bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD47 Glycoprotein Interacts with p4.1R and p55 in the Erythrocyte.
Authors: Avent Neil D. , Plummer Zoe E. , Head David J. 
Published: October 2019

Abstract
CD47 is a 47–50kDa membrane glycoprotein with 5 known isoforms. The role of CD47 within the erythrocyte membrane remains the subject of much research and debate though we recently provided evidence that CD47 may function as an inducer of eryptosis (Head et al, 2005). As both a cytoskeletal linked fraction and a smaller membrane diffuse fraction of CD47 exists, it is most likely that there are a number of protein species that are able to bind to CD47 at its cytoplasmic face. Our research has focused on a study of the molecular interactions of erythrocyte CD47 with erythrocyte membrane skeletal proteins protein 4.1R (p4.1R), protein 4.2 (p4.2) and p55. Here we demonstrate the ubiquitous expression of all CD47 isoforms in haemopoietic cells and tissues using basic and real-time PCR. Via immunoprecipitation of CD47 from mature erythrocyte membranes using the anti-CD47 mAb BRIC-126, yeast two-hybrid analysis and in vitro co-immunoprecipitation of 35[S] labelled peptides in a cell-free translation procedure, a novel ternary complex involving CD47, p55 and p4.1R has been indicated. More specifically, the potential interaction between p55/p4.1R and the cytoplasmic face of CD47 has been localised to the PDZ and FERM domain of these proteins respectively. Though research suggests p4.2 provides the major cytoskeletal attachment of CD47 (Bruce et al, 2002; Mouro-Chanteloup et al, 2003), a direct interaction between CD47 and p4.2 was not suggested by our study and remains undemonstrated. We continue to further the evidence for a functional role of CD47 in eryptosis. We propose p4.1R links CD47 to the apoptotic machinery of the cell and suggest a mechanism whereby cytoskeletal rearrangement and PS exposure occurs. 4.1null cells have been obtained and are currently being investigated. Further characterisation of the eryptotic pathway may offer insight into potential therapies for erythroleukaemia characterised by resistance of the erythroid lineage to apoptosis. This work has equal significance in the stabilization of red cell preparations in the blood transfusion setting.
Gaucher Disease Psychosine Mis-Traffics Endocytosed Cargo to a Non-Functional, Pre-Lysosomal Compartment.
Authors: Roos Jonathan C. , Wang Susan Z. , Cox Timothy M. 
Published: October 2019

Abstract
Gaucher’s disease is a rare metabolic disorder resulting from a deficiency in glucocerebrosidase activity. It is characterized by the formation of large multinucleate macrophages which accumulate enzyme substrate - principally glucosylsphingosine (psychosine) and glucosylceramide - as well as unrelated products such as cholesterol, lipofuscin, glycopeptides and oligosaccharides. The mechanism of this storage remains obscure. Psychosines have been shown to inhibit cytokinesis (Kanazawa et al., 2000) in cells expressing a G-protein linked receptor: GPR65 (Im et al., 2001). Signalling via GPR65 was shown to affect phosphorylation and calcium release as well as result in the formation of intracellular actin clots. Could such psychosine-triggered signalling account for the secondary accumulation of lipids that are not substrates for glucocerebrosidase in Gaucher disease? We investigated the effects of psychosines and other lysosphingolipids on human macrophages and monocyte-related cell lines expressing GPR65. Trafficking was evaluated by pulse-chase experiments using fluorescent lipids and markers of pH. Changes induced by lysosphingolipids were visualized by electron microscopy and compared with Gaucher cells from affected spleen. Confocal fluorescence and electron microscopy were used to identify sub-cellular compartments. GPR65 expression in Gaucher spleen and other cells was confirmed by RT- PCR.
At sub-pathophysiological concentrations (1-5μM), psychosines but not ceramides disrupt endocytosis: mis-trafficked membrane and ingested materials accumulate in a large vesicular compartment. This multivesicular body bears most lysosomal markers including LAMP1 and LAMP2 but has a neutral pH and does not contain acid phosphatase or Cathepsin D -consistent instead with late endosomes. The ultrastructure of the psychosine-treated monocytic cells closely resembles naturally occurring splenic Gaucher cells and both are also of course multinucleate. This suggests that psychosines dysregulate intracellular trafficking in Gaucher disease. Recycling is perturbed and membrane lipids and other cargo accumulate in a pre-lysosomal compartment where, with glucosylceramide, they may be preferentially concentrated. Since related lysolipid metabolites are present in several other sphingolipidoses, we propose that a common trafficking mechanism accounts for the cellular pathology seen in this class of diseases.
In addition, an active role for psychosines in pathogenesis may also begin to suggest mechanisms to explain why Gaucher patients carry a high relative risk for developing myeloma and other B-cell dyscrasias (Rosenbloom et al., 2005). The majority of these malignancies are associated with mutations at position 14q32. Receptor GPR65 is also located at that position and its regulation may predispose to mutation and eventual transformation.
Intracellular HMGB1 Transactivates the Human IL1B Gene Promoter through Association with an ETS Transcription Factor PU.1.
Authors: Tsukada Junichi , Mouri Fumihiko , Mizobe Takamitsu , Higashi Takehiro , Izumi Hiroto , Kohno Kimitoshi , Tanaka Yoshiya 
Published: October 2019

