bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Rapid and Sustained Increase in Neutrophil Elastase Levels Correlates Temporally with Decreased Coagulation Factor Activity during E.coli-Sepsis in Baboons.
Authors: Samis John A. , Stewart Kenneth A. , Taylor Fletcher B. , Nesheim Michael E. 
Published: April 2020

Abstract
Infection is the most common pathology resulting in disseminated intravascular coagulation (DIC) in humans. Disturbances of the cell interactions between neutrophils and endothelial cells are an integral part of the pathophysiology of E.coli-sepsis. Although neutrophil elastase (NE) is required for the destruction of gram negative bacteria, dysregulated NE activity towards coagulation factors may contribute to the coagulopathies that accompany infections if its activity is not properly regulated by alpha-1-protease inhibitor (A1PI). The temporal relationships between these variables were studied in plasma samples from three baboons before and after infusion of either two sublethal doses (2.50x106 cfu/kg and 1.10x108 cfu/kg) or a lethal dose (6.50x108 cfu/kg) of E.coli. In comparison to pre-infusion levels, the A1PI/NE complex levels increased by up to 17-fold after 2hrs and remained 6-fold above baseline after 53hrs as the amount of E.coli was increased. In contrast, the A1PI activity and specific activity both decreased by up to 30% during the 2–6hrs following the lethal dose of E.coli. Large transient decreases, of between 60–96%, in the factor V and VIII activity and fibrinogen levels occurred only during the 2–6hrs following infusion of the lethal dose. In addition, there was a 70–80% decrease in the factor II, VII, IX, and X clotting activity after 6–24hrs as the dose of E.coli was increased. In conclusion, there is a temporal association between the changes in A1PI activity, the appearance of the A1PI/NE complex, and the decrease of coagulation factor activity during E.coli-sepsis in baboons. Further, the sustained elevation of the A1PI/NE complexes beyond 48hrs, even after sublethal challenge with E.coli, indicates that a persistent increase in the neutrophil activity occurs as part of the inflammatory response to bacterial infection in this animal model. This study demonstrates that the neutrophil/endothelial interaction continues far beyond the first margination stage into the second ischemia/reperfusion stage during E.coli-sepsis. The results raise the possibility that clotting factor degradation/inactivation by NE as well as consumption via the coagulation cascade occurs following infusion of sublethal and lethal doses of E.coli.
At 90 Micrograms Per Kilogram, Recombinant Factor VIIa Partially Corrects Abnormal Hemostatic Function in Blood from Factor IX Deficient Patients.
Authors: Carr Marcus E. , Martin Erika J. , Kuhn Janice G. , Nolte Melinda 
Published: April 2020

Abstract
Reports of the potential clinical utility of recombinant Factor VIIa (rFVIIa, NovoSeven®, NovoNordisk) in a variety of bleeding disorders have raised the possibility that this agent may have broad spectrum hemostatic properties. We have previously demonstrated that rFVIIa produces dose dependent correction of hemostatic status in patients with Factor VIII deficiency and/or Factor VIII inhibitor. In this study we measured the dose dependent effects of rFVIIa on Platelet Contractile Force (PCF), Clot Elastic Modulus (CEM) and Thrombin Generation Time (TGT) (Hemodyne® Analyzer, Hemodyne, Inc.) in patients with factor IX deficiency. Results were compared to those seen with recombinant factor IX (BeneFIX® [GI, Wyeth]) and factor nine concentrate (Mononine® [Aventis-Behring]). Blood from three patients with factor IX deficiency was collected via aseptic venipuncture into evacuated tubes containing 3.2% sodium citrate. The samples were spiked with increasing amounts of each hemostatic agent so as to yield levels ranging from 25 to 200% of the recommended dose. At baseline, each of the factor IX deficient patients had prolonged TGT, decreased PCF and decreased CEM. Despite the fact that each patient had less than 1% of normal factor IX activity, there was significant variability between the patients in their baseline values. TGT for the three patients varied from 587 to 1200 seconds. Baseline PCF ranged from 0.9 to 1.4 Kdynes, while CEM varied from 1.6 to 5.4 Kdynes/cm2. Normal TGT, PCF and CEM values for asymptomatic controls (mean ± SD, n=25) were 392±100 seconds, 7.8±1.3 Kdynes and 18.8±5.6 Kdynes/cm2 respectively. At the 100% of recommended dose, each of the factor IX agents produced shortening of the TGT, and increases in PCF and CEM. The degree of normalization of these parameters did not appear to depend on the baseline values. The patient with the worst baseline parameters completely normalized his values at 100% doses of BeneFIX® and Mononine®. The patient with the best baseline values also normalized his parameters with BeneFIX® but not with Mononine®. The patient with intermediate baseline abnormalities improved all parameters with both factor IX products but appeared to respond better to BeneFIX® at the 100% dose level. The response to NovoSeven®, at 100% of the dose recommended for treatment of factor VIII inhibitor patients, was detectable but clearly inferior to the response to the factor IX agents. These results indicate significant variability in hemostatic status between factor IX deficient patients both at baseline and in response to factor replacement. While rFVIIa has activity in these patients, the dose previously shown to correct TGT in factor VIII deficient patients may not be appropriate for factor IX deficient patients.
Estrogen Replacement Therapy (ERT) Is Not Associated with an Increased Risk of Leukemia.
Authors: Ross Julie A. , Sinner Penny J. , Blair Cindy K. , Cerhan James R. , Folsom Aaron R. 
Published: April 2020

