bluuebunny/crossref_metadata_2025_split huggingface dataset:
Functional Analysis of CD9 during Megakaryopoiesis
Authors: Kono Tomoko , Mukai Harumi , Kozuma Yukinori , Kojima Hiroshi , Ninomiya Haruhiko 
Published: October 2019

Abstract
CD9 is a four transmembrane protein belonging to a tetraspanin family and regulates cell motility and adhesion. Several reports have indicated that CD9 form complexes with integrin including platelet fibrinogen receptor integrin aIIb-b III and involve in platelet function. We have previously reported that the c-myb knock down (KD) mice exhibited anemia and thrombocytopenia, and the expression level of CD9 mRNA was markdly increased in the c-myb KD mice. Reverse correlation of c-Myb expression with the CD9 gene expression was verified and agonistic antibody of CD9 stimulated megakaryocytic colony formation. These observations suggested that the CD9 expression was downregulated by c-myb. In our current study, we investigated the role of CD9 during megakaryopoiesis and platelet function by using CD9-null mice. Numbers of megakaryocytes and platelets, CFU-Meg, and ploidy were not different between wild-type and CD9-null mice. However, proplatelet formation (PPF) was significantly impaired in CD9-null megakaryocytes, and the size of proplatelets was smaller than those generated by wild-type megakaryocytes. Furthermore, after the bone marrow suppression with 5-fluorouracil (5-Fu), the recovery phase of platelet counts were delayed in the CD9-null mice. To clarify the reason of this platelet- recover delay, the number of bone marrow megakaryocyte was investigated using anti-vWF antibody staining, but the serial measurement of megakaryocyte number in CD9-null mice was not changed compared with wild-type mice. And also megakaryocyte ploidy was not changed in CD9-null mice compared with wild-type mice. Previous reports revealed that the cytoskeleton reorganization has a key role for the PPF formation, and Rac1-PAK1 signaling is important for actin restructure and aggregation prior to the cytoskeleton reorganization. The PPF formation was suppressed by Rac1 inhibitor. Although the mRNA expression levels of Rac1 in CD9-null mice were almost same as that in wild-type mice, PAK1 activation, major target of Rac1, was delayed in CD9-null mice compared with wild-type mice using thrombin-stimulating platelets. These results suggested that the delay of PAK1 activation cause the suppression of PPF formation in CD9-null mice.
Our previous and current studies demonstrate that c-Myb suppresses the CD9 expression in a steady-state condition, while in the stress megakaryocytosis, CD9 is upregulated and acts to induce megakaryopoiesis and platelets production through Rac1-PAK1 signaling pathway. Elucidation of c-Myb-CD9 regulatory function seems to be important to understand the stress megakaryopoiesis.
Significance of Initial Lumbar Puncture on Day8 of Remission Induction Therapy in Childhood Acute Lymphoblastic Leukemia: The Results from the Tokyo Children’s Cancer Study Group L99-15 Protocol.
Authors: Hasegawa Daisuke , Manabe Atsushi , Ohara Akira , Koh Katsuyoshi , Ogawa Chitose , Hanada Ryoji , Tsuchida Masahiro 
Published: October 2019

Abstract
Traumatic lumbar puncture with the presence of leukemic blasts (TLP+) in the cerebrospinal fluid (CSF) has been shown to adversely affect the outcome of children with acute lymphoblastic leukemia (ALL). In the Tokyo Children’s Cancer Study Group (TCCSG), we adopted a strategy to defer the initial lumbar puncture (LP) and intrathecal (IT) chemotherapy until 1 week after prednisolone (PSL) monotherapy so that circulating blasts were substantially reduced when the initial LP/IT was performed. The result of L89-12 study was previously reported, in which the incidence of TLP+ was significantly diminished but the impact on the rate of central nervous system (CNS) relapse was not determined because over 80% of patients received cranial irradiation (CRT). We analyzed the result of L99-15 study in which CRT was employed in only 16% of patients. Seven hundred and fifty-five children with newly diagnosed ALL were enrolled onto the TCCSG L99-15 study between April 1999 and June 2003. Patients received the initial IT administration of methotrexate, hydrocortisone, and cytarabine on day 8 following PSL monotherapy. Patients with CNS2 (<5 leukocytes/microL of CSF with leukemic blasts) received additional IT therapy. Patients with CNS3 (>5 leukocytes/microL of CSF with leukemic blasts) or CNS1s (negative CSF finding, with cranial nerve palsy) received additional IT therapy and CRT. Patients with TLP+ did not require any reinforcement of CNS-directed therapy. The frequency of CNS1s, CNS2, CNS3, or TLP+ was 0.5%, 1.1%, 0.5% and 0.8%, respectively. These figures were almost equal to the results from L89-12 study and much lower than those reported by other study groups such as BFM. Of 22 patients (2.9% of all) classified as CNS positive group (CNS-1s, -2, -3 and TLP+), only one patient had a CNS relapse concurrently with bone marrow (BM) relapse, whereas 22 of 723 patients classified as CNS negative had CNS relapse (isolated 12, combined with BM 10). Overall, 4-year event-free survival rate was 78.2 +/− 1.6%, and 4-year cumulative incidence of overall CNS relapse was 3.3 +/− 0.7%. Our strategy may underestimate the frequency of CNS leukemia at diagnosis, however, unnecessary intensification of CNS treatment may be avoided while overall CNS relapse rate is quite low in spite of restricted indication for cranial irradiation.
Arsenic Trioxide Entered Cerebrospinal Fluid with the Help of Mannitol Overwhelm the Meningeal Relapse of Acute Promyelocytic Leukemia.
Authors: Zhou Jin , Meng Ran , Sui Xinhua , Feng Guosheng , Li Baoyu 
Published: October 2019

