bluuebunny/crossref_metadata_2025_split huggingface dataset:
Conservative Whole Blood Extended Differential Method.
Authors: Paul Ronald , Lopez Lidice L. , Careaga Diana B. , Rubio Oilda , Mylvaganam Ravindra 
Published: April 2020

Abstract
The neutrophilic precursors (excluding bands), blast cells and erythroblasts normally reside in the bone marrow. The presence of any of these precursors as well as atypical/variant lymphocytes in peripheral blood is indicative of a diseased or disordered state. Therefore, it is of clinical importance to identify and enumerate these atypical cell populations. In general, commercial hematology systems do not have the ability to enumerate all of the nucleated atypical cells that are traditionally identified by manual differential. However, the manual differential is labor intensive and can be highly variable between qualified readers resulting in significant cost and quality assurance issues for the laboratory. We have developed a unique fluorescent based Extended Differential method that uses a minimum number of monoclonal antibodies in combination with a cell permeant metachromatic nucleic acid dye to detect circulating atypical cells. Data will be presented comparing the results of this approach with the manual differential. The current study was comprised of 579 clinical samples containing a variety of atypical cell types as determined by morphological examination. Analysis of difference plots comparing the Extended Differential method with the manual differential revealed good agreement between the percent of immature granulocytes (IGs) and the number of nucleated red blood cells (NRBCs) per 100 white blood cells (WBCs). The same analysis applied to the determination of blast percent exposed a greater frequency of disparate results. These disparate results were further investigated using a comprehensive monoclonal antibody based reference method incorporating a variety of specificities including anti-CD45 and anti-CD34. The data demonstrated that cells classified as atypical/variant lymphocytes or smudge cells by morphological examination were classified as blasts utilizing CD45 and/or CD34 expression with the Extended Differential and flow reference methods. In addition to the difference analyses, the results obtained with manual differential and Extended Differential methods were subjected to Receiver Operating Characteristics (ROC) analysis in order to determine the overall analytical sensitivity and specificity using a threshold of 1% or greater as the criteria for the presence of atypical cells. The ROC analysis demonstrated areas under the curve (AUC) of 0.9743, 0.9831 and 0.9643 for IGs, blasts and NRBCs, respectively showing good agreement for the presence or absence of the respective atypical cell populations (p &lt; 0.0001).
Analyses of Immune Defense Mechanisms of Human Polymorphonuclear Neutrophils Co-Cultivated with Aspergillus fumigatus Germ Tubes.
Authors: Loeffler Juergen , Mezger Markus , Einsele Hermann 
Published: April 2020

Abstract
Invasive fungal infections with the opportunistic pathogen Aspergillus fumigatus show an increasing incidence due to a higher number of patients with hematological malignancies. Polymorphonuclear neutrophils (PMNs), as part of the innate immunity, recognize fungal pathogens at an early stage after infiltration. Besides phagocytotic mechanisms, PMNs kill pathogens by the release of reactive oxygen species (ROS). Human PMNs were isolated from blood of healthy donors and co-cultivated with A. fumigatus germ tubes for defined time points. Oxidative burst was determined in a kinetic measurement by the use of dichlorfluorescein. In parallel, PMNs were co-cultivated with A. fumigatus germ tubes, followed by whole genome expression analyses (Affymetrix U133 Plus 2.0 Array). We could demonstrate that A. fumigatus germlings of the clinical relevant strain ATCC 9197 represented a strong stimulus for the release of ROS. PMNs actively tracked germlings and directly attached to fungi as demonstrated by real-time microscopy. In addition, co-cultivation of PMNs with A. fumigatus germ tubes resulted in a strong upregulation of genes involved in self-protection against radicals (hämoxygenase, heat shock 70kDa protein HSPA8, thioredoxin, HSPA1B, HSP90AB1, Ferritin). After 6h of co-cultivation, 195 genes showed an at least 4fold altered gene expression. Therein, 4 genes encoding for cytokines and chemokines (IL-8, CCL3, CXCL2, IL1RN) were significantly upregulated. Luminex ELISA analyses confirmed array data and revealing IL-8 to be strongly released (5fold) by PMNs after fungal co-culturing. In conclusion, A. fumigatus had a substantial effect on the activity of human PMNs. In consequence, various defence strategies were activated, including phagocytosis, ROS release and mobilization of other immune effector cells by secretion of chemoattractant cytokines. A better understanding of innate immune defense mechanisms may provide new directions for antifungal therapies.
The Value of CT-Guided Percutaneous Lung Biopsy in the Diagnosis of Pulmonary Fungal Infections in Patients with Hematologic Diseases.
Authors: Cai Zhen , Shi Jimin , Huang He , Lin Yu , Ye Xiujin , He Jingsong , Xie Wanzhuo , Zhang Jie , Li Li , Zheng Weiyan , Wei Guoqing , Luo Yi , Zhang Buwen , Lin Maofang 
Published: April 2020