Abstract
Recent studies have reported an increased risk of breast, endometrial, ovarian, and gall bladder cancer associated with ERT. One proposed mechanism by which ERT increases risk is through the binding of estrogen to receptors at the tissue site. Estrogen receptors are also present on certain hematopoietic cells as well as on myeloid leukemia cells. Further, downregulation of the estrogen receptor through DNA methylation has been associated with increased acute myeloid leukemia (AML) survival. We evaluated whether ERT is associated with an increased risk of leukemia, particularly AML, in the Iowa Women’s Health Study. A cohort of 37,172 post-menopausal Iowa women ages 55 to 69 years with no history of prior cancer was linked annually to the population-based state cancer registry through 2001. In addition to other self-reported cancer risk factors, participants were asked about current and former use of ERT in 1986 and on four subsequent follow-up questionnaires. A total of 201 cases of leukemia were identified over 16 years of follow-up including 64 AMLs and 87 chronic lymphocytic leukemias (CLLs). Compared to never users of ERT at study baseline, current [multivariate relative risk (RR), 1.09; 95% Confidence Interval (CI) 0.70–1.72) and former users (RR=0.81, 95% CI=0.58–1.14) were at no increased risk of developing leukemia. For AML, current users also had no increased risk (RR=0.86, 95% CI=0.39–1.92) and there was a suggestion that former users had a slight decreased risk (RR=0.68, 95% CI=0.38–1.22). For CLL, all results were around unity. Duration of ERT or time-dependent use had no appreciable effect on the RRs. We conclude that ERT use is unlikely to be a risk factor for leukemia, including AML. Supported by NCI R01 CA39741.
Effect of Age on Prognosis in Childhood Acute Myeloid Leukemia.
Authors: Razzouk Bassem I. , Estey Elihu , Pounds Stanley B. , Lensing Shelly , Pierce Sherry , Brandt Mark , Rubnitz Jeffrey E. , Ribeiro Raul C. , Rytting Michael , Pui Ching-Hon , Kantarjian Hagop M. , Jeha Sima C. 
Published: April 2020

Abstract
Age affects the treatment outcome in adults with acute myeloid leukemia (AML) but its effect in pediatric AML is less certain. We reviewed outcome in 423 patients, 21 years of age or younger, who were treated for newly diagnosed AML (excluding acute promyelocytic leukemia) at St. Jude Children’s Research Hospital (n=288) or M.D. Anderson Cancer Center (n=135) between 1983–2002. Patients were divided into those treated between 1983–1989 and 1990–2002. During these two time periods, distinct sets of protocols were used in both institutions, with higher intensity treatment used in the more recent era. After accounting for the effects of cytogenetics, Down syndrome (favorable), increased WBC (unfavorable), FAB type M7 (unfavorable) and early treatment era (unfavorable), every additional year of age conveyed a 4.4 % increase in the risk of death and a 3.3% increase in the risk of any adverse event (death, failure to achieve complete remission, or relapse), with p&lt; 0.001 for the effect of age on survival or event-free survival (EFS). When patients were divided into those above or below the median age (10 years), there was a significant interaction between age and treatment era (p=.05), with patients below 10 having better EFS when treated in the recent era. For patients below 10, the relative risk of an event was 1.6 folds higher in the early treatment era than in the recent era ( p=.016). For patients 10 and older, the improvement in EFS according to treatment era was not significant. Treatment site did not influence the EFS or survival, and the effects of age were similar at both sites. These results suggest that age is an independent prognostic factor in childhood AML with younger children benefiting from more intensive treatment than their older counterparts.
Rapid and Quantitative Detection of Mutations of the FLT3 Gene in AML.
Authors: Gilkes Amanda F. , Walsh Valerie , Sweeney Marion C. , Austin Steven J. , Gale Rosemary E. , Mills Kenneth I. 
Published: April 2020

