bluuebunny/crossref_metadata_2025_split huggingface dataset:
Investigating the Mechanisms Underlying FVIII Antibody Production in Hemophilic Mice Following Recombinant and Plasma-Derived FVIII Infusion
Authors: Qadura Mohammad , Waters Braden , Burnett Erin , Chegeni Rouzbeh , Othman Maha , Lillicrap David 
Published: April 2020

Abstract
The formation of Factor VIII (FVIII) antibodies after FVIII treatment in hemophilic patients is currently the major treatment-related complication faced by hemophilic patients. It has been reported that the use of plasma-derived FVIII (pdFVIII) in patients and animal models of hemophilia A will result in reduced anti-FVIII antibody formation compared to recombinant FVIII (rFVIII). In an attempt to explain this phenomenon, an initial hypothesis relating to the uptake of FVIII by antigen presenting cells has been formulated. In contrast, here we examine the hypothesis that the cytokine microenvironment induced by pdFVIII has a critical influence on the reduction of anti-FVIII antibody titers in hemophilia A mice. Micro-array and confirmatory quantitative RT-PCR experiments performed on CD11c+ dendritic cells, isolated 24 hours after rFVIII or pdFVIII infusion, show that pdFVIII causes a different immune gene expression profile than rFVIII administration. Both treatments caused the up regulation of pro-inflammatory gene expression. However, the rFVIII and pdFVIII treatments caused the up regulation of genes that induce Th1 and Th2 responses, respectively. Moreover, after administering rFVIII or pdFVIII concentrates to mice, we observed that each treatment induced a distinct T cell splenic cytokine micro-environment. Recombinant FVIII induced the release of Th1 cytokines and IL10, while pdFVIII induced the release of Th2 cytokines and TGF-α. In this mouse model, we observed the formation of high titers of anti-human VWF antibodies in the pdFVIII-treated mice and we propose that in these animals, VWF competes with FVIII for antigenic presentation. We further investigated the potential of antigenic competition by treating mice with FVIII and increasing concentrations of another unrelated protein, recombinant human factor IX (FIX). Our studies have shown an inverse relationship between increasing concentrations of FIX and the production of anti-FVIII antibodies. In summary, these studies allude to new and additional mechanisms that may contribute to the reduction of anti-FVIII antibody development in hemophilia A mice treated with pdFVIII.
Quality of Life Scores Pre and Post Autologous Stem Cell Transplant for Patients with Lymphoid Malignancies
Authors: Dabney Jane , Rybicki Lisa , McLellan Linda , Foster Larry , Visnosky Michelle , Curtis Julie , Bolwell Brian 
Published: April 2020

Abstract
We performed prospective quality of life assessments on 304 patients with non-Hodgkin’s lymphoma (NHL), Hodgkin’s disease (HD), or Multiple Myeloma (MM), undergoing an autologous stem cell transplant (ASCT) from June 2003 through December 2007. Patients underwent QOL assessment one to two weeks prior to stem cell mobilization for ASCT, and 40–44 days (Day 42) post ASCT. We used the Functional Assessment of Cancer Therapy–Bone Marrow Transplant (FACT-BMT) assessment tool which includes five component scores: physical well being (PWB), social well being (SWB), emotional well being (EWB), functional well being (FWB), and additional concerns (AC). The total scores the sum of the score of the five components. We also administered a Profile of Mood State (POMS) scale which measures depression, vigor, anger, tension, confusion, and fatigue. Of 304 total patients, 65% had NHL, 21% MM, and 14% HD; median age was 53; 60% were male; 25% received prior radiation therapy; preparative regimens were busulfan based or melphalan; all patients received peripheral stem cells alone, median inpatient length of stay was 21 days. Using the FACT scale, there was a significant change from baseline data to day 42 scores. 218/304 patients completed baseline and Day 42 evaluations. Compared to baseline, patients had a significantly increased day 42 EWB score (p &lt; 0.001), which occurred despite the fact that the functional well being scores decreased in a statistically significant fashion (p = 0.002). Using the POMS tool, statistically significant changes from baseline to day 42 were a reduction in depression (p &lt; 0.001), a reduction in anger (p &lt; 0.001), a reduction in tension (p &lt; 0.001), and a reduction in confusion (p &lt; 0.001), which occurred despite the fact that patients also had a reduction in vigor (p &lt; 0.001). We conclude that using these two QOL tools despite a reduction in functional well being and vigor one month after ASCT, other parameters of coping, including emotional well being, depression, anger, and tension, improve as patients are recovering from the transplant experience. We hypothesize that this reflects resolution of pre transplant fears and anxiety. Whether these scores can be further improved with additional intervention requires further study.
A Comparison of IEF and MSMS for Clinical Hemoglobinopathy Screening in 40,000 Newborns
Authors: Daniel Yvonne , Turner Charles , Farrar Lisa , Dalton R. Neil 
Published: April 2020

