bluuebunny/crossref_metadata_2025_split huggingface dataset:
Receptor-Based Identification of Novel Peptide Ligands as Inhibitors of Blood Stage Malaria.
Authors: Li Xuerong , Lauterbach Sonja B. , Lanzillotti Roberto , Chen Huiqing , Kane Ravi , Coetzer Theresa L. , Chishti Athar H. 
Published: October 2019

Abstract
Malaria parasite Plasmodium falciparum uses multiple receptors on the surface of human red blood cells to attach and invade host cells during blood stage infection. Glycophorins, including glycophorin A, B, and C have been implicated as host receptors and play an important role during Plasmodium falciparum invasion in human erythrocytes, particularly in the sialic acid-dependent parasite strains. To identify the parasite proteins that could bind to human glycophorins, we screened a phage display cDNA library of P. falciparum (FCR3 strain, a sialic acid-dependent strain) using human glycophorins and native intact human erythrocytes as bait. After four rounds of library screening and panning, 35 phage clones were identified that bound to purified glycophorins immobilized on the plastic surface. DNA sequencing of 12 phage clones revealed that they encode the same 7-amino acid sequence, ETTLKSF. Using immobilized intact human erythrocytes, a similar screening strategy led to the isolation of additional 20 phage clones, and the DNA sequencing of 6 such clones again revealed the same 7-amino acid sequence, ETTLKSF. In vitro binding of synthetic ETTLKSF peptide to purified glycophorins and intact erythrocytes was confirmed by ELISA and indirect immunofluorescence assays. Pull-down experiments demonstrated that the ETTLKSF peptide specifically interacts with glycophorin C, but not with glycophorin A and B, on human erythrocytes. The synthetic ETTLKSF peptide blocked merozoite invasion of P. falciparum in human erythrocytes in a dose-dependent manner, whereas the control peptides were without any effect. We have named the ETTLKSF peptide as GBL-1, the Glycophorin Binding Ligand-1. We propose that further characterization of GBL-1, the first peptide ligand that specifically binds to human glycophorin C, could lead to the development of novel anti-malarial agents that prevent P. falciparum invasion in human erythrocytes via the glycophorin C receptor.
Quantitative Bioluminescent Method Appropriate for Bimodality Analysis To Improve and Follow a Syngenic Murine Model of B-Cell Lymphoma Expressing Human CD20.
Authors: Daydé David , Dommange Florence , Lerondel Stéphanie , Golay Josée , Le Pape Alain , Paintaud Gilles , Bardos Pierre , Watier Hervé , Cartron Guillaume 
Published: October 2019

Abstract
The anti-CD20 monoclonal antibody rituximab (C2B8) has shown promising results in the clinical treatment of patients with B-cell Non-Hodgkin’s Lymphoma (B-NHL). However, its therapeutic effect is variable whereas a better knowledge of factors affecting rituximab response could lead to improve its efficacy. It has been suggested that tumour burden could influence rituximab exposure and response in human. The lack of method to assess precisely tumour burden has however prevent to confirm such influence. Study of factors affecting antibody exposure requires bimodality analysis allowing to precise factors related to antibody from those related to tumour. We purposed to develop a quantitative method of tumour burden using bioluminescence imaging (BLI) and simultaneously scintigraphic study of monoclonal antibody biodistribution and exposure (indium-111 labelled ibritumomab, 2B8). Firstly, we developed a murine model of T-lymphoma cells transduced with human CD20 cDNA (EL4-hCD20) and transfected with luciferase plasmid (EL4-hCD20-luc). C57Bl6J mice and C57Bl6J TyrC2-J mice (albino phenotype) were injected with 8.10^3 EL4-hCD20-luc lymphoma cells by i.v. route. All mice died within 30 days with liver, spleen, bone marrow and lymph nodes involvement confirmed by immunohistochemistry and PCR analysis. In order to evaluate tumour growth and dissemination, potassium luciferin (2 or 4 mg) were injected bi-weekly intra peritoneally and tumour burden was analysed using Hamamatsu® CCD imaging system. Background noises and cosmic rays, which disturb the signal of bioluminescence, were eliminated from merging image before bioluminescent activity analysis. The sensitivity of analysis was below than 10^2 El4-hCD20-luc cells and there was proportionality between bioluminescent activity and cells number from 10^3 to 10^9 cells. The use of different binning allowed the follow-up of tumour burden evolution from 9 days after inoculation until the death of mice. We validated the used of black phenotype C57Bl6J compared to C57Bl6J TyrC2-J phenotype and demonstrated that the amount of luciferin could influence tumour growth. Regions of interest were delimited by segmentation method and we developed a reliable quantification method of bioluminescence activity taking into account absorption coefficients of the cells light emitted according to their localization. Such model and quantification method makes possible the follow-up of tumour growth and monoclonal antibody efficacy and exposure studied by bimodality analysis. D.D. is supported by a grant from Région Centre.
Direct and Enhanced Cytotoxicity of the Bcl-2 Family Inhibitor GX15-070 on Rituximab-Sensitive and Rituximab-Resistant B-NHL Clones.
Authors: Martinez-Paniagua Melisa A. , Vega Mario I. , Huerta-Yepez Sara , Martinez-Miguel Bernardo , Hernandez-Pando Rogelio , Gonzalez-Bonilla Cesar R. , Bonavida Benjamin 
Published: October 2019

