bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeting Pim Kinases in Hematological Malignancies.
Authors: Gourley Eric S. , Liu Xiao-Hui , Lamb Jeremy D. , Vankayalapati Hariprasad , Grand Cory L. , Bearss David J. 
Published: October 2019

Abstract
The three Pim kinases represent a small subfamily of serine/threonine kinases known to be involved in a number of signaling pathways as downstream effectors and potent inhibitors of apoptosis. Unlike most other kinases, Pim kinases lack a regulatory domain which means they are controlled largely at the transcriptional level and that the mRNA expression levels of Pim kinases in cells correlate with their activity. While normal expression of Pim-1 kinase is seen in cells of hematopoietic origin, examination of gene expression of Pim-1 in different malignancies using cDNA microarray analysis suggests that Pim-1 is overexpressed in a large percentage of ALL, AML, CML, DLBCL, Prostatic Adenocarcinoma, Bladder, and Oral cancers. Pim-2 is also largely expressed in ALL, AML, Squamous Cell Lung and Adenocarcinoma of the Lung while Pim-3 is restricted to Melanoma, Pancreatic Adenocarcinoma, Gastric cancers. This implicates Pim-1 and Pim-2 in the onset and progression in several of hematological malignancies and therefore they represent interesting potential targets for drug development. To evaluate the potential of Pim-1 and -2 as a drug targets we have identified and synthesized a series of Pim kinase inhibitors using our proprietary CLIMB™ drug discovery process. Through the use of CLIMB™, the published Pim-1 kinase crystal structure was used to build several models that were then used to predict potential small molecule inhibitors of Pim-1 and Pim-2 from a large virtual library. A subset of leads, based on calculated binding energies as well as additional physical chemical properties, were screened using a number of in silico physicochemical and ADMET prediction algorithms to determine which compounds were most likely to be successful in a biological context. Lead candidates were initially screened using biochemical enzyme-based and cell-based assays. Cell-based activity was determined in HEL (acute megakaryocytic leukemia), K562 (chronic myeloid leukemia), MO7e (myeloid leukemia), and other human leukemia and lymphoma cell lines. From several lead candidates a series of analogs were produced with improved inhibitory activity and pharmacokinetic characteristics. In the Pim-1 and Pim-2 in vitro kinase assay and in the cell-based assay a number of leads exhibited inhibitory activity with IC50 concentrations in the low nanomolar range. Here we present the details of the biochemical and cell-based assay results as well as the activity in tumor xenograft models of our Pim kinase inhibitors.
Targeting the JAK2 Kinase in Hematological Malignancies.
Authors: Olsen Colin E. , Gourley Eric S. , Lui Xiao-Hui , Walker Jeff , Grand Cory L. , Warner Steven L. , Vankayalapati Hariprasad , Bearss David J. 
Published: October 2019

Abstract
JAK2 is an intracellular protein tyrosine kinase whose dysregulation has been implicated in leukemia, lymphoma, and myeloproliferative disorders (MPD). Increased kinase activity of JAK2, caused by point mutation of the JH2 autoinhibitory region or formation of JAK2 fusion proteins, causes increased activation of downstream signaling pathways affecting cell differentiation, proliferation, migration, and apoptosis. Through the use of CLIMB™, our proprietary drug discovery process, the published JAK2 crystal structure was used to build several models that were then used as a substrate for in silico docking of 2.3 million virtual small molecule compounds to generate a subset of leads based on calculated binding energies. These leads were further screened using a number of in silico physicochemical and ADMET prediction algorithms to determine “druggable” leads which were most likely to be successful in a biological context. Lead JAK2 inhibitor candidates exhibit low nanomolar IC50 activity against the JAK2 and JAK2 V617F mutant enzymes. Cancer cell lines expressing either the wild-type or mutant JAK2 enzyme demonstrate sensitivity to these inhibitors resulting in IC50 values in low micromolar to nanomolar range. Consistent with the inhibition of the JAK2 enzyme, activity of downstream signaling partners are severely decreased. The phosphorylation level of STAT5, a downstream modulator of JAK2 signaling, in treated HEL cell lysates was analyzed by western blot analysis. These results showed that lead JAK2 candidates caused an inhibition of STAT5 phosphorylation at a low nanomolar EC50. This series of compounds are currently being tested in in vivo xenograft models. Evaluation of lead candidates in biochemical assays against the hERG and CYP450 enzymes showed that these compounds have little inhibitory activity against these enzymes. SuperGen’s lead selective JAK2 inhibitors exhibit potent inhibition of wild-type and mutant JAK2 kinase activity translating into potent inhibition of cellular signaling pathways and cancer cell proliferation.
Evidence for HLA Class II Susceptible and Protective Haplotypes for Osteomyelitis in Pediatric Patients with Sickle Cell Anemia.
Authors: Almawi Wassim Y. , Mahdi Najat , Al-Ola Khadija , Ali Muhallab E. , Al-Subaie Abeer M. 
Published: October 2019