Abstract
Introduction: Patients with hematologic diseases are at high risk for invasive fungal infections (IFI).The diagnosis of IFI is difficult with specificity as low as 20%–30%. The currently used diagnostic methods for IFI such as CT scan, fungal cultures, and antigen detection are insufficient in reaching a definitive diagnosis. Histopathology with tissue diagnosis is the method of choice. CT-guided percutaneous lung biopsy (CT-PCLB) can provide pathological diagnosis and hence significant guidance for clinical management.
Objectives: To evaluate the sensitivity and safety of CT-PCLB for the diagnosis of pulmonary fungal infections in patients with hematologic diseases.
Patients and Method: We retrospectively studied 16 patients hospitalized in our department from April 2005 to December 2006 who were clinically regarded as having pulmonary fungal infections by clinical symptoms and imaging and who subsequently underwent CT-PCLB. Among the 16 patients, 10 had AML, 2 had ALL, 2 had ITP, 1 had MDS and 1 had multiple myeloma. At the time of percutaneous lung biopsies, median WBC was 3.2 k/ul (range 0.7–7.2 k/ul), median platelets were 82 k/ul (range 32–453 k/ul). Peripheral blood cultures were performed in all patients. Eight patients who had symptoms of cough and expectoration also received sputum culture for at least three times. Of the 16 patients, 10 were diagnosed with fungal infections (8 aspergillus’s and 2 mold fungi), 4 with chronic organizing pneumonitis, 1 with tuberculosis, and 1 with pneumocystis carinii. All tissue diagnoses were from histological examination with samples obtained from CT-PCLB. However, the results of blood cultures and sputum cultures were negative in these 8 patients. CT-PCLB diagnosed 10 out of 16 patients for a fungal infection rate of 63%. The biopsy-induced complications encountered were pneumothorax in 3/16 (19%) and hemoptysis in 6% (1/16). No surgical interventions were needed for the 3 patients with pneumothoraxes that resolved spontaneous under close observation. Antifungal agents (such as voriconazole) had been given to 8 patients diagnosed as aspergillus infection and all patients had a complete or partial response by CT imaging. However, 2 patients who received broad-spectrum antibacterial antibiotics showed clinical improvement, which indicated possible false negative diagnoses for fungal infections by CT-PCLB.
Conclusion: CT-guided percutaneous lung biopsy (CT-PCLB) was a safe, sensitive and specific method for the diagnosis of pulmonary fungal infections in patients with hematologic diseases. Its definitive value should be tested in a prospective clinical trial.
Iron Chelator ICL670 Inhibits HIV-1 Transcription.
Authors: Debebe Zufan , Ammosova Tatyana , Nick Hanspeter , Niu Xiaomei , Jerebtsova Marina , Ray Patricio , Gordeuk Victor R. , Nekhai Sergei 
Published: April 2020

Abstract
HIV-1 replication is induced by the excess of iron and iron chelation by desferrioxamine (DFO) inhibits viral replication in HIV-1 infected CEM T cells [1]. Treatment of cells with DFO or 2-hydroxy-1-naphthylaldehyde isonicotinoyl hydrazone inhibits expression of proteins that regulate cell-cycle progression, including cycle-dependent kinase 2 (CDK2) [2]. HIV-1 transcription is activated by Tat protein, which recruits transcriptional co-activators to the HIV-1 promoter. Elongation of HIV-1 transcription is mediated by the interaction of HIV Tat with host cell cycle-dependent kinase 9 (CDK9)/cyclin T1, which phosphorylates the C-terminal domain of RNA polymerase II. Our recent studies showed that CDK2 participates in HIV-1 transcription by phosphorylating Tat [3]. Thus inhibition of CDK2 by iron chelators might present a new approach to inhibit HIV-1 transcription. We evaluated the effect of a clinically approved orally effective iron chelator, 4-[3,5-bis-(hydroxyphenyl) -1,2,4-triazol-1-yl]-benzoic acid (ICL670 or deferasirox) on HIV-1 transcription. ICL670 inhibited Tat-induced HIV-1 transcription in CEM, 293T and HeLa cells at concentrations that did not induce cytotoxicity. The chelator decreased cellular activity of CDK2 but not its protein level and reduced HIV-1 Tat phosphorylation by CDK2. ICL670 did not decrease CDK9 protein level but significantly reduced association of CDK9 with cyclin T1 and reduced phosphorylation of Ser-2 residues of RNA polymerase II C-terminal domain. In conclusion, our findings add to the evidence that iron chelators may inhibit HIV-1 transcription by deregulating CDK2 and Cdk9. Further consideration should be given to the evaluation of ICL670 for future anti-retroviral therapeutics and to the development of iron chelators specifically as anti-retroviral agents.
A Multi Center Trial of hyperCVAD+Rituxan in Patients with Newly Diagnosed Mantle Cell Lymphoma.
Authors: Epner Elliot M. , Unger Joseph , Miller Thomas , Rimzsa Lisa , Spier Catherine , Leblanc Michael , Fisher Richard 
Published: April 2020