Abstract
Rituximab (chimeric anti-CD20 mAb) has been used for the treatment of Non Hodgkin B cell lymphomas (B-NHL), alone or in combination with CHOP. However, a subset of patients does not respond to treatment or develops refractoriness to further treatments. Therefore, there is an urgent need to develop new alternatives to treat these patients. We have reported that treatment of B-NHL cell lines with rituximab inhibits anti-apoptotic survival pathways and down regulates the expression of anti-apoptotic Bcl-2 family proteins (i.e. Bcl-2/Bcl-xl) resulting in sensitization to chemotherapeutic drugs. Further, rituximab-resistant clones showed over-expression of anti-apoptotic gene products (Jazirehi et al., Cancer Research1:1270–81, 2007). Therefore, we hypothesize that inhibitors of anti-apoptotic Bcl-2 family may reverse the resistance to apoptotic stimuli. We examined chemical inhibitors that mimic natural ligands of the anti-apoptotic BH3-only proteins. GX15–070 (Gemin X Biotechnologies, Inc., Canada) inhibits Bcl-2 protein-protein interactions resulting in Bak and Bax oligomerization, release of cytochrome C, and activation of caspases (Shore and Viallet, Hematology, 2005; ASH, 226–230). Treatment of B-NHL cell lines (Raji, Ramos, 2F7, DHL-4) with subtoxic concentrations of GX15–070 (20–50 uM) resulted in inhibition of cell proliferation and subsequently induction of apoptosis as determined by TUNEL. There was a time and concentration-dependent effect of GX15–070 on cytostasis and apoptosis. Analysis of cells treated with GX15–070 by western blotting revealed that Bcl-2, Bcl-xl, and Mcl-1 protein expressions were significantly inhibited as compared to controls. The protein inhibition by GX15–070 was not expected and needs further investigation. We also examined the effect of combination treatment of GX15–070 with the chemotherapeutic drug CDDP and there was an additive or synergistic cytotoxic effect. Treatment of rituximab-resistant clones (generated from 2F7, Raji and Ramos) treated with GX15–070 resulted in significant inhibition of cell growth and apoptosis. The cytotoxicity of GX15–070 in the B-NHL cell lines was tumor specific, because treatment of human peripheral blood leukocytes from different donors did not show any cytotoxic effect. Likewise, treatment of nude mice with different concentrations of GX15–070 did not show any detectable toxicity. These findings demonstrate that GX15–070 is cytotoxic to various drug/rituximab-resistant B-NHL cell lines and is not toxic to normal human leukocytes. This study suggests that combination of GX15–070 with subtoxic concentrations of chemotherapeutic drugs may have additive/synergistic effects. The present findings support the potential therapeutic application of GX15–070 in the treatment of patients with B-NHL that are resistant to current therapies.
Hematopoiesis in Regenerated Bone Marrow on the Hydroxyapatite Scaffold.
Authors: Fujita Atsushi , Migita Makoto , Ueda Takahiro , Fukunaga Yoshitaka , Shimada Takashi 
Published: October 2019

