bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Review Of Computed Tomography Pulmonary Angiograms In Three Teaching Hospitals: The Rise Of Sub-Segmental Pulmonary Emboli and Their Management
Authors: Goy Jennifer , Lee Justin Y. , Levine Oren , Chaudhry Salman , Crowther Mark A. 
Published: October 2019

Abstract

Background
The availability of Computed Tomography Pulmonary Angiography (CTPA) has led to an increase in the number of investigations for Pulmonary Embolism (PE). With more widespread use of these high resolution scans, the frequency of identification of isolated Small Sub-segmental Emboli (SSPE) is also expected to increase. Current clinical practice guidelines do not make any treatment distinctions for SSPE, though the benefits of anticoagulation for SSPE have not been established.

Aims
To review the frequency of Pulmonary Embolism and Sub-segmental Pulmonary Embolism identified through CTPA as well as their management

Methods
Retrospective review of 2213 patient charts who underwent CTPA in three Hamilton teaching hospitals from 2009-2011. In depth chart review of patients with SSPE was undertaken to determine the frequency with which patients who received anticoagulation therapy for SSPE. The frequency of bleeding complications and recurrent thrombosis were also investigated in this detailed SSPE chart review.

Results
Our patient population (mean age 65) consisted of 1099 medical inpatients (50%), 702 surgical inpatients (32%) and 412 (18%) emergency department patients.
PE was identified in 26 % of scans (n=576). Of these, SSPEs were the only identified thrombus in 82 patients (4% of total scans and 14% of identified PEs). In 55 of these 82 SSPEs, in addition to the SSPE, an alternative diagnosis that might explain the PE symptoms was found.
Fifty-two percent (n=43) of the patients with an SSPE received therapeutic anticoagulation. In these life threatening bleeding occurred in 2 patients. There was no documented recurrent thrombosis or thrombosis-related deaths in three month follow-up among the 39 patients who did not receive anti-coagulation for SSPE.
Of the 1,608 CTPAs that did not identify PE, an alternative diagnosis to account for the patient’s symptoms was identified on CT in 1078 (67%) and no alternative cause was found in 531 (33%).

Summary/Conclusions
Our study demonstrated a much lower frequency of pulmonary embolism in comparison to approximate 50 % pre-test probability of a positive scan seen in studies which validated CTPA for the diagnosis of PE. Isolated SSPEs accounted for 14% of all PEs found in our study population – and were present in 4% of all patients undergoing CTPA. A substantial proportion of patients were anti-coagulated SSPE (52%) and two developed life-threatening bleeding complications. No recurrent VTE was documented in patients who were not anticoagulated for PE, though follow-up was limited to hospital records. Randomized controlled trial data is needed to further investigate the risks and benefits of anticoagulation in patients with SSPE.

Disclosures:
No relevant conflicts of interest to declare.
Upregulation Of Hepcidin Expression By Lactoferrin Administration To Pre-Weanling Mice
Authors: Elif Memisoglu , Cooper Jeffrey A. , Fleming Robert E. 
Published: October 2019

Abstract

Background
Hepcidin is a peptide hormone produced in hepatocytes which regulates dietary iron absorption and systemic iron distribution. The hepatic expression of the hepcidin gene is regulated by signals which reflect body iron status and erythropoietic activity. The regulation of hepcidin by iron status includes a signal from the circulating transferrin via hepatocellular transferrin receptor (TfR2). Lactoferrin is a member of the transferrin family of iron-binding glycoproteins found in high concentrations in milk. Lactoferrin has a well described role in host defense; however, its in iron metabolism is less clear. Like transferrin, lactoferrin will deliver iron to hepatocytes but unlike transferrin, lactoferrin cannot deliver iron to erythroid cells. Lactoferrin does not interact with TfR1 or TfR2. We took advantage of these properties of lactoferrin to determine if delivering iron to the liver (but not erythron) in a form which is independent on transferrin and its receptors could regulate hepcidin.

Methods
To take advantage of the naturally-occuring low iron and low hepcidin states at this developmental stage, experiments were performed on pre-weanling mice. Mice at 12-14d (pre-weanling) were administered human lactoferrin equivalent to 2 mg/kg iron (n=10) or carrier (n=10) intraperitoneally (IP) and sacrificed 6 h later. Other mice were administered a comparable IP dose of transferrin (n=5) or carrier (n=4). Additional groups of mice were delivered Lf (n=8) or carrier (n=7) enterally (by orogastric tube), and the sacrificed 6h or 24h later. Liver hepcidin (Hamp1) mRNA was measured by real-time RT-PCR, normalized to beta actin expression, and analyzed by commercial software (Rest 2007, Qiagen). Liver iron distribution was determined histochemically in tissue sections by modified Perls' staining.