Abstract
Currently, newborn haemoglobinopathy screening is carried out using HPLC or isoelectric focusing (IEF). We have previously described a rapid and specific electrospray mass spectrometry–mass spectrometry (MSMS) technique, using multiple reaction monitoring (MRM) based peptide analysis, for simultaneous detection of the clinically significant haemoglobinpathies; haemoglobin (Hb)S, HbC, HbE, HbDPunjab and HbOArab. Here we report the results of a comparison of 40,000 newborn blood spots screened by both IEF and MSMS.
For both IEF and MSMS analysis, blood spots (3.2mm) were punched into separate 96 well plates. IEF was performed using the Resolve haemoglobin test kit (PerkinElmer Life Sciences, Waltham, USA) and Isoscan imaging system. For MSMS analysis, the blood spots were digested for 30min at 37°C with a trypsin reagent, and diluted in mobile phase (acetonitrile: water, 50:50, with 0.025% formic acid. Sample, 2μl, was injected directly into the mobile phase (flow rate 80μl/min) and analysed, in positive ion mode, using a Sciex API4000 (Applied Biosystems, Warrington, UK). Specific MRM transitions for HbS, HbC, HbE, HbDPunjab, HbOArab, normal beta, alpha, gamma and delta chains were acquired; total acquisition time per sample was 60 sec. This enabled identification of sickling disorders and thalassaemia major, as well as assessment of transfusion state and potential identification of HbLepore and HbBarts.
40,000 blood spot samples for routine newborn haemoglobinopathy screening were analysed in parallel. HbS was detected in 199 samples; 8 were HbS/HbF only and 3 HbSC. HbC was detected in 39 samples, HbDPunjab in 52, HbE in 48. No HbOArab or HbLepore mutations were detected by either method. There have been no discrepancies between the analytical techniques. Using MSMS, mutation positive samples can be re-run in product ion scan mode to provide peptide sequence and hence unequivocal confirmation of the haemoglobin variant. In addition, 5,000 samples were analysed on a Sciex API4000 Q trap; using the information dependent acquisition facility provided “real time” peptide sequencing thus removing the requirement for re-injection.
Sample preparation is very quick and simple for both methods, but the consumable costs associated with the MSMS technique are &lt;10% of those for IEF. The capital cost of MSMS can be offset by high throughput and/or integration with current inherited metabolic disease screening by MSMS. The specificity of the MSMS analysis implies that haemoglobinopathy detection can be limited to specified conditions, based on agreed screening policy, and can eliminate the need for costly and time consuming second line testing.
This study demonstrates that newborn haemoglobinopathy screening can be carried out rapidly, easily, and cost effectively using MSMS technology. It enables rationalisation of technology platforms in newborn screening by consolidating screening for haemoglobinopathies and inherited metabolic diseases onto MSMS.
Engineering Kunitz Domain of Human Tissue Factor Pathway Inhibitor-2 to Selectively Inhibit Fibrinolysis: An Antifibrinolytic Agent with Potential to Replace Aprotinin
Authors: Bajaj Madhu S , Ogueli Godwin I , Schmidt Amy E , Shanker Sreejesh , Bajaj S. Paul 
Published: April 2020