Abstract
Background: In bone marrow (BM), osteoblastic cells lining the endosteal surface are a key component of the niche to promote and regulate hematopoietic stem cells (HSCs). Anatomical architecture of BM could be regenerated by culturing BM derived stromal cells on the hydroxyapatite (HA) scaffold. In the present study, we examined whether the regenerated BM has the ability to support HSCs in vivo.
Methods: Our study was designed as follows;
BM stromal cells from C57/BL6 mice (Ly5.2) were cultured on the HA scaffold with numerous small pores for 3 days in vitro and the scaffold with attached cells was implanted subcutaneously onto the back of C57/BL6 recipient mice. 4.0x105 Lineage negative (Lin−) Ly5.1 BM cells transduced with a lentiviral vector containing the luciferase (Luc) gene were intravenously administered into the recipient mice after lethal irradiation. Eight weeks after transplantation, the scaffolds were removed from the first recipient mice and subcutaneously transplanted into the lethally irradiated second recipient mice. The mice also received fresh Ly5.2 BM cells for survival. Biodistribution and kinetics of Luc+ Ly5.1 cells were monitored by in vivo bioluminescence imaging and FACS.
Results: In the secondary transplanted mice, Luc+ hematopoitic cells were detected in the scaffolds for at least 6 months after transplantation. Subcutaneous injection of G-CSF resulted in wide distribution of bioluminescence signals from the original scaffolds to whole body including the head, extremities, chest, and abdomen. The presence of Ly5.1 B and T lymphocytes in the circulation was confirmed by FACS analysis 5 months after secondary transplantation.
Conclusions: The regenerated BM on the HA scaffold is capable of supporting HSCs in vivo suggesting that the functional niche is reconstituted. Hematopoiesis in the regenerated BM may have a significant impact for development of new therapeutic strategies for various hematopoietic diseases.
Three Restricted Forms of Epstein-Barr Virus Latency Counteracting Apoptosis in c-Myc Expressing Burkitt Lymphoma Cells.
Authors: Kelly Gemma L. , Stylianou Julianna , Bell Andrew I. , Wei Wenbin , Rowe Martin , Rickinson Alan B. 
Published: October 2019

Abstract
Epstein-Barr virus (EBV) is aetiologically linked with Burkitt Lymphoma (BL) but its contribution to lymphomagenesis, versus that of the chromosomal translocation activating c-myc expression, remains unclear. This is in part because the full virus growth transforming programme that is expressed when EBV infects normal resting B cells, is not expressed in BL. Instead EBV in BL normally exhibits a restricted Latency I form of infection characterised by expression of only one latent antigen EBNA1 from the BamHI Q promoter. Here we describe an endemic BL, Awia, which uniquely is heterogeneous at the single cell level for EBV gene expression. Analysis of single cell clones of Awia-BL revealed cells displaying three forms of restricted EBV latency:
classical Latency I, ’Wp-restricted latency’ expressing EBNAs 1, 3A, 3B, 3C and -LP, and a novel ’EBNA2+/LMP1- latency’ in which all EBNAs including EBNA2 are expressed without the latent membrane proteins LMPs 1 and 2.
Comparison with rare EBV-negative clones from the same tumour showed that each form of infection provides the c-myc-expressing BL cells with a specific degree of protection from apoptosis. Microarray analysis was carried out on the isogenic Awia-BL clones to determine the influence of EBV gene expression on cellular transcription. Interestingly it was found that expression of the pro-apoptotic Bcl2 family protein BIM, which has previously been implicated in c-myc induced apoptosis, was downregulated in the apoptosis resistant BL cells. Our work suggests that EBV acts as an anti-apoptotic rather than a growth-promoting agent in BL by downregulating expression of the cellular BIM protein. In addition the microarray analysis on these isogenic Awia-BL clones showed that the EBNA profile influences the differentiation status of the BL cell on the germinal centre to plasmacytoid differentiation pathway. These findings may reflect viral functions that are important not just for BL pathogenesis but also for EBV’s normal strategy of persistence in the B cell system.
SB1317, a Potent and Orally Active FLT3-CDK Inhibitor with High Anti-Tumor Efficacy in Models of Hematological Malignancies.
Authors: Goh Kee C. , Ong Wai C. , Hu Changyong , Liang Ai L. , Stunkel Walter , Tan Yong C. , Sangthongpitag Kanda , Goh Siok K. , Bonday Zahid Q. , William Anthony D. , Dymock Brian W. , Kantharaj Ethirajulu , Wood Jeanette M. 
Published: October 2019

