bluuebunny/crossref_metadata_2025_split huggingface dataset:
Activation of G12/13 Pathway Is Essential for the Dense Granule Release in Platelets.
Authors: Jin Jianguo , Kim Soochong , Kunapuli Satya 
Published: October 2019

Abstract
Platelet secretion is an important physiological event in hemostasis. A number of agonists such as thrombin and thromboxane A2 induce platelet secretion. ADP does not cause dense granule release in aspirin-treated platelets, although ADP induce platelet shape change, aggregation and alpha granule release through Gq and Gi pathways. The protease activated receptors 1 and 4, and the thromboxane receptor activate the G12/13 pathways in addition to the Gq pathways. We postulated that the platelet dense granule release reaction depends on both Gq and G12/13, and in the absence of signaling through either G protein abolishes secretion. In other words, co-stimulation of Gq and G12/13 pathways is a major requirement for dense granule release in platelets. We rationalize that because U46619 and thrombin can activate both Gq and G12/13, they cause platelet dense granule release, whereas ADP fails to cause platelet secretion from dense granules because it does not activate G12/13. As a first step towards testing this hypothesis, we supplemented ADP signaling in aspirin-treated platelets with selective activation of G12/13 pathways using YFLLRNP, a partial agonist of PAR-1. YFLLRNP selectively active G12/13 signaling pathway without activating Gq or Gi pathways at low concentrations. YFLLRNP (60 μM) or 2MeSADP (100 nM) failed to cause dense granule release. However, addition of YFLLRNP (60 μM) and 2MeSADP (100 nM) together caused dense granule release. We proceeded to confirm these results in Gq null mice by selectively activating phospholipase C (PLC), a downstream signaling molecule from Gq. In aspirin-treated Gαq knockout mouse platelets 80 μM m-3M3FBS, a direct PLC activator, causes calcium mobilization from intracellular stores and platelet shape change, but does not cause dense granule secretion. We have previously shown that Gq/PLC pathways downstream of ADP are not sufficient for aggregation and require concomitant signaling from Gi for platelet aggregation. Thus, lack of aggregation by PLC activation alone in G?q knockout mouse platelets is consistent with our previous observations. In G?q null mouse platelets, m-3M3FBS (80 μM) or AYPGKF (500 μM) alone failed to cause dense granule release. However, addition of m-3M3FBS (80 μM) and AYPGKF (500 μM) together caused dense granule release. In addition, consistent with our previous findings, co-stimulation of G12/13 pathways and Gi (AYPGKF + 2MeSADP) in G?q knockout mouse platelets caused aggregation, but failed to cause dense granule release. We conclude that supplemental signaling from G12/13 is required for Gq-mediated dense granule release and that ADP fails to cause dense granule release because the platelet P2Y receptors, although activate PLC, do not activate G12/13 pathways.
Effect of Adding Rituximab to Three Cycles of CHOP Plus Invoved-Field Radiotherapy for Limited-Stage Aggressive Diffuse B-Cell Lymphoma (SWOG-0014).
Authors: Miller Thomas P. , Unger Joseph M. , Spier Catherine , Stea Baldassarre , Cantu Emilia , LeBlanc Michael , Fisher Richard I. 
Published: October 2019

Abstract
The 5-year survival rate of patients with limited disease (LD) and aggressive histologies of non-Hodgkin lymphoma (NHL) who have at least one adverse risk factor is about 70% after treatment with three cycles of CHOP followed by involved-field radiotherapy (CHOP(3) plus RT). We tested the effect of adding four infusions of rituximab to CHOP(3) plus RT in 62 evaluable patients. Patients had an aggressive histology of diffuse B-cell NHL (diffuse large cell or Burkitt’s-like) and had LD. LD is defined as stage I with at least one adverse risk factor (age > 60 years, elevated serum LDH, or a performance status of 2) OR non-bulky stage II disease. Adverse risk factors are defined using a stage-modified IPI system and include: non-bulky stage II, age >60 years, elevated serum LDH and a performance status of 2. Patients with bulky stage II disease were excluded as their prognosis is similar to advanced disease, and patients with stage I disease and no risk factors were excluded as their prognosis exceeds 90% survival at 10 years with CHOP(3) plus RT. Rituximab was infused at 375 mg/m2 on days −7, 1, 22, and 43, and CHOP was given at standard doses on days 3, 24, and 45 (CHOP(3) plus R plus RT). Involved-field RT was given as previously described (N Engl J Med1998;339:21–26). With a median follow-up of 2.4 years, the progression-free survival (PFS) and overall survival (OS) measured at 2 years was 94% and 95%, respectively. In order to estimate whether the addition of rituximab adds benefit to CHOP(3) plus RT, and therefore, would be worth testing in a comparative trial, we identified 68 patients from SWOG study 8736 treated with CHOP(3) plus RT using the same criteria to include LD and aggressive diffuse B-cell histologies. These previously reported patients treated with CHOP(3) plus RT (ref above) have a median follow-up of 12.0 years. The PFS and OS for this group measured at 2 years was 85% and 93%, respectively. Four patients have relapsed in the rituximab-treated group and 10 patients had relapsed in the CHOP(3) plus RT alone treated group within the first 2 years. Three patients have died in the rituximab- treated group and five patients had died on the CHOP(3) plus RT alone treated group within the first 2 years (see Table). Short-term toxicity appeared similar. We conclude that the addition of rituximab to standard treatment with CHOP(3) plus RT results in outcome which compares favorably to our historical experience and merits further study.