Abstract
Background and Objective: Osteomyelitis (OM) is a common complication and significant cause of morbidity in patients with sickle cell anemia (SCA); however, its mechanisms are poorly understood. In view of their link to inflammatory diseases and autoimmune disorders, we investigated the association of HLA class II alleles and haplotypes with the pathogenesis of OM in Bahraini SCA patients.
Patients and Methods. SCA patients comprised 42 with and 150 patients without OM; HLA-DRB1* and -DQB1* genotyping was done by PCR-SSP. HLA allele frequency was determined by the gene counting method, and haplotype frequency was determined by the maximum-likelihood method.
Results. At the allele level, only DRB1*100101 (0.229 vs. 0.082, Pc = 0.003) was positively associated with OM after applying the Bonferrni correction factor. At the haplotype level, DRB1*070101-DQB1*0201 (Pc = 0.001), and DRB1*100101-DQB1*050101 (Pc = 0.001) were more prevalent among patients, while DRB1*030101-DQB1*0201 (Pc = 0.020) and DRB1*040101-DQB1*0302 (Pc = 0.039) was more prevalent among controls, thereby conferring disease susceptibility or protection to these DRB1*-DQB1* haplotypes, respectively.
Conclusion. These results demonstrate that specific HLA halpotypes influence osteomyelitis risk in SCA, thereby suggesting that specific HLA types may serve as a marker for identifying SCA patients at high risk for osteomyelitis.
HLA-DRB1* and DQB1* Haplotype Frequencies Haplotype OM Patients Controls P Pc OR DRB1*030101-DQB1*0201 0.015 0.141 0.002 0.020 0.07 DRB1*030201-DQB1*0401 0.042 0.016 0.212 0.908 2.95 DRB1*040101-DQB1*0302 0.016 0.126 0.004 0.039 0.08 DRB1*070101-DQB1*0201 0.219 0.060 <0.001 0.001 4.58 DRB1*110101-DQB1*030101 0.056 0.108 0.278 0.962 0.53 DRB1*100101-DQB1*050101 0.208 0.063 <0.001 0.001 4.03 DRB1*130101-DQB1*060101 0.042 0.034 0.773 1.000 1.45 DRB1*150101-DQB1*060101 0.085 0.068 0.774 1.000 1.27 DRB1*160101-DQB1*0201 0.064 0.020 0.121 0.725 3.10 DRB1*160101-DQB1*050101 0.011 0.099 0.016 0.149 0.11
Evaluation of a Dye Binding Method for Determining the Solution Concentration of Low Molecular Weight Heparins.
Authors: Kim Yumi , John Boggs , Megan Hunt , Seneviratne Janaki , Yamamoto Ralph 
Published: October 2019

Abstract
Low Molecular Weight Heparins are carbohydrate-based therapeutics with anticoagulant activities. Common brands include: Lovenox, Fragmin and Arixtra, which are currently used to treat Deep Vein Thromobosis. Unlike protein or peptide-based drugs, easy test methods for measuring the concentration of carbohydrates in solution are not available. In this report, we extend the use of commercially available dye-binding reagents and protocol (Biocolor Ltd, Ireland) for rapidly measuring the solution concentration of LMWHs. The assay was evaluated according to typical validation parameters: Linearity, Accuracy, Precision (Reproducibility and Intermediate), Solution Stability and Specificity. The assay is also capable of monitoring LMWH “stability” as changes in the dye binding response appears associated with changes in Anti-fXa and anti-fIIa activity, Molecular Weight and Free Sulfate levels. The assay was also used to measure the concentration of two Products, Lovenox (99.6% of claim) and Fragmin (94% of claim), from pre-filled syringes.
A 21-Year-Old Woman with Thrombotic Thrombocytopenic Purpura as Initial Symptom of Systemic Lupus Erythematosus and a Literature Review.
Authors: Zhou Rongfu , OuYang Jian , Zhou Ming , Zhang Qiguo , Xu Yong 
Published: October 2019

