bluuebunny/crossref_metadata_2025_split huggingface dataset:
Single Dose Administration of rFVIIa and NN1731 in Two Hemophilia A Dogs.
Authors: Ezban Mirella , Frost Lone , Viuff Dorthe , Møss Judi , Kloos Mark , Dillow Aaron , Merricks Elizabeth , Raymer Robin , Nichols Timothy C. 
Published: October 2019

Abstract
Introduction: The objective of this pilot study was to evaluate and compare the pharmacokinetic and pharmacodynamic (PK/PD) profile of rFVIIa and NN1731 in two hemophilia A dogs. In addition, it was the aim to evaluate the use of TEG for monitoring rFVIIa/NN1731 activity after in vivo administration and to compare with ex vivo spiking data from a previous study. NN1731 is a new rFVIIa analoge with enhanced activity (Allen et al. Arterioscler. Thromb. Vasc. Biol.2007;27:683–689). In hemophilia patients as well as hemophilia dogs the clot formation is impaired and reflected in coagulation assays such as thromboelastography (TEG) and APTT. The choice of hemophilia dogs is based upon the knowledge that the pharmacokinetics of human coagulations factors (FVIII, FIX and rFVIIa) as well as the effective dose is similar to that in humans. In normal dogs, it is not possible to evaluate the effect of these procoagulant proteins in coagulation assays as no impaired clotting is observed.
Methods: rFVIIa and NN1731 (280 μg/kg IV) were administered to two hemophilia dogs on separate days and plasma samples collected at different time points. FVIIa activity was measured by the FVIIa clot assay and values were used for pharmacokinetic assessment. The same pharmacokinetic models, a non-compartmental method and a two compartment model, respectively, were used as was the case in the First Human Dosing (FHD) trial of NN1731 (NN1731–1639). Analysis of PD markers in dogs included: APTT, PT and whole blood thromboelastography analysis, recently developed for use in hemophilia dogs.
Results: Based on the FVIIa activity profile in the two dogs it was observed that the values obtained at the first time point (C5 min), were higher after treatment with NN1731 than after rFVIIa. All activity based assays including TEG demonstrated that NN1731 was cleared faster than rFVIIa., FVIIa activity (FVIIa clot assay), showed a rapid initial distribution and/or elimination of FVIIa activity (t1/2α:0.3 h) followed by a less rapid elimination phase (t½β:3.5 h). Similar profile and values were obtained for NN1731 in the FHD dose study (J. Møss et al, ISTH, 2007)
Conclusions: This study indicates that in hemophilia A dogs, NN1731 and rFVIIa have distinct PK profiles and very similar to what is observed in man. All activity assays show the same qualitative profile, the FVIIa clot assay being the most sensitive assay. The TEG data obtained in vivo are in accordance with the values obtained after in vitro spiking. The data support the use of hemophilia dogs for evaluating the pharmacokinetic and pharmacodynamic profiles of FVIIa related proteins.
Correlation between Two ADAMTS-13 Activity Assays Based on Different Principles.
Authors: Geiter Sabine , Graf Markus , Vetr Helga 
Published: October 2019

Abstract
Defects in ADAMTS-13, the von Willebrand Factor (vWF) cleaving protease, are thought to be the main cause for the microvascular thrombotic disorder TTP (thrombotic thrombocytopenic purpura) that is in more than 90% of cases fatal if not treated early and appropriately. Usually this disease is clinically diagnosed, but in recent years the need for rapid and reliable diagnostic tests for ADAMTS-13 levels has increased. We present here the comparison of two commercially available assays for quantification of ADAMTS-13 activity both suitable for routine analysis but based on different principles. The two assays differ in their test principle and the readout system as follows: Assay 1 is a fluorogenic assay using a FRETS-vWF73 substrate and a kinetic measurement (TECHNOZYM®ADAMTS-13 ELISA); with this assay ADAMTS13 Antigen can also be determined in a second step. Assay 2 is a chromogenic assay and detects the cleaved vWF73 substrate by a specific monoclonal antibody (TECHNOZYM®ADAMTS-13 Activity ELISA). Citrated plasma of normal donors (n=7), of pooled normal plasma (n=15) and of TTP patients (n=14) were tested in both assays. Results are reported in both assays as percentage of normal activity. The standard for both assays is prepared from a pool of 100 normal donors and defined as 100%. The samples comprised a range from 0.2% up to 107% activity. The overall correlation coefficient between the two different activity assays was 0.96. 5 samples were found to have less than 5% activity in both assays. These results show that data obtained by the new TECHNOZYM®ADAMTS-13 Activity ELISA correlate very well with the fluorogenic assay (TECHNOZYM®ADAMTS-13 ELISA) in spite of the fact that these assays are based on very different principles.
Age-Associated Difference in Gene Expression of Pediatric Myelo-Monocytic and MLL-Rearranged AML.
Authors: Jo Aoi , Tsukimoto Ichiro , Ishii Eiichi , Asou Norio , Igarashi Takashi , Hayashi Yasuhide , Ichikawa Hitoshi 
Published: October 2019

