic Aesthetic reception in media tools Was widespread massively, then reading Qur’an in this time using Rythm isn't a matter to be coached directly. By phenomenological approach, this research tries to mapping the typology as well as the history of the reader (Qori) who changes to use social media as a listener and appreciator by uploading his/her reading. The result shows that three aspects are influencing the reader's acts in social media, 1) to show the existence of the Quran 2) religious narcism, 3) authoritative freedom. Keyword: Performative, Aesthetic Reception, Qari’, Social Media