bluuebunny/crossref_metadata_2025_split huggingface dataset:
Functional Activity of In Vivo Primed Granulocytes: A Comparative Study.
Authors: Ribeiro Daniel , Laufs Stephanie , Buss Eike B. , Zeller W. Jens , Ho Anthony D. , Fruehauf Stefan 
Published: October 2019

Abstract
Recombinant granulocyte colony-stimulating factor (G-CSF) has been widely used in the treatment of chemotherapy-induced neutropenia as well as in mobilization of peripheral blood stem cells in context with autologous bone marrow transplantation. Recombinant G-CSF expressed in E.coli (Filgrastim) and G-CSF expressed in CHO-cells (Lenograstim), are in clinical use. Here we study the effects of the different G-CSF on functional activity of granulocytes including chemotaxis, oxidative burst and antigen expression.
Granulocytes were obtained from patients with hematological malignancies before and after the administration of one of the three G-CSFs and isolated using a polymorphprep density gradient. Chemotactic properties were assessed using a Boyden chamber assay in combination with an under agarose assay, both using fMLP as chemotactic stimulus. Release of superoxide anions served as measure of the oxidative burst after stimulation with PMA using a chemiluminescence assay. The viability and surface antigen expression were assessed by FACS.
FACS analysis showed a decrease in CD10, CD11b and CD62L of the selectin family contrary to an increased expression of the VLA-5 alpha chain CD49, the LPS-receptor CD14 and the IgG receptor FcγRI (CD64). A stronger effect of lenograstim on CD11b and CD14 could be assessed contrary to filgrastim showing a stronger effect on CD62 and CD64. We observed a decrease of chemotactic activity and non-directed random migration in patients receiving filgrastim, as opposed to the results in patients receiving lenograstim. No obvious differences were found in production of superoxide anions. Whether the superior chemotactic activity and migration of Lenograstim- vs. Filgrastim-primed granulocytes translates into improved antibacterial activity and improved clinical endpoints warrants further study in the clinical setting.
Early Diagnosis of Invasive Aspergillosis in Hematological Patients Using a PCR-ELISA Assay: A Prospective Validation.
Authors: Cornet Muriel , Katsahian Sandrine , Vekhoff Anne , Levy Vincent , Rio Bernard , Marie Jean-Pierre , Bouvet Anne , Florent Martine 
Published: October 2019

Abstract
Purpose: Prospective evaluation of a PCR-ELISA method for the early diagnosis of invasive aspergillosis (IA) in hematological patients.
Patients and Methods: The PCR-ELISA assay was evaluated using 1,494 sera samples from 201 hematological adult patients. Monitoring was performed twice weekly during 256 consecutive high-risk treatment periods.
Results: Underlying diseases were acute leukemia 52.7%, lymphoma 20.9%, multiple myeloma 12.4%. Eighty four patients (41.8%) were stem cell transplant recipients. Fifty-five patients were diagnosed with IA. PCR-ELISA positivity (i) preceded the empirical antifungal therapy in 20/31 patients (median 10.5 days), (ii) preceded or was simultaneous to the radiological diagnosis by chest CT in 21/31 patients (median 8 days), (iii) preceded the mycological/histological diagnosis in 7/12 patients (median 70 days) and, (iv) anticipated or was simultaneous to the galactomannan (GM) antigen positivity in 50% (median 16.5 days) and in 89 % of the patients (median 33 days) using a 0.5 and a 1.5 optical density index (ODI) cut-off for the GM assay, respectively. For at least two positive samples the sensitivity, specificity, positive and negative predictive values of the PCR-ELISA assay, were 63.6%, 82.2%, 57.4% and 85.7%, respectively. When the PCR-ELISA and GM detections were combined, values of the combined test for at least two positive samples with either assay were 80%, 47.3%, 36.4%, 86.2% using the 0.5 GM ODI cutoff, and 67.3%, 82.2%, 58.7% and 87% using the 1.5 GM ODI cutoff.
Conclusion: In addition to serial screening with GM antigenemia and chest CT surveillance, the PCR-ELISA assay may improve the early diagnosis of IA.
Leukemogenic MLL Fusion Proteins Promote MLL Association with HOX Loci Resulting in Persistent Expression.
Authors: Milne Thomas A. , Martin Mary Ellen , Slany Robert K. , Hess Jay L. 
Published: October 2019