Abstract
Acute myelogenous leukemia (AML) is a heterogeneous disease with a variety of genetic alterations including the MLL gene rearrangement. Microarray-based gene expression profiling has been applied to diagnose AML patients and to explore their underlying molecular pathology. In this study, we focused on pediatric myelo-monocytic leukemia and analyzed 40 pediatric AML patients with FAB M4 and M5 subtypes [excluding inv(16) and t(8;21) cases] using Affymetrix HG-U133plus2.0 microarray. Patients consisted of 14 infants (<1 year old) and 26 children (>=1 year old), and 23 patients possessed MLL gene rearrangements. Unsupervised analysis revealed that 10 of the 14 infants display very distinct gene expression and large set of genes were over-expressed among the infants. Among these highly expressed genes, ZFHX1B, ARHGAP26 (GRAF), and FOXO3A (AF6) were included. With the use of these distinctively expressed genes, we were able to divide the 40 pediatric patients into three subgroups: 10 infants with very high expression as Group A, 2 infants and 10 children with medium expression as Group B, and 2 infants and 16 children with low expression as Group C. The average age of Group A was 0.3 year old, Group B was 3.1 years old, and Group C was 6.6 years old. All subgroups included MLL gene rearrangements as well as normal and other complex karyotypes. FAB subtypes were equally distributed among each subgroup. The comparison of the outcomes among the subgroups showed a striking difference where Group C presented extremely poor outcome (3-year EFS 28%). Genes specifically over-expressed among Group C included poor prognostic factors such as WT1 and KIT. In addition, we found that gene expression signature of MLL rearrangement was different among the subgroups as well. For example, EVI1, SOCS2 and MEF2C were up-regulated in MLL-rearranged AML of Group C but not in the others. HOXA9 and HOXA10 were regulated independently of MLL-rearrangement in pediatric myelo-monocytic AML. These results suggest that age is an important factor contributing to the biology of pediatric myelo-monocytic AML and sub-grouping procedures can provide proper stratification so that the poor prognostic subgroup can be distinguished to receive suitable therapy.
MicroRNA Expression Profiles in Acute Myeloid Leukemia with Common Translocations.
Authors: Li Zejuan , Lu Jun , Sun Miao , Mi Shuangli , Zhang Hao , Luo Roger T. , Qian Zhijian , Neilly Mary Beth , Wang Yungui , Jin Jie , Zhang Yanming , Bohlander Stefan K. , Larson Richard , LeBeau Michelle , Thirman Michael J. , Golub Todd R. , Rowley Janet D. , Chen Jianjun 
Published: October 2019

Abstract
Acute myeloid leukemia (AML) is the most common type of acute leukemia in adults. It is estimated that 13,410 cases will be diagnosed and 8,990 will die of AML in the United States in 2007 (http://seer.cancer.gov). AML is a genetically diverse hematopoietic malignancy with variable response to treatment. Expression profiling of protein-coding genes using DNA microarray in AML has resulted in inconsistent data from different laboratories. Therefore, further validation of these observations in large cohorts and in independent studies is definitely required before clinical application becomes feasible. Recently, Golub and colleagues described a new, bead-based flow cytometric microRNA (miRNAs, miRs) expression profiling method that could successfully classify tumors. MiRNAs are endogenous ∼22 nucleotide non-coding RNAs, which can function as oncogenes and tumor suppressors. To provide new insights into the complex genetic alterations in leukemogenesis and to identify novel markers for diagnosis and treatment of AML, we performed a genome-wide analysis of miRNA expression profiles using the bead-based method on 54 AML samples with common translocations including t(15;17), t(8;21), inv(16), and 11q23 rearrangement, along with normal controls. In both unsupervised and supervised hierarchical cluster analyses, we observed that t(15;17) samples grouped together as one cluster, as do the 11q23 rearrangement samples. Interestingly, t(8;21) and inv(16), both CBF (core-binding factor) AMLs, grouped together as a unique cluster. Forty-one miRNAs exhibited significantly differential expression between different subtypes of AMLs, and/or between AMLs and normal controls. Notably, expression signature of a minimal number of two, three, and seven miRNAs could be used for class prediction of CBF, t(15;17), and 11q23 rearrangement AMLs, respectively, with an overall diagnostic accuracy of 94–96%. We further showed that overexpression of the two discriminatory miRNAs in CBF AML is associated with epigenetic regulation, rather than DNA copy number amplification. Moreover, several important target genes of these discriminatory miRNAs have also been validated. We are currently exploring the role of these discriminatory miRNAs and their critical target genes in the development of AML using in vitro and in vivo models. This work will enhance our understanding of the biological role of these miRNAs and their targets in leukemogenesis.
Deregulation of RARγ and RARγ-Specific miRNAs in AML.
Authors: Barrett Angela N. , Gupta Rajeev , Glasow Annegret , Grimwade David , von Lindern Marieke , Waxman Samuel , Mills Ken , Burnett Alan , Enver Tariq , Zelent Arthur 
Published: October 2019

