bluuebunny/crossref_metadata_2025_split huggingface dataset:
A New and Rapid Immunoassay for the Detection of 11-Dehydro-Thromboxane B2.
Authors: Geske F. Jon , Hurley Beth , Whittier Amy , Tew Daniel , Ens Gordon , Lopez Luis 
Published: April 2020

Abstract
Aspirin inactivates cyclooxygenase, inhibiting synthesis of thromboxane A2 (TxA2) and other molecules relevant in the development of atherothrombotic vascular events. There is recent evidence suggesting that many individuals are resistant to aspirin’s effects, resulting in increased TxA2 levels. Current methods for detecting aspirin resistance include a number of blood-based assays that measure in vitro platelet aggregation. However, these methods are not quantitative and can be affected by factors that are unrelated to aspirin sensitivity. There is a quantitative commercial immunoassay available for 11-dehydro-thromboxane B2 (11-dh-TxB2), a stable metabolite of TxA2. However, this assay uses a polyclonal antibody to 11-dh-TxB2 and requires an overnight incubation. In order to more quickly and accurately detect aspirin resistance in at-risk patients, we are developing a rapid, quantitative competitive immunoassay for 11-dh-TxB2 using a monoclonal antibody. This new assay can be completed in three hours and utilizes urine as the sample matrix. We compared the monoclonal-based assay with the polyclonal assay using twelve urine controls run multiple times over a period of three months. The controls were derived from aspirin users as well as non-users. The %CV range in the monoclonal assay was 0.3 −12.9% with an average of 4.0%. The %CV range in the polyclonal assay was 0–35.8% with an average of 10.1%. In comparing standard curves, the R2 value of the monoclonal assay averaged 0.9967 compared to 0.9894 for the polyclonal assay. Additionally, although the polyclonal assay was slightly more sensitive (limit of detection was 11.9pg/ml vs. 64.5pg/ml in the monoclonal assay), the monoclonal’s sensitivity is sufficient to detect the lower levels of 11-dh-TxB2 in patients that respond well to aspirin. To validate this, an additional urine panel was established consisting of 10 samples from both aspirin users and non-users, and 11-dh-TxB2 values from the two assays were compared. The correlation (r) between the methods was 0.944, confirming the clinical relevance of the monoclonal assay. Thus, this sensitive, urine-based monoclonal ELISA for 11-dh-TxB2 detection eliminates the potential for analytical variables that can occur in the blood-based assays while offering the sensitivity and quantitation of the ELISA.
The Prevalence and Clinical Significance of IgA Antiphospholipid Antibodies (aPL).
Authors: Shen Yu-Min , Dyszkiewicz-Korpanty Anna , Lee Ray , Balani Jyoti , Frenkel Eugene , Sarode Ravindra 
Published: April 2020

Abstract
AIMS: We investigated the prevalence and clinical relevance and significance of the various immunoglobulin (Ig) isotypes of the different aPLs and lupus anticoagulant (LAC).
METHOD: Our study registry for the year 2003 included 472 patients who had a complete antiphospholipid antibody laboratory work up and clinical evaluation. 137 had the presence of LAC, detected by the dilute Russell’s viper venom time (Dade Behring, Marburg, Germany), PTT-LA (Diagnostica Stago, Asnieres, France), or hexagonal phospholipid neutralization test (Diagnostica Stago, Asnieres, France); 211 had elevated titers of antibodies of IgG, IgM or IgA isotypes against cardiolipin (aCL), phosphatidylserine (aPS), or β2-glycoprotein-I (aβ2GPI), detected by ELISA (Corgenix, Inc, Westminister, CO, USA). 204 had no evidence of either LAC or aPL.