Abstract
Rearrangements of the mixed lineage leukemia gene MLL are associated with aggressive lymphoid and myeloid leukemias. The resulting MLL fusion proteins enforce high-level expression of HOX genes including HOX A7 and HOX A9 and the HOX cofactor MEIS1, which is pivotal for leukemogenesis. The mechanism by which this occurs and the relationship to normal MLL function is unknown. To address this we performed a detailed study of where MLL and MLL fusion proteins bind at target genes in hematopoietic cells using quantitative chromatin immunoprecipitation (ChIP). In addition we characterized the histone modifications at the HOX A9 locus that occur with normal down modulation of HOX A9 expression during hematopoietic differentiation and how this is perturbed by induction of conditionally transforming MLL fusion proteins. These studies suggest that a major mechanism of regulating MLL, which is expressed throughout hematopoiesis, is through modulating it’s binding to target promoters. MLL binds directly to the promoters and coding regions of HOX A7, HOX A9, and MEIS1 only in myeloblasts and not in neutrophils, indicating MLL is physically associated with genes only when they are actively transcribed. Expression of A cluster HOX loci and MEIS1 is transiently increased by the addition of IL-3 but remains persistently elevated when MLL-ENL or dimerized MLL fusion proteins are expressed. Expression of either fusion protein is associated with increased binding of wild type MLL accompanied by increases in histone acetylation and histone H3 lysine 4 methylation, marks that are normally almost completely erased during myeloid differentiation. In addition MLL-ENL induces increased lysine 79 methylation. MLL extensively colocalizes with RNA polymerase II, which shows abnormal pausing in the coding regions of HOX genes in Mll null cells. These findings suggest MLL fusion proteins maintain expression of HOX genes critical for leukemogenesis through increased MLL binding resulting in both MLL dependent histone modifications and lysine 79 methylation and ultimately promotion of transcriptional elongation.
Incorporation of the Non-Human Sialic Acid Neu5Gc into Human Leukemic Cells and Targeting by Natural Human Anti-Neu5Gc Antibodies.
Authors: Nguyen Dzung H. , Tangvoranuntakul Pam , Varki Ajit 
Published: October 2019

Abstract
Humans are incapable of synthesizing the common mammalian cell surface sialic acid N-glycolylneuraminic acid (Neu5Gc), due to an irreversible genetic mutation that occurred after our last common ancestor with great apes. Despite this, we found trace levels of Neu5Gc in certain normal human tissues, and higher levels in fetal and malignant tissues, apparently due to incorporation from dietary sources. Circulating anti-Neu5Gc antibodies also occur in most normal humans, with marked individual variations in levels. We now show that while normal human blood cells metabolically incorporate very little free Neu5Gc from the culture medium, human leukemic cell lines do so efficiently, displaying it on their cell surfaces, as detected by flow cytometry using a monospecific polyclonal chicken antibody, and confirmed chemically by HPLC. Specific deposition of IgG from Neu5Gc-reactive human sera onto Neu5Gc-expressing leukemic cells could be demonstrated. This was associated with lytic cell killing, apoptosis, and antibody-dependent cell-mediated cytotoxicity. Human sera with low levels of anti-Neu5Gc antibodies did not mediate such effects. These data show for the first time that the “natural” anti-Neu5Gc antibodies found in normal humans can be functionally active. The selective incorporation of Neu5Gc into leukemic cells could provide an approach for targeting leukemic cells in vivo, via naturally occurring Neu5Gc-specific antibodies. However, in contrast to unstimulated blood cells, activated human T-lymphocytes also incorporated some Neu5Gc, albeit to a lesser degree than leukemic cells, allowing IgG binding and complement deposition. Thus, exposure of rapidly dividing activated T cells to Neu5Gc could potentially affect ongoing immune responses. Since incorporation into leukemic cells was most efficient, it may still be possible to define a therapeutic window to selectively target malignant cells.
Hemostatic Effects of Fibrinogen-γ Chain Dodecapeptide-Conjugated Polymerized Albumin Particles In Vitro and in Vivo.
Authors: Okamura Yosuke , Watanabe Naohide , Takeoka Shinji , Suzuki Hidenori , Murata Mitsuru , Ikeda Yasuo , Handa Makoto 
Published: October 2019