Abstract
All-trans-retinoic acid (ATRA) plays important regulatory roles in hematopoiesis and is successfully used in differentiation therapy of acute promyelocytic leukemia (APL). Although ATRA effectively inhibits growth and stimulates myelomonocytic differentiation of myeloid progenitors, it is equally potent in causing expansion of multipotent hematopoietic stem cells. Results of studies utilizing mice that lack expression of a specific RAR and/or RAR subtype specific retinoids, as well as work addressing the molecular pathogenesis of acute promyelocytic leukemia (APL), indicate that the effects of ATRA on hematopoietic stem cells and myeloid progenitors are differentially mediated via the RARγ and RARα, respectively. Using qPCR we have now shown that RARγ is expressed in human cord blood derived stem cells but not in more mature myeloid progenitors or myelomonocytic cells. This change in the RARγ expression levels is paralleled by a reciprocal change in expression of RARγ specific miRNA, which we have identified and validated using multiple experimental strategies, including RARγ 3′UTR based reporter assays. RARγ is also expressed in blasts derived from non-APL AML patients (over 80% of cases examined) and AML cell lines, but is not expressed in ATRA responsive APL cell lines. The expression of RARγ miRNA, on the other hand, is markedly decreased in AML blasts when compared to the levels detected in cord blood derived CD34+ myeloid progenitor cells. Taken together our results suggest that finely tuned and miRNA mediated down-regulation of RARγ expression in the myelomonocytic lineage provides a switch from pro-proliferation to RARα mediated pro-differentiation effects of ATRA. We predict that use of a RARα specific agonist, possibly in conjunction with a strategy that negatively targets RARγ (as with RARγ selective antagonist or siRNA), would be most effective in retinoid based differentiation therapy of non APL-AML.
Truncations in CCND1 mRNA 3′ UTR Alters microRNA Regulation in Mantle Cell Lymphoma.
Authors: Chen Robert W. , Bemis Lynne , Amato Carol , Diane Birks , Han Myint , William Robinson A. 
Published: October 2019

Abstract
Mantle Cell Lymphoma (MCL) represents only 5–10% of all non-Hodgkins lymphomas, making it an uncommon but difficult form of lymphoma to treat. It has a poor prognosis among the B cell lymphomas with median survival of three years. The genetic hallmark of MCL is the t(11,14) translocation causing amplification of cyclin D1 (CCND1), a known cell cycle regulator which is overexpressed in many other cancers. MicroRNAs (miRNA) are a new class of abundant small RNAs that play important regulatory roles at the post transcriptional level. They act by binding to the 3′ untranslated region (UTR) of mRNAs and block either their translation or initiate their degradation. Recent reports have shown truncations in the CCND1 3′ UTR occur in MCL and indicate a worse prognosis. We hypothesized that truncations in 3′ UTR of CCND1 alter it’s regulation by microRNAs. Based on bioinformatics, we identified microRNA 16 with putative docking sites in the 3′UTR of CCND1. Mir-16 has been implicated as a cell cycle regulator. We identified 2 cell lines (Jeko-1 and Z138) with truncations in CCND1 3′ UTR and demonstrated increased CCND1 mRNA expression by qRT-PCR, increased protein expression by western blot, and higher proliferative potential by cell cycle. We prepared a reporter construct by ligating the full length 3′ UTR of CCND1 to GFP. We then co-transfected this construct with mimics of mir-16 into a cancer cell line and demonstrated downregulation of CCND1 protein expression by flow cytometry. In the MCL cell line Granta-519 with non-truncated CCND1, transfection with mimics of mir-16 deminstrated decreased expression of CCND1 mRNA. These studies suggest that the overexpression of CCDN1 In MCL may result from altered regulation of gene expression from loss of a miRNA regulatory site and may give new clues into the patho-biology of this disease and insights into possible new therapies.
The Risk of Thrombotic Events in Neonates Treated with Recombinant Factor VIIa.
Authors: Puetz John , Darling Ginger , Brabec Petr , Blatny Jan , Mathew Prasad 
Published: October 2019

