bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Difference in the Incidence and the Trend of Malignant Lymphomas in Japan and the United States.
Authors: Chihara Dai , Ito Hidemi , Matsuda Tomohiro , Shibata Akiko , Sobue Tomotaka , Matsuo Keitaro 
Published: October 2019

Abstract
Abstract 2695

Background:
Malignant lymphomas (ML) are heterogeneous groups that the detailed classification is evolving dramatically. An incidence of malignant disease in certain population reflects cumulative exposure to environment, genetics and their combination overtime. Therefore, a comparison of incidences in various population and their secular trends is very helpful to understand etiology of disease. The aim of this study is to assess the incidence and the trend of each ML subtypes and to evaluate the difference between Japan and US.

Materials and Methods:
We used the data from a population-based cancer registry in Japan and from the Surveillance Epidemiology and End Results (SEER) program 9. Registry data of the US, SEER 9 included 95,155 cases and the data of Japan included 48,658 cases. The period covered in this analyses was 1993 to 2006 in Japan and 1993 to 2008 in the US. Rates of sex-specific, age-standardized incidence with 95% confidence intervals were estimated and standardized by age-adjustment according to the world standard population. We also estimated the annual percent change using joinpoint regression analysis and evaluated the significance of the trend.

Results:
The overall age-standardized incidence rate of all malignant lymphomas per 100,000 in 2006 was 22.4 for males and 16.0 for females in the US, 7.4 for males and 4.9 for females in Japan. The incidence is higher in the US than Japan with most of the subtypes especially for the nodular sclerosis HL, CLL/SLL and FL. In general, B-cell lymphomas showed large difference in incidence while T-cell lymphomas (TCL) showed similar incidence between Japan and the US. The total numbers of ML are constantly increasing in Japan but not in the US as shown in the figure {annual percent change (95%CI), Japan; +2.6% (2.1, 3.1), US; +0.2% (−0.0, 0.4)}. As for details, follicular lymphoma, mantle cell lymphoma, Burkitt lymphoma and the total numbers of TCL are constantly increasing in both countries.

Conclusion:
In conclusion, we showed the marked difference in the incidence and the trend of hematologic malignancies between Japan and the US. The incidence of hematologic malignancies is lower in Japan than the US, but is increasing significantly. The most remarkable difference in the incidence was seen with nodular sclerosis HL, CLL/SLL and FL which was consistent with previous reports. Even with the TCL, the incidence seems to be similar to higher in the US except for the ATLL. The improvement in the quality of cancer registry systems and the organization of these registries between countries enables us to unite the data worldwide that will bring us new insights.
Solid line and circle are the data of the US. Dashed line and hollow circle are the data of Japan.

Disclosures:
No relevant conflicts of interest to declare.
Thrombus Development Processes Are Dependent On Endothelial Injuries: Examined By In Vivo Molecular Imaging
Authors: Nishimura Satoshi , Eto Koji , Nagai Ryozo 
Published: October 2019

Abstract
The thrombotic cellular mechanisms associated with cardiovascular events remains unclear, largely because of an inability to visualize thrombus formation. In addition, the contribution of endothelial cell (EC) injuries to thrombus formation processes are unclear, and we developed in vivo imaging technique based on single- and multi-photon microscopy to revealed the multicellular processes during thrombus development (Figure a,b). We visualized the cell dynamics including single platelet behavior, and assessed dynamic cellular interplay in two thrombosis models. First, we visualized that rapidly developing thrombi composed of discoid platelets without EC disruption was triggered by ROS photochemically induced by moderate power laser irradiation (Figure c). In this model, thrombus consisted by discoid platelet aggregations without leukocyte recruitment. The second model is, thrombus with EC disruption. High power laser induced EC erosion and extravasations of circulating leukocytes with thrombus development. Inflammatory cytokine, adhesion molecules dynamically control these two processes. (Figure d)Figure.Figure.
Using this technique, we elucidated that Lnk (adapter protein) regulates integrin signaling leading to stabilization of developing thrombus without EC disruption. Specifically, adhesion molecules dynamically control these processes. Thrombus formation was initiated by the binding of platelet GPIb-alpha to endothelial von Willebrand Factor in this model, and actin linker talin-dependent activation of alphaIIb-beta3 integrin in platelets was required for late phase thrombus stability.
As for the thrombus formation with EC disruption, chemokine expressions in endothelium and leukocyte (especially neutrophils) recruitment played a significant role in these processes. TLR4 signaling also contributed to these steps.
In sum, using our imaging system can be a powerful tool to analyze thrombus formation and evaluate the therapeutic strategies.

