bluuebunny/crossref_metadata_2025_split huggingface dataset:
10 Year Musculoskeletal Outcomes with Tailored Primary Prophylaxis: The Canadian Hemophilia Prophylaxis Study.
Authors: Blanchette Victor S. , Rivard Georges E. , Pai Mohan K. , Israels Sara J. , McLimont Marjorie , Feldman Brian M. 
Published: October 2019

Abstract
High-dose prophylaxis for severe hemophilia A [25–40 factor (F) VIII Units (u) kg−1 on alternate days] reduces arthropathy, but does so at great cost. (NEJM2007; 357: 535–544). Less intensive prophylaxis regimens might be cost-effective while reducing the need for indwelling venous access devices and their attendant complications. Tailored prophylaxis starts with FVIII, 50 u kg−1 weekly (Step1). If bleeding frequency is unacceptable (4 joint/soft tissue bleeds or 5 bleeds into a joint on a step of protocol), or if a target joint develops (≥3 bleeds into a single index joint-ankle, elbow or knee) over a consecutive 3-month period the dose of FVIII is escalated to 30 u kg−1 twice weekly (Step 2); a third escalation to 25 u kg−1 on alternate days if bleeding frequency remains unacceptable (Step3).
Objectives: To assess the degree of joint damage in the 6 index joints for subjects receiving tailored prophylaxis.
Methods: We studied an inception cohort of boys ages 1–2.5 years with severe hemophilia A (<2% FVIII) and no inhibitors treated with tailored prophylaxis. Patients were followed at 6 monthly intervals and parents have kept detailed diaries. Joint damage was measured using a validated physiotherapy joint score and function was assessed using the Child Health Assessment Questionnaire (CHAQ)
Results: 54 boys were enrolled from 11 Canadian hemophilia treatment centers. The total follow-up is 3423 patient months (mean 62.2 months, median 54 months and range 5–117 months). 16 (29.6%) boys have been escalated to Step 2 and 14 (25.9%) have escalated to Step 3. 11 of the escalations were due to target joint bleeding. Subjects on study had a mean of 1.4 joint bleeds/patient-year. Physiotherapy Scores Maximum Possible Score Median Observed Range Ankles 56 2 0–20 Elbows 48 0 0–13 Knees 56 0 0–15 All Joints Swelling 18 0 0–4 Muscle Atrophy 18 0 0–3 CHAQ 3 0 0–2 89% of participants were compliant with the primary prophylaxis protocol. Non compliant subjects had the poorest outcomes. No treatment related adverse events have occurred.
Conclusion: Tailored primary prophylaxis should be considered as a valid option for long term prophylaxis in young boys with severe hemophilia A.
Maximum Possible Score Median Observed Range Ankles 56 2 0–20 Elbows 48 0 0–13 Knees 56 0 0–15 All Joints Swelling 18 0 0–4 Muscle Atrophy 18 0 0–3 CHAQ 3 0 0–2
The mTOR Inhibitor RAD001 (Everolimus) Improves Survival in Preclinical Models of Primary Human ALL.
Authors: Crazzolara Roman , Cisterne Adam , Thien Marilyn , Hewson John , Bradstock Kenneth Francis , Bendall Linda J. 
Published: October 2019