Abstract
Background: In recent years, recombinant factor VIIa (rFVIIa) has been used in non-hemophilia bleeding situations (factor VII deficiency, trauma, liver disease, uremia, surgical bleeding, platelet disorders, and intracranial hemorrhage) for achievement of hemostasis. Although, the risk of thrombosis in hemophilia patients with inhibitors receiving rFVIIa is quite low, its use in other clinical situations has been complicated by some reports of thrombotic events. Recently, rFVIIa has been used to treat coagulopathic and/or bleeding neonates with good success. However, the prevalence of thrombotic events in these neonates is completely unknown. This study was initiated to determine the risk of thrombotic events associated with rFVIIa use in neonates.
Methods: We reviewed all published literature in neonates receiving rFVIIa. In addition, we reviewed all data submitted to the SeveN Bleep Registry, a database developed by the scientific standardization subcommittee on pediatric and neonatal hemostasis of the International Society on Thrombosis and Haemostasis (ISTH) to record all uses of rFVIIa in pediatric non-hemophilic patients. As the baseline prevalence of thrombosis for bleeding and/or coagulopathic neonates is also unknown, we also reviewed the records of 100 consecutive neonates from a single institution who received fresh frozen plasma (FFP) alone to treat their coagulopathy and/or bleeding.
Results: A total of 98 non-hemophilic neonates received rFVIIa. The majority of these neonates received rFVIIa only after failing to achieve hemostasis with standard care (FFP, cryoprecipitate, platelet transfusions). Of those receiving rFVIIa, 7 had a thrombotic event reported. In the control group that received FFP alone, 7 neonates also suffered a thrombotic event. Although the risk of thrombosis in these two groups is similar, neonates receiving rFVIIa tended to have indwelling line related thrombosis, while those receiving FFP tended to have strokes or myocardial insults. Overall the prevalence of thrombotic events in bleeding and/or coagulopathic neonates appears to be 7%, whether or not they received rFVIIa.
Conclusions: In this study, the overall prevalence of thrombotic events was similar in the rFVIIa and FFP group. As data for this study was collected in a retrospective manor, and thereby subject to publication and submission bias, a more accurate determination of the prevalence of thrombosis in neonates will require a prospective study.
Thrombelastographic Whole Blood Coagulation Parameters Demonstrate Improved Clot Strength Induced by Tranexamic Acid during Cardiopulmonary Bypass.
Authors: Sørensen Benny , Christensen Thomas D. , Jepsen Mogens Harrits , Folkersen Lars , Severinsen Inge , Hjortdal Vibeke E. , Ingerslev Jørgen 
Published: October 2019

Abstract
Background: Tranexamic acid reduces the fibrinolytic activity, and may hereby reduce bleeding associated with surgery. However, to date no studies have been published that monitor and display the effect ex vivo. We hypothesized that tranexamic acid improves clot strength and overall coagulation capacity following cardiopulmonary bypass, and the aim of this study was to assess this effect ex vivo using thrombelastography.
Methods: Patients (N=8) undergoing elective coronary artery bypass grafting using cardiopulmonary bypass were randomly assigned to receive either preoperatively 1 gram of tranexamic acid or no tranexamic acid (control group). Blood specimens were collected before and immediately after cardiopulmonary bypass, and whole blood coagulation analysis was performed on a ROTEG® thrombelastograph employing activation with a reaction mixture of tissue factor (Innovin®, final dilution 1:50000 ∼0.15 pM) and single chain recombinant tissue plasminogen activator (4 nM). The coagulation signal was processed to obtain dynamic parameters of clot formation such as maximum velocity (MaxVel), time to maximum velocity (t, MaxVel), minimum velocity (MinVel), and time to minimum velocity (t, MinVel). Overall haemostatic capacity was evaluated using a calculated parameter denoted overall coagulation quality (OCQ) = (maxvel/t, MaxVel)*(t, MinVel - t, MaxVel)).
Results: The integrated mean difference in time (seconds (sec)) to minimal velocity was consistently positive in the tranexamic acid group compared to the control group (1139 sec (95% CI: 463 to 1667) vs −1474 sec (95% CI: −2454 to −588). The overall coagulation quality was compromised in both groups, but less in the tranexamic acid group (−6.98 (95% CI: −25.68 to 11.71) vs −15.31 (95% CI: −37.28 to 6.66)).
Conclusion: Ex-vivo monitoring of the effect of tranexamic acid during cardiopulmonary bypass is feasible, and patients receiving tranexamic acid produce a significantly more lysis resistant clot and a less compromised overall coagulation quality as compared to control patients.
Functional Platelet Counting in Patients with Renal, Haematological and Cardiological Disease Using Blood Collecting Bottles Coated with Collagen, Ristocetin and Arachidonic Acid.
Authors: Smith Graham , Sheridan-Smith Loida , Smith Margaret A. , Pambakian Sam , Campbell Jim 
Published: October 2019