Results
Parenteral administration of lactoferrin in pre-weanling mice was associated with a 32-fold increase in liver Hamp1 expression after 6 hours (P<0.001). This compares with an only 4.5 fold increase in Hamp1 expression 6h after a comparable dose of transferrin. Enteral lactoferrin likewise resulted in a ∼30-fold increase in Hamp1 expression (P<0.005) after 24 hours. Iron delivered to the liver from lactoferrin was distributed in sinusoidal lining cells as well as hepatocytes.

Conclusions
Exogenously administered iron-rplete lactoferrin increases liver Hamp1 expression in pre-weanling mice. These observations demonstrate that iron delivery to the liver in a form not dependent upon transferrin or its receptors upregulates hepcidin expression. These findings moreover raise the possibility that lactoferrin-mediated changes in hepcidin expression may contribute to reported benefits of supplemental lactoferrin on host defense.

Disclosures:
No relevant conflicts of interest to declare.
The HU Inducible SAR1 Gene Acts Through GαI /JNK/JUN Pathway To Activate γ-Globin Expression
Authors: Zhu Jianqiong , Chin Kyung , Aerbajinai Wulin , Kumkhaek Chutima , Li Hongzhen , Rodgers Griffin P. 
Published: October 2019

Abstract
Hydroxyurea (HU) has been effectively used in the treatment of β-hemoglobinopathies by augmenting γ-globin chain to compensate for the impaired or defective β-globin chain, but the molecular mechanism underlying HU-mediated γ-globin regulation remains unclear. We previously reported that over-expression of the HU-induced SAR1 gene increased γ-globin expression in primary erythroid cells, and SAR1 is required for HU's effects in this process. Here we present further mechanistic details underlying SAR1’s effects. First, we examined the role of COPII pathway in HbF expression by genetically silencing the key downstream COPII components Sec12/Sec23 or by chemical inhibition of vesicle protein transport from ER to Golgi with BFA. We found that blocking this pathway through siRNA Sec12/23 or BFA only reduced HU-induced γ-globin expression by 32% and 28%, respectively, without a change in basal HbF level (as determined by QT-PCR or flow cytometry). These data suggested that other signaling pathways might account for HU- and SAR1-mediated HbF induction. Additional studies also indicated that stable tranfected SAR1 was able to activate Gαi /JNK/JUN signaling in erythroid cell. Inhibition of JNK/JUN activation by the JNK inhibitor SP600125 was able to reduce by 8.79% and 38.63% HbF positive cells in vector- and SAR1-transfected-cells, respectively. Silencing of Gαi2 or Gαi3, but not Gαi1 or Gαq expression in SAR1-stable transfected cells could significantly eliminate SAR1-stimulated JUN protein phosphorylation, and resulted in the 54% and 46% reduction of γ-globin expression, respectively. Reciprocal co-immunoprecipitation assays revealed a direct association between Sar1 and Gα (Gαi and Gαq) protein in both erythroid and non-erythroid cells. These data taken together with our previous findings indicate that HU induces SAR1 which in turn activates γ-globin expression predominantly through the Gαi/JNK/JUN pathway. These findings illuminate an alternative approach to targeted therapy for the beta-globin disorders.

Disclosures:
No relevant conflicts of interest to declare.
Treatment of Relapsed ACUTE Leukemia after ALLO-HSCT By Salvage Chemotherapy with Low-Dose Cytarabine and Aclarubicin Combined with G-CSF Priming: More Effective and LESS Toxic?
Authors: Zhou Hua , Ge Ling , Zhu Xiaming , Qiu Huiying , Jin Zhengming , Miao Miao , Fu Chengcheng , Ma Xiao , Chen Feng , Xue Shengli , Ruan Changgeng , Depei Wu , Tang Xiaowen 
Published: October 2019

Abstract
Background: Recurrence is a major cause of treatment failure after allogeneic hematopoietic stem cell transplantation (allo-HSCT) for acute leukemia, and subsequential treatment options are very limited.
Aims: We evaluate the efficacy and toxicity of cytarabine and aclarubicin combined with granulocyte colony-stimulating factor priming (CAG regimen), consisting of concurrent use of granulocyte colony-stimulating factor (G-CSF) with low-dose cytarabine and aclarubicin, as a salvage therapy for acute leukemia patients who relapsed after allo-HSCT.
Methods: Fifty-nine patients (32 male and 27 female) with acute leukemia, with a median age of 27 years, relapsed post allo-HSCT and received salvage chemotherapy. Twenty-seven patients received CAG regimen while 32 patients received non-CAG regimen such as intensive chemotherapy.
Results: 1.The overall response rate (ORR) of CAG was significantly superior than that of non-CAG groups (55.6% vs 28.1%, P= 0.033). Especially for AML patients (64.3% vs 20%, P= 0.025). However, ORR of ALL in CAG group was similar with that in non-CAG group (50% and 35.2% respectively; P = 0.471).
2. Median overall survival (OS) from the starting of CAG chemotherapy and the estimated 2-year OS of CAG group were 9 (1-27) months and 16.1%. Meanwhile, median OS and the estimated 2-year survival of non-CAG group were 4 (1-49) months and 8.8% (Figure1). For the patients who received donor lymphocyte infusion (DLI) as a subsequential therapy, two-year OS of CAG and non-CAG group were 17.2% and 12.5% respectively (P=0.577).
3. The median duration of neutropenia and thrombocytopenia of CAG group were significantly shorter than that of non CAG group, 6 (1-12) vs 11 (5-28) days (P=0.000) and 8 (1-14) vs 14 (7-35) days (P=0.000). Treatment related mortality (TRM) was found in 2 cases in CAG group compared with 10 cases in non-CAG group.
4. For CAG group, univariate analysis results showed that lower leukocyte level and lower medullar blast percentage at relapse post-transplant were significantly associated with a higher overall response rate (P=0.005 and P=0.000).Furthermore, multivariate analysis results showed that the response to chemotherapy was the only factor that correlated with better survival (HR 0.426, P=0.016)
Conclusion: CAG regimen as a salvage chemotherapy for relapsed acute leukemia post allo-HSCT could effectively reduce tumor burden with mild toxicity, especially for hypoplastic acute leukemia patients. For certain type of relapsed acute leukemia patients post allo-HSCT, CAG regimen may be an optimal choice as the bridge therapy followed by DLI or second allo-HSCT.