Abstract
The Akt/mammalian target of rapamycin (mTOR) signaling pathway is a potential target in acute lymphoblastic leukemia (ALL) due to its constitutive expression and critical role in regulating leukemic cell growth and survival. In this study, we investigated whether RAD001 (Everolimus, Novartis), is effective as a therapeutic agent in a murine model of human ALL. Leukemic cells from 5 separate patient samples were injected into sub-lethally irradiated NOD/SCID mice and treatment was started after leukemia was clearly detectable in the blood (>5% peripheral blasts) and continued for 4 weeks. In 2 of 3 cases examined, animals treated with RAD001 as a single agent (5mg/kg, 3 times weekly) showed 2 to12-fold reduction in the number of circulating blasts and 2.5 to 30-fold decrease in splenomegaly after only 7 days of treatment (p<0.01). In 1 case, blasts completely disappeared by 14 days of treatment accompanied by recovery of normal hematopoiesis. In contrast vehicle treated mice showed progressive leukemia. In all 5 cases examined, decreased leukemic burden was associated with increased survival, with a median survival of 52 days in the RAD001 treated mice compared to 24 days in the control group (p<0.02 for all samples). Combination treatment with RAD001 and Vincristine (0.15–0.25 mg/kg, weekly) was more effective than either agent alone in reducing leukemic cell count and prolonging survival (p<0.02 compared to Vincristine alone). In one case, all mice treated with either RAD001 alone or RAD001 plus Vincristine survived for greater than 10 weeks following the completion of treatment and only one animal (RAD001 treated) showed signs of disease on elective sacrifice. Cell cycle analysis of cells recovered from the spleens of RAD001 treated animals revealed an arrest in the G0/G1 phase. Furthermore, electron microscopy demonstrated the predominant appearance of autophagy as well as limited apoptosis in cells within the sternums of treated animals. In conclusion, we have demonstrated the efficacy of RAD001 alone or with a subtherapeutic dose of Vincristine in an in vivo leukemia model. These preclinical results support further clinical development of mTOR inhibitors for the treatment of ALL patients. RAD001 may provide significant therapeutic benefit when used alone, or in combination with standard chemotherapy agents such as Vincristine.
Wolinella Asparaginase as an Immunologically Distinct, Efficacious, and Potentially Less Toxic Asparaginase for Use in Lymphoid Malignancies.
Authors: Watt Tanya C. , Keller Frank G. , Durden Donald L. 
Published: October 2019

Abstract
L-asparaginase targets the protein translational mechanism and is an essential component in the treatment of acute lymphoblastic leukemia (ALL) and lymphoid malignancies. The current available preparations, E. coli and Erwinia have significant toxicities, often limiting their clinical use. In order to provide patients with a novel form of asparaginase, a recombinant form of Wolinella (rWS) asparaginase was developed. C3H mice were transplanted with 2 × 106 6C3HED lymphoma cells. Mice treated with native or rWS asparaginase demonstrated a significant decrease in tumor volume, and they remained tumor free for greater than 30 days after treatment. Human ALL cell lines were tested for sensitivity to rWS asparaginase and the molecular basis for rWS asparaginase sensitivity was examined in these cells. We hypothesized that rWS asparaginase would be immunologically distinct to that of E. coli. The sera of 6 patients previously identified to have high titer inhibitors to E. coli asparaginase were tested for antibodies to rWS asparaginase. The antibodies to E. coli did not bind to the rWS asparaginase, therefore demonstrating that there is no cross-reactivity to rWS asparaginase. Thus, antibodies or hypersensitivity reactions to previous forms of asparaginase will not prohibit therapy with rWS asparaginase. Therefore rWS asparaginase represents the first recombinant form of asparaginase and shows promise as a novel form of asparaginase which is immunologically distinct from E. coli and Erwinia asparaginase. In addition, it appears to be equally efficacious, and in previously published data, may have less toxicity. Asparaginase is an important component of treatment regimens for lymphoid malignancies, and therefore future studies will include a Phase I/II trial utilizing rWS asparaginase in relapsed lymphoid malignancies.
Non-Compliance Is the Predominant Cause of Aspirin Resistance.
Authors: Schwartz Kenneth A. , Schwartz Dianne E. , Barber Kimberly , Reeves Matt , DeFranco Anthony 
Published: October 2019