Effect of Rituximab in Treating Limited Disease
MEASUREMENT CHOP(3) + R + RT (0014) CHOP(3) + RT (8736) Results of SWOG study 0014 compared to SWOG study 8736 No. patients 62 68 2-year PFS 94% 85% 2-year OS 95% 93% No. relapses 2 yrs 4 10 No. deaths 2 yrs 3 5
Red Blood Cells Facilitate Reverse Cholesterol Transport.
Authors: Kuypers Frans A. , Larkin Sandra , Beckstead Jenifer , Oda Michael , Ueda Kazumitsu , Ryan Robert O. 
Published: October 2019

Abstract
Lecithin:cholesterol acyl transferase (LCAT)-dependent conversion of cholesterol (CH) to cholesteryl ester (CE), a key component of the reverse cholesterol transport (RCT) pathway, is essential for cholesterol processing. We hypothesized that red blood cells (RBCs) function in this pathway by facilitating phosphatidylcholine (PC) re-generation from LCAT-derived lysophosphatidylcholine (LPC). Addition of 14C-oleate to fresh RBCs resulted in an ATP-dependent incorporation of radiolabel into PC via the Lands pathway. Prior depletion of red cell LPC content reduced the incorporation of 14C-oleate into PC, which was restored by the addition of LPC. Reconstituted PC/cholesterol/apolipoprotein A-I (apoA-I) high-density lipoprotein (rHDL) was used as substrate for LCAT dependent conversion of CH to CE. Addition of LPC to this reaction mix inhibited CE production. The inhibition was overcome, however, by inclusion of RBCs, suggesting rHDL-generated LPC migration to RBCs. RBCs depleted of LPC increased their ability to generate 14C-PC from 14C-oleate in the presence of rHDL with LCAT, indicating that LCAT-derived LPC can be utilized as a substrate for PC production in RBCs. Radio-labeled CH associated with RBCs was recovered as CE only when rHDL substrate and LCAT were present, indicating RBC-associated CH migration to rHDL. When RBCs containing 14C-PC were incubated with rHDL and LCAT, PC transferred from RBCs to the rHDL particles. The interaction of apoA-I with the membrane lipid transporter ATP-Binding Cassette A1 protein (ABCA1) in cell membranes has been shown to play an essential role in the formation of HDL and facilitates RCT. Using monoclonal antibodies, we were able to show that RBCs contain ABCA7, closely related to ABCA1. Our data show that fluorescently labeled apoA-I binds to RBCs, suggesting that the interaction between ABCA7 and apoA-I may be important for HDL/RBC interaction. Together, our data supports an active role for RBCs, the major cell type in blood, in LCAT-mediated lipoprotein remodeling. Thus, RBCs represent a hitherto underappreciated component of the RCT pathway.
CCAAT Enhancer Binding Protein-α (C/EBPα) Determines Myeloid Versus Erythroid Cell Fate in Multipotential Progenitors.
