bluuebunny/crossref_metadata_2025_split huggingface dataset:
Alemtuzumab (Campath IH) in Conditioning Therapy with Cyclophosphamide and Total Body Irradiation in Matched and Mismatched Unrelated Donor Transplantation of Children with Acute Lymphoblastic Leukaemia: A Report from 3 U.K. Centres.
Authors: Gharib Maged I. , Greenfield Hayley M. , Wynn Robert F. , Veys Paul , Naik Paru , Will Andrew M. , Cornish Jacqueline M. , Oakhill Anthony , Goulden Nicholas J. 
Published: October 2019

Abstract
Bristol has previously published extensively on the use of Campath antibodies during the unrelated donor cell transplantation of children with relapsed or otherwise high risk Acute Lymphoblastic Leukaemia (A.L.L.). These former reports (Br J Haematol1996;94(3):574–8; Blood1999; 94(7) 2236–46) concerned the use of Campath IG (a rat monoclonal antibody directed at CD52) given pre-transplant to the patients by intravenous infusion during conditioning therapy that also included cyclophosphamide (120 mgs/kg) and fractionated total body irradiation (1440 Gy). During this treatment the unrelated donor marrow was T cell depleted in vitro using Campath IM (also monoclonal and of rat origin). The results of such treatment demonstrated results comparable with HLA-matched sibling BMT, with low levels of acute graft versus host disease. In this communication transplant centres in 3 UK hospitals (Bristol, Great Ormond Street, London and Manchester) report their combined experience of using Campath IH (Alemtuzumab, a humanised monoclonal antibody - also directed at CD52) in the conditioning therapy of children with relapsed or otherwise high risk A.L.L.. The conditioning was otherwise again with Cyclophosphamide and fractionated total body irradiation. In this protocol a T cell replete graft was given and there was no in vitro T cell depletion of the unrelated donor marrow.
We report 34 successive patients where follow up for at least 12 months was available. The median age was 6.5 years and 28 patients received fully matched grafts (matched at HLA-A, -B, -C, -DRB1 and DQB1). The remaining patients received a graft that was mismatched at one of these class I loci. All patients engrafted and the median time to neutrophil count sustained above 0.5x10e9 / l was 20 days (range 12–58 days). Only tow patients experienced grade III acute GvHD and no patient experienced grade IV acute GvHD. 22 patients survive at a median follow up of 23.8 months including 4 of the patients who received a mismatched graft. There were only 3 deaths in the first 100 days following transplant (2 due to disseminated, invasive adenovirus and one to aspergillus). Beyond 100 days 3 further patients have died of adenovirus, 3 have relapsed, 2 have died of chronic GvHD and one died of a pulmonary haemorrhage of uncertain aetiology.
We therefore conclude the combination of Aletuzumab with cyclophosphamide and TBI is an effective and safe conditioning therapy for children with relapsed A.L.L. All patients engraft and there are low rates of acute and chronic GvHD.
Hypoxia-Independent Regulation of Hepatic Erythropoietin Production by Vascular Endothelial Growth Factor.
Authors: Tam Betty Y.Y. , Hoffman Jana , Park Sang-ki , Lee Jeng-Shin , Sundram Uma , Wu Grace , Schrier Stanley L. , Johnson Randall , Mulligan Richard , Kuo Calvin J. 
Published: October 2019

Abstract
Erythropoietin (Epo) is an essential regulator of red blood cell production. While adult Epo synthesis occurs almost exclusively in kidney, extrarenal Epo synthesis has been described in liver and brain in response to hypoxia or anemia. We have used adenoviruses to express soluble ectodomains of the VEGF receptors (sVEGFRs) Flk1 and Flt1 in the circulation of mice, achieving high-level VEGF sequestration, potent conditional ablation of VEGF function in adult animals, and inhibition of tumor growth and angiogenesis (Kuo et al. PNAS (2001). 98: 4605). Intriguingly, adenoviral expression of soluble Flk1 or Flt1 ectodomains, or a recently described soluble receptor VEGF Trap, is associated with profound stimulation of erythropoiesis and polycythemia, with hematocrit as high as 75% from basal levels of ~45%, and elevation of plasma Epo levels. sVEGFR expression results in a profound >100-fold induction of hepatic erythropoietin synthesis while renal Epo production is completely repressed. This effect is specific for adenoviruses expressing soluble VEGF receptors and not other receptor ectodomains, does not require liver transduction or hypoxia, and is not affected by Ad Cre-induced deletion of the hepatocyte HIF-1a gene in HIF-1aflox/flox mice. Selective inhibition of hepatocyte-produced VEGF in Ad Cre-treated VEGFflox/flox mice is sufficient to re-activate hepatocyte Epo production, likely via hepatocyte × endothelium crosstalk. These data suggest VEGF may function physiologically to repress hepatic Epo synthesis, while VEGF blockade leads to de-repression of hepatic Epo production via a novel hypoxia- and HIF-1a-independent pathway.
Sequential Enzymatic Down-Regulation and Degradation of the Lyn Tyrosine Kinase in Erythropoietin-Stimulated Cells.
Authors: Ingley Evan , Schneider Jessica R. , McCarthy David J. , Harder Kenneth W. , Hibbs Margaret L. , Alexander Warren S. , Hilton Douglas J. , Klinken Svend Peter 
Published: October 2019