Abstract
High mobility group box chromosomal protein 1 (HMGB1) was recently identified as a late proinflammatory mediator of endotoxin lethality, which amplifies and sustains the inflammatory processes via macrophage/monocyte activation by extracellular release of HMGB1. However, it was originally described as a nonhistone nuclear protein to bind and distort the structure of DNA and to regulate transcription through association with several transcription factors. Here we investigated the effect of intracellular HMGB1 on transcriptional gene regulation of an immediate early proinflammatory cytokine IL-1β in transient transfection studies using HMGB1 expression vector (pcDNA3HMGB1) and murine macrophage cells RAW264.7. When pcDNA3HMGB1 was introduced into RAW264.7 cells with pGL3HT reporter containing the -131 to +12 minimal IL-1β gene promoter, IL-1β promoter activity was significantly induced by expression of HMGB1. The gene promoter possesses two important transcription factor binding motifs, one for PU.1, a myeloid and B cell-specific transcription factor that belongs to the ETS family, and the other a binding site for NF-IL6. To verify the functional role of intracellular HMGB1 in transactivation of the IL-1β promoter, a PU.1 expression vector and/or pcDNA3HMGB1 were cotransfected into PU.1-deficient murine thymocytes EL4 cells along with pGL3HT reporter. As a result, HMGB1 synergized with PU.1 to transactivate the IL-1β promoter, but not HMGB1 alone. This argument was supported by our GST-pulldown data, which demonstrated direct physical interaction of HMGB1 with PU.1. In addition, deletion of the PU.1 winged helix-turn-helix DNA-binding domain significantly inhibited the association of PU.1 with HMGB1. To determine whether HMGB1 could affect PU.1 DNA-binding affinity, we performed electrophoretic mobility shift assay using a radiolabeled IL-1β -59 to +12 promoter element (DT), recombinant PU.1 (rPU.1) and GST-HMGB1. Two complexes with slower and faster mobilities were generated by the addition of HMGB1 to a mixture of rPU.1 and DT probe. The two complexes were abrogated by preincubation with anti-PU.1 Ab, while anti-HMGB1 Ab reacted only with the complex with a slower mobility, indicating that the complex with a slower mobility formed by addition of HMGB1 contained both HMGB1 and PU.1, while the band with a faster mobility contained only PU.1. From the present study, we propose that intracellular HMGB1 might function as a coactivator in PU.1-mediated transcriptional activation, which facilitate access of PU.1 to specific DNA targets. The fact that PU.1 is a transcription factor essential to macrophages/monocyte-specific proinflammatory cytokine genes further raise the possibility that interaction of PU.1 with HMGB1 may play an important role in the inflammation cascade in macrophages/monocytes.
Thrombopoietin (TPO) Enhances Heamatopoietic Differentiation of Human Embryonic Stem Cells.
Authors: Srivastava Anand S. , Mishra Rangrath , Carrier Ewa 
Published: October 2019