Abstract
FLT3 is the most common mutated gene in acute myeloid leukemia (AML), and FLT3 mutations are strongly correlated with poor prognosis. Small-molecule inhibitors of FLT3 kinase have been evaluated in clinical trials but with limited success due to the emergence of activating mutants other than FLT3. CDK1, 2, 4 and 6 are well-established anti-cancer targets due to their direct role in cell cycle control. CDK7 and 9 are transcriptional CDKs that are emerging anti-cancer targets as a result of recent advances in the understanding of their effects on apoptotic regulators. Recent evidence suggests that the combined targeting of CDK1, 2 and 9 enhances apoptotic killing of tumor cells. Most CDK active compounds currently in clinical development do not target all of these CDKs and have poor pharmacokinetic/pharmacodynamic properties. Compounds targeting these CDKs, in addition to FLT3, may be more effective in AML and also be effective against other hematological as well as solid tumors. SB1317 is a novel potent inhibitor of FLT3 kinase (IC50 = 45 nM) and CDK2 (IC50 = 11 nM). The CDK spectrum also includes potent inhibition of CDK1 and 9 (IC50 = 19 and 10 nM respectively). SB1317 inhibits proliferation of a broad panel of tumor cell lines, and is particularly potent against the mutant FLT3-dependent AML cell line, MV4-11 (proliferation IC50 = 18 nM). SB1317 reduced phospho-Rb and phospho-FLT3 levels in MV4-11 cells in a dose-dependent manner. It induced apoptosis in MV4-11 cells as well as in primary AML cells from patients. The pharmaceutical, metabolic and pharmacokinetic properties of SB1317 render it amenable to oral dosing. In a nude mouse subcutaneous model of AML (MV4-11), SB1317 induced complete tumor regression after oral daily dosing (40 mg/kg for 21 days). In an orthotopic model of AML (HL60), SB1317 significantly prolonged median survival time (59 days versus 40 days) after treatment at 100 mg/kg p.o. 2d on/5d off. It also showed significant anti-tumor activity in a nude mouse model of B-cell lymphoma (Ramos) with a tumor growth inhibition of 63% (15 mg/kg i.p. 5d on/5d off). These results demonstrate the therapeutic potential of this novel kinase inhibitor for the treatment of hematological malignancies.
Use of Enoxaparin Versus Unfractionated Heparin in the Prevention of Venous Thromboembolic Events in Acutely Ill Medical Patients: A Meta-Analysis of Individual Patient Data.
Authors: Mismetti Patrick , Liotier Jérôme , Kleber Franz-Xaver , Sherman David G. , Moulin Nathalie , Laporte Silvy 
Published: October 2019

Abstract
Background The ACCP guidelines recommend prophylaxis with low-molecular-weight-heparin (LMWH) or unfractionated heparin (UFH) to prevent venous thromboembolism (VTE) in acutely ill medical patients. However, there is a paucity of data on the relative benefits of using LMWH versus UFH. A meta-analysis was performed using individual patient data to assess the efficacy of the LMWH enoxaparin versus UFH for VTE prophylaxis in hospitalized medical patients.
Methods A systematic literature review was conducted to identify randomized clinical trials comparing enoxaparin 4000 IU S.C. once-daily versus UFH administered S.C. twice (2 × 5000 IU) or three-times daily (3 × 5000 IU) for VTE prophylaxis. Data from electronic database searches were supplemented by information from trial registries, abstract books, meeting proceedings and the trial sponsors. All investigators or sponsors were contacted to provide individual patient data. Recorded outcomes were VTE, major bleeding, and deaths at day 14 as adjudicated by the adjudication committee of each trial. All trial analyses were performed on an intention-to-treat basis. The pooled analysis was based on the logarithm of the relative risk (RR) weighted by the inverse of the variance.
Results Individual patient data were obtained from 3600 patients enrolled in 4 randomized clinical trials, 3 in stroke patients and 2 in patients with various acute illnesses. The data from 1 additional trial is currently unavailable (100 patients). The median age of evaluated patients was 71 years; 51% are male; 32% presented with a creatinine clearance <50 mL/min. Meta-analysis of the overall incidence of VTE (including asymptomatic DVT, symptomatic VTE and PE-related death) showed a 38% significant reduction in favor of enoxaparin (RR=0.62 [95% CI 0.50–0.77], p<0.001, heterogeneity test pHet=NS). A non-significant 13% increase in major bleeding was also observed with enoxaparin (RR=1.13 [95% CI 0.53–2.44], p=0.75, pHet=NS). Finally, there was a non-significant 16% reduction in death with enoxaparin (RR=0.84 [95% CI 0.63–1.10], p=0.20, pHet=NS).
Conclusion The results of this meta-analysis support superior efficacy of enoxaparin, with a non-significant increase in major bleeding, compared with UFH for VTE prophylaxis in acutely ill medical patients.
Bleeding and Thromboembolic Events after Termination of Therapy with Idraparinux for Prevention of Recurrent Venous Thromboembolism-Observation after the van Gogh Trials.
Authors: Harenberg Job , Joerg Ingrid , Hagedorn Antje , Giese Christina 
Published: October 2019