Abstract
Erythropoietin (Epo) is a key molecule in the development of red blood cells. We have shown previously that the tyrosine kinase Lyn is involved in differentiation signals emanating from an activated Epo-receptor. We have recently identified Cbp (Csk binding protein, a negative regulator of Src) as a novel interactor with Lyn. The proline-rich region of Cbp associates with the SH3 domain of Lyn, which then phosphorylates Cbp on multiple tyrosine residues forming additional binding sites for the Lyn SH2 domain. Phosphorylated Y314 of Cbp, in turn, recruits the negative regulators Csk/Ctk, which phosphorylate Lyn and down-regulate its kinase activity. A phosphotyrosine-specific yeast two-hybrid system we developed, confirmed that the SH2 domain of Csk associated with Y314 of Cbp. Significantly, another well characterized negative regulator, SOCS1, also bound this phosphorylated tyrosine residue, resulting in elevated ubiquitination and degradation of Lyn. Thus, Cbp co-ordinates a two step down-regulation of Lyn. In Epo-responsive J2E cells, Cbp is rapidly phosphorylated by Lyn and the enzymatic activity of Lyn is suppressed by Csk/Ctk after 10 minutes of Epo stimulation. Subsequently, SOCS1 is involved in the turn over of Lyn protein two hours post-Epo induction. These data demonstrate for the first time a two phase process regulated by Cbp for inactivating and degrading Lyn after Epo stimulation.
Janus Kinases Interact with and Phosphorylate Rack-1 (Receptor for Activated C Kinase-1), a WD Motif Containing Protein.
Authors: Haro Takashi , Shimoda Kazuya , Kakumitsu Haruko , Kamezaki Kenjirou , Numata Atsuhiko , Harada Mine 
Published: October 2019

Abstract
We recently reported that Tyk2 was essential for IFN-a-induced B lymphocyte growth inhibition, although Stat1 is not required for this IFN-a-mediated inhibition. This means that other signaling molecules besides Stat1, and which are activated by Tyk2, are thought to transduce the IFN-a signal inhibiting B lymphocyte growth. We performed a yeast two-hybrid screen for proteins that interact with Tyk2, and identified Rack-1, originally described as a receptor for activated C kinase beta, associated with Tyk2. Receptor for activated C kinase (Rack)-1 is a protein kinase C interacting protein, and contains a WD repeat but has no enzymatic activity. In addition to protein kinase C, Rack-1 also binds to Src, phospholipase C gamma, and ras-GTPase-activating proteins. Thus, Rack-1 is thought to function as a scaffold protein that recruits specific signaling elements. In a cytokine signaling cascade, Rack-1 has been reported to interact with the IFN-alpha/beta receptor and Stat1. In addition, we show here that Rack-1 associates with a member of Jak, tyrosine kinase 2 (Tyk2). Rack-1 interacts weakly with the kinase domain and interacts strongly with the pseudo-kinase domain of Tyk2. Rack-1 associates with Tyk2 via two regions, one in the N-terminus and one in the middle portion (a.a.138–203) of Rack-1. In addition, not only Tyk2 but other Jak kinases associate with Rack-1, and each Jak activation causes the phosphorylation of Tyrosine 194 on Rack-1. After phosphorylation, Rack-1 is translocated from cytoplasm or membrane toward the perinuclear region. In addition to functioning as a scaffolding protein, these results raise the possibility that Rack-1 functions as a signaling molecule in cytokine signaling cascades.
Comparison of the Structure, Binding, and Activity of Cell Surface TFPIα and TFPIβ.
Authors: Piro Orlando , Broze George J. 
Published: October 2019