Abstract
The most common single mutation observed in approximately one-third of AML cases, is an activating mutation of the tyrosine kinase receptor FLT3. The presence of the mutation is associated with adverse prognosis in terms of increased risk of disease relapse. The majority of mutations (25–35% of AML patients) occur as an internal tandem duplication (ITD) within the juxtamembrane region. A further 3–7% of patients have a mutation within the tyrosine kinase domain (TKD) of FLT3. The type of mutation, and its population size, may have prognostic implications. We have used the Agilent 2100 Bioanalyser to assay PCR reactions for the ITD and the TKD mutations. The “chips” used within the Bioanalyser separate RNA or DNA through glass etched micro channels filled with gel matrix, nucleic acid is stained and laser detected. The PCR primers and protocol for the ITD analysis were as previously published (Kottaridis et al. Blood. 98, 2001), but within our laboratory the reactions were analysed on the Bioanalyser using a DNA 500 chip. We detected the presence of an ITD in 19.9% of the 291 AML samples, with percent of mutant RNA varying from 2.9% to 97.5% (median 27%) and with the size of the ITD varying from 15–212 bp (median 41 bp). A separate PCR reaction using primers flanking the TKD domain was used, in combination with an Eco RV or Hinf I (Smith et al, MTAL Workshop, 2003) digestion, to detect and quantitate the different mutations. Distinctive digestion and/or heteroduplex patterns were observed that identified D835, D835del, I836del or D839G mutation. TKD mutations were observed in 12.9% (10.1% D835 mutations, 1.4% I836del, 1.4% D839G) of samples examined, with the mutant RNA ranging from 1.0% to 52.8%. The D835/6 mutation rate was higher than previously reported and may be due to the efficiency and sensitivity of the 2100 Bioanalyser for the analysis of FLT3 mutations. This gave an overall FLT3 mutation rate in the AML population studied of approximately 32.8%; females had a higher overall incidence of 39.1% compared to 27.2% for males. The samples were derived from two NCI trials, AML 14 for patients over 60 years old and AML15 for younger patients. The rate of ITD mutations was 15.3% in AML14 patients compared to 27.5% in AML15, although the incidence of TKD mutations was similar (13.5% v 14.3%). The incidence of ITD mutations was over 45% in patients in the 30–40 year age group at diagnosis. A large percentage of these patients have also been gene expression profiled and the molecular effect of the FLT3 ITD mutations across the age groups has also been examined.
Loss of Heterozygosity in Childhood Acute Lymphoblastic Leukaemia Detected by Genome-Wide Microarray Single Nucleotide Polymorphism Analysis.
Authors: Hall Andrew G. , Bloodworth Lisa C. , Hogarth Linda A. , Bown Nick P. , Irving Julie A. 
Published: April 2020