Abstract
BACKGROUND: A Phase II Study multinstitutional study by the lymphoma group within the Southwest Oncology Group (SWOG 0213) was performed to determine the 1 year progression free survival of patients with newly diagnosed mantle cell lymphoma (MCL). Previously, this regimen was utilized in a similar group of newly diagnosed MCL patients in a single institution study by Romaguera et al (JCO200523:7013) at the MD Anderson Cancer Center. They reported a 97% response rate, a 87% CR rate, and a 3 year failure free survival rate of 73%.
PATIENTS AND METHODS: This study was a prospective phase II trial of rituximab plus fractionated cyclophosphamide, vincristine, doxorubicin, and dexamethasone (hyperCVAD; considered one cycle) alternating every 21 days with rituximab plus high-dose methotrexate-cytarabine (considered one cycle) for a total of eight cycles per the MD Anderson protocol. Newly diagnosed MCL patients under age 70 with adequate organ function and measurable disease were eligible. All cases were histologically confirmed.
RESULTS: 49 eligible patients were enrolled. There was one probable treatment-related death due to colitis. Grade 4 hematologic toxicity was reported for 87% of patients. 40 patients have been evaluated for response. 3 patients had inadequate assessment and are assumed to be non-responders.16 patients (40%) had complete responses, 7 patients (18%) had unconfirmed complete responses, and 12 patients (30%) had partial responses for an overall response rate was of 88% (35/40; 95% CI, 73%–96%). Median follow-up among patients still alive is 1.6 years (range: 0.3–4.1 years). The progression-free survival estimate at 1-year is 89% (95% CI: 80%–98%) and at 2 years is 64% (95% CI: 46%–82%). The overall survival estimate at 1 year is 91% (95% CI: 82%–99%) and at 2 years is 76% (95% CI: 60%–91%). 13 of the 49 eligible patients have progressed or died. One year overall survival was 91% and one year progression free survival is 89%. Two year PFS is 63% (95% CI: 46%–82%). Two year OS is 76% (95% CI: 60%–91%). The 1-year progression-free survival estimate is 89% (95% CI: 80%–98%). The 1-year overall survival estimate is 91% (95% CI: 82%–99%). The overall response rate was 88% (35/40; 95% CI, 73%–96%).
CONCLUSIONS: The hyperCVAD + rituximab regimen is active as reported in MCL with a one year progression free survival of 89%. However, similar to the reported MD Anderson experience, this regimen is toxic and a continuous pattern of relapse over time is observed.
The Impact of P2Y-Mediated Activation on Release of Angiogenic Proteins by Platelets from Healthy Individuals.
Authors: Bambace Nadia M. , Pace Thomas R. , Schneider David J. , Holmes Chris E. 
Published: April 2020