Abstract
TFPI is mainly produced by endothelial cells and alternative mRNA splicing generates two forms, TFPIα and TFPIβ. A small proportion of expressed TFPI remains associated with the cell surface through both direct (TFPIβ) and indirect (TFPIα) glycosylphosphatidylinositol (GPI)-mediated anchorage. TFPIβ lacks the C-terminus and Kunitz-3 (K3) domains of TFPIα and, instead, contains an unrelated C-terminus that directs the attachment of a GPI anchor. TFPIα and TFPIβ migrate with similar MWs (~45,000) on SDS-PAGE due to differing levels of glycosylation; removal of carbohydrate produces protein MWs of 36,000 and 28,000, respectively. Stable clones of ECV304 cells, a human cell line with some endothelial properties, that express reduced levels of TFPIα, TFPIβ, or both were produced using a plasmid-based small interfering RNA gene-silencing technique (siRNA) (Table 1); empty vector transfected clones served as controls. Total TFPI on the cell surface and in the medium of the selected clones was evaluated by flow cytometry (expressed as mean fluorescence intensity) and immunoassay, respectively (Fig. 1). Surface TFPI activity was determined by a two stage chromogenic assay based on the ability of each form to inhibit FX activation by FVIIa on cells with comparable levels of tissue factor (TF). Although TFPIβ accounted for only ~20% of the cell surface TFPI, it was responsible for most of the anti-FVIIa/TF activity (Fig. 2), suggesting a potential alternative role for surface TFPIα. At physiologic concentrations, rTFPIα expressed by ECV304 and mouse C127 cells binds to ECV↓TFPIα cells, but rTFPIα produced by mammalian A-293, HepG2, and CHO cells, and by E. coli does not, implying that post-translational modifications are important for the cell binding of TFPIα. Experiments in C127 cells show that optimal cell binding of TFPIα requires its basic C terminus and the P1 residue (R199) of its K3 domain; mutations within the putative heparin binding region of K3 (K232A, K240A, K241A) do not affect binding. Additional preliminary studies suggest that the cell binding of TFPIα requires N-linked glycosylation and is also reduced by mutation of serine 2, which is phosphorylated in C127 cells. Overall, these results call into question the physiologic relevance of previous cell binding experiments that relied on the use of high concentrations of rTFPIα expressed in E. coli.
table 1
Surface TFPIα (ng/107 cells) Surface TFPIβ (ng/107 cells) Each result represents the average of five independent experiments Cont 2.3 0.5 ECV↓TFPIα 0.4 0.5 ECV↓TFPIβ 2.4 0.04 ECV↓TFPIαβ 0.3 0.05
Influence of Hemoglobin Response to Epoetin Alfa on Quality-of-Life in Anemic Children with Cancer Receiving Myelosuppressive Chemotherapy.
Authors: Razzouk Bassem I. , Hockenberry Marilyn , Hinds Pamela S. , Feusner James , Rackoff Wayne , Hord Jeffrey D. 
Published: October 2019

Abstract
In a double-blind, placebo (PBO)-controlled study of children age 5–18 years (y) with cancer and anemia who received myelosuppressive chemotherapy (CT), the use of epoetin alfa (EPO) increased hemoglobin (Hb) levels (Hb increase ≥1 g/dL from baseline after 4 weeks [wks], independent of RBC transfusion in the previous 28 days; 65.0% for EPO vs 50.0% for PBO; P=.034) and reduced transfusion requirements (transfusion after wk 4 = 51.4% for EPO vs 69.4% for PBO; P=.008). However, no difference in the primary end point, self-reported quality of life (QOL) using the Pediatric Quality of Life Inventory (PedsQL-I™), was observed overall between the 2 treatment arms (ProcASCO 23:abstract 8527, 2004). In adults, increases in Hb during EPO therapy are associated with improved QOL (Cancer95:888, 2002). Given these findings, this post-hoc analysis of the pediatric study was undertaken to determine if a difference in QOL outcomes is observed when the Hb response of children is considered. The study included patients (pts) who were receiving myelosuppressive CT for malignant solid tumors (ST), Hodgkin’s disease (HD), acute lymphocytic leukemia (ALL), or non-Hodgkin’s lymphoma (NHL) and who were anemic (Hb <12 g/dL for boys >12 y, Hb <11 g/dL for girls >12 y, and Hb <10.5 g/dL for children 5–12 y) at study entry. Pts were stratified by tumor type (ST/HD or ALL/NHL) and randomized 1:1 to receive intravenous EPO 600 IU/kg or PBO weekly for 16 wks. The dose was increased to 900 IU/kg weekly after 3–4 wks if Hb increased by <1 g/dL. A total of 222 pts (111 EPO, 111 PBO) were included in primary analysis (98 ST, 75 ALL, 27 HD, and 22 NHL). For this analysis, Hb and PedsQL-I data beyond week 4 were available for 97 EPO pts and 89 PBO pts. PedsQL-I scores improved significantly from baseline among Hb responders but not among Hb nonresponders in both treatment arms (TABLE). The results of this analysis demonstrate that among pediatric cancer pts receiving myelosuppressive CT, Hb response is associated with improvement in QOL. The fact the Hb nonresponders reported higher QOL at baseline in the PBO group than the EPO group may explain, in part, why no difference in final QOL was observed overall in the primary analysis. The significant improvement in QOL among Hb responders suggests that additional studies are warranted to predict and optimize Hb response, QOL, and long-term outcomes with EPO treatment in anemic children with cancer.
Hb Response and PedsQL-I Score
Groups Hb Responder Baseline Final Change P-Value Data presented as mean ± SD (no. of patients) EPO Yes 69.9±17.15 (65) 79.1±15.53 (66) 9.2±20.20 (65) <.001 No 67.1±19.15 (29) 70.0±15.79 (31) 2.6±16.52 (29) .407 PBO Yes 70.0±15.68 (46) 78.0±17.38 (49) 8.5±13.10 (46) <.001 No 72.9±15.76 (40) 75.6±16.51 (40) 2.7±16.96 (40) .315
Molecular Serotype Analysis of Adenoviruses: Clinical Implications.
Authors: Lion Thomas , Ebner Karin , Matthes-Martin Susanne , Watzinger Franz 
Published: October 2019