Abstract
Classifying patients as resistant to aspirin is an arbitrary designation with up to 60% of patients reported to be aspirin resistant. Our previous publication showed that 16 or 9% of 192 patients with a history of M.I. could be labeled as aspirin resistant and that all of these patients were aspirin resistant because of non-compliance. This report compares the relative frequency of aspirin resistance between known compliant and non-compliance subjects to demonstrate that non-compliance is the predominant cause of aspirin resistance. Platelet prostaglandin agonist (PPA) stimulated light aggregometry was used to measure amount of aspirin induced platelet inhibition and arachidonic acid (AA) stimulated platelet aggregation to classify platelets as either aspirin inhibited or not. After not taking aspirin for 7 days blood for aggregation studies was drawn, off aspirin time point, which was followed by observed ingestion of 365mg of aspirin. Two hours after observed aspirin ingestion aggregation studies were repeated, on aspirin time point. The difference in the slopes of the PPA light aggregation curves on and off aspirin was defined as the net aspirin response. After supposedly refraining from aspirin for 7 days, 45 subjects were judged non-compliant with the protocol and removed from the study. Of the remaining 185 subjects 39 were normals and 146 had a past history of myocardial infarction (MI). There was no difference in net aspirin response between normal and MI subjects. The net aspirin response in known compliant patients was statistically normally distributed. Only 4% of complaint subjects (2 normals and 5 MI) had a net aspirin response of less than one standard deviation which would qualify as a conservative designation of aspirin resistance. Of the 230 enrolled subjects 16 were noncompliant with their daily prescribed aspirin, 45 were non-compliant with the protocol not to take aspirin for 7 days and one patient had NANSAID interference with aspirin’s effect. In total a maximum of 30% of the 230 patients could be classified as aspirin resistant and or non-compliant with 9% aspirin resistant because of non-compliance 18% non-compliant with the protocol and only 3% because of a decreased net aspirin response in known compliant patients. We conclude that:
the predominant cause of aspirin resistance is noncompliance; net aspirin response in compliant patients is normally distributed; in compliant patients no difference in net aspirin response is observed between normals and MI patients; and PPA stimulated light aggregometry measured off aspirin and 2 hours after observed aspirin ingestion is a useful technique for measuring net aspirin response.
Rearrangement and Deletion of the PAX5 Gene in Pediatric Acute B-Cell Lineage Lymphoblastic Leukemia.
Authors: Kawamata Norihiko , Ogawa Seishi , Zimmermann Martin , Sanada Masashi , Hemminki Kari , Yamatomo Go , Nannya Yasuhito , Koehler Rolf , Flohr Thomas , Miller Carl W. , Harbott Jochen , Ludwig Wolf-Dieter , Stanulla Martin , Schrappe Martin , Bartram Claus R. , Koeffler Phillip H. 
Published: October 2019

Abstract
Pediatric acute lymphoblastic leukemia (ALL) is the most common malignant disease in children. The disease results from accumulation of mutations of tumor suppressor genes and oncogenes. Recently, higher resolution SNP-chips (50,000–500,000 probes) have been developed allowing us to identify genes involved at the start sites of deletions/duplications. This permitted us both to identify unbalanced translocations involving t(1;19)(q23;p13) (TCF3/PBX1) and t(12;21)(p13;q11) (ETV6/RUNX1), as well as, to find novel fusion genes involving PAX5 in B-cell lineage ALL. PAX5 gene was rearranged to a variety of partner genes including ETV6, FOXP1, AUTS2 and C20orf112. In each case, tthe C-terminal end of the PAX5 gene was replaced by the partner gene. The PAX5 fusion gene products suppressed transcriptional activity of PAX5 in a dominant negative fashion. We also found a point mutation of PAX5 at codon 26 (Val 26 Gly); and this mutated PAX5 had attenuated transcriptional activity. Expression of PAX5/C20orf112 fusion gene in a B-cell line suppressed endgenous expression of PAX5 target genes including BLK1 and CD19. Furthermore, deletion of PAX5 was common in B-cell lineage ALL (34/339 cases). PAX5 gene is localized on chromosome 9p and concurrent deletion of PAX5 and INK4A genes were frequently detected in B-cell linage ALL. PAX5 gene may behave as a tumor suppressor gene during early development of B-cells and its alteration by either fusion to another gene, point mutation, or deletion may be associated with leukemogenesis of B-cell lineage ALL.
Placental Growth Factor Is Elevated in Patients with Sickle Cell Disease and Associated with Pulmonary Hypertension, Hemolysis, Inflammation, Iron Overload, and Hepatic Dysfunction.
Authors: Mendelsohn Laurel , Tailor Anitaben , Kato Gregory J 
Published: October 2019