Abstract
Recently, it was demonstrated that TPO enhances hematopoietic differentiation of primate ES cells, but its role in differentiating human ES cells is unknown. We sought to investigate the regulatory mechanism of TPO induced signals mediated by the c-mpl cytoplasmic domain during human embryonic stem (hES) cells hematopoietic commitment. We hypothesize that in human embryonic stem cells, binding of TPO to its c-mpl receptor causes three-dimensional alterations which bring the c-mpl cytoplasmic domain and Janus Kinase into close-proximity and thus induces the phosphorylation and dimerization of STAT5 molecule. Dimerized STAT5 molecules detach from the receptors and migrate to the nucleus where they bind GAS site and induce transcription of a set of target, hematopoiesis-related genes. NIH human ES cell lines (WI01) were used in this experiment. In brief, to induce EB formation, cells were incubated in differentiation medium, which consisted of knockout DMEM medium (GIBCO/BRL, Carlsbad, USA), supplemented with 20% non-heat-inactivated fetal bovine serum (FBS, Hyclone, USA), 1% nonessential amino acids, 1 mM L-glutamine, and 0.1 mM β-mercaptoethanol. Subsequently, DMEM was replaced by IMDM (GIBCO/BRL, USA) with the same supplements and additional two cytokines (100 ng/mL SCF and 100 ng/mL Flt-3 ligand (Flt-3L)) (control group). To investigate the role of TPO and VEGF, cells were additionally treated with 100 ng/mL TPO alone or in combination with 100 ng/mL rhVEGF. All cytokines were from the R&D systems (USA). Significant increase in the numbers of embryoid bodies (EBs) formation in TPO (125/105), TPO/VEGF (150/105 cells) when compared to controls (10/105 planted ES cells) was documented. This corresponded with the increase in CFU-C and the number of CD31/CD34 positive and CD34-positive progenitors. Analysis of gene expression during hematopoietic development demonstrated that TPO/VEGF combination increased mRNA expression of the TPO receptor (TPO-R) and VEGF (VEGF-R) receptors in hematopoietic progenitors obtained from human ES cells. We are in the process of determining the role of JAK/STAT pathway in this process; functional studies involve blocking of TPO/c-mpl using TPO-R-specific antibodies and determining its impact on human ES-derived hematopoiesis.
The Detection and Estimation of Fetal Hemoglobin by Means of the Alkali Denaturation Test
Authors: JONXIS J. H. P. , HUISMAN T. H. J. 
Published: October 2019

Abstract
This study illustrates that the value of both alkali denaturation methods is somewhat limited. At high percentages of fetal hemoglobin, the results obtained with the method of Singer and coworkers, are about 10 per cent too low. The Brinkman and Jonxis method is not accurate when the percentage of fetal hemoglobin is lower than 10 per cent. The alkali resistant fraction found with the method of Singer and associates in adult blood, is a mixture containing fetal hemoglobin. The presence of COHb-A will interfere with the determination of fetal hemoglobin, especially when the optional alkali denaturation method is used.
Polycythemia Associated with Disturbed Function of the Respiratory Center
Authors: PARE PETER , LOWENSTEIN LOUIS 
Published: October 2019

Abstract
1. An unusual case of absolute polycythemia associated with hypoxemia, probably resulting from abnormal function of the respiratory center has been presented.
2. The mechanism of development of this type of polycythemia has been discussed.
3. A classification of polycythemia on the basis of blood volume and arterial oxygen saturation has been suggested.
The Effect of Chloramphenicol and Other Antibiotics on Leukocyte Respiration
Authors: FOLLETTE JAMES H. , SHUGARMAN PETER M. , REYNOLDS JOHN , VALENTINE WILLIAM N. , LAWRENCE JOHN S. 
Published: October 2019

Abstract
1. Leukocyte homogenates show active, reproducible respiratory activity in a fortified system.
2. Chloramphenicol inhibits leukocyte respiration in vitro when used in concentrations similar to other commonly used inhibitors.
3. Chlortetracycline, oxytetracycline, penicillin and streptomycin have no significant effect on leukocyte respiration.
4. Urethane and atabrine produce inhibition of leukocyte respiration, but the latter is more active.
5. The possible sites of action of chloramphenicol in cellular metabolism are discussed.
Studies on Urinary Excretion of Vitamin B12Co60 in Pernicious Anemia for Determining Effective Dosage of Intrinsic Factor Concentrates
Authors: BEST WILLIAM R. , WHITE WILFRID F. , ROBBINS KENNETH C. , LANDMANN WENDELL A. , STEELMAN SANFORD L. 
Published: October 2019