Abstract
Adenovirus (AdV) infections are a major cause of morbidity and mortality in allogeneic stem cell transplant (SCT) recipients. Human adenoviruses represent a large family, currently including 51 serotypes, which are divided into 6 species (A-F). We have recently demonstrated by a species-specific RQ-PCR approach covering the entire spectrum of human adenoviruses that molecular detection of AdV in peripheral blood precedes the onset of life-threatening virus disease, and provides a basis for early preemptive treatment (Lion et al., Blood102(3):1114–20, 2003). An association of AdV species with clinical manifestation and response to antiviral therapy has been reported, but little is known about the clinical role of individual AdV serotypes. In immunocompromised patients, the use of serological testing for identification of adenovirus serotypes is limited due to the impaired immune response. We have therefore determined the nucleotide sequence of the complete AdV hexon gene in all 51 human serotypes, and identified regions permitting rapid serotyping at the molecular level. Serotypes belonging to the species A,B, C, E, and F, can be determined by fragment length analysis of a single PCR product, respectively. Serotype identification within the largest AdV species D requires sequencing of a single 300bp PCR amplicon. In view of the great predominance of species C in our region, we have also established real-time PCR tests permitting identification of its four serotypes. Analysis of all AdV C positive cases within more than 6.000 clinical specimens investigated at our center over the past years revealed the highest prevalence for serotype 2 (57%), followed by the serotypes 1 (39%) and 5 (4%). In some instances, two different AdV C serotypes were present simultaneously. We have demonstrated that the identification of specific virus strains within individual AdV serotypes, which may be required for investigation of possible transmission of infections within the hospital, can be achieved by sequencing of PCR products derived from an appropriate AdV target region within the hexon gene. The possibility of rapid molecular serotype and strain analysis provides a basis for studies on adenovirus epidemiology, and may in future have implications for the selection of the most appropriate antiviral treatment.
T to Myeloid Lineage Reprogramming by the Synergistic Action of C/EBP and PU.1.
Authors: Laiosa Catherine V. , Xie Huafeng , Ye Min , Graf Thomas 
Published: October 2019