RESULTS: Of the patients with aPL, 67 (32%) had elevated titers of IgG, 126 (60%) had IgM, and 118 (56%) had IgA. Thrombotic events occurred in 61% (41 of 67), 52% (65 of 126), and 61% (72 of 118) of the patients with IgG, IgM, and IgA respectively. Patients with IgG and IgA isotypes of aCL, aPS and aβ2GPI had higher rates (58–67%) of thrombotic events (see table 1) than patients with IgM isotypes (46–50%). Stepwise logistic regression analysis identified elevated titer of IgA of any aPL as an independent risk factor for thrombosis (see table 2), even in the absence of LAC. Thrombotic events observed include deep venous thrombosis (25%), pulmonary embolism (12%), cerebrovascular accident (50%), myocardial infarction (8%), and peripheral arterial occlusion (3%); pregnancy complication, retinal vascular thrombosis, and dialysis access thrombosis were also seen.
CONCLUSION: Our study demonstrates that aPL of IgA isotype is prevalent and significantly associated with thrombotic events.
Proportions of Patients with Thrombosis amongst various aPLs and Ig Isotypes aCL aPS aβ2GPI Total * Two-sided P &lt; 0.05 by McNemar’s test for equality of two dependent proportions IgG 28/43 (65%) 23/36 (64%) 13/21 (62%)* 41/67 (61%) IgM 17/37 (46%)* 28/58 (48%) 45/90 (50%)* 65/126 (52%) IgG 15/26 (58%)* 22/33 (67%) 54/91 (59%)* 72/118 (61%) Total 44/79 (56%) 58/105 (55%) 85/159 (53%) 118/211 (56%) Risk Factors for Thrombosis Antibody Odds Ratio 95% Wald Confidence Limit * p &lt; 0.05 by the Wald chi-square test. Hosmer and Lemeshow goodness-of-fit test was used to assess the model fit. N=472 for the 1st four analyses, N=335 for the last three analyses. LAC 1.627 1.073–2.466* IgG regardless of LAC 1.364 0.781–2.384 IgM regardless of LAC 0.874 0.569–1.344 IgA regardless of LAC 1.587 1.012–2.490* IgG LAC negative 1.790 0.863–3.714 IgM LAC negative 0.878 0.515–1.499 IgA LAC negative 1.812 1.035–3.172*
Comparative Genomic Analysis of the 5′-Upstream Sequences of von Willebrand Factor and 27 Other Endothelial-Specific Genes Identifies Conserved Noncoding Sequences That Are DNase Hypersensitive.
Authors: Bernat John A. , Kondrashov Alexey S. , Crawford Gregory E. , Collins Francis S. , Ginsburg David 
Published: April 2020

Abstract
Von Willebrand factor (VWF), a key blood clotting protein, is a standard histologic marker for endothelial origin. Previously published studies characterizing tissue-specific regulatory elements have concentrated on the proximal promoter region (between −2,645 and the end of murine Vwf intron 1). Transgenic mouse models using these sequences have found elements needed for VWF expression in some, but not all, populations of endothelial cells. To identify potential novel VWF distal regulatory elements, as well as sequences contributing to the more general endothelial-specific expression program, we aligned human and mouse orthologous genomic sequences from VWF and 27 other endothelial-specific genes, extending from the start of transcription to the neighboring 5′-upstream gene (up to 100 kb of sequence), excluding the core promoter, and including intron 1. We analyzed the resulting 262 endothelial cell-specific gene conserved noncoding sequences (EC-CNSs) using a specific quantitative PCR assay for DNase I hypersensitive sites, an indicator of an open chromatin state which suggests the presence of a transcription regulatory complex. 33% of the EC-CNSs (including 6 of 9 upstream of VWF) were hypersensitive (HS) in human umbilical vein endothelial cells (HUVECs), while only 16% were HS in control human fibroblast cells. 20% of the EC-CNSs were HS in HUVECs only, suggesting that they may identify novel endothelial-specific transcriptional elements. When the 262 EC-CNSs were compared to CNSs from non-endothelial genes, a single motif, AGGAA(A/G), was identified as overrepresented in EC-CNSs. This association was replicated in an independent set of 28 additional genes with high HUVEC expression. The motif was also overrepresented in the subset of HS EC-CNSs. The AGGAA(A/G) motif includes the core binding sequence [GGA(A/T)] of the Ets family of transcription factors, which contains several members implicated in tissue-specific gene expression. These results suggest that the AGGAA(A/G) motif may represent the target sequence for a specific Ets family member critical for determining the endothelial-specific gene expression program. We are currently testing this hypothesis by analyzing this motif in Vwf BAC transgenic mice containing 177 kb of upstream sequence, including all ten identified EC-CNSs. Additionally, we have expanded our analysis of the VWF genomic region to include over 700 kb of sequence surrounding and including the gene, searching for additional EC-CNSs conserved across eleven mammalian species. This comparative genomic approach, coupled with functional analysis of putative regulatory elements, should provide new insight into the endothelial cell gene expression program and the more general problem of tissue-specific gene regulation.