Abstract
Studies were made in pernicious anemia patients on the urinary excretion of B12Co60 after a small oral dose followed by a large parenteral injection of nonradioactive vitamin B12.
(1) Increasing doses of intrinsic factor concentrates give increasing excretions of radioactivity at low doses; little additional increase at moderate doses; and at times a subsequent diminution at excessive doses. Data on 34 tests of a particular intrinsic factor concentrate in 18 pernicious anemia patients tend to support an excretion proportional to the logarithm of intrinsic factor dosage at low to moderate levels, but do not exclude the possibility of a linear approach to a plateau.
(2) Assay by hematologic response was compared with the urinary excretion tests in 13 pernicious anemia patients. This data shows a relation between the two tests though the correlation is far from complete.
(3) Methods are outlined for testing all intrinsic factor preparations with the same amount of tracer vitamin B12 as will be incorporated commercially. More sensitive comparative tests of similar intrinsic factor preparations may be made using smaller amounts of B12Co60.
(4) The literature is reviewed to determine which variations in technic might lead to the most reliable quantitation of intrinsic factor activity.
Experimental Research on the Proliferative and Differentiative Activity of the Erythroblast in Chronic Erythremia, the Chronic Form of Di Guglielmo’s Disease
Authors: BERNARDELLI E. , MELE V. , BANI M. 
Published: October 2019

Abstract
Using the method of culture in vitro, the authors have studied the proliferative and differentiative activities of the erythroblastic tissue in chronic erythremic myelosis.
The results of the research allowed the authors to reach the following conclusions:
1. the erythroblastic cells of chronic erythremic myelosis reveal indisputable differentiative activity in vitro;
2. the rate of maturation of the erythremic erythroblasts is slower than normal;
3. the proliferation of the erythroblastic tissue in chronic erythremic myelosis is normal;
4. in the genesis of the erythroblastic hyperplasia in chronic erythremic myelosis, the most evident factor is a defect of maturation of the erythroblasts.
Rivaroxaban — An Oral, Direct Factor Xa Inhibitor — Prevents Arterial Thrombotic Occlusion in Electrolytically Injured Rat Carotid Arteries.
Authors: Haertlein Barbara , Parry Tom J. , Chen Cailin , Perzborn Elizabeth , Andrade-Gordon Patricia , Damiano Bruce P. 
Published: October 2019

Abstract
Introduction: Rivaroxaban is an oral, direct Factor Xa (FXa) inhibitor in advanced clinical development for the prevention and treatment of thromboembolic disorders, including arterial indications such as acute coronary syndromes (ACS). The aim of this study was to determine whether rivaroxaban can prevent arterial thrombotic occlusion in electrolytically injured rat carotid arteries.
Methods: Anesthetized, male Sprague-Dawley rats were equipped with an intravenous (i.v.) catheter to administer drug or vehicle and an intra-arterial catheter for arterial pressure monitoring and blood sampling. The left carotid artery was exposed and a bipolar electrode placed proximally around it. A pulsed Doppler flow probe was then placed distally around the vessel. Blood was obtained before drug or vehicle administration to determine activated clotting time (ACT), prothrombin time (PT), Russell’s viper venom time (RVVT), activated partial thromboplastin time (aPTT) and thrombin-antithrombin (TAT) complex concentration; baseline carotid blood flow was recorded for 20 minutes. Vehicle (polyethylene glycol:ethanol:water or saline, 0.5 mL/kg/min), rivaroxaban (0.3, 1 and 3 mg/kg; dosing volume 0.5 mL/kg/min), or the low molecular weight heparin enoxaparin (10 mg/kg) were infused i.v. over 30 seconds. After infusion, direct current (3 mA) was applied to the left carotid artery for 5 minutes. Blood flow was monitored for 30 minutes after injury. Blood was sampled again 14.5 and 30 minutes after injury for ACT determination, and after 30 minutes for PT, RVVT, TAT and aPTT determination.
Results: Rivaroxaban dose-dependently increased the median time to arterial thrombotic occlusion (TTO), and was significantly more effective than vehicle at doses of 1 and 3 mg/kg (vehicle, 13.2 minutes; rivaroxaban 1 and 3 mg/kg, >30 minutes; P<0.05 by Kaplan-Meier analysis). Compared with enoxaparin (10 mg/kg), rivaroxaban inhibited TTO at doses of 1 and 3 mg/kg more effectively. Rivaroxaban (0.3–3 mg/kg i.v.) dose-dependently prolonged ACT, PT and RVVT, and substantially decreased the formation of TAT complexes, but did not substantially affect aPTT. Enoxaparin prolonged ACT, aPTT and RVVT, and decreased TAT complex formation, but had no effect on PT.
Conclusions: Rivaroxaban potently inhibited electrolytically induced arterial thrombotic occlusion in rat carotid arteries, resulting in substantial changes in coagulation parameters. The coagulation parameter changes were consistent with inhibition of FXa activity as indexed by the RVVT. These findings suggest that rivaroxaban may be an effective anticoagulant in arterial thrombotic disorders, such as ACS.
Final Update of Phase I-II Study of the Farnesyltransferase Inhibitor Tipifarnib in Combination with Idarubicin and Cytarabine for Patients with Newly Diagnosed Acute Myeloid Leukemia or High-Risk Myelodysplastic Syndrome.
Authors: Delmonte John , Kantarjian Hagop M. , Garcia-Manero Guillermo , Estrov Zeev , Ravandi Farhad , Verstovsek Srdan , O’Brien Susan , Koller Charles A. , Faderl Stefan , Thomas Deborah A. , Wright John J. , Cortes Jorge 
Published: October 2019