Abstract
Platelet function is a vital factor in preventing bleeding irrespective of circulating platelet number. As a result, patients often have their surgery cancelled due to taking Aspirin or related medications. Similarly, patients bleed in the ITU setting when their counts and humoral coagulation are normal. This raises the concept of exhausted platelets which is often difficult to prove. The availability of a bedside technique which enumerates patients’ "functional" platelets, which is reliable and reproducible, represents a major step forward in determining whether these patients require platelet transfusion. Helena Laboratories, Beaumont, Texas, US have produced blood collection bottles which are coated with platelet agonists equine collagen (C), risocetin (R) or arachidonic acid (AA) in addition to 3.2mg sodium citrate. Our group is the first in Europe to use these bottles in association with point of care testing instrumentation (Horiba ABX Pentra, Chicksands, Beds, England). Samples were taken from normal, renal dialysis, cardiac catheterisation, haematological and ITU patients into K2EDTA and the above bottles and treated identically prior to counting. Samples taken into K2EDTA served as baseline controls whilst free platelets in the synchronously taken agonist samples represented non functional platelets (the normal functional ones having aggregated onto the inside wall of the tube). Data on 54 normal subjects showed that, when using C, only 6.2+/−3.1% of platelets were non functional (NF). Similar figures were found for R and AA. In dialysis patients with eGFR 4–25mls/min (n=32), the R NF% was 58.07+/−17.59%. In a similar group of renal patients on dialysis and Aspirin (n=12), the R NF% was 60.55+/−21.86% which was not statistically different from the non Aspirin group (p=0.955). In cardiac patients, all of whom were on Aspirin, C NF% was 35.23+/−23.12%, AA NF% 51.94+/−23.5% and R NF% 51.53+/−19.04%. C vs AA showed a significant p-value of 0.003and C vs R p=0.002. AA vs R was not significant at p=0.94. These data show that ristocetin is a surprisingly sensitive method of detecting non functional platelets in renal and Aspirin users. Classically, ristocetin is not thought of being a detector of Aspirin effect although data from Sloand et al. JASN (1997) 8(5),799 shows that ristocetin may detect renal effect on platelet GpIb-IX (CD42b). These data demonstrate the potential value of ristocetin aggregation ex vivo in determining the need for platelet transfusion and may provide clinicians with an improved and more logical rationale for such an expensive therapy.
Treatment of Established Graft-Versus-Host Disease by PG490-88.
Authors: Chen Benny J. , Liu Congxiao , Chao Nelson J. 
Published: October 2019

Abstract
Graft-versus-host disease (GVHD) is a major complication following hematopoietic cell transplantation. While prophylaxis of GVHD has improved, treatment for GVHD remains unsatisfactory. PG490–88, a semi-synthetic derivative of a novel compound purified from a Chinese herb, is very effective in prevention of both acute and chronic GVHD. In this study, we tested whether PG490–88 is effective in treating established GVHD. The study was performed in a clinically relevant MHC-matched and minor mismatched model (B10.D2 to BALB/c). BALB/c (H2d, Mls-2a, Mls-3a) mice were lethally irradiated (8.5 Gy) and transplanted with 1x107 bone marrow cells and 1x108 spleen cells obtained from B10D.2 (H2d, Mls-2b, Mls-3b) mice. PG490–88 was administered intraperitoneally (i.p.) daily at a dose of 0.0535 mg/kg/day starting on day +15 post transplantation when the clinical signs of GVHD were first observed. Therefore, all animals in both vehicle control and PG490–88-treated groups developed GVHD before the treatment was started. After treatment, there was no difference in the severity of clinical GVHD between the treated and the control groups. However, PG490–88-treated mice survived significantly longer than the mice in the vehicle control group (Figure, median survival time: 50 vs. 65 days, P<0.0001). These data suggest that PG490–88 as a single agent is active in the therapy of established GVHD. Further preclinical studies will determine optimal schedules and combinations that could be utilized in clinical studies.
Figure Figure