Abstract
Loss of heterozygosity (LOH) is detectable in many forms of malignancy, including leukaemia, using techniques such as microsatellite analysis and comparative genomic hybridisation. However, these techniques are laborious and require the use of relatively large amounts of DNA if the whole genome is to be examined. Here we describe the use of oligonucleotide microarrays to characterise single nucleotide polymorphisms (SNPs) in lymphoblasts isolated from children with acute lymphoblastic leukaemia for the pan-genomic mapping of LOH with a resolution of 100–200kb. Results were compared with DNA obtained during remission and on relapse. Abnormalities were seen in 8 of 10 cases. The two cases with no abnormalities and one case which showed identical changes affecting whole chromosomes at relapse and presentation remain in remission 1–9 years following retreatment. The 7 cases which showed LOH not affecting entire chromosomes died following relapse, suggesting that partial LOH may be associated with a poor prognosis. In 4 cases LOH was only detectable at relapse suggesting that progressive LOH may be a cause of disease progression and/or drug resistance. This was supported by detailed analysis of one case in which LOH involving the glucocorticoid receptor (GR) was associated with mutation of the remaining allele. In cell line models the loss of a functional GR is associated with profound resistance to steroids. The most frequent abnormality detected in this series involved chromosome 9p. In each of the four cases where this was observed LOH included the INK4 locus. In three of the four cases INK4 loss was only observed at relapse (see figure), suggesting that this abnormality may be commonly associated with treatment failure, supporting previous reports that 9p abnormailities are associated with a poor prognosis. One case was reported as showing monosomy 20 as the sole cytogenetic aberration but LOH analysis identified 9p LOH and loss of 20q, with retention of heterozygocity for 20p. These findings strongly implicate unbalanced translocation der(9)t(9;20),-20 as described by Clark et al (Leukaemia, 2000, 14:241). Our observations demonstrate that SNP array analysis is a powerful new tool for the analysis of allelic imbalance and unbalanced translocations in leukaemic blasts.
Generation of Anti-Leukemia Reactive T-Cells by Costimulation with Established B-ALL Cell Lines Expressing Costimulatory Molecules.
Authors: Yan Ying , Steiherz Peter , Ruan Jianda , Chen Yibang , Abhyankar Sunil , Guan Xiuqin , Williams Casey , Belt Robert , McGuirk Joseph 
Published: April 2020

Abstract
Activated B cells expressing the B7 family of molecules are efficient antigen presenting cells. However, most of B-lineage leukemia cells do not fullfill these criteria, having lost their immunostimulatory potential both in vivo and in vitro. We have established 3 human B-ALL(L3) cell-lines (BA-25, BA-78 and BA91) which in comparison with an established pre-B-ALL cell line (BA-127) and other patients-derived pre-B-ALL blast cells do express the costimulatory molecules CD80 and/or CD86, as well as other critical costimulatory molecules for T-cell stimulation such as: CD40, ICAM-I, LFA-3 and CD72. Flow cytometry analysis demonstrate that these B-ALL cells have an activated B-cell phenotype (CD19+, CD20+, CD22+, CD38+, CD71+, CD75+, DR+, HLA-class-I and HLA-class-II). Chromosomal translocations consistent with B-ALL (L3) were determined in BA-25 [t(2;28)(p12;q24)], BA-78 [t(8;14)(q24,q32)] and BA-91 [t(8;14) (q24,q32)]. No EBV DNA sequence was found in these B-ALL cell lines by Southern blot analysis with a probe for EBV genome. In mixted lymphocyte reaction (MLR) assay, Allogeneic T cells were stimulated by BA-25 and BA-91 cells at a B : T cells ratios of 1/30, 1/100, 1/300 and 1/1000. The BA-25 and BA-91 demonstrated 1-2 logs higher ccpm than the control pre-B-ALL cells (BA-127) which with no expression of B7 family molecules. The dose response curves for the B-cell mixture ratio display a linear pattern between 217-213 log2 ccpm. To determine T cell stimulation potential of the B-ALL cells, we co-cultured allogeneic PBMC with irradiated (3,000 Rad x-ray) BA-25 cells at 30:1 effector/stimulator ratio and achieved a 3–4 fold expansion of T cells after one round 7-day stimulation. Stimulation of allogeneic PBMC with BA-25 elicited proliferation of the T-cells and lysis of BA-25 as an target in a 4h Cr51 release assay, demonstrating BA-25 as an antigen presenting cell. Studies also show that the BA-25 cells could stimulate allo-cytolytic reactions by CTLs against other human leukemic target cells which lack these molecules and fail to stimulate T cell responses by themselves. Thus, BA-25 and other similar lines may prove useful for the provision of costimulatory signal needed to stimulate T-cell response against leukemias lacking these molecules.
Use of Plasma RNA for Real-Time Quantitative RT-PCR to Monitor Imatinib Therapy in Patients with Chronic Myeloid Leukemia.
Authors: Thomazy Vilmos A. , Kantarjian Hagop M. , Imam Maha , Davies Peter J. , Aguilar Christine , Jilani Iman , Cortes Jorge , Giles Francis J. , Albitar Maher 
Published: April 2020