Disclosures:
No relevant conflicts of interest to declare.
The Use Of Platelet Transfusions In The Intensive Care Unit and Impact On Platelet Count: A 30,000 Patient Registry Study
Authors: Ning Shuoyan , Barty, MLT Rebecca , Liu Yang , Heddle Nancy , Arnold Donald 
Published: October 2019

Abstract

Background
Thrombocytopenia is a common complication of critical illness and an independent risk factor for bleeding and death in the intensive care unit (ICU). Platelet transfusions are commonly used to improve platelet counts; however, the expected platelet increment from a transfusion in this setting has not been established. The objective of this study was to describe the frequency of platelet transfusion administration and their effect on platelet count increments in a large cohort of non-oncology critically ill adults.

Methods
We performed an analysis of a registry database, which was developed to capture clinical and laboratory data on all blood transfusions administered in 3 academic hospitals in Hamilton, Ontario, Canada. We included all patients ≥18 years who received one or more platelet transfusion during an ICU admission. Data validation was done by integrity checks with medical records and laboratory information system performed by a biostatistician. Non-transfused ICU patients were used as controls. The absolute increment in platelet count was calculated for each single platelet transfusion using the closest platelet count taken within 24 hours before the transfusion and 4-24 hours after the transfusion.

Results
Between April 2006 and October 2012, 33,222 patients were admitted to ICU, including 29,511 (88.8%) who did not have a diagnosis of cancer. Of those, 4,502 (15.3%) received one or more platelet transfusion during any ICU admission (n=4,690); 31.9% were female and median age at the time of first admission was 69 years (IQR 59-77). Among the 25,009 non-transfused patients admitted to ICU during the same period, 38.1% were female and the median age was 65 years (IQR 52–76). Median pre-transfusion platelet count was 87 x109/L (IQR 59-131) and a single platelet transfusion resulted in a median platelet count increment of 21 x109/L (IQR 6-40) as measured 6.7 hours (IQR 5.1-9.8) after the transfusion. There were 277 (25.4%) transfusions that yielded a platelet count increment of 5 x109/L or less. ICU mortality was 562/4,690 (12.4%) for patients who received a platelet transfusion, compared with 2,251/33,033(6.8%) for patients who were not transfused during their ICU stay.

Summary/Conclusion
Among this large cohort of non-oncology ICU patients, platelet transfusions were commonly administered for thrombocytopenia that was generally mild. In this setting one platelet transfusion resulted in a median platelet count rise of 21 x109/L. Many transfusion episodes yielded no appreciable increase in platelet count. Further studies are needed to determine the clinical effects of platelet transfusion in this setting controlling for confounding.

Disclosures:
Heddle: CIHR: Research Funding; Canadian Blood Services: Membership on an entity’s Board of Directors or advisory committees; Health Canada: Research Funding; Macopharma: Consultancy; ASH: Honoraria.
Procoagulant Activity In Stored Units Of Red Blood Cells
Authors: Butenas Saulius , Aleshnick Maya , Foley Jonathan H , Keating Friederike K. 
Published: October 2019