Downregulation of P-Selectin from Activated Platelets Involves Both Receptor Internalization and Shedding upon Binding to PSGL-1.
Authors: Dole Vandana S. , Bergmeier Wolfgang , Eichenberger Sarah C. , Wagner Denisa D. 
Published: April 2020

Abstract
P-selectin is present in the alpha granules of platelets and is translocated to the cell surface where it mediates adhesion to leukocytes via binding to PSGL-1. We have previously shown that P-selectin is rapidly lost from the platelet surface when the activated platelets are infused into mice or incubated in vitro with whole blood. This downregulation of surface expression correlates with increased levels of soluble P-selectin in the plasma indicating that some of the protein is shed. The data presented here indicate that the interaction of platelet P-selectin with leukocyte PSGL-1 plays a role in P-selectin shedding. Firstly, P-selectin expression on activated platelets is retained significantly longer (1h) when the cells are infused into PSGL-1−/− mice whereas in the WT recipients, as early as 15 min after infusion P-selectin cannot be detected on the cell surface. Secondly, the concentration of soluble P-selectin in plasma of PSGL-1−/− mice is several fold lower compared to WT mice. In addition, we demonstrate that surface expression of platelet P-selectin is also downregulated by receptor internalization and that internalization is the predominant mechanism in the absence of PSGL-1. When analyzed 4h after infusion into WT mice, approximately 50% of the activated platelets which no longer express P-selectin on the surface can be reactivated to express P-selectin on the cell surface. In contrast, when infused into PSGL-1−/− recipients, most of the platelets can be reactivated to express P-selectin on the cell surface. In summary, engagement with leukocyte PSGL-1 is important for platelet P-selectin shedding and in its absence surface P-selectin is downregulated by receptor internalization. The enzymatic mechanism of P-selectin shedding and its significance needs to be elucidated.
Canonical Notch Signaling Is Dispensable for the Maintenance of Adult Hematopoietic Stem Cells.
Authors: Maillard Ivan , Pross Seth E. , Shestova Olga , Sai Hong , Aster Jon C. , Bhandoola Avinash , Pear Warren S. 