Abstract
The long acting polymethylated antithrombotic compound Idraparinux has been investigated in the vanGogh trials for prevention of recurrent venous thromboembolism (VTE) using once weekly subcutaneous injections for 3–6 months (vanGogh PE and DVT trial) and for additional 6 months (vanGogh Extension trial). After termination of the studies Idraparinux was eliminated with a half live of 60 days or longer (Harenberg et al, submitted). We followed-up the patients (n=23) at our center for bleeding events, recurrent VTE and other severe events over 15 months. During the study period 2 major bleeding complications occurred in patients randomized to warfarin (days 70 and 186). During follow-up, 2 patients initially randomized to Idraparinux suffered from major bleeding events (days 65 and 140 during follow-up). Bleeding was related to an additional therapy of warfarin or low-molecular-weight heparin. At that time, the long elimination half life time of Idraparinux was unknown. One patient required continuation of anticoagulation due to homocygous factor V Leiden mutation and received warfarin starting one week after termination of idraparinux. At day 65 he was hospitalized because of a major intramuscular bleeding. One patient underwent total hip replacement at day 140 of follow-up receiving nadroparin for prophylaxis of VTE and developed a major postoperative bleeding with reoperation and transfusion. The concentrations of Idraparinux were 0,16μg/ml in each patient. Thereafter the long elimination time of Idraparinux was taken into consideration. In 1 of the patients warfarin was started at a 0.1μg/ml Idrparinux and titrated slowly into an INR range of 1.6 to 2.0 until Idrapaninux decreased to 0.05μg/ml. In the other patient postoperative prophylaxis of VTE was performed with LMWH at 0,01μg/ml Idraparinux (day 571 of follow-up). No adverse events occurred in both patients. During follow-up VTE occurred in 2 patients each initially randomized to warfarin (days 58 and 406) and Idraparinux (days 266 and 381), respectively. The concentrations of idraparinux were <0.01μg/ml in both patients. The data show that the very long half life of Idraparinux may lead to serious bleeding complications, if anticoagulation is given on top of the circulating compound within 3 or 4 months after termination of therapy. Cautious anticoagulation during this period may avoid bleeding. Recurrent VTE occurs late after termination of Idraparinux but in probably to the same frequence as after termination of warfarin.
Development and Performance of the GTI Diagnostics ADAMTS-13 Activity Assay (ATS-13™) and ADAMTS-13 Inhibitor Assay (ATS-13™ INH).
Authors: Pressman Melissa A. , Wuitschick Elizabeth , Chance Suzette C. 
Published: October 2019

Abstract
Thrombotic Thrombocytopenic Purpura (TTP) is a disease characterized by the sudden onset of a classical pentad of symptoms: thrombocytopenia, fever, renal insufficiency, neurologic deficit, and microangiopathic hemolytic anemia. ADAMTS-13 is the protease responsible for cleaving von Willebrand Factor (VWF); decreased levels or deficiency of ADAMTS-13 activity has been demonstrated in the plasma of TTP patients. The lack of ADAMTS-13 activity results in the accumulation of multimers of VWF in the plasma and ultimately intravascular platelet aggregation resulting in the clinical symptoms associated with TTP. The presence or absence of inhibitory antibodies directed against ADAMTS-13 influences the treatment protocol for these patients. GTI has developed a rapid, quantitative assay for ADAMTS-13 activity that is based on the cleavage of a von Willebrand Factor substrate and subsequent fluorescent detection. A method comparison was done of GTI’s ADAMTS-13 Activity Assay (n=397) using normal plasma, plasma from suspected TTP patients, and plasma from patients suspected to have other coagulation diseases (ITP, HUS, HIT, APS). All samples were also run on the collagen-binding (n=197) or the Furlan method (n=200) for comparison. This data demonstrated good correlation. Assay precision (with-in run, n=20 and assay-to-assay, n=40) was evaluated and there was less than 20% cv total imprecision for samples with levels of activity across the assay range. In addition, GTI has developed an assay for the semi-quantitative detection of inhibitory antibodies directed against the ADAMTS-13 protease. This assay is similar in format and principle to our ATS-13™ kit; however inhibition of the ADAMTS-13 protease activity is determined in the patient’s heat-inactivated sample following mixing with a provided control at a single dilution. Assay precision and preliminary method comparison data will be presented.