Abstract
It has been observed that the procoagulant activity of stored units of red blood cells (RBC) increases over time, which is, in part, related to the expression/exposure of tissue factor (TF) on blood cells and microparticles (Keating et al, Transfusion 2011;51:1086). However, in many instances there is a discrepancy between the levels of TF measured in stored units of RBC and changes in procoagulant activity observed over the storage time. We hypothesized that, in addition to TF, an exposure of an acidic phospholipid, phosphatidylserine (PS), on blood cells (Whelihan et al, Blood 2012;120:3837), could be the cause of an elevated procoagulant activity.
Four healthy donors were recruited. RBC units were prepared and stored at 4°C for 30 days. On selected days, aliquots were removed, reconstituted with autologous plasma and recalcified, and were tested in the presence of corn trypsin inhibitor in the thromboelastography assay for procoagulant activity. For all 4 donors, the clotting time decreased from ∼3000-4000 s on day 1 to ∼1000-1600 s on day 30, with the most dramatic changes occurring between days 1 and 10. Addition of an inhibitory anti-TF antibody slightly prolonged clotting times, suggesting the presence of endogenous TF in RBC units. The concentration of TF, quantitated in an activity-based assay, did not change significantly over time for 3 of 4 donors and was within the range of 40-130 fM. For the fourth donor, TF concentration reached a maximum of 310 fM on day 10 and then decreased steadily to 60 fM on day 30.
In an attempt to identify other components in RBC units responsible for the procoagulant activity, the effect of lactadherin, which binds to the phosphatidylserine exposed on cell membranes, was investigated. An addition of 100 nM lactadherin substantially prolonged the clotting time of the reconstituted RBC. Thus, in days 3-5 the clotting time in the presence of lactadherin was by 2.4-3.4-fold longer than in the absence of it. With increasing age of the RBC units, the effect of lactadherin diminished and at day 30, a prolongation of the clotting time did not exceed 1.3-1.8-fold. The addition of anti-TF monoclonal antibody in the presence of lactadherin further enhanced the clotting time of the reconstituted RBC.
In conclusion, our data indicate that in stored units of RBC, the procoagulant activity is dependent upon the presence of active monocyte TF and phosphatidylserine exposed on blood cells and platelets. However, the failure of lactadherin in combination with the anti-TF antibody to completely inhibit the procoagulant activity in RBC units suggests that other components of blood (most likely platelets) support this activity as well.

Disclosures:
No relevant conflicts of interest to declare.
The Transcription Factor IRF8 is a Key Transcription Factor for Basophil Development
Authors: Kurotaki Daisuke , Sasaki Haruka , Osato Naoki , Sasaki Izumi , Kaneda Chika , Sato Hideaki , Nishiyama Akira , Kaisho Tsuneyasu , Aburatani Hiroyuki , Morse Herbert C. , Ozato Keiko , Tamura Tomohiko 
Published: October 2019

Abstract
Basophils are the rarest granulocytes circulating in the peripheral blood. They play critical roles in anti-parasite Th2-type immune responses and chronic allergic disorders. The developmental pathway for basophils has been recently demonstrated; myeloid progenitors pass through common myeloid progenitors, granulocyte-monocyte progenitors, granulocyte-committed progenitors (GPs), and basophil-committed progenitors (BaPs) in the bone marrow. BaPs then give rise to mature basophils. However, our understanding of how this pathway is regulated remains still elusive. Interferon Regulatory Factor-8 (IRF8), a hematopoietic cell-specific IRF transcription factor, is essential for the development of monocytes, dendritic cells, and eosinophils, while it inhibits neutrophil differentiation. Its role in the development of basophils has yet to be analyzed.
In this study, we investigated whether IRF8 has any role in the development of the basophil lineage. We found that Irf8–/– mice displayed a severe reduction of basophil counts in the bone marrow, peripheral blood and spleen compared to wild-type (WT) mice. Irf8–/– mice retained GPs but lacked BaPs. Cell transfer experiments revealed that the defect of basophil development in Irf8–/– mice resides in bone marrow cells. We utilized IRF8-GFP chimera knock-in mice to examine IRF8 protein expression in the basophil lineage at a single cell level. We found that GPs, but not BaPs and mature basophils, expressed IRF8. Furthermore, purified Irf8–/– GPs failed to efficiently give rise to basophils in vitro. These results indicate that IRF8 acts at the stage of GPs in a cell-intrinsic manner. To understand the mechanism by which IRF8 promotes basophil development, we performed transcriptome analysis of purified GPs from WT and Irf8–/– mice by microarray. Because IRF8 is no more expressed in BaPs, we envisaged that IRF8 acts by inducing downstream transcription factors in GPs. The expression of several transcription factor genes such as Gata2 and Spib was reduced in Irf8–/– GPs compared to WT GPs. Analysis of DNA motifs in the promoter regions of genes downregulated in Irf8–/– GPs predicted that GATA transcription factor(s) may act downstream of IRF8. Indeed, retroviral transduction of GATA2, known to be essential for basophil development, into Irf8–/– hematopoietic progenitor cells rescued basophil differentiation in vitro. On the other hand, Spib–/– mice showed no obvious defects in basophil development. Taken together, these results suggest that the IRF8-GATA2 axis in GPs critically regulates basophil development.