Published: April 2020

Abstract
Canonical Notch signaling operates through a highly conserved pathway that regulates the differentiation and homeostasis of hematopoietic cells. Ligand-receptor binding initiates proteolytic release of the Notch intracellular domain (ICN) which migrates to the nucleus, binds the transcription factor CSL/RBPJk and activates target genes through the recruitment of transcriptional coactivators of the Mastermind-like family (MAML). Notch signaling is essential for the emergence of hematopoietic stem cells (HSCs) during fetal life, but its effects on adult HSCs are controversial. In gain-of-function experiments, activation of Notch signaling in adult HSCs increased their self-renewal potential in vitro and in vivo. However, loss-of-function studies have provided conflicting results as to the role of physiological Notch signaling in HSC maintenance and homeostasis. To address this question, we expressed DNMAML1, a GFP-tagged pan-inhibitor of Notch signaling, in mouse HSCs. We have shown previously that DNMAML1 interferes with the formation of the ICN/CSL/MAML transcriptional activation complex and blocks signaling from all four Notch receptors (Notch1-4) (Maillard, Blood 2004). Transfer of DNMAML1-transduced bone marrow (BM) as compared to control GFP-transduced BM into lethally irradiated recipients gave rise to similar long-term stable expression of GFP for at least 6 months after transplant. DNMAML1 and GFP-transduced cells contributed equally to all hematopoietic lineages, except to the T cell and marginal zone B cell lineages, which are Notch-dependent. Expression of DNMAML1 did not affect the size of the BM progenitor compartment (Lin negative, Sca-1 positive, c-Kit high, or LSK cells), or the proportion of LSK cells that were negative for Flt3 and L-Selectin expression (containing long-term HSCs). The stem cell function of DNMAML1-transduced LSK cells was further assessed with in vivo competitive repopulation assays in lethally irradiated recipients. DNMAML1 and GFP-transduced LSK cells competed equally well with wild-type BM, as judged by their contribution to the myeloid lineage up to 4 months post-transplant, through two successive rounds of transplantation. Our data indicate that canonical Notch signaling is dispensable for the maintenance of stem cell function in adult HSCs.
Results of a Phase II Study of 506U78 (Nelarabine) in Refractory Indolent B-Cell or Peripheral T-Cell Lymphoma.
Authors: Thompson Michael A. , Pro Barbara , Sarris Andreas , Hagemeister Fredrick B. , Goy Andre , Bleyer Archie , Cabanillas Fernando F. , Samaniego Felipe , Fayad Luis E. 
Published: April 2020

Abstract
BACKGROUND: Compound 506U78 (Nelarabine), a water soluble pro-drug of 9-β-d-arabinofuranosyl-guanine, is demethoxylated by adenosine deaminase to ara-G in lymphoblasts. Intracellular ara-G is then phosphorylated via deoxycytidine kinase and mitochondrial deoxyguanosine kinase to its active 5′-triphosphate, ara-GTP. Ara-GTP, a potent inhibitor of DNA polymerase, also incorporates into DNA resulting in cell death. Studies in children and adult T-cell leukemia and non-Hodgkin’s lymphoma (NHL) have shown activity, but with reversible neurotoxicity as the most prominent adverse event.
METHODS: We report here the results of a phase II study with 506U78 in adults (median age 64, range 33–81) with relapsed or refractory indolent B-cell or peripheral T-cell NHL. Patients had received a median of 2 prior courses of chemotherapy. 506U78 was given at 1.5 g/m2/day IV on days 1, 3, and 5 every 28 days to a maximum of 6 cycles unless toxicity or progressive disease. If after 4 cycles a complete response (CR) or partial response (PR) was not achieved, then treatment was discontinued.
RESULTS: There were 23 patients enrolled including 13 with T-cell and 10 with indolent B-cell NHL. Among 19 assessable for response, the overall response rate (ORR) in T-cell NHL was 4/9 (44%) with 2 CR and 2 PR, while in indolent B-cell NHL the PR was 3/9 (33%) with no CR. In responders, the median time to progression (TTP) was 8 months (range 2–22 months). Sixteen of 22 (73%) patients evaluable for toxicity had grade 3 or 4 adverse events. Neurotoxicity included one grade 3 and one grade 4 event.
CONCLUSION: Compound 506U78 is active as a single agent in T-cell and B-cell NHL. Our response rate was higher than the ORR of 10.5% reported previously in the Czuczman et al. (2004) Blood 104(11) abstract in adults with T-cell NHL and there was less neurotoxicity in our study. This may be related to different dosing regimens (every 28 days vs. every 21 days). Compound 506U78 should be further evaluated in clinical trials.