Abstract
We describe a 21-year-old woman with systemic lupus erythematosus (SLE) who first presented with an thrombotic thrombocytopenic purpura (TTP), and compare the clinical manifestations and prognosis between SLE patients with thrombotic thrombocytopenic purpura in the reported literature. At the time of admission, she was suffering from petechia, purpura and had neurological symptoms. Laboratory findings demonstrated haemolytic anaemia, thrombocytopaenia and high levels of fragmentocytes. Serological test results were highly positive for antinuclear antibodies (ANA) with a particle fluorescent pattern, and also detected antibodies against Smith antibody, RNP antibody and SSA antibody but was negative for dsDNA. Direct Coombs-test was positive and D-dimer was negtive. The activity of ADAMTS13 was significantly reduced even after 4 times plasmapheresis. TTP as presenting sign in the patient with SLE was diagnosed. After 5 times plasmapheresis treatments and immunosuppressive therapy she recovered and abnormal laboratory tests were gradually returned to normal. With a follow-up of 20 months, she had a normal life. Compared with other reports, TTP can be differentiated from other thrombotic microangiopathic syndromes by its normal levels of prothrombin time, partially activated thromboplastin time (APTT), fibrinogen and direct Coombs-test. But TTP might also be a complication of SLE and the manifestations of TTP are similar to those in SLE. The detection of the fragmentation of peripheral red blood cells helped the early diagnosis of TTP. Coomb’s test might be positive when patient with TTP had SLE. ADAMTS13 and its inhibitory antibody had an important role in the pathogenesis of TTP. Plasma exchange combined with corticosteroid and cyclophsphamide should be used as early as possible in TTP patient with SLE. The prognosis was related to the treatment time and methods including plasmapheresis.
Determination of ADAMTS13 Antigen and Activity Levels in Patients with Acute Myocardial Infarction and Acute Ischemic Stroke.
Authors: Dong Ningzheng , Liu Fang , Ji Shundong , Ruan Changgeng 
Published: October 2019

Abstract
von Willebrand factor-cleaving protease (ADMTS13) is a metalloproteases regulating von Willebrand factor multimers size in plasma, its activity decrease has been reported in many physiological and pathological conditions. But information on ADAMTS13 in arterial thrombosis disease and in the general population is limited. In this study, we investigated the ADAMTS13 antigen and activity levels in 87 healthy control subjects of different ages, in 93 patients with acute myocardial infarction (AMI) and in 30 patients with acute ischemic stroke (AIS). ADAMTS13 Antigen levels were measured using a sandwich ELISA, and ADAMTS13 activity levels were analyzed by the FRETS-VWF73 method. A pool of plasma obtained from 40 blood donors was used as reference and was defined to contain 100% of the protease. The individual subject of the pool was not included in the study. Compared with 50 sex- and age-matched control subjects, ADAMTS13 antigen and activity levels were significantly lower in patients with arterial thrombosis diseases. Antigen levels were 87.8±19.8%, 61.8±18.8%, and 70.2±15.5% respectively in control, AMI and ASI patients. ADAMTS13 activity were 81.7±13.9%, 59.2±22.1% and 65.4±15.8% respectively. In the 87 healthy subjects, the range of ADAMTS13 activity is from 46.3% to 119.0%, and antigen is from 60.8% to 141.9%. In different age groups (17 to 20 years old, 21 to 40 years old, 41 to 60 years old, and older than 60 years old), ADAMTS13 antigen levels were 76.0%, 109.7%, 99.3%, and 74.5% respectively, and activity levels were 80.8%, 81.1%, 77.9%, and 87.3% respectively. It seems the antigen reached the highest level in people from 21 to 40 years old, then gradually decreased with age. The antigen and activity mean in males (87.1% and 80.8%) was lower than that in females (94.7% and 86.1%), but the difference is not significant (p=0.145 and p=0.124). In conclusion, the decrease of ADAMTS13 antigen and activity in AMI and AIS patients indicated ADMATS13 might contribute to arterial thrombosis, and the plasma ADAMTS13 antigen level in the general population was significantly affected by age.
Rituximab in Warfarin Resistance Treatment in Patients with Thrombophilia Due to Primary Antiphospholipid Syndrome: A Pilot Study.
Authors: Chavez Jaime Garcia , Mendoza-Torres Moises L. , Vela-Ojeda Jorge , Reynoso Eduardo E. 
Published: October 2019