Disclosures:
No relevant conflicts of interest to declare.
Cyclosporine A May Affect the Expression of Foxp3 by Regulation of Akt and Stat3 Phosphorylation in CD4+CD25+ Regulatory T Cells from Mouse Model of Severe Aplastic Anemia
Authors: Lin Zenghua , Liu Hong , Wang Xuli , Zhang Yaping , Cai Yifeng , Qian Yu 
Published: October 2019

Abstract
Although Cyclosporine A(CsA) is an effective immunosuppressant in the treatment of aplastic anemia(AA), its affects on CD4+CD25+ regulatory T cells(Tregs) from patients with AA still remains controversial. We have first developed a novel mouse model of severe aplastic anemia(SAA) induced by administration with IFN-γ and busulphan. In this study, the SAA model female mice were intragastrically administered with CsA at daily dose of 5 mg/kg for 10 days(the CsA-treated group, n=60), the SAA group(n=60) and the un-treated group (n=60) were as control. All mice selected to set up the SAA model developed the typical clinical and pathological patterns of SAA from day 10 post dosing. Most of them presented bleedings in association with anemia and infections. At day 15 after treatment, a mortality of 35% was found in the SAA group, while a mortality of 20% was found in the CsA-treated group because of impaired liver function and infections. In order to explore the effect of CsA on CD4+CD25+ Tregs from the novel mouse SAA model, the mononuclear cells of the peripheral blood and spleen were subjected to assess the percentage of CD4+CD25+ Tregs and the intracellular Foxp3 expression in CD4+CD25+ Tregs by flow cytometry(FCM). In the SAA group, the FCM analysis showed decreased ratios of CD4+CD25+ Tregs in CD4+ T cells and down-expression of Foxp3 in CD4+CD25+ Tregs compared with the un-treated group(P<0.05, respectively). These results were in accordance with our previous observations. However, after treatment with CsA, the expression of Foxp3 in CD4+CD25+ Tregs was increased(P<0.05) but not the ratios of CD4+CD25+ Tregs in CD4+ T cells(P>0.05). In addition, after being pured by immunomagnetic beads, the splenic CD4+CD25+ Tregs was subjected to assess the Akt and Stat3 phosphorylation using Western blot. Compared with the un-treated group, CD4+CD25+ Tregs with increased Akt phosphorylation accompanied by increased Stat3 phosphorylation was found in SAA group(P<0.05, respectively). On the contrary, CsA-treated group exhibited CD4+CD25+ Tregs with a reduction in both Akt phosphorylation and Stat3 phosphorylation compared with the SAA group(P<0.05, respectively). These results indicated that CsA may increase the expression of Foxp3 by up-regulation the levels of Akt and Stat3 phosphorylation in CD4+CD25+ Tregs from the novel mice model of severe aplastic anemia.

Disclosures:
No relevant conflicts of interest to declare.
Activation Of Akt Enhances Ribosomal RNA Synthesis In AML Cells Through a Novel Isoform Of TIF-1A and Inhibition Of Filamin A Cleavage
Authors: Truong Nguyen Le Xuan , Huang Min , Chan Steven M. , Majeti Ravi , Mitchell Beverly S. 
Published: October 2019