Abstract
Abstract 1531
Poster Board I-554
Placental Growth Factor (PlGF) is a functional cytokine in the vascular endothelial growth factor (VEGF) family that generally promotes angiogenesis, depending on the specific context, and can also promote atherogenesis. Produced in erythroid cells, its level in patients with sickle cell disease (SCD) has been previously related to the rate of erythropoiesis. We evaluated PlGF plasma levels in SCD patients by ELISA, and related it to biomarkers of pulmonary hypertension (PH), an emerging and serious complication of SCD linked to early mortality. We find that PlGF levels are significantly higher in SCD (n=95) than healthy African American control subjects (n=19) (median 16.6 vs. 2.1 pg/mL, p<0.001). PlGF levels were higher in SCD patients with elevated pulmonary pressure (normal pulmonary pressure vs. mildly elevated vs. highly elevated: medians 13.7 vs. 16.7 vs. 19.8 pg/mL, p<0.0001). Supporting a linkage to rate of hemolysis, PlGF correlated with LDH (p=0.001) and inversely with hemoglobin level (p<0.0001). Suggesting a link to inflammation, PlGF correlated significantly with C-reactive protein (p=0.001) and erythrocyte sedimentation rate (p<0.001). PlGF correlated with markers of iron overload, including ferritin, transferrin saturation and inversely with transferrin (all p<0.001). Finally, PlGF correlated with markers of hepatic dysfunction, including low albumin and high direct bilirubin (p<0.001).
We found significantly higher PlGF levels in SCD patients taking hydroxyurea compared to those not taking it (median 17.4 vs. 14.0 pg/ml, p<0.01). Confirming that hydroxyurea increases PlGF levels, in a separate cohort of seven patients, PlGF levels rose significantly from their baseline values after initiating hydroxyurea (median approx 22 vs. 27, p<0.05). Our data suggest that elevated PlGF level is associated with PH in patients with SCD, and PlGF is correlated with severity of hemolysis, inflammation, iron overload and hepatic dysfunction. Considering the variable evidence in the literature for either stimulating or inhibiting angiogenesis, it is not clear whether pathologic elevation of PlGF may be mediating pulmonary hypertension, or perhaps conversely providing an adaptive response to vascular damage. It has been suggested by Perelman et al. that PlGF may mediate leukocyte activation that might promote disease severity in SCD. However, hydroxyurea, which tends to ameliorate SCD complications, stimulates PlGF level in an unexpected manner, possibly related to the ability of hydroxyurea to stimulate erythropoietin production, which might in turn induce PlGF. Further research is needed to reconcile the role of PlGF in PH in SCD.

Disclosures
Tailor: Mesoscale: Employment.
Peroxisome Proliferator-Activated Receptor (PPAR)-Gamma Agonists Inhibit Dectin-1 mediated Activation of Human Dendritic Cells (DC).
Authors: Kock Grethe , Bringmann Anita , Held Stefanie , von Schwarzenberg Karin , Brossart Peter 
Published: October 2019

Abstract
Abstract 1645
Poster Board I-671
Human Dectin-1 (hDectin-1) is a member of the C-type lectin-like receptor family that was shown to be the major receptor for fungal beta-glucans and to play an important role in the cellular responses mediated by these pathogens. Activation of Dectin-1 via extracts such as zymosan or more specifically with curdlan can lead to DC activation characterized by upregulation of surface molecules or secretion of cytokines. Futhermore, Dectin-1 is important for recruitment of leukocytes and production of inflammatory mediators at the site of infection.. Peroxisome proliferator-activated receptor (PPAR) and its ligands, cyclopentenone prostaglandins or thiazolidinediones, have been shown to have profound modulatory effects on B and T lymphocytes as well as DCs. Cyclopentenone prostaglandins are produced during the late phase of inflammation due to up-regulation of cyclooxygenase 2 (COX2), a key enzyme for the synthesis of cyclopentenone prostaglandins, that mediate their effects by activation of PPAR-gamma dependent and independent pathways. In our study we analyzed the effects of troglitazone (TGZ), a high affinity synthetic ligand of PPAR-gamma, on the Dectin-1 mediated activation of antigen presenting cells (APC). TGZ was added to the cell culture medium during the differention of monocyte derived DC. On day 5-6 of culture the cells were additionally treated with curdlan or zymosan and phenotypical and functional analyses were performed. Activation of PPAR-gamma resulted in a reduced stimulation of APC via Dectin-1 characterized by down-regulation of CD1a, CD83 and costimulatory molecules on the cell surface and reduced secretion of cytokines and chemokines involved in T-lymphocyte activation and recruitment including IL-1beta, TNF-a, IL-6, IL-12, MIP-1a and Rantes. Western blot analyses revealed that these inhibitory effects were mediated by the inhibition of the mitogen-activated protein (MAP) kinase pathways characterized by a reduced phosphorylation of ERK1/2 and p38. Furthermore, we found that Dectn-1 induced c-Jun phosphorylation was reduced by pretreatment with TGZ. In addition, Dectin-1 induced nuclear translocation of NF-kappaB family members RelB and cRel was dramatically reduced in APC incubated with TGZ. The observed inhibition of signaling pathways was not due to a reduced expression of Dectin-1, phosphorylation of Syk or increased secretion of IL-10 and TGF-beta. TGZ had no effect on the Dectin-1 induced phosphorylation and nuclear expression of NFAT. Our data demonstrate that PPAR-gamma ligands inhibit Dectin-1 mediated activation by interfering with the MAP kinase, c-Jun and NF-kappaB signaling pathways, thus confirming their important role as a negative feedback mechanisms in the regulation of potentially harmful inflammation signaling pathways.