Abstract
Introduction: The inability to obtain a therapeutic value of the INR regardless of the warfarin high doses administration represents a significant problem to resolve in the oral anticoagulation therapy, this phenomena is called “Warfarin Resistance”. Based on the experience and success of Rituximab in the treatment of autoimmune diseases like Immune Thrombocytopenic Purpura, we treated 7 patients with Primary Antiphospholipid Syndrome (PAS) and Warfarin Resistance.
Objective: To assess if Rituximab can revert the warfarin resistance in patients with thrombophilia associated to PAS
Material and Methods: We evaluated 7 patients with PAS who were receiving 15 mg of warfarin and had persistent low INR values. Rituximab was given weekly at dose of 375 mg/m2 for 4 doses. Effectiveness and safety were registered weekly during 2 months since the start of treatment with clinical evaluation and INR, aPTT, warfarin serum levels and CD20 blood lymphocytes count determinations.
Results: All patients were women with a mean of 35 years old (37– 48) and INR of 1.11 (0.79–1.50). All the patients (100%) reached the therapeutic level of INR between days 8 to 15, but the response lasted only a mean of 24 days (8 – 40). The maximum INR values were observed through the 2nd to the 3rd doses of Rituximab. The warfarin serum levels did not change during the assessment period.
Conclusion: Rituximab was effective in reverting resistance to Warfarin therapy in patients with thrombosis for primary Antiphospholipid Syndrome, however it lasted a very short time and eventually all patients relapsed.
Hemoglobin Levels Prior to Blood Transfusions in Oncology Patients Receiving Chemotherapy and Erythropoiesis-Stimulating Agents (ESAs): Observational Data from the DOSE Registry.
Authors: Larholt Kay M. , Burton Tanya , Pashos Chris L. , Bookhart Brahim K. , Piech Catherine Tak , McKenzie Scott R. 
Published: October 2019

Abstract
Objective: Blood transfusions in oncology patients (pts) occur at various hemoglobin (Hb) levels depending on each pt’s characteristics and situation, including underlying malignancy, treatment and comorbidities. Minimal data on the Hb levels at which transfusions are given in these pts are available. To characterize those Hb levels, or transfusion triggers, in oncology pts receiving ESAs, observational data from an ongoing registry were analyzed.
Methods: Real-world data on ESA-treated oncology pts in United States oncology clinics were analyzed from an ongoing prospective, observational registry (Dosing and Outcomes Study of Erythropoiesis-Stimulating Therapies [DOSE}). Data were collected from participating hospital- and community-based outpatient practices between December 2003 and July 2007. Hb values within 3 days prior to transfusion were assessed and summarized. If multiple Hb values were recorded, the Hb value closest to the transfusion was reported. Similarly, if two Hb values occurred on the same day, the lower of the values was reported.
Results: Of 334 transfusions, 314 had a Hb reported within 3 days. While the mean transfusion trigger was 8.55g/dL (SD 1.1), the trigger ranged from <7 to >10g/dL. Sixty-nine percent of transfusions occurred with a Hb trigger of ≥8g/dL, including 9% with a transfusion trigger >10g/dL, as described in the table below. Subgroup analyses based on age (< 65 v ≥65) and payer type (Medicare v other) reported similar findings.
Conclusion: Data from this ongoing observational registry found that the Hb transfusion trigger in oncology pts treated with ESAs varies considerably, with the majority of transfusion events occurring at Hb ≥8.0g/dL and approximately 9% occurring at Hb>10g/dL. Further analyses should explore pt characteristics as well as clinic-specific or other factors that may help to explain this variation.
Transfusion trigger <7g/dL 7–7.9g/dL 8–8.9g/dL 9–9.9g/dL >10g/dL All patients Number of transfusions (%) (N=314) 12 (4%) 89 (28%) 140 (45%) 46 (15%) 27 (9%) Subset by age Age≥65 (n=155) 7(5%) 45 (29%) 70 (45%) 19 (12%) 14 (9%) Age <65 (n=159) 5 (3%) 44 (28%) 70 (44%) 27 (17%) 13 (8%) Subset by payer type Medicare (n=150) 7 (5%) 45 (30%) 62 (41%) 19 (13%) 17 (11%) Other (n=164) 5 (3%) 44 (27%) 78 (48%) 27 (16%) 10 (6%)
Augmentation of Vasculogenesis by Injected Human Hematopoietic Stem Cells in Response to Ischemia Follows Early Activation of Innate Immune Response.
Authors: Finney Marcie R. , Fanning Laura R. , Kozik Margaret , Matsuyama Shigemi , Joseph Matthew E. , Goldberg Johnathan L. , Greco Nicholas J. , Haviernik Peter , Fu Pingfu , Pompili Vincent J. , Laughlin Mary J. 
Published: October 2019