Abstract
We have studied a prototype of platelet substitutes focusing on a dodecapeptide, HHLGGAKQAGDV (H12)1), which is specific for fibrinogen γ-chain carboxy-terminal sequence (γ 400–411). In this study, we conjugated H12 to the surface of polymerized albumin particles (polyAlb) as biocompatible and biodegradable carriers to produce particles having hemostatic ability, and evaluated their in vitro and in vivo effects.
H12 (H12; 9.6 x 103 /particles) containing cysteine to the N-terminal was conjugated to polyAlb (260 ± 60 nm), and the effect of H12-polyAlb on platelet thrombus formation was evaluated in vitro with thrombocytopenic whole blood ([platelet] = 2.0 x 104 /μL, anticoagulated with PPACK) under flow conditions (shear rate; 150 s−1). Thrombocytopenic rats were made by busulphan injection at a dose of 20 mg/kg, and a 2.5 mm length x 1.0 mm depth template-guided incision (QuikheelTM, Becton-Dickinson, San Jose, CA) was made 1 cm from the tip of tail. The tail was immersed in a 50 mL cylinder of saline and the time taken to stop bleeding was measured.
When thrombocytopenic blood in the presence of H12-polyAlb ([rHSA]=0.14 mg/mL) was flowed on collagen-plate, the surface coverage of DiOC6-labeled platelets evaluated by fluorescence microscopy was increased to 3.9 ± 1.1 % (n=3) from 2.1 ± 0.4 % (n=3) obtained in the absence of H12-polyAlb. In the same experiments, rhodamine-labeled H12-polyAlb was found to be involved in platelet-platelet interactions by binding to activated platelet surface, thus enhancing platelet thrombus formation. Next, in vivo hemostatic effect was tested by measuring tail bleeding time of thrombocytopenic rats 5 minutes after the intravenous administration of H12-polyAlb. The bleeding times of normal ([platelet] = 8.1 ±0.9 x 105 /μL) and thrombocytopenic rats ([platelet] = 2.0 ±0.3 x 105 /μL) were 187 ± 51 s and 609 ± 153 s (n=6), respectively. H12-polyAlb administration at a dose of 4 mg/kg significantly shortened the bleeding time to 342 ± 73 s (n=10), whereas polyAlb was without effects (553 ± 104 s, n=6). These results indicate that H12-polyAlb can be a suitable candidate for an alternative to human platelet concentrates infused into thrombocytopenic patients.
Long Distance Trans DNA Hypomethylation in Cyclin D1 Deregulated B Cell Malignancies.
Authors: Liu Hui , Wang Jin , Huang Jing , Epner Elliot M. 
Published: October 2019

Abstract
Cyclin D1 is a common target gene in B cell malignancies. Cyclin D1 is deregulated by translocation in most patients with mantle cell lymphoma (MCL) and 15–20% of patients with multiple myeloma (MM). Cyclin D1 is not expressed in normal lymphocytes. Gene targeting experiments in cyclin D1 overexpressing MCL and MM were carried out and genetic variants isolated which had lost the translocated or inserted 11:14 chromosomes. These clones no longer expressed cyclin D1 but expressed high levels of cyclin D3. Analysis of DNA methylation patterns in these clones demonstrated that the translocated chromosome exerts a long distance trans DNA hypomethylating effect on the normal chromosome (transvection) at the cyclin D1 locus and at least 100 kilobases upstream. Thus, in the absence of the translocated chromosome, the normal chromosome is densely DNA methylated. This long distance trans hypomethylating effect was also demonstrated in a MM cell line (U266) which contains an insertion rather than a translocation of IgH sequences. Combined FISH/ immunofluorescent antibody labelling studies have shown the presence of both the normal and translocated chromosome 11’s at the outer nucleolar membrane, where they are tethered by the proteins CTCF and nucleophosmin, as demonstrated using chromatin immunoprecipitation assays. Tethering of the translocated and nontranslocated chromosomes by CTCF/nucleophosmin provides a mechanism for pairing and long distance DNA transhypomethylation.
Correlation between Real-Time Microvascular Abnormalities and Whole Blood Viscosity (WBV) in Type-2 Diabetes Mellitus (T2DM) Patients.
Authors: Cheung Anthony T. , Ramanujam Sahana , Barbosa Michelle A. , Asfour Violet K. , Medina Maria V. , Miller Joshua W. 
Published: October 2019