Abstract
The storage and release by platelets of the pro-angiogenic protein, vascular endothelial growth factor (VEGF) and the anti-angiogenic protein, endostatin may influence not only wound healing but also tumor angiogenesis and metastasis. The intra-platelet mechanisms and platelet activators that control angiogenic protein release are incompletely elucidated. Previous work demonstrated that the proteinase-activated receptors (PARs), PAR-1 and PAR-4 differentially impact platelet release of these angiogenic proteins (Ma et al., PNAS 102:216). We determined the influence of the adenosine diphosphate (ADP) receptors, P2Y1 and P2Y12 on platelet release of VEGF and endostatin. Minimally altered whole blood from healthy volunteers was incubated with ADP alone [12.5 μM] or in combination with a reversible P2Y1 antagonist [MRS2179 (1.25 mM)] or a reversible P2Y12 antagonist [cangrelor (1mM)]. VEGF and endostatin release was measured with the use of an ELISA assay. ADP-induced activation of platelets increased the concentration of VEGF released by an average of 19.7 +/− 14 ng/mL (p= 0.019) but did not change the concentration of endostatin. Inhibition of the P2Y12 receptor prevented the ADP mediated release of VEGF and decreased VEGF release by 27.9 +/− 14 ng/ml (p=0.005) whereas inhibition of the P2Y1 receptor did not significantly reduce the release of VEGF (p=0.3, mean decrease 14.3 +/− 30 ng/ml) relative to ADP stimulated control. In summary, ADP-induced activation of platelets leads to the release of VEGF but not endostatin. Release of VEGF is attenuated by a P2Y12 but not a P2Y1 receptor antagonist. Accordingly, activation of platelets by ADP appears to be pro-angiogenic and inhibition by a P2Y12 antagonist should attenuate this effect.
G-Quartet Oligonucleotide T70923 Blocks GP Ib-VWF Mediated Platelet Aggregation under Static and Flow Conditions.
Authors: Zhou Zhou , Bernado Aubrey , Guan Yongli , Ball Chalmette , Jing Naijie , Dong Jing-fei 
Published: April 2020

Abstract
Hemostasis at site of the vessel injury is initiated by platelets being tethered to exposed subendothelium. The tethering events are mediated by interaction between subendothelial von Willebrand factor (VWF) and its platelet receptor GP Ib-IX-V complex. If the ligand-receptor interaction, when it occurs in intact vascular lumen or at the site of ruptured atherosclerotic plaques, results in life-threatening thrombosis found in patients with thrombotic thrombocytopenic purpura and myocardial infarction. The development of these thrombotic events suggests that the VWF-GP Ib interaction may be a potential target for therapeutic intervention. To explore such potentials, we have developed and examined the effects of a G-quartet oligonucleotide (T30923, GGGTGGGTGGGTGGGT) on the VWF-GP Ib interaction. G-rich DNA sequences, from which the G-quartets derive, were originally identified in the telomeres DNA and have the ability to form rigid G-quartet structures that interact with proteins. The T30923 structure was designed and determined by nuclear magnetic resonance to form an intramolecular G-quartet that is composed of two identical G-quartets in the center and two GTGT loops on the top and bottom of stacked G-quartets, respectively. Platelet aggregation was induced by four agonists: ristocetin, botrocetin, collagen, and ADP in the presence of either 20 mM of T30923 or the control oligonucleotide. We found that T30923 specifically and dose-dependently blocked platelet aggregation induced by ristocetin, but not by other agonists, with a maximal inhibition achieved at 20 mM. Computer-simulation based on the crystal structure of the VWF A1 domain and the N-terminal VWF-binding domain of GP Iba suggested that the G-quartet binds to multiple sites on VWF-A1. Since the action of ristocetin mimics that of fluid shear stress, we tested the ability of T30923 to block VWF-GP Ib interaction under flow condition. Shear-induced platelet aggregation at 100 dyn/cm2 was blocked 59.3%±11.8% by 20 mM of T30923, but not a control oligonucleotide. Furthermore, the G-quartet T30923 blocked over 80% of platelet adhesion to immobilized VWF, but not collagen under a constant wall shear stress of 30 dyn/cm2. These results demonstrate the potential of T30923 to serve as a VWF antagonist. Furthermore, alterations of DNA sequences and tertiary structure provide a large number of G-quartet variants to be generated and then screened for specific phenotypes. The approach may lead to a new class of potential G-quartet based antithrombotic agents.
Quantitative Analysis of Angiogenesis Inhibition by Cleaved High Molecular Weight Kininogen Using VESGEN.
Authors: Ribita Daniela , Zlotoff Benjamin , McKay Terri L. , Gedeon Daniel J. , Hylton Alan G. , Rasin Arkady , McCrae Keith R. , Parsons-Wingerter Patricia 
Published: April 2020