Abstract
Background and objective Central nervous system (CNS) relapse of acute promyeloytic leukemia (APL) is increasingly reported after treatment with all-trans retinoic acid (ATRA) and arsenic trioxide (As2O3), the optimal therapy for these cases remains unclear. As2O3 highly effective in APL marrow relapse, but its efficacy in APL CNS relapse is undefined. Recent researches showed that little As2O3 could pass through the intact blood- brain-barrier (BBB), which limits its use in prevention and treatment of central nervous system acute promyelocytic leukaemia (CNSAPL). After discovered the different tolarance between NB4 cell line and human cortex neuron to As2O3in vitro, and primarily cleared the safe range of As2O3 level in central nervous system (CNS), we created a non-invasive method to help As2O3 enter into CNS effectively.
Methods Five volunteers with isolated meningle APL relapse received the new As2O3 regimen (mannitol helped As2O3 penetration) after accepted the conventional treatment: As2O3 0.16mg/kg/d diluted with 5%glucose 250ml common speed intravenous infusion with intensive intrathecal chemotherapy (MTX 12mg/dose + cytarabine 50mg / dose + Dexamethasone 10mg/dose) and showed chemotherapy resistance. The new regimen included 125ml of 20% mannitol bolus through medial cubital vein with the speed of 12„d`30ml / min, followed by As2O3 0.08mg/kg/d diluted with 250ml-20% mannitol intravenous infusion with the speed of 6ml /min, and then followed by As2O3 0.08mg/kg/d diluted with 5%glucose 250ml intravenous infusion with the speed of 0.5ml/min, meanwhile, associated with intensive intrathecal chemotherapy. The elemental arsenic levels in CSF and plasma during the new regimen performance were dynamic monitored with atomic fluorescence, and the differentiation and apoptosis of the blast cells in CSF were detected with flow cytometry assay.
Results The elemental arsenic level in the CSF treated with mannitol helped As2O3 penetration was fluctuated between 0.04 mg /L (0.2 micromol /L) and 0.05mg / L(0.25 micromol /L), which was much higher than that with common speed As2O3 intravenous infusion, and reached to the As2O3 effectively therapeutic level. The blast cells in CSF showed significant differentiation and apoptosis after treated with mannitol helped As2O3 penetration. Three of the five patients obtained complete remission, two of them reached partial remission.
Conclusions The new regimen of mannitol helped As2O3 penetration promoted the entrance of elemental arsenic to CNS, which might be benefit to prevent and cure CNS APL, especially to chemotherapy resistant CNS APL.
One Antigen HLA-Mismatched Related and 8/8 Allele Matched Unrelated Donors Are Associated with Similar Survival after Hematopoietic Cell Transplantation for Acute Leukemia.
Authors: Valcarcel David , Kan Fangyu , Wang Tao , Lee Stephanie J. , Spellman Stephen R , Hale Gregory A. , Marino Susana , Marks David , McCarthy Philip L. , Oudshoorn Machteld , Petersdorf Effie W , Ringdén Olle , Setterholm Michelle , Sierra Jorge 
Published: October 2019