Authors: Suh Hyung-Chan , Gooya John , Renn Katie , Friedman Alan , Johnson Peter , Keller Jonathan 
Published: October 2019

Abstract
C/EBPα is a bZip transcription factor, which is required for granulocyte development, and loss of C/EBPα function is associated with the development of acute myelogenous leukemia and myelodysplastic syndrome. While the precise mechanisms that regulate cell fate decisions during hematopoietic development are largely unknown, expression of transcription factors (PU.1 and GATA-1) can induce lineage conversion. In this regard, C/EBPα can drive the differentiation of B cells into macrophages, and bi-potential cell lines into granulocytes at the expense of macrophages. C/EBPα can also promote the transdifferentiation of myoblasts into adipocytes. We have recently found that there are increased numbers of erythroid cells in the fetal liver of C/EBPα −/ − mice. Also, C/EBPα is expressed in more primitive progenitor cells than granulocyte/macrophage progenitors (GMP) including hematopoietic stem cells. Therefore, we initiated experiments to evaluate whether C/EBPα has a functional role in regulating cell fate decisions in progenitors more primitive than GMP where it may promote a myeloid versus erythroid cell fate decision. To test this hypothesis, we over-expressed C/EBPα using retroviral vectors in 1) bone marrow cells (BMC) and evaluated their growth and differentiation in vitro, and in vivo when transplanted into mice; 2) purified multipotential progenitors with erythroid and myeloid potential, and erythroid restricted progenitors; and 3) murine erythroid leukemia (MEL) cells. We found that there was a marked decrease in erythroid lineage cells and an increase in myeloid cells in mice transplanted with BMC that over-expressed C/EBPα. We also observed a decrease in erythroid cell growth in vitro with BMC that expressed C/EBPα. Furthermore, when infected with retroviral vectors that express C/EBPα, erythroid restricted progenitors acquired myeloid cell morphology and myeloid specific cell surface markers. In addition, MEL cells that over-express C/EBPα showed increased myeloid gene expression including GM-CSFR, PR3 and myeloid specific esterase, while they showed decreased expression of β-globin and Epo receptor (EpoR) which is required for erythroid cell differentiation and survival. We detected high levels of EpoR in C/EBPα −/ − suggesting an inverse relationship between C/EBPα and EpoR expression. Thus, C/EBPα is a dual function transcription factor that can repress erythroid specific genes while enhancing myeloid lineage gene expression. Consequently, C/EBPα acts as a switch to drive hematopoietic progenitor cells toward myeloid cell development at the expense of erythroid maturation, and can reprogram erythroid cells into myeloid cells. # Funded in part by DHHS #NO1-CO-12400
PU.1 and pRb Bind GATA-1 on DNA and Recruit a Histone H3K9 Methyl Transferase-Containing Complex to Repress the Erythroid Transcription Program.
Authors: Stopka Tomas , Amanatullah Derek F. , Skoultchi Arthur I. 
Published: October 2019

Abstract
Current work indicates that transcriptional repression is at least as important as transcriptional activation in normal development. Inappropriate or untimely transcriptional repression in immature hematopoietic cells is often the basis for a block to differentiation in hematologic malignancies. Activation of PU.1, a myeloid and B-cell specific transcription factor, in erythroid cells plays a key role in Friend virus-induced mouse erythroleukemia (MEL). Previous results from our laboratory showed that PU.1 blocks the erythroid differentiation-promoting activity of GATA-1 by binding directly to GATA-1 on DNA and inhibiting its transcriptional function. PU.1-mediated repression of GATA-1 on transiently transfected GATA-1 target genes is dependent on the corepressor pRb that also binds to PU.1 (Rekhtman et al., Genes & Dev 1999 and Mol Cell Biol 2003). To further investigate the mechanism of PU.1-mediated repression of GATA-1 in chromatin, we examined the occupancy of several GATA-1 target genes by PU.1 and pRb, as well as the state of core histone modifications at these loci in MEL cells by quantitative chromatin immunoprecipitation. These studies included both endogenous GATA-1 target genes and an exogenous GATA-1 target gene (alpha globin) integrated at a specific locus in MEL cells by Recombinase-Mediated Cassette Exchange. We found that GATA-1 sites at both the exogenous, integrated gene as well as at endogenous genes (including the regulatory regions of the alpha globin, beta globin, alas-e, eklf, p45 nf-e2) are occupied by a GATA1 - PU.1 - pRb complex in undifferentiated MEL cells. The presence of all three components of the complex is dependent on intact GATA-1 binding sites in the exogenous, integrated gene. The histone methyltransferase Suv39H1 and the histone H3MeK9 binding protein, HP1alpha, are also present at the repressed loci. During induced differentiation of MEL cells, PU.1, pRb, Suv39H1 and HP1alpha occupancy at these sites declines but GATA-1 continues to be present at its binding sites. The disruption of the repression complex at these loci during differentiation as well as during siRNA-mediated PU.1 knock down is associated with conversion of methylated H3K9 to acetylated H3K9 and significant transcriptional derepression of these GATA-1 target genes. These findings support a model for repression of GATA-1 by PU.1 at endogenous loci through recruitment of the corepressor pRb and associated histone methyltransferase (Suv39H1) and H3MeK9 binding (HP1alpha) activities.
Pharmacogenetics of Outcome in Children with Acute Lymphoblastic Leukemia.