Abstract
PI3Kinase/Akt signaling is frequently activated in acute myelogenous leukemia (AML) and inhibitors of this pathway have shown some promise in both pre-clinical and early clinical studies. In particular, much attention has been focused on components of the mTOR complex as downstream effectors of Akt activation that promote cell proliferation and survival in AML and other malignancies. We have discovered a direct effect of activated and phosphorylated Akt in increasing ribosomal RNA (rRNA) synthesis in both leukemic cell lines and primary AML cells by a novel pathway that is independent of mTOR. We have defined TIF-90 as a splice variant of TIF-1A, a molecule that tethers RNA Polymerase I to the ribosomal DNA promoter and is required for rRNA synthesis. TIF-90 is localized in the nucleolus and directly co-locates with Pol I in imaging, pull down, and rDNA promoter occupancy studies. Activation and phosphorylation of Akt correlates with high levels of TIF-90 expression and increased TIF-90 occupancy of the rDNA promoter in primary AML cells, while overexpression of constitutively activated Akt (Akt-Myr) increases rRNA synthesis in a TIF-90-dependent fashion in cell lines. Activation of Akt increases the expression of TIF-90 by inhibiting its ubiquitination and proteasomal degradation by the p53 ubiquitin ligase, Mdm2. Furthermore, activated Akt directly inhibits the cleavage of the actin-binding protein, Filamin A, into a 90 kDa fragment that interacts with TIF-90 and prevents it from occupying the rDNA promoter. Expression of the 90 kDa Filamin A cleavage fragment alone is sufficient to inhibit rRNA synthesis by binding to TIF-90. Thus, inhibition of Filamin A cleavage by Akt is associated with increased rRNA synthesis. The Akt inhibitor AZD8055, but not the mTORC1 inhibitor Rapamycin, promotes the cleavage of filamin A, decreasing both TIF-90 occupancy of the rDNA promoter and rRNA synthesis. Examination of 21 primary AML samples reveals an inverse correlation between the phosphorylation of Akt and the cleavage of Filamin A; conversely, AZD8055 inhibits both Akt phosphorylation and rRNA synthesis in these samples while promoting the cleavage of Filamin A. These data demonstrate a novel pathway by which Akt directly enhances rRNA synthesis. This pathway is active in primary AML cells and in cell lines that have high levels of phosphorylated Akt. Inhibition of rRNA synthesis has been shown to result in nucleolar disruption, cell cycle arrest, and ultimately cell death. Our data support an mTOR-independent effect of Akt activation on cell proliferation that would require direct targeting of PI3Kinase or activated Akt to have a major therapeutic effect in AML.

Disclosures:
No relevant conflicts of interest to declare.
Absence Of The E3 Ubiquitin Ligase SKP2 Attenuates Notch-Induced T-Cell Leukemogenesis
Authors: Rodriguez-Rodriguez Sonia , Wang Lin , Zhang Huajia , Zollman Amy , Cardoso Angelo , Carlesso Nadia 
Published: October 2019

Abstract
Acute Lymphoblastic Leukemia (ALL) is the most common malignancy in childhood, accounting for almost 30% of pediatric cancers. In pediatric T-cell acute lymphoblastic leukemia (T-ALL), around 30% of the children still undergo disease relapse, which is associated with poor prognosis. Hence, novel therapeutic strategies for the treatment of T-ALL are necessary, since conventional therapies still fail to cure a significant number of patients. Notch signaling contributes to the regulation of normal T-cells homeostasis, whereas oncogenic, activated form of Notch is present in the majority of the T-ALL cases.
Previous studies in our laboratory demonstrated that Notch1 signaling induces transcriptional activation of SKP2, the F-box protein of the SCF E3-ubiquitin ligase complex. SKP2 targets the major cell cycle inhibitors, the CKIs p21Cip1, p27Kip1, p57Kip2 and p130, for proteasome-mediated degradation. Increased Skp2 expression accelerates cell cycle progression in hematopoietic cells, and its overexpression is frequently found in cancers, in particular lymphomas and leukemias, in which is associated with poor prognosis.
We found that Skp2 expression is dynamically regulated during T-cell differentiation coinciding with the Notch expression pattern. Moreover, primary thymocytes cultured in vitro responded to Notch stimulation by the Delta1 ligand increasing their Skp2 expression and their cell cycle activity. As anticipated, we found that Skp2 expression is increased in T-ALL patient samples. Our hypothesis is that Notch activation promotes T-cell leukemogenesis by altering the cell cycle control through upregulation of SKP2. To better define the role of SKP2 in T-ALL, we analyzed the Skp2 expression in a murine model of Notch-induced T-cell leukemia. Mice transplanted with stem/progenitor cells transduced with the constitutive active form of Notch (ICN), developed T-ALL and died by week 12 after transplant. Analysis of T-ALL cells revealed a 5 fold upregulation of Skp2 expression compared to controls. Next, we tested whether SKP2 was required for T-ALL development. To this end, we evaluated leukemia development in mice transplanted with SKP2 deficient cells overexpressing ICN. Stem/progenitor cells derived from Skp2+/+, Skp2+/- or Skp2-/- mice were transduced with ICN and then transplanted into WT recipients. Loss of SKP2 significantly delayed the development of T-cell leukemia in transplanted mice and increased their survival by 60% at 12 weeks.
Taken together, these results demonstrate a previously unrecognized role for SKP2 in the initiation and progression of T-ALL and its potential role as a therapeutic target.

Disclosures:
No relevant conflicts of interest to declare.