Abstract
Tissue factor pathway inhibitor-2 (TFPI-2) inhibits factor XIa, plasma kallikrein and factor VIIa/tissue factor; accordingly, it has been proposed for use as an anticoagulant. Full-length TFPI-2 or its isolated first Kunitz domain (KD1) also inhibits plasmin and therefore it has been proposed for use as an antifibrinolytic agent. However, the anticoagulant properties of TFPI-2 or KD1 would diminish its antifibrinolytic function. In this report, structure based investigations and analysis of the serine proteases profiles revealed that coagulation enzymes prefer a hydrophobic residue at the P2′ (nomenclature of Schechter and Berger, BBRC, 27:157–162, 1967) position in their substrates/inhibitors, whereas plasmin prefers a positively charged arginine residue at the corresponding position in its substrates/inhibitors. Based upon this observation, we changed the P2′ residue Leu17 (bovine pancreatic trypsin inhibitor/aprotinin numbering) in KD1 to Arg (KD1-L17R) and compared its inhibitory properties with the wild-type KD1 (KD1-WT). Both WT and KD1-L17R were expressed in E. Coli, folded and purified to homogeneity. Amino-terminal sequences and mass spectra revealed proper folding of the KD1-WT and KD1-L17R. As compared to KD1-WT, the KD1-L17R neither prolonged the activated partial thromboplastin time of normal plasma nor it inhibited factor XIa, plasma kallikrein or factor VIIa/tissue factor. Further, KD1-L17R inhibited plasmin with ~4-fold increased affinity. In a mouse liver laceration model of bleeding from small vessels, KD1-L17R reduced total blood loss by 84% compared with KD1-WT, which reduced total blood loss by 10%. Moreover, in this bleeding model, KD1-L17R was more effective than aprotinin (70% reduction), which has been used as an antifibrinolytic agent to decrease blood loss during major surgery. In this model, KD1-L17R was also more effective than the lysine analogue tranexamic acid (52% reduction). In additional studies, in a tail transection model of bleeding from a large vessel, KD1-L17R reduced total blood loss by 70% and was more effective than KD1-WT (46% reduction), aprotinin (43% reduction) and tranexamic acid (67% reduction). Notably, as compared to aprotinin, renal toxicity manifesting as multifocal tubular necrosis by histopathology was not observed with KD1-L17R or KD1-WT. In conclusion, KD1-L17R is a specific inhibitor of plasmin without anticoagulant properties and is more effective in reducing blood loss compared with known antifibrinolytic agents in clinical use.
The Small GTPase Rap1A Is Central for Migration and Adhesion of Mesenchymal Stromal Cells (MSCs)
Authors: Deak Erika , Ruester Brigitte , Göttig Stephan , Puccetti Elena , Ruthardt Martin , Seifried Erhard , Scheele Jürgen , Henschler Reinhard 
Published: April 2020

Abstract
MSCs are currently intensively investigated for their tissue regenerative and immunomodulatory properties, and for many applications are applied via the intravenous route. We have previously shown that MSCs roll and adhere on endothelial cells both in vitro and in vivo, using part of the array of the adhesion receptors which are known to mediate adhesion of hematopoietic stem and progenitor cells during mobilization from bone marrow and homing after transplantation (Blood108:3938, 2006). To identify further gene products which are potentially relevant for coordinated circulation behaviour of MSCs, we kept human MSCs under shear stress by continuous stirring for 0, 3, 6 and 12 h, isolated RNA and performed cDNA microarrays. We found approx. 500 out of 21.000 sequences regulated by shear stress. Interestingly, differentiation-associated genes (osteogenic, lipogenic) were generally downregulated. Several adhesion receptors, including integrins α4 and α7 were time-dependently upregulated. Among G proteins and -associated factors, the Guanosin Nucleotide Exchange Factors (GEFs) for the small GTPase Rap1, RAP-GEF2 and PDZ-GEF1 were both strongly upregulated by shear stress, whereas Rho GTPase regulator RGS-3 was downregulated. We next isolated MSCs from murine bone marrow from Rap1A deficient or wild type mice. Wild type MSCs were found to express Rap1A RNA and protein. However, it was not possible in 5/5 cases to isolate MSCs by the culture expansion method from Rap1A−/− mouse bone marrow. Yet we were able to derive and analyze MSCs from Rap1A+/− mice, and found that their binding to immobilized HUVEC endothelial cells in laminar flow chambers was approximately threefold reduced compared to wild type controls. Similarly, shear stress resistant adhesion to both, Vascular Cell Adhesion Molecule (VCAM)-1 and Intercellular Adhesion Molecule (ICAM)-1 was about 50% decreased compared to wild type MSCs isolated from littermate mice. Moreover, SDF-1-induced adhesion strengthening on VCAM-1 and ICAM-1 was also decreased in the Rap1A+/− MSCs. Transfection of MSCs with an expression vector encoding dominant negative Rap1 (Rap1N17) decreased, whereas pretreatment of MSCs with the Rap1 specific EPAC activator, 8-Br-(4PCT)-cAMP increased adhesion of MSCs to VCAM-1. These data demonstrate an essential role of the GTPase Rap1A for integrin-mediated adhesion of MSCs. Interference with Rap1 signalling thus provides a means to modulate adhesion, and potentially, homing behaviour of MSCs.
Id1 Provides a Proper Hematopoietic Progenitor Niche Function
Authors: Suh Hyung Chan , Ji Ming , Gooya John , Lee Michael , Klarmann Kimberly , Keller Jonathan R. 
Published: April 2020