Authors: Rocha Jose Claudio , Cheng Cheng , Liu Wei , Kishi Shinji , Das Soma , Cook Edwin H. , Sandlund John T. , Rubnitz Jeffrey , Ribeiro Raul C. , Pui Ching-Hon , Evans William E. , Relling Mary V. 
Published: October 2019

Abstract
The acquired genetic characteristics of acute lymphoblastic leukemia (ALL) blasts are often used to guide the intensity of therapy, whereas the germline host genetic characteristics of the patient generally have not been considered. Multiple common, functionally important polymorphisms affect genes whose products determine the pharmacokinetics and pharmacodynamics of antileukemic agents. It is not yet known how genetic polymorphisms may interact to affect the outcome of antileukemic therapy. Combining classification and regression tree with failure time analysis, we assessed whether 16 genetic polymorphisms, alone or in combination, predicted relapses in 246 children with ALL, 116 of whom were treated on the lower-risk (LR) and130 on the higher-risk (HR) arms of the St Jude protocol Total XIIIB. Genotyping was performed for the following polymorphic loci: CYP3A4*1B and CYP3A5*3; GSTP1 313A>G, GSTM1 and GSTT1 deletions; MDR1 exon 21 (2677G>T/A) and MDR1 exon 26 (3435C>T); MTHFR 677C>T and MTHFR 1298A>C; NR3C1 1088A>G; SLC19A1 80G>A; TPMT 238G>C, 460G>A and 719A>G; TYMS enhancer repeat; UGT1A1 promoter repeat polymorphism; VDR intron 8 G>A and VDR FokI (start-site) T>C. In all children with available RNA in their diagnostic ALL blasts, gene expression levels of prognostic genotypes were analyzed using the Affymetrix genechip array HG_U95Av2. Among the HR group, the glutathione S-transferase M1 (GSTM1) non-null genotype was associated with the risk of hematological relapse (5-year cumulative incidence, 17.1%±4.5% compared to 5.1%±2.9% for GSTM1 null genotype, p = 0.03), and among the non-null genotypes, the thymidylate synthetase (TYMS) 3/3 genotype was associated with a further increase in hematologic relapse risk (5-year cumulative incidence, 29.2%±9.5% compared to 10.9%±4.7% for TYMS 2/3 or 2/2 genotypes, p = 0.02). Increased expression levels of these two target genes (p < 0.0001 and p = 0.09, respectively) were consistent with resistance to the drugs interacting with these gene products. For central nervous system relapse, among the HR group, the vitamin D receptor (VDR) start site (p = 0.02) and intron 8 genotypes (p = 0.04) predisposed, whereas for LR patients the TYMS 3/3 genotype predisposed (p = 0.04). The genotypes associated with outcome have pharmacologic plausibility: e.g., high GST activity (GSTM1 non-null) could cause anticancer drug resistance; high TYMS activity (TYMS 3/3) would be less inhibited by antifolates. In conclusion, germline polymorphisms influence the outcome of antileukemic therapy, and therefore represent determinants of response that can be used to optimize therapy.
Prognostic Factors and Long-Term Outcome for Patients with Hodgkin’s Lymphoma Who Relapse after an Autologous Stem Cell Transplantation.
Authors: Constans Mireia , Sureda Anna , Arranz Reyes , Caballero Maria Dolores , Lahuerta Juan Jose , Hernandez-Boluda Juan Carlos , Vidal Maria Jesus , Garcia-Laraña Jose , Rifon Jose , Ribera Josep Maria , Fernandez-Abellan Pascual , Moraleda Jose Maria , Bernal Maria Teresa , Mateos Maria Victoria , Martin-Mateos Maria , Cordoba Rafael , Garcia-Conde Javier , Sierra Jorge , Conde Eulogio ,   for the Spanish Cooperative Group GEL/TAMO
Published: October 2019

Abstract
We have analyzed overall survival (OS-REL), progression-free survival (PFS-REL) and the clinical variables influencing the long-term outcome for patients with Hodgkin′s lymphoma (HL) who relapse after an autologous stem cell transplantation (ASCT). From 639 patients autografted for HL [383 males (60%) and 256 (40%) females, median age of 30 (1–66) years] reported to the GEL/TAMO Cooperative Group between January/1984 and January/2003, 175 patients (27%) relapsed at a median time of 10 (4 – 125) months [median (range)] after ASCT. They were 97 males (56%) and 78 (44%) females with a median age of 28 (10 – 66) [median (range)] years at transplantation. Sixty-three patients were autografted in complete remission, 73 in sensitive relapse and 31 in resistant relapse. One hundred and sixty three patients (94%) received different chemotherapy (CT) protocols as conditioning regimen and 12 patients (6%), CT plus total body irradiation. At relapse, 94 patients (53%) presented