Abstract
Patients in need of an allogenetic hematopoietic cell transplant but who lack an HLA genotypically identical sibling donor, are faced with the decision to consider a single HLA antigen mismatched related donor, or a search for a suitable 8/8 matched unrelated donor. We compared the outcomes of adult patients (≥18 years old) receiving a transplant for the treatment of AML or ALL in first or second remission from either a one-antigen mismatched related donor (MMRD group, N=89) reported to the CIBMTR or an 8/8 HLA-A, B, C and DRB1 allele matched unrelated donor (UD group, N=700) facilitated by the NMDP between 1995–2005. MMRD group was typed by serological or DNA-based methods for HLA-A, -B and –DR with all results verified by lab report review. The UD group was retrospectively typed for HLA-A, B, C and DRB1 by high resolution typing methods. Most received myeloablative conditioning regimens (77%), calcineurin inhibitor-based GVHD prophylaxis (100%) and T cell replete grafts (100%). 13% received ATG with the conditioning regimen. Median follow-up was 54 and 38 months in the MMRD and UD groups, respectively. The MMRD group was younger (median age 35 vs 43, p=0.002), had more ALL patients with low-risk cytogenetics (43% vs 18%, p=0.005), had older donors (median age: 38 vs 34, p=0.047), were more likely to receive methotrexate for GVHD prophylaxis (89% vs 77%, p=0.014) and were more likely to be transplanted prior to 2001 (62% vs 24%; p<0.001). There were no differences in patient or donor gender, diagnosis, disease-status, cytogenetic-risk of AML, time from diagnosis to transplant, stem cell source, conditioning regimen, use of ATG and Karnofsky index. Univariate comparisons (MMRD vs. UD) showed: 3-year OS (42% vs 44%, p=0.647), 3-year DFS (41% vs 41%, p=0.931), 3-year TRM (39% vs 31%, p=0.136), 3-year incidence of relapse (20% vs 28%, p=0.094), grade III–IV acute GVHD by 100 days (22% vs. 15%, p=0.147), chronic GVHD by 1 year (35% vs 47%, p=0.029). All multivariate analyses were adjusted for patient and transplant characteristics and are shown in the table below.
In summary, transplants utilizing one-antigen mismatched related and 8/8 allele-matched unrelated donors did not significantly differ in overall survival or disease free survival, but chronic GVHD was more frequent after UD transplantation.
Outcome RR (MMRD vs. UD) 95% CI p-value Survival 0.99 0.73–1.34 0.94 Disease-free survival 1.06 0.80–1.41 0.69 Treatment related mortality 1.14 0.77–1.69 0.52 Relapse 0.81 0.50–1.30 0.38 Acute GVHD III–IV 1.53 0.91–2.57 0.11 Chronic GVHD 0.58 0.39–0.85 0.006
Risks and Benefits of Unrelated Donor Peripheral Blood Progenitor Cells (PBPC) in Children and Adolescents with Acute Leukemia.
Authors: Eapen Mary , Ringden Olle , Locatelli Franco , Frangoul Haydar , Remberger Mats , Laughlin Mary J. , Rocha Vanderson 
Published: October 2019

Abstract
Although PBPC is an acceptable alternative to bone marrow (BM) for transplanting children with leukemia, there are no published studies describing outcomes after unrelated donor PBPC transplants. We compared the results of 385 unrelated donor BM transplants that were allele-matched (n=186) or mismatched (n=199) at HLA A, B, C, DRB1 and 110 PBPC transplants that were matched (n=60) or mismatched (n=50) at HLA A, B, C, DRB1 in patients younger than 18 years of age. All patients had acute leukemia and were transplanted in 2000–2006. Median follow up of surviving patients was 2 years in both treatment groups. There were no significant differences in patient and disease characteristics, transplant conditioning regimen, graft-versus-host disease (GVHD) prophylaxis and donor-recipient HLA disparity by graft type. Though the early probability of neutrophil recovery (³500/ul) was faster after transplantation of PBPC (31% vs. 10% at day-14, p<0.001) the probability of recovery by day-28 was similar after PBPC and BM transplants (94% and 91%, p=0.391). In contrast, platelet recovery (³20,000/ul) was better after PBPC transplants (86% vs. 76% at day-60, p=0.022). Risks of grade 2–4 (hazard ratio [HR] 1.24, p=0.147) and grade 3–4 (HR 1.07, p=0.785) acute GVHD were similar after PBPC and BM transplants. The risk of developing chronic GVHD was significantly greater after PBPC transplants compared to BM transplants (HR 2.36, p<0.001). After adjusting for disease status, donor-recipient HLA disparity and age, the of transplant-related mortality (TRM) relapse, treatment failure (relapse or death from any cause; inverse of leukemia-free survival and overall survival were similar after PBPC and BM transplants. The Table below shows the day-100 probability of grade 2–4 acute GVHD and the 3-year probabilities of chronic GVHD, TRM, relapse, leukemiafree survival and overall survival by graft type. These results differ from transplantation of PBPC from HLA-matched siblings where higher chronic GVHD translated into higher TRM and lower LFS. It remains to be seen whether the observed higher chronic GVHD after PBPC transplants will eventually result in the long term in higher mortality or fewer leukemia recurrence.
PBPC BM Grade 2–4 acute GVHD 53% 49% Chronic GVHD 58% 33% TRM 20% 24% Relapse 34% 28% Leukemia-free survival 46% 48% Overall survival 49% 49%
Signal Transduction Inhibitors in Lymphoproliferative Diseases (LPD).
Authors: Grupp Stephan A. 
Published: October 2019