Disclosures
No relevant conflicts of interest to declare.
A Reduced Rate of Leukaemia Relapse After HLA-Identical Sibling Stem Cell Transplantation for Acute Myeloid Leukaemia but Not Other Haematological Malignancies Is Associated with Donor KIR Genes 2DL5A, 2DS1 and 3DS1.
Authors: Stringaris Kate , Adams Sharon , Uribe Marcela , Eniafe Rhoda , Wu Colin , Savani Bipin N. , Barrett A. John 
Published: October 2019

Abstract
Abstract 2282
Poster Board II-259
Stem cell transplantation (SCT) from a healthy donor can be curative for patients with haematological malignancies resistant to other treatments. Elimination of malignant cells through a graft-versus-leukaemia (GVL) effect involves donor T- and NK-cells but their relative contribution to this process is poorly defined. NK-cell alloreactivity and GVL effects are controlled by the nature of the interaction of NK activation receptors and killer-immunoglobulin-like-receptors (KIR) with major-histocompatibility locus class I antigens on thetarget cell. We performed KIR-genotyping of HLA-matched donors in two hundred and forty eight T-celldepleted SCTs to identify genetic factors affectingtransplant outcome (transplant-related mortality (TRM), leukaemic relapse and survival). Transplant-related factors and donor KIRgenotyping significant in univariate analysis were entered into a multivariateanalysis to identify independent variables predictive of outcome for differentforms of leukaemia.
Patients studied were transplanted at a single center between 1993-2008.They included 70 AML, 99 CML, 40 ALL, 39 MDS patients and 13 patients with CLL, NHL or Myeloma. Median age was 36 years (range 9-66). All received aT cell depleted SCT from an HLA identical sibling. Patients were conditioned with totalbody irradiation 12-13.5Gy (4Gy for patients over 55y) and cyclophosphamide120mg/kg +/- fludarabine 125mg/m2. Patients received 0.2-5 × 10 5 CD3 cells /kg and amedian CD34 cell dose of 3 × 10 6/kg. Cyclosporine was used as the sole GVHDprophylaxis.
Multivariate analysis confirmed known predictive factors; for survival: highCD34 cell dose(RR 0.51, p <0.001) and standard rather than high risk disease (RR 3.1, p <0.001), for TRM: poor lymphocyte recovery at day 30 (RR 0.4, p < 0.02), for relapse: high risk disease (RR 4.65, p<0.002). In addition, the presence of donor B haplotype KIR genes: 2DL5A, 2DS1 and 3DS1was associated with significantly less relapse in patients with AML (13% vs 57% ) but not inpatients with other myeloid or lymphoid malignancies. AMLpatients receiving SCT from donors with these KIR genesspan style=“font-size: 10pt;”>relapsed five times less frequently than patientstransplanted from donors with other KIR genotypes.No other factors relating to NK alloreactivity including (missing donor KIR ligand, presence of donor haplotype B, total number of inhibitory KIR and total number of activatory KIR) was foundto be predictive for transplant outcomes. Thesefindings suggest specific, genetically determined,interactions between NK-cells and AML cells whichfacilitate the graft-versus-leukaemia effect and haveimplications for donor selection for AML patients.

Disclosures:
No relevant conflicts of interest to declare.