Abstract
Novel technologies were used to study the microvascular abnormalities in Type-2 diabetes mellitus (T2DM) patients and to correlate these real-time pathological complications with whole blood viscosity (WBV) over a wide range of shear rates (1–1,000s−1). Computer-assisted intravital microscopy (CAIM) was used to non-invasively and objectively quantify the microvascular abnormalities in the conjunctival microcirculation in T2DM patients (n=12; age range = 45 to 68) and healthy non-diabetic control subjects, with the medical history of each subject blinded to the investigators. Fifteen recognizable microvascular abnormalities existed in T2DM patients and not in the control subjects, though not all the abnormalities were found in each patient. A severity index (SI) -- the arithmetic sum of microvascular abnormalities in each patient quantified via CAIM -- was computed to give a score to correlate with medical history, WBV and shear rates. T2DM SI (6.9±1.7) differed significantly (P<0.01) from control SI (0.6±0.7) and correlated significantly with disease severity, but not with the duration of the disease since diagnosis. The results, together with results from a previous study, lend support to the hypothesis that diabetic complications may have occurred in the pre-diabetic period; indicative of the existence of a “time window” before the onset of clinically detectable hyperglycemia. To adequately assess WBV to correlate with T2DM SI, a computer-assisted scanning capillary viscometer, the Rheolog™, was used to generate a viscosity profile for each patient or control subject over a range of shear rates (1 to 1,000s−1) using one 3-ml citrated venous blood sample from each subject. Based on the viscosity profiles generated, WBV of T2DM patients at various shear rates differed significantly from control values. For example, averaged WBV of T2DM patients at a shear rate of 300s−1 (4.01±0.33cp) differed significantly (P<0.02) from averaged WBV of control subjects (3.34±0.05cp). WBV correlated with SI of T2DM patients, disease severity and medical records. This correlation may have unique clinical implications - it can be used to study shear-stress induced pathogenesis on endothelial dysfunction in various vascular diseases and in early detection of T2DM in carriers (e.g. siblings of T2DM patients) before clinical diagnosis of T2DM during the pre-diabetic “time window” of early detection. Follow-up studies on the correlation between real-time microvascular abnormalities (SI), disease severity, WBV and shear rates in Alzheimer’s Disease patients and in early detection of T2DM are in progress.
Platelet-Derived Microparticles Induce Angiogenesis and Stimulate Post-Ischemic Revascularization.
Authors: Dashevsky Olga , Brill Alexander , Rivo Julia , Varon David 
Published: October 2019

Abstract
Platelet attachment to the subcellular matrix at injured sites of the vasculature is followed by their activation and release of microparticles. Platelet-derived microparticles (PMP) have been shown to be involved in the regulation of hemostasis. However, little is known about the role of PMP in the regulation of angiogenesis and related clinical conditions. We have recently demonstrated that platelets as a cellular system induce angiogenic responses both in vitro and in vivo. In the present study, we investigated the potential role of PMP in angiogenesis. A strong dose-dependent pro-angiogenic effect of PMP in the rat aortic ring model (5.3±2.1 mm2 surface covered with sprouting vessels versus 0.24±0.2 mm2 in the control, p<0.001) was observed. This effect was reversed by selective inhibition of VEGF, bFGF and PDGF (surface covered with vessels 0.7±0.5 mm2, 1.7±1.5 mm2, and 2.4±1.2 mm2, respectively, p<0.02 versus control), but not by inhibition of heparanase (5.1±0.8 mm2, p>0.5 versus control). PMP exert their stimulatory effect via PI3-kinase, Src kinase and ERK, whereas protein kinase C seems not to be involved, as judged by the aortic ring sprouting model. Using confocal and electron microscopy, we also demonstrate that PMP bind to non-activated endothelial cells. In addition, PMP markedly increased invasion of human endothelial cells through a layer of matrigel. This effect was abolished by an inhibitor of VEGF receptor tyrosine phosphorylation or laminaran sulfate (heparanase inhibitor). It was also partially reduced by PDGF blocking mAb, whereas blocking of bFGF had no effect. Furthermore, we have demonstrated that PMP induce angiogenesis in an in vivo model, in which beads (30 μl) of 4% agarose gel containing the substances under study were transplanted subcutaneously into mice. Image analysis of the capillary area revealed the following: control beads − 0.2±0.05 mm2, VEGF + bFGF containing beads − 4.8±1.1 mm2, PMP (100 μg/ml) containing beads − 5.1±1.3 mm2, p<0.001 versus control. The latter finding was further supported by immunohistochemical staining of the skin in the vicinity of the beads for von Willebrand factor, a marker of endothelial cells (control − 4.0±3.2, VEGF+bFGF − 12±4.4, PMP − 17±6.5 capillaries per view field, p<0.05 versus control). Finally, we explored the potential effect of PMP in a rat myocardial infarction model. Ischemia was induced by LAD ligation followed by injection of either PMP or PBS into the ischemic region. Preliminary evaluation of the LAD myocardial territory in sham-operated animals revealed 157±42.0 capillaries per view field. In contrast, number of capillaries observed 3 weeks after induction of ischemia was reduced to 34±21.5. When PMP were injected into the ischemic region, there was an increase in capillary number up to 97±27.3. In conclusion, PMP induce angiogenesis in both in vitro and in vivo models. Local injection of PMP into the ischemic myocardium may improve revascularization.
The Proadhesive Properties of Multimerin in Supporting Cellular Adhesion: Evidence for RGD and Non-RGD Dependent Adhesive Mechanisms.
Authors: Adam Frederic , Zheng Shilun , Joshi Nilesh , Suehiro Youko , Kelton David S. , Sandhu Amin , Jeimy Samira , Hayward Catherine P.M. 
Published: October 2019