Abstract
Monitoring minimal residual disease (MRD) in chronic myeloid leukemia after imatinib therapy can help in determining therapeutic strategy. Sensitive methods based on the detection of the BCR/ABL fusion gene or its fusion transcript by FISH or RT-PCR, respectively, are used to monitor residual leukemia in peripheral blood or bone marrow cells. While sensitive, these methods require an invasive procedure to obtain bone marrow. The patchy nature of the residual disease may cause some of the reported variation in detecting MRD. We hypothesized that plasma RNA level reflects total body disease and is unaffected by the tendency of some myeloid cells not to circulate. We measured the BCR/ABL fusion transcript by real-time quantitative RT-PCR using RNA isolated from blood plasma. We used a quantitative RT-PCR assay capable of measuring both the b2a2 and b3a2 BCR/ABL fusion transcripts by targeting exon 13 of BCR with the 5′ and exon 2 of ABL by the 3′ primer. RNA isolated from 10–30 μl of frozen plasma was subjected to real-time quantitative RT-PCR using the Taqman technology. Quantification was achieved by extrapolation from a standard curve generated from a ten-fold dilution series of a synthetic oligonucleotide corresponding to the (+) strand of the target sequence. To normalize the fusion mRNA levels we also measured β-actin mRNA from the same samples. Assays were run in duplicate. The results of plasma RT-PCR were correlated with results of conventional cytogenetic analysis, FISH, and quantitative RT-PCR of bone marrow suspension (“bone marrow PCR”). Plasma and bone marrow samples were obtained at 3, 6, 9, 12 and 15 months after imatinib therapy. Ninety-five plasma samples were analyzed. There was a strong positive correlation between the results of plasma RT-PCR vs. positive cytogenetics (n=95, p&lt;0.00001), FISH (n=88, p&lt;0.00001), and bone marrow RT-PCR (n=74, p&lt;0.00001). At 12 months after imatinib therapy all but 2 of the 29 patients were in complete hematologic and cytogenetic remission (CR); one patient was in partial remission (PR) and one in hematologic, but not cytogenetic, remission. These two non-CR patients were positive by both bone marrow and plasma-based RT-PCR assay for the fusion transcript. Three of the four CR patients positive by both plasma and bone marrow PCR at 12 months progressed and converted to PR/minimal response at 15 months post-therapy. The level of plasma BCR/ABL fusion mRNA at 12 months was significantly higher (p=0.04) in these patients than in those who did not progress. Our results show that, by using 10–30 μl of plasma per assay, RT-PCR is at least as sensitive as cytogenetic or FISH analysis of bone marrow cells in monitoring MRD.
Minimal Residual Disease (MRD) Evaluation by Flow Cytometry in Acute Leukemia Patients Undergoing Allogeneic Stem Cell Transplantation.
Authors: Vidriales Maria-Belen , Pérez Jose J. , López-Berges Consuelo , Gutierrez Norma C. , Ciudad Juana , Vazquez Lourdes , del Cañizo Consuelo , Díaz-Mediavilla Joaquín , Fernández-Calvo Javier , Ramos Fernando , Rodriguez María Jesús , Calmuntia María José , Orfao Alberto , Miguel Jesús San 
Published: April 2020

Abstract
Investigation of minimal residual disease (MRD) in acute leukemias by immunophenotyping is increasingly used for disease monitoring. Most MRD studies using flow cytometry techniques have focused on patients treated with conventional chemotherapy, while information on its value in patients undergoing allogeneic transplantation is scanty. The aim of the present study is to evaluate whether or not immunophenotypical assessment of MRD could also be a valuable tool in patients undergoing allogeneic transplantation for acute leukaemia. For that purpose we have analysed the level of MRD before and after (month +3) transplantation, by multiparameter flow cytometry, in a series of 38 acute leukaemia patients (26 ALL cases -20 cases in 1st CR and 6 in 2nd CR- and 12 AML cases -all of them in 1st CR), that showed an aberrant immunophenotype at diagnosis. Although the level of MRD in the BM obtained before transplantation showed a tendency to predict RFS, differences did not reached statistical significance. By contrast, the evaluation of the BM obtained 3 months after allo-transplantation had significant prognostic value. Thus, patients with low MRD levels (&lt;0.05% for ALL and &lt;0.2% for AML) (n=17) had a significantly longer RFS than patients with MRD positive (median RFS: 40 vs 16 months) (p=0.001). Multivariate analysis showed that the immunological evaluation of MRD after transplantation had independent prognostic value for RFS in acute leukaemia, together with age, WBC count, and cytogenetics. Our results suggest that immunological analysis of MRD should be included in the follow-up evaluation of acute leukaemia patients undergoing allogeneic stem cell transplantation.