Abstract
Angiogenesis requires specific molecular regulation of complex vascular branching morphology. The cleaved form of high molecular weight kininogen (HKa) has been shown to cause apoptosis of proliferating endothelial cells, and inhibit angiogenesis in vivo. However, critical morphological effects of HKa on the complex branching morphology of angiogenic vascular trees have not been defined. We used the quail chorioallantoic membrane (CAM) angiogenesis assay, in which angiogenesis agonists or antagonists cause uniform perturbation of angiogenesis throughout the entire vascular tree, to assess the effects of HKa on branching morphology. HKa (1.56 to 50 micrograms) was applied uniformly to the CAM, and its effect on angiogenesis determined after 24 hours of incubation. VESGEN vascular analysis software, currently under development at NASA Glenn Research Center, was used to quantify major vessel parameters in low-magnification images of vascular trees automatically extracted by VESGEN. Results derived from qualitative observation and initial quantification of two experiments demonstrates that HKa significantly inhibits vascular growth. Vessel length density (Lv) and branch point density (Br) in CAM specimens treated with 25 micrograms of HKa decreased by 23% and 31% respectively (24 ± 1 cm/cm2 and 441 ± 43 /cm2, compared to 31 ± 0 cm/cm2 and 677 ± 51 /cm2 in vehicle-treated controls; mean +/− S.E. for n = 3). The fractal dimension (Df) of skeletonized vascular images is a highly sensitive indicator of vessel density that ranges from 1.35 for strong angiogenesis inhibition, to 1.40 and 1.48 for controls and strong angiogenesis stimulation, respectively. Df decreased to 1.35 ± 0.01 in the HKa-treated specimens, relative to 1.39 ± 0.00 in controls. However, thickening of large vessels in response to HKa resulted in a virtually equivalent vessel area density (Av) of 0.167 ± 0.012 cm2/cm2, relative to 0.174 ± 0.015 cm2/cm2 in controls. Further studies of the site-specific effects of HKa on branching morphology within the vascular tree are in progress. These results demonstrate that the antiangiogenic effects of HKa result in a unique antiangiogenic ’fingerprint’ vascular morphology in comparison to other angiogenesis inhibitors that include transforming growth factor-β1 and angiostatin.
Protein Phosphatase 2B Inhibitor, Cyclosporine A, Stimulates Endothelial Cell Exocytosis of Ultra-Large VWF Multimeric Strings by Regulating Munc18c Phosphorylation.
Authors: Nolasco Lettica , Gushiken Francisca C. , Patel Vimal , Turner Nancy , Moake Joel , Vijayan K. Vinod 
Published: April 2020

Abstract
Exocytosis of Weibel Palade bodies (WPBs) containing ultra-large Von Willebrand factor (ULVWF) plays an essential role in the processes of inflammation and thrombosis. The exocytosis of ULVWF involves the fusion of WPBs to the endothelial cell plasma membrane via vesicular trafficking families of proteins. Reversible post-translational modification, including the phosphorylation of tyrosine (Tyr) and/or serine/threonine (Ser/Thr) residues or S-nitrosylation of cysteine residues on one or more vesicular trafficking proteins tightly regulates the exocytotic process. The phosphorylation of any trafficking protein is determined by the interplay between protein kinases and protein phosphatases and any alteration in the activity of either enzyme can affect exocytosis. In comparison to agonist-induced, protein kinase-dependent signaling pathways that regulate exocytosis in endothelial cells, there is scant information about any possible role for the regulation of protein phosphatases in this process. Inhibition of protein phosphatase 2B (PP2B) also called calcineurin by the immunosuppressive drug, cyclosporine (CsA), is associated with thrombotic microangiopathy in a subset of transplant patients. Calcineurin is the target of a cyclophilin-CsA complex. We detected both calcineurin and cyclophilin A in resting human umbilical vein endothelial cells (HUVECs). Because CsA is a potent inhibitor of PP2B, we hypothesized that CsA induces ULVWF secretion and thrombotic events at least in part, by facilitating the phosphorylation of vesicle trafficking proteins that promote the exocytosis of ULVWF from endothelial cells. In vitro, we found that nanomolar concentrations of CsA or a cell permeable PP2B auto inhibitory peptide stimulated the release of ULVWF from HUVECs. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) viability assays revealed that PP2B inhibitors did not damage the HUVECs. In vivo, plasma levels of VWF were increased after intraperitoneal injection of CsA. These studies suggest that PP2B negatively regulate ULVWF exocytosis. Microscopic studies revealed that the ULVWF strings secreted by HUVECs treated with CsA or PP2B auto-inhibitory peptide induced the adhesion of platelets. CsA-treated HUVECs exhibited an increased serine phosphorylation of Munc18c, a vesicle trafficking associated protein that promotes granule exocytosis. These findings support a mechanism whereby inhibition of PP2B by CsA enhances cell secretion, including endothelial cell secretion of ULVWF by facilitating Munc18c phosphorylation.