Phase I/II Study of Oral FodosineTM, a PNP Inhibitor in Refractory Cutaneous T-Cell Lymphoma Patients.
Authors: Duvic Madeleine , Breneman Debra , Cooper Maureen , Fonseca Gustavo , Bennett J. Claude , Kilpatrick J. Michael , Bantia Shanta 
Published: April 2020

Abstract
Suppression of T-cell immunity seen in children with inherited purine nucleoside phosphorylase (PNP) deficiency supports the potential application of PNP inhibitors for the therapy of T-cell malignancies. Forodesine (FodosineTM) is a specific PNP inhibitor that raises plasma 2′-deoxyguanosine (dGuo) and intracellular dGTP levels, leading to T-cell apoptosis. IV administration of forodesine has shown activity in cutaneous T-cell lymphoma (CTCL) patients. To determine the safety, pharmacokinetics and pharmacodynamics of oral forodesine in patients with refractory CTCL, as well as preliminary evidence of efficacy, five cohorts of patients (≥ 3patients per cohort) were evaluated sequentially, receiving forodesine at 40, 80, 160, 320, and 480 mg/m2 once per day for 28 days. At the end of the treatment cycle (Day 28), all patients with stable or improved disease could enter a long-term, follow-up period. To date, 12 patients have been treated, 11 completing the 28-day cycle. Forodesine is orally absorbed (40%–50%), mean terminal half-life of 12 to 20 h. dGuo levels were elevated in all patients (0.8–2.8 μM, predose levels ≤ 0.004 μM). Forodesine was generally safe and well tolerated in this population. Safety data is available for first 7 patients. No serious adverse events possibly related to the drug occurred. The most common adverse event possibly drug related was nausea (2 patients). Of the 11 patients completing the treatment cycle, 9 began long-term treatment. Of these 9 patients, 4 have received &gt;3 months of treatment, while 1 patient continues to receive for &gt;8 months. Efficacy data are available for 8 patients: 2 with partial response, 3 with minor response, 2 with progressive disease, and 1 with stable disease (Table). Forodesine showed preliminary evidence of clinical activity in refractory CTCL patients. Additional clinical and pharmacodynamic data will be presented.
Clinical and Pharmacodynamic Activity of Oral Forodesine In CTCL Patients Patient Diagnosis Dose (mg/m2) Treatment Plasma dGuo (Cmax, M) μ Clinical Response PR = partial response; MR = minor response; SD = stable disease; PD = progressive disease; N/A = not available 2001 Sezary syndrome (IVA) 40 Completed 0.6 PR 3001 Mycosis fungoides (IVB) 40 Completed 1.7 PD 3002 Mycosis fungoides (IIB) 40 Completed 1.2 MR 3003 Mycosis fungoides (IB) 80 Completed 1.2 MR 3004 Sezary syndrome (IVA) 80 Completed 1.0 MR 3005 Sezary syndrome 80 Completed 1.6 PD 2002 IIA 80 Completed 1.1 PR 2003 III 80 Completed 1.7 N/A 3006 Mycosis fungoides/Sezary syndrome (IVB) 160 Completed 2.8 SD 4001 III 320 Completed 1.8 N/A 1001 Mycosis fungoides (IB) 320 Completed 1.3 N/A 3012 Mycosis fungoides/Sezary syndrome (IVA vs B) 320 Not Completed N/A Not evaluable
Lenalidomide and Novel Immunomodulatory Drugs (IMiDs®): A New Approach to the Regulation of Erythropoiesis and Hemoglobin Synthesis in Anemia.