Background: Tipifarnib (Zarnestra, Z) is a non-peptidomimetic farnesyltransferase inhibitor (FTI) with clinical activity in hematologic malignancies, including acute myeloid leukemia (AML), myelodysplastic syndrome (MDS) and chronic myeloid leukemia. Preclinical data suggest that tipifarnib may be synergistic with some chemotherapeutic agents.
Methods: We designed a phase I/II study in patients (pts) age 15–70 years, with previously untreated AML or high-risk MDS (blasts ≥ 10%), who received induction with idarubicin (Ida) 12 mg/m2/day on days 1–3, cytarabine (ara-C) 1.5 g/m2 IV over 24 hours daily on days 1–4 (days 1–3 only if age ≥ 60 years) and Z, with first cohort (n=6) receiving 200mg PO BID and all others 300 mg PO BID x 21 days every 28 days. Pts achieving a complete remission (CR) received consolidation (5 courses) with Ida 8 mg/m2/day on days 1–2, ara-C 0.75 g/m2/day on days 1–3, and Z 300 mg PO BID x 14 days every 4–6 weeks. Maintenance was with Z 300 mg PO BID x 21 days every 4–6 weeks for 6 months.
Results: We treated 95 pts, median age 50 yrs (range, 17–61 yrs), and all are evaluable for response with a median followup of 61 weeks (range, 35–138). Seventy pts (73%) responded: 61 (64%) achieved a CR and 9 (9%) a CRp. Median CR duration was 72 weeks, (range, 4–121) with median OS 70 weeks (range, 1–138). Response by cytogenetics was: 34/41 (83%) for diploid, 13/19 (69%) with −5/−7, 1/2 (50%) with t(8;21), and 21/32 (65%) with other abnormalities. Response by age was 31/43 (72%) for < 50 yrs, 12/18 (67%) for ≥ 50 yrs with diploid karyotype, and 18/34 (53%) for ≥ 50 yrs with abnormal karyotype. Response by FLT3 status was 12/16 (75%) for mutated, 51/66 (78%) for unmutated. Overall response rate (CR+CRp) in a similar historical population treated with the same chemotherapy regimen, idarubicin/cytarabine (IA), without Z was 72% (p=0.847), median CR duration 52 weeks (range, 2–319) (p=0.493), median OS 65 weeks (range, 3–322) (p=0.698). The most common grade ≥ 3 adverse events have included diarrhea (39%), hypokalemia (27%), rash (21%), and hepatic dysfunction (18%). Fifty-three (56%) pts have required treatment dose reductions during induction, 21 pts (40%) during consolidation, and 3 pts (18%) during maintenance.
Conclusion: We conclude that Z combined with IA induces a high rate of CR in high-risk MDS or AML, but the outcome may not be superior to what is obtained with IA alone.