Abstract
Multimerin is a large soluble protein, with an uncertain function, found in platelets, megakaryocytes, endothelium and extracellular matrix fibers but not in plasma. The observation that multimerin contains structural features of an adhesive protein, including an Arg-Gly-Asp (RGD) sequence, led us to investigate its ability to support adhesion of platelets, megakaryocytes, endothelial cells and other cell types. Multimerin had the ability to support the adhesion of both platelets and megakaryocytes and this required cellular activation and the multimerin RGD site. Studies of normal and Glanzmann platelets indicated that multimerin interacted with the major platelet integrin receptor, αIIbβ3 and radioimmunoprecipitation analyses confirmed that multimerin bound to αIIbβ3. Multimerin also supported adhesion of endothelial cells, neutrophils and other cells including smooth muscle cells, fibroblast cells, human embryonic kidney (HEK293) and epithelial cells. Unlike platelets, these cells do not express αIIbβ3; this indicated that other integrin or non-integrin receptors could be involved in cellular adhesion to multimerin. Comparisons of cell adhesion to wild-type and RGE-multimerin indicated that unlike platelets and megakaryocytes, some other cell types (e.g. endothelial cells, smooth muscle cells and neutrophils) were capable of adhering to RGE-multimerin. This suggested that cellular adhesion to multimerin occurs by both RGD and non-RGD dependent mechanisms. Finally, unlike platelets, megakaryocytes and neutrophils, adhesion of other cell types to multimerin did not require cellular activation. In conclusion, our data indicate multimerin has fairly broad proadhesive properties, involving RGD and non-RGD dependent mechanisms, and that cellular receptors including αIIbβ3 interact with multimerin to mediate its binding to activated platelets, endothelial cells and potentially other cell types.
ADP Signaling Defect in Children with Severe Acquired Aplastic Anemia: A Potential Common Pathway for Development of Aplastic Anemia.
Authors: Ford Jason C. , Wadsworth Louis D. , Schultz Kirk R. 
Published: October 2019

Abstract
Severe Acquired Aplastic Anemia is a rare disease with an incidence in British Columbia of 6.9 per million/year in Eastern/Southeastern Asians, 7.3 per million/year in South Asians and 1.7 per million/year in those of White/mixed ethnic descent. Two children presented with mucosal bleeding >4 years after achieving complete remission with immune suppressive therapy (antithymocyte globulin, cyclosporin A and prednisone). Evaluation revealed normal routine coagulation profiles and normal platelet numbers but mildly prolonged bleeding times (patients 8.5 - 11.0 minutes; control 3.0 – 8.0 minutes) and abnormal platelet aggregation as measured by aggregometry (optical and luminescence) after ADP stimulation. The patients had normal aggregation and ATP secretion after stimulation with thrombin (ATP secretion only), collagen, epinephrine, arachidonic acid, and ristocetin (1.125, 1.0, and 0.5 mg/mL concentrations, optical aggregation only) but showed abnormal aggregation with ADP (2.5, 5.0, and 10 μM). An additional 4 aplastic anemia patients were tested >1 year after successful immune suppressive therapy with no bleeding history: 1 of the 4 had the identical defect in platelet aggregation and ATP release after ADP stimulation, with no other defects on aggregometry. All 3 patients with abnormal platelet aggregation were of South or East Asian background, and the 3 with normal aggregation were of Caucasian/mixed ethnicity. Of the patients with an abnormal ADP platelet response, one has subsequently developed ulcerative colitis. Based on these observations we hypothesize that some patients with severe acquired aplastic anemia have an underlying defect in a signaling pathway shared by platelets, T cells, and B cells. ADP-induced signaling in platelets is induced through the surface molecule P2Y12, which is not expressed on lymphocytes. P2Y12 signaling occurs in association with a G protein Galphai2, an important signaling molecule in T and B cells and associated with development of ulcerative colitis in Galphai2 knock out mice. The specific signaling defect in platelets is currently being elucidated in these patients, and the incidence of these defects in aplastic anemia patients must be characterized.