There are a wide variety of diseases characterized by abnormal proliferation of lymphocytes, with differing clinical manifestations and in a spectrum from benign to malignant. Underlying all of these disorders, however, are subversions of normal lymphocyte signaling pathways that contribute to the pathologic process, can be characterized, and can potentially be targeted. Acute lymphoblastic leukemia (ALL) is the most common cancer of childhood and an example of malignant LPD, while patients with nonmalignant LPD, such as autoimmune lymphoproliferative syndrome (ALPS), can be difficult to diagnose, suffer from chronic infections that can be life-threatening, have a decreased life expectancy, and often require long-term treatment with immunosuppressive agents, including steroids, which can result in long-term debilitating consequences. In order to improve outcome and potentially reduce side effects and long-term sequelae from current cytotoxic therapy, the development of novel, biologically relevant, molecular, and cellular targeted therapies is critical. Potential targeted therapies can be identified and tested using in vitro assays, in vitro bone marrow stromal cell culture systems, murine models of ALL and LPD, and xenografts of primary human ALL. These pre-clinical assays have indentified promising and clinically relevant targets in the mTOR, IL-7, and Notch pathways, some which are now being tested in clinical trials. These include
rapamycin, being tested in both ALL and ALPS; temsirolimus, a second generation rapamycin analogue that shows promise in combination with cytotoxic chemotherapy in ALL; inhibitors of IL-7 signaling; and DAPT, a gamma-secretase inhibitor that has shown pre-clinical activity against both lymphoproliferation and autoimmune manifestations including nephritis in ALPS.
GATA Factor Mechanisms and Globin Gene Regulation.
Authors: Bresnick Emery H. , Kim Shin-Il , Bultman Scott J. , Lee Sherry , Boyer Meghan E. , Fujiwara Tohru , Wozniak Ryan J. 
Published: October 2019

Abstract
Key steps in hematopoiesis and the expression of genes encoding hemoglobin subunits are critically dependent upon specific members of the GATA factor family of transcription factors. Our recent efforts have focused on elucidating how GATA factors select functional sites in chromatin and how they function combinatorially with additional regulatory factors. GATA motifs are often arranged in close proximity to E-boxes, and such composite elements commonly mediate GATA factor- and Scl/TAL1-dependent transcriptional responses. Only a small fraction of these composite elements in chromatin are occupied by GATA factors and Scl/TAL1, and a specific epigenetic signature distinguishes occupied versus unoccupied elements genome-wide. In the context of hemoglobin synthesis, we are using genetic and molecular approaches to dissect the multistep mechanism underlying the control of β-globin transcription. GATA-1-containing complexes assemble at the β-globin Locus Control Region (LCR) prior to the murine adult βmajor promoter. Though the LCR physically interacts with the βmajor promoter, this interaction is not required for the binding of several trans-acting factors to the LCR or the promoter. A hypomorphic mutation of the chromatin remodeler BRG1 limits the extent to which RNA Polymerase II (Pol II) is recruited to the promoter and also abrogates the LCR-promoter interaction. Whereas looping is not required for assembly of the full complement of promoter complex components, looping is linked to the establishment of maximal levels of Pol II at the promoter. Collectively, these results provide insights into the relationship between, and importance of, individual steps in the multi-step activation mechanism. I will discuss progress on unraveling mechanisms underlying GATA-1-mediated activation of the adult β-like globin genes as well as fundamental aspects of GATA factor function, which have broad relevance in diverse systems. promoter. Though the LCR physically interacts with the β promoter, this interaction is not required for the binding of several -acting factors to the LCR or the promoter. A hypomorphic mutation of the chromatin remodeler BRG1 limits the extent to which RNA Polymerase II (Pol II) is recruited to the promoter and also abrogates the LCR-promoter interaction. Whereas looping is not required for assembly of the full complement of promoter complex components, looping is linked to the establishment of maximal levels of Pol II at the promoter. Collectively, these results provide insights into the relationship between, and importance of, individual steps in the multi-step activation mechanism. I will discuss progress on unraveling mechanisms underlying GATA-1-mediated activation of the adult β-like globin genes as well as fundamental aspects of GATA factor function, which have broad relevance in diverse systems.
DNA Methylation and Globin Gene Expression.
Authors: Saunthararajah Yogenthiran , Lavelle Donald 
Published: October 2019