Abstract
Development of hematopoietic stem cells (HSC) and their progeny is maintained by the interaction with cells in the microenvironment. In addition to hematopoietic cells, Id1 is expressed in stromal cells known to support hematopoiesis, and is involved in cell proliferation, differentiation and senescence. Therefore, to investigate the role of Id1 in hematopoiesis, we examined hematologic phenotypes of Id1−/− mice. In this study, we found increased neutrophils and macrophages, and decreased B cells and platelets in peripheral blood, and decreased BM cellularity. While the percentages of hematopoietic stem cells (HSC) in Id1−/− mice were increased relative to the Id1+/+ mice, their total numbers and function appeared normal. For example, Id1 was not required for self-renewal or repopulation of HSC. In contrast, we found that there were increased numbers of hematopoietic progenitor cells (HPC) in S phase of cell cycle in Id1−/− mice BM, suggesting that the loss of Id1 within HPC promotes proliferation. However, purified Id1−/− HPC had the same proliferation potential as Id1+/+ HPC when cultured in vitro. In transplantation experiments, we proved that BM microenvironment in Id1−/− mice is defective by showing that the Id1+/+ HSC showed impaired hematopoietic development in Id1−/− mice, while the Id1−/− HSC had normal repopulation potential in an Id1+/+ microenvironment. In agreement with these findings, Id1−/− BM stromal cell cultures supported enhanced proliferation of hematopoietic progenitors. Furthermore, quantitative PCR showed that SCF, M-CSF, OPN, SDF-1 and TGF-α mRNA expression was decreased in Id1−/− stromal cells relative to Id1+/+ stromal cells, while G-CSF, GM-CSF, and VEGF mRNA expression was significantly increased. Id1−/− BM showed decreased number of mesenchymal stem/progenitor cells. Thus, Id1 does not play a role in maintaining HSC, but is involved in regulating hematopoietic progenitor niche. Funded by NCI contract No. N01-CO-12400.
Asthma in Pediatric Sickle Cell Acute Chest Syndrome: In An Inner City London Hospital
Authors: Bartram Jack L , Fine-Goulden Miriam R , Green Dido , Ahmad Rahail , Inusa Baba PD 
Published: April 2020

Abstract
Acute Chest Syndrome (ACS) is the second most common cause of hospitalisation in patients with Sickle Cell Disease (SCD) and up to 25% of those admitted will require intensive care management. ACS is a leading cause of death in SCD. It may also play a role in the development of chronic lung disease in SCD patients and the prevalence of Asthma in SCD patients is high. The pathogenesis of ACS is complex. Previous work has suggested a relationship between asthma and higher risk of ACS in children with SCD. Data in the UK is limited. Our aim therefore was to describe the presentation, course and outcome of ACS in our local SCD pediatric population, compared with those children who had ACS with SCD and physician diagnosed Asthma (Asthma).
Methods: The data collection took place at The Evelina Children’s Hospital, which is part of St Thomas’ Hospital, a large teaching hospital in Central London, England. There are over 400 children with SCD registered, and around 30 new SCD births per year. A retrospective analysis of patient hospital electronic and paper records was performed of 63 ACS presentations over a three year period from 2003 to 2006. Inclusion in the study required a new infiltrate on chest radiograph plus acute respiratory symptoms in a patient with SCD under the age of 16 years. The group included 16 (25%) presentations in children with SCD and Asthma.
Results: No Known Asthma 47 Presentations; Mean age 6.2 yrs (range 1–15yrs); HbSS 87%, HbSC13%; Previous ACS 26% (n=12); Mean length of stay 5.4 days (range 1–27); Mortality 0; Mean C-Reactive protein (CRP) on admission 70 (normal &lt;5); Mean oxygen saturations on presentation 92% in air (40% of patients presented with saturations &lt;92% in air)
Physician Diagnosed Asthma 16 Presentations; Mean age 4.6 (range 1–15yrs); HbSS 94%, HbSC 6%; Previous ACS 63% (n=10); Mean length of stay 5.4 (range 2–14); Mortality 0; Mean CRP on admission 41; Mean oxygen saturations on presentation 92% in air (50% of patients presented with saturations &lt;92% in air)
DISCUSSION: Demographics: Comparable in terms of age and haemoglobin genotype. Presentation: Patients with asthma were more likely to have had previous ACS. Children with asthma presented with a lower CRP. Treatment: The treatment in both groups including the use of blood transfusion, and need for transfer to intensive care were comparable. However there was an observed difference in the use of inhaled bronchodilators (non asthma 21% v asthma 50%). Steroids were rarely used (4%) to treat the patients who did not have a pre-existing diagnosis of asthma, however were used to treat most (94%) of those patients with asthma. Outcome: Length of stay was comparable, no deaths in either group.
CONCLUSION: Although patients in our study group with asthma had a higher frequency of previous ACS episodes, we did not demonstrate that patients with asthma suffer a more severe course of illness.
Exercise-Induced Pulmonary Hypertension in Sickle Cell Anemia: a Study with Exercise Stress Echocardiography
Authors: Nunes Newton , Campos Orlando , Figueiredo Maria Stella , Cancado Rodolfo Delfini , Vicari Perla , Carvalho Antonio C C , Bordin Jose Orlando 
Published: April 2020