Abstract
The immunocompromised NOD.SCID murine model of hindlimb ischemia has been used extensively to test in vivo the impact of human hematopoietic stem cell (HSC) therapy on neovascularization. Although NOD.SCID lack T and B cell adaptive immune responses, innate responses are intact. We utilized this model to test the hypothesis whether injection of human HSC following femoral ligation impacts neovascularization within the ischemic bed and whether early murine innate cellular responses differ in animals treated with HSC. NOD.SCID underwent femoral ligation and were divided into 3 study groups: controls (media alone), human HSC (CD133+ HSC selected from umbilical cord blood-UCB), or human peripheral blood mononuclear cells. Laser Doppler studies day +28 after femoral ligation showed significantly improved blood flow ratio in animals treated with human HSC vs. controls, 0.55 ± 0.06 (n=9) and 0.37 ± 0.03 (n=12, p<0.05) respectfully. Histology day +42 confirmed increased capillary density in animals treated with human HSC (320±18 cells/mm2) when compared to control animals (131±.6.9 cells/mm2; p<0.0001). Gene expression microarrays (Affymetrix Murine GeneChip 430 2.0) of RNA extracted from tissues harvested from the ischemic limbs 3 days following femoral ligation and injection of cells (n=3 each experimental group) comparing control mice to those receiving human HSC vs. controls revealed 414 genes with 2-fold or greater change. Included in these were genes regulating innate immunity: monocyte chemoattractant protein-5 precursor (MCP-5), CCL11, CXCL5, CXCL9, IL4 receptor, CXCL10, CCL5 and toll-like receptor 3 (TLR3). Of note large expression differences were noted comparing animals treated with human HSC including kruppel-like factor 15 (4 fold increase), IL6 receptor (3.25 fold increase), cyclin A2 (4.25 fold decrease). No significant increases in genes regulating angiogenesis were noted comparing animals treated with human HSC vs controls, including VEGF, angiopoietin, or bFGF. In summary, NOD.SCID treated with human HSC after femoral ligation demonstate significantly improved capillary density and blood flow recovery at later time points (day 28–42). Proteins regulating innate immunity are significantly higher at early time points in animals treated with human HSC suggesting an important role for inflammatory responses leading to augmentation of vasculogenesis after acute vascular injury.
Peripheral Localization and Regulation of the Murine β-Globin Locus.
Authors: Ragoczy Tobias , Telling Agnes , Byron Rachel , Bender M.A. , Groudine Mark 
Published: October 2019

Abstract
The interphase cell nucleus is structurally and functionally compartmentalized, making the subnuclear position of genes an important determinant of their activity. During cellular differentiation, as tissue-specific expression programs unfold, dynamic reorganization of the nucleus positions sets of genes in active or repressive compartments. The nuclear periphery has emerged as an unusually complex compartment in this process. While it is marked by facultative heterochromatin and has been considered primarily as a repressive compartment, recent work suggests that active genes may also associate with the periphery. Moreover, the nuclear envelope is riddled with nuclear pore complexes, the gateways for molecular exchange between the cytoplasm and the nucleus, resulting in substantial traffic through this compartment. Using murine erythropoiesis as a model system, our recent three dimensional analysis of the positioning of the β-globin locus revealed that, when inactive in undifferentiated embryonic stem cells and erythroid progenitor cells, the locus is positioned predominantly at the nuclear periphery and appears to contact the lamina. This association is lost with progressing erythroid maturation, and the locus is repositioned towards the nuclear interior concomitant with increasing β-globin gene expression. Importantly, however, β-major globin expression begins at the nuclear periphery prior to relocalization, suggesting that associations of the locus at the periphery may shift from repressive to activating complexes. We are investigating the interactions of the β-globin locus with the nuclear periphery by two approaches:
Using enhanced imaging permitted by Cryo-ImmunoFISH, we are analyzing the position of the locus relative to specific components of the nuclear lamina and nuclear pore complexes at distinct differentiation stages. In addition, we are probing the physical interactions of the locus with the periphery by biochemical means.
To this end we are using ChIP-chip to identify lamina associated proteins binding the β-globin locus and to determine what sequence elements within the locus mediate these interactions. Ultimately these experiments will shed further light on mechanisms regulating β-globin gene expression during erythropoiesis and how stage-specific nuclear localization contributes to this process.