Abstract
During hematopoiesis, transcription factors can not only direct differentiation of committed progenitors but also drive lineage commitment decisions from uncommitted multipotent progenitors. To further study their instructive role, we asked whether critical steps in the hematopoietic differentiation pathway could be reversed, and more specifically, whether myeloid transcription factors, C/EBP alpha and C/EBP beta, could re-direct committed T lymphoid progenitors into a myeloid fate. Previously in our laboratory, it was shown that C/EBP alpha efficiently reprograms CD19+ B cells to Mac1+ Gr1+ functional macrophages (Xie et al. (2004) Cell 117:663). To address lineage conversion of T to myeloid cells, we purified T cell subsets by MACS enrichment or FACS sorting and infected these cells with retroviruses expressing C/EBP alpha or C/EBP beta. Cells were cultured on myeloid supporting stroma with myeloid cytokines and analyzed for expression of antigens and marker genes by flow cytometry and RT-PCR. To rule out the presence of contaminating myeloid progenitors in our cultures, we started with a marked population of T lymphoid progenitors derived from Lck-Cre Rosa-EYFP reporter mice in which T cells are irreversibly marked with EYFP upon T cell commitment. T cell origin was further verified using PCR to detect TCRbeta DJ rearrangements. Using double negative DN3 and DN4 T cells, we show that a large proportion of EYFP+ DN cells infected with C/EBP alpha or C/EBP beta are induced to express surface antigens Mac1 and Gr1, express myeloid cytokine receptors and Fc receptors, and exhibit a myeloid morphology, compared to control infected cells. The reprogrammed cells are also functional as they can bind and phagocytose sheep red blood cells. In addition, the pan-T cell marker Thy1 and other T lineage genes including Lck, Rag2, and preT alpha are downregulated in reprogrammed cells. Furthermore, using a conditional knockout approach developed in Daniel Tenen’s laboratory, we show that endogenous expression of PU.1 is required for activation of Mac1 during this process. In conclusion, C/EBP alpha acts alone to extinguish the T lymphoid program while, in concert with PU.1, it activates the myeloid program leading to reprogramming of the T cell phenotype.
Health-Related Quality of Life (HRQL) in Adult Long-Term Hematopoietic Cell Transplant (HCT) Survivors: A Report from the Bone Marrow Transplant Survivor Study.
Authors: Bhatia Smita , Francisco Liton , Carter Andrea , Baker K. S. , Forman Stephen J. , Gurney James G. , Robison Leslie L. , Weisdorf Daniel J. , Grant Marcia 
Published: October 2019

Abstract
We describe HRQL reported by adult survivors of autologous and allogeneic HCT and the risk factors associated with poor HRQL. Eligible subjects were individuals undergoing HCT at either City of Hope or the University of Minnesota between 1974 and 1998, and surviving two or more years after HCT. We analyzed the data from 1003 HCT survivors older than 21 years enrolled in a retrospective cohort study who had completed a validated City of Hope HCT-QOL tool. Of these, 54.5% were males, 80.2% were Caucasians, and 55% had received allogeneic HCT. The primary diagnoses included CML (n=233), AML (n=241), NHL (n=200), ALL (n=99), HD (n=88), and other (n=142). The median age at HCT was 35.4, median age at study participation: 43.4 years, and the median length of follow-up: 9.4 (2 to 28 years). Survivors’ responses were compared with those of the population norms in Physical, Psychological, Social and Spiritual domains, and overall quality of life (QOL). Poor QOL was defined as QOL scores less than two standard deviations below that reported for the tool norms. Using this definition, 9.2% of this cohort was identified to have poor overall QOL; 13.7% with poor Physical; 10% with poor Psychological; 6.7% with poor Social; and 4.3% with poor Spiritual well-being. Table below shows the results of the risk factors identified for poor QOL by multivariate analysis. Age, sex, type of HCT, and time since HCT were not associated with poor QOL. Thus, overall 9% of the long-term HCT survivors report poor overall QOL. Certain vulnerable sub-populations exist, specifically, low household income, difficulty in obtaining health insurance, inability to return to work, and the presence of pain and anxiety that impact upon the HRQL of this population, and require additional attention and possible intervention.
Predictors of Poor
Risk Factors for Poor QOL Overall QOL Physical Psychological Social Spiritual Relative Risk (p-value) Relative Risk (p-value) Relative Risk (p-value) Relative Risk (p-value) Relative Risk (p-value) Non-white race 2.1 (0.06) 2.4 (0.005) 1.6 (0.1) 2.0 (0.06) 0.5 (0.2) Inability to return to work 3.4 (0.003) 4.9 (<0.001) 3.6 (<0.001) 3.6 (0.002) 0.8 (0.7) Inability to return to school 2.4 (0.2) 4.6 (0.005) 5.2 (0.002) 1.1 (0.8) 0.9 (0.9) Difficulty in obtaining health insurance 1.7 (0.1) 1.9 (0.04) 0.9 (0.9) 3.6 (<0.001) 2.2 (0.05) Absence of health insurance 2.6 (0.05) 1.3 (0.6) 0.7 (0.5) 1.6 (0.4) 4.2 (0.005) Household income <$20k 3.0 (0.04) 0.9 (0.9) 4.4 (0.006) 2.2 (0.1) 3.1 (0.05) CGVHD 2.1 (0.02) 1.8 (0.02) 1.9 (0.04) 1.4 (0.3) 1.5 (0.3) Severe pain 8.8 (<0.001) 12.6 (<0.001) 2.4 (0.07) 5.4 (0.001) 2.8 (0.07) Severe anxiety 240.2 (<0.001) 5.3 (0.001) 60.2 (<0.001) 7.8 (<0.001) 19.7 (<0.001)