Authors: de Parseval Laure Moutouh , Brady Helen , Verhelle Dominique , Corral Laura G. , Glezer Emilia , Morris Christopher L. , Chen Roger , Chan Kyle 
Published: April 2020

Abstract
Clinical trial results have demonstrated that lenalidomide (Revlimid®) reduces or even eliminates the need for red blood cell transfusions in some anemic myelodysplastic patients. We have examined whether lenalidomide and Actimid™, members of a new class of immunomodulatory drugs (IMiDs®), which are currently under evaluation for the treatment of hematological cancers could regulate erythropoiesis and hemoglobin synthesis. For this purpose, we used an in vitro culture model to differentiate human erythroid progenitors from bone marrow or peripheral blood CD34+ cells. We demonstrate that lenalidomide and AztimidTM modulate erythropoiesis and increase proliferation of immature erythroid cells. In addition to the regulation of erythroid differentiation, lenalidomide and ActimidTM are potent inducers of fetal hemoglobin. Unlike other inducers of fetal hemoglobin such as 5-aza-cytidine that are cytotoxic, IMiDs® promoted survival of erythroblast cultured with known cytotoxic drug. Gene expression profiling of erythroid differentiated cells showed that IMiDs® regulate specific erythroid transcription factors and genes that participate in hemoglobin synthesis, and genes invoved in cell cycle and cellular differentiation. Globin gene expression is controlled by IMiDs® during erythroid differentiation by inducing fetal hemoglobin synthesis. Our results support the hypothesis that IMiDs® restore effective erythropoiesis in myelodysplastic patients and protect erythroid cells from the cytotoxic effect of chemotherapeutic agents. In conclusion, IMiDs® may represent an interesting new therapy for cancer-related anemia and β-hemoglobinopathies.
GA Binding Protein (GABP) Is Required for Myeloid Cell Development and Differentiation.
Authors: Drumea Karen C. , Yang Zhong-fa , Rosmarin Alan G. 
Published: April 2020

Abstract
GABPα is an ets transcription factor that regulates genes that are required for innate immunity, including CD18 (β2 leukocyte integrin), lysozyme, and neutrophil elastase. GABP consists of two distinct and unrelated proteins that, together, form a functional transcription factor complex. GABPα binds to DNA through its ets domain and forms a multimeric complex by recruiting its partner, GABPβ, which contains the transactivation domain. GABPα is a single copy gene in both the human and murine genomes and it is the only protein that can recruit GABPβ to DNA. We cloned GABPα from a murine genomic BAC library and prepared a targeting vector in which the GABPα ets domain is flanked by loxP recombination sites (floxed allele, designated fl). Mice that bear one intact (and one disrupted copy) of GABPα, i.e. hemizygous mice, are phenotypically normal. Intercrossing of hemizygous mice yielded no nullizygous mice, indicating that homozygous loss of GABPα causes an embryonic lethal defect. To determine the effect of GABPα deletion on myeloid cell development, we bred heterozygous and homozygous floxed mice to mice that bear the interferon-responsive Mx1-Cre transgene, which express Cre in response to injection of the synthetic polynucleotide, poly I-C. Bone marrow cells underwent efficient deletion of GABPα following poly I-C injection; in contrast, other somatic tissues did not efficiently delete the floxed allele. Bone marrow, peripheral blood, and other tissues were examined for cellular morphology and flow cytometry. We compared mice that lack GABPα in bone marrow (i.e. fl/fl Mx1-Cre mice injected with poly I-C) to littermate controls (i.e. fl/fl mice injected with poly I-C). Mice that lack GABPα exhibited a striking and statistically significant decrease in granulocytes and monocytes in bone marrow and peripheral blood, compared with controls; in contrast, there was an increase in erythroid cells in GABPα null bone marrow. This indicates that the loss of GABPα has lineage-specific effects on myeloid cell development. Morphologic analysis indicates that mice which lack GABPα possess more immature granulocytes compared to control mice. Thus, GABP disruption causes a striking loss of myeloid cells in the bone marrow and peripheral blood of mice in a lineage-specific manner. Furthermore, the maturation block of murine granulocytes that is caused by GABPα disruption demonstrates the crucial role of GABP in myeloid differentiation.