Understanding how the human γ-globin gene is regulated has important clinical implications because increased levels of fetal hemoglobin (HbF) are beneficial to patients with sickle cell disease and β-thalassemia. DNA methylation is strongly implicated in developmental silencing of the γ-globin gene based on:
an inverse correlation between DNA methylation of the γ-globin gene and its expression, the acquisition of CpG residues within the γ-globin 5’ region during evolution as the γ-globin gene was recruited to fetal stage expression, and the ability of pharmacological inhibitors of DNA methyltransferase (5-azacytidine; decitabine) to reactivate high-level expression of the γ-globin gene in experimental primates that led to clinical trials demonstrating that decitabine increased HbF to therapeutic levels in patients with sickle cell disease.
Decitabine treatment in vivo decreases DNA methylation of the γ-globin gene and increases association of RNA polymerase II, acetyl Histone H3 and H4, and Histone H3 (lys4) trimethyl with the γ-globin gene, strongly suggesting that decitabine increases γ-globin gene transcription. These results are consistent with the hypothesis that γ-globin expression in adults is repressed by the binding of methyl DNA binding proteins to the methylated γ-globin promoter with subsequent recruitment of co-repressor complexes that actively repress γ-globin transcription. The reduction of γ-globin gene DNA methylation induced pharmacologically in adults by decitabine is linked to high level γ-globin expression, as is the complete loss of γ-globin gene methylation attained physiologically during erythroid differentiation of fetal liver hematopoietic progenitor cells. The mechanism of action of the drug has not been definitively established, however, and the role of DNA methylation in regulation of γ-globin gene expression remains an active area of investigation. High level γ-globin expression in baboon erythroid progenitor cell cultures without a reduction of γ-globin gene DNA methylation suggests the existence of alternative mechanisms of activation. In addition to reducing DNA methylation, decitabine activates the p38 MAP kinase pathway, increases p21WAF, and accelerates terminal erythroid differentiation. The role of these effects remains to be investigated. Increased understanding of the role of DNA methylation in γ-globin gene regulation is likely to impact the design of future therapies to increase HbF levels.
Selectins, Heparins, and Cancer: Rationale for Clinical Trials.
Authors: Varki Ajit P. , Varki Nissi M. , Borsig Lubor 
Published: October 2019

Abstract
Heparin has been in use as an anticoagulant for many decades, based on its ability to enhance antithrombin activity. However, heparin is a natural product, comprising a complex polydisperse mixture of highly sulfated glycosaminoglycan chains, only a fraction of which bind antithrombin. We reported that heparin has an unrelated biological effect in inhibiting P- and L-selectin binding to their natural ligands. Meanwhile, animal studies and analyses of human clinical data suggest that heparin therapy increases survival in various cancers. In contrast, clinical trials using Vitamin K antagonists showed no major effect on survival in most studies. Thus, the heparin effect on cancer may not be due primarily to its anticoagulant function, but rather to inhibition of P- and L-selectin. Indeed, we have demonstrated that the selectins play critical roles in animal models of the hematogeneous phase of carcinoma metastasis and of the platelet-rich microthromboses typical of classical Trousseau’s syndrome. This involves interactions of selectins with pathological ligands produced by carcinoma cells, particularly sialylated, fucosylated, and sulfated mucins. The critical selectin-mediated interactions blocked by heparin occur early in the metastatic cascade. Also, inhibition occurs at concentrations in the clinically acceptable range. Furthermore, there are many other salutary actions of heparin in cancer, including adsorption of cytokines and growth factors, inhibition of thrombin generation, and inhibition of heparanases. We, therefore, suggest that heparin therapy for metastasis prevention in humans be revisited, with these new paradigms in mind. Most of the above studies used unfractionated heparin. We found that selectin-inhibiting effects of low molecular weight heparins (LMWH) are varied and that the synthetic heparinoid pentasaccharide that specifically binds antithrombin does not block at clinically relevant levels. Optimal dosing to sustain clinically acceptable levels of all these drugs in mice gave congruent results. These data suggest a rational clinical trial comparing an optimal LMWH with the pentasaccharide in the pre-/peri- and immediate post-operative periods of newly diagnosed carcinomas. Notably, heparin can also attenuate inflammatory pathologies in animal models by blocking L- and P-selectin.