with advanced stages (III-IV), 49 patients (28%) with B symptoms and 19 (11%) with bulky disease. Relapse was extranodal in 45% of the patient population. Ten percent of the patients had a poor performance status (ECOG ≥2) and 37 patients (22%) had a hemoglobin (Hb) level < 100 g/l at relapse. Thirty four patients (20%) received no further therapy, 107 (60%) received different CT +/− radiotherapy and the remaining 34 patients (20%) were treated with a second stem cell transplantation (13 patients, an ASCT and 21 an allogeneic transplantation). OS-REL and PFS-REL were of 35% ± 4% and 23% ± 4% at 3 years, respectively. Advanced clinical stage (III-IV) at relapse [relative risk (RR) 4.4, 95% confidence interval (CI) (1.7 – 10.8), p = 0.002] and an interval between ASCT and relapse < 12 months [RR 2.4, 95%CI (1.1 – 5.0), p = 0.03] were independent adverse prognostic factors for PFS-REL. Advanced clinical stage at relapse [RR 4.4, 95%CI (1.4 – 14.4), p = 0.012], extranodal disease [RR 2.4, 95%CI (1.3 – 15.4), p = 0.02] and a Hb level < 100 g/l at relapse [RR 3.4, 95%CI (2.0 – 10.4), p = 0.03] were significant adverse prognostic factors for OS-REL. Although the long-term outcome of HL patients who relapse after ASCT is poor, results are better in patients with late relapses and localized disease. Moreover, a minority of the patients can become long-term progression-free survivors after relapse. The identification and construction of prognostic sub-groups according to the number of risk factors may be useful to develop risk-adapted therapeutic strategies.
Early Response to Induction Is Predictive of Survival in Childhood Philadelphia Chromosome Positive Acute Lymphoblastic Leukaemia: Results of the Medical Research Council ALL 97 Trial.
Authors: Roy Anindita , Bradburn Mike , Moorman Anthony V. , Burrett Julie , Mitchell Chris , Kinsey Sally E. , Vora Ajay J. , Lilleyman John , Eden Osborn , Hann Ian , Saha Vaskar 
Published: October 2019

Abstract
We report the outcome of children with Philadelphia positive acute lymphoblastic leukemia (Ph+ ALL) treated on the national trial for childhood ALL, MRC ALL 97/99, between Jan 1997 and Jun 2002. A diagnosis of Ph+ ALL was made by karyotyping and/or fluorescent in situ hybridisation. Outcome was evaluated according to NCI risk criteria and treatment response. Statistical methods included Kaplan Meier analysis, logrank and stratified logrank tests. Of the 1937 patients treated on the MRC ALL 97/99 protocol, 1836 were successfully screened for Ph chromosome and 42 (2.3%) were positive. This subgroup of patients was heterogeneous at the cytogenetic level with 27 (77%) out of 35 assessable cases having one or more abnormality in addition to the Ph translocation. Four recurrent secondary abnormalities were observed: an extra Ph chromosome, loss of 9p (del 9p), high hyperdiploidy and monosomy 7. The median age at diagnosis and presenting white cell count were 6.8 years and 32.5 × 109/L respectively. 19 (45%) had <25% bone marrow blasts within the first two weeks of treatment and were defined as a Good-response group (GRG), the others were classified as a Poor-response group (PRG). 6/10 (60%) of the patients with an extra Ph chromosome were in the GRG, whereas 8/10 (80%) of those with del 9p belonged to the PRG. 36/42 (86%) achieved complete remission (CR1) at the end of induction of whom 28 underwent bone marrow transplantation (BMT). The median follow up was 41 months (range 21–84 months). The 3-year estimate for event free survival (EFS) (with 95% confidence intervals) was 57% (41–70%). The EFS for the GRG and PRG were 74% (48–88%) and 44% (22–64%) respectively. Significant predictors for survival were presenting white cell count (p=0.02) and early response to treatment (p=0.03), but not age at diagnosis or gender. BMT in CR1 appeared to reduce the risk of a subsequent bone marrow relapse. These results are a significant improvement on previous results, particularly for the PRG. The EFS for the PRG in this report is 44% compared to a 4 yr. EFS of 10% in previously published data from the BFM/AIEOP group. We believe that the early introduction of intensive chemotherapy as in the MRC trial improves early response rates and overall outcome in Ph+ ALL, especially for the PRG. BMT in CR1 is recommended in children with Ph+ ALL, nonetheless newer alternative treatment strategies, such as the use of imatinib mesylate need to be evaluated for children with Ph+ ALL.