Abstract
Background: Pulmonary hypertension (PH) at rest is a risk factor for death in patients with sickle cell anemia. Exercise treadmill stress echocardiography (ETSE) is able to detect latent PH.
Objective: To investigate the occurrence of exercise-induced PH in patients with sickle cell anemia and normal pulmonary pressure (PP) at rest, and its relationship with clinical and echocardiographic variables.
Design: observational transversal study.
Patients and Methods: We studied 44 patients (22 men, mean age = 25.3 y.o.) with sickle cell anemia and normal PP at rest (tricuspid regurgitant jet velocity: TRV &lt; 2.5 m/s). They were divided into 2 groups: (G1) normal PP after ETSE (TRV ≤ 2.7 m/s); G2: exercise-induced PH (TRV &gt; 2.7 m/s). The cut off points of TRV at rest and exercise were derived from healthy control subjects, matched for age, sex, and body surface area. Data obtained from ETSE were correlated with clinical, echocardiographic and ergometric variables, including left atrial volume indexed for body surface area and E/E’ ratio derived from pulsed wave and tissue Doppler. Student’s t test e chi-square test were used for statistical analysis. Multivariate logistic analysis was used to determine the independent predictors of abnormal PP response to exercise. Statistical significance was defined as a p &lt; 0.05.
Results: The exercise-induced PH occurred in 57% of patients (G2), with levels of TRV ranging from 2.8 to 4.5 m/s (3.4 ± 0.4m/s), significantly higher than those observed in G1 (2.5 ± 0.3m/s, p &lt; 0.001). Exercise-induced PH was significantly related to higher creatinine levels, larger right ventricular diastolic area, TRV ≥ 2.3m/s at rest, left atrial volume index ≥ 41ml/m2, and E/E’ ratio ≥ 6.3. When the multivariate analysis was performed, TRV at rest [OR 6.5 (1.3–33.6), p = 0.03], left atrial volume index [OR 6.4 (1.2–33.5), p = 0.03]; and E/E’ ratio [OR 5.6 (1.2–26.5], p = 0.03) remained independently related to exercise-induced PH.
Conclusion: Patients with sickle cell anemia and normal PP at rest may exhibit exercise-induced PH which was independently related to increased baseline PP levels, increased left atrial volumes end indirect parameters of left ventricular and diastolic pressure. The clinical relevance of these findings deserves future studies.
Suppressed CEBPA Function Is Associated with Favorable Prognosis in Normal Karyotype AML Patients
Authors: Fos Jose , Bonadies Nicola , Pabst Thomas , Mueller Beatrice U 
Published: April 2020

Abstract
The myeloid transcription factor CEBPA is crucial for normal differentiation of granulocytes. In addition, genomic mutations and deregulated expression of CEBPA have been reported in specific subsets of patients with acute myeloid leukemia (AML). We here investigated the prognostic significance of CEBPA mRNA, CEBPA protein, and CEBPA function in 105 consecutive de novo AML patients with a particular focus on AML patients with a normal karyotype. We found that the DNA binding activity of CEBPA in normal karyotype AML patients as determined by an ELISA-based assay conferred significant prognostic information: normal karyotype AML patients with suppressed CEBPA function showed a better OS (p=0.0068) and DFS (p=0.0138) compared to patients with conserved CEBPA function. In addition, suppressed levels of the 42kDa CEBPA protein in these patients tended to predict favorable outcome, whereas differences in p30 CEBPA protein levels and in mRNA expression did not affect the outcome. Finally, AML patients with suppressed CEBPA function had more frequently FLT3-ITD and an unmutated nucleophosmin status identifying CEBPA function as an independent prognostic marker for normal karyotype AML. This is the first study comprehensively assessing CEBPA transcription factor function in leukemic cells from AML patients. Our data suggest that suppressed CEBPA function is associated with favorable prognosis in normal karyotype AML patients independently of other molecular markers, and we propose assessment of CEBPA binding activity to be integrated into clinical practice. Moreover, our results highlight the importance of posttranscriptional mechanisms of CEBPA modulation in AML.