Disclosures
No relevant conflicts of interest to declare.
Third Party Invariant Natural Killer T Cells Protect from Graft-Versus-Host Disease Lethality through Expansion of Donor CD4+FoxP3+ Regulatory T Cells
Authors: Schneidawind Dominik , Baker Jeanette , Buechele Corina , Meyer Everett H. , Negrin Robert S. 
Published: October 2019

Abstract
Graft-versus-host disease (GVHD) is driven by extensive activation and proliferation of alloreactive donor T cells causing significant morbidity and mortality following allogeneic hematopoietic cell transplantation (HCT). Invariant natural killer T (iNKT) cells are potent regulators of immune responses in both humans (TCRα Vα24-Jα18) and mice (TCRα Vα14-Jα18). As the iNKT cell receptor and the glycolipid-presenting molecule CD1d interaction is highly conserved, we explored the role of adoptively transferred third party CD4+ iNKT cells in a murine model of allogeneic HCT. BALB/c (H-2Kd) recipient mice were irradiated with 8 Gy and transplanted with T cell-depleted bone marrow together with 1x106 CD4+/CD8+ T cells (Tcon) from C57BL/6 (H-2Kb) donor mice. Adoptive transfer of purified (>95%) 5x104 CD4+ iNKT cells from FVB/N (H-2Kq) third party mice resulted in a significant survival benefit (p<0.001) while retaining Tcon mediated graft-versus-tumor (GVT) effects against A20 lymphoma cells (p=0.002). Consistently, weight and GVHD scores improved in mice that received a single injection of third party CD4+ iNKT cells as compared to animals that received Tcon alone. Notably, CD4+ iNKT cells from third party mice were as protective as CD4+ iNKT cells from donor mice (p=0.50). Signal intensity deriving from expanding luciferase expressing alloreactive Tcon was significantly lower in animals treated with third party CD4+ iNKT cells (p=0.003). Interestingly, inhibition of Tcon proliferation was similar to animals that received CD4+ iNKT cells from donor mice (p=0.90). In addition, adoptive transfer of third party CD4+ iNKT cells promoted a Th2-biased cytokine response of alloreactive donor T cells. Although we found that third party CD4+ iNKT cells were rejected by day +10 after allogeneic HCT, adoptive transfer of these cells resulted in a robust expansion of luciferase expressing donor CD4+FoxP3+ regulatory T cells (Treg) as measured by bioluminescence imaging (p=0.006). Using FoxP3DTR C57BL/6 donor mice, depletion of Treg from the graft abrogated both donor Treg expansion and protection from GVHD lethality through third party CD4+ iNKT cells. We conclude that low numbers of highly purified and adoptively transferred third party CD4+ iNKT cells protect from lethal GVHD through activation and expansion of donor Treg with retained GVT effects. Despite the fact that iNKT cells are a rare cell population, the in vivo activity of small numbers of cells and feasibility of in vitro expansion provide the basis for clinical translation.

Disclosures
No relevant conflicts of interest to declare.
Safety of Peripheral Hematopoietic Stem Cell Mobilization with Lenograstim in Allogeneic Healthy Donors ≥ 50 Years Old: A Single-Centre Experience
Authors: Martino Massimo , Dattola Antonella , Fedele Roberta , Pontari Antonella , Moscato Tiziana , Cuzzola Maria , Martinello Tiziana 
Published: October 2019

Abstract
Background and Objectives
The use of allogenic hematopoietic stem cell transplantation (HSCT) is increasing in older patients because of reduced ablative regimens and improved supportive care. An old patient usually has a old sibling healthy donors (HDs). The safety of G-CSF for mobilization in elderly HDs has not been provided. In our institution a long-term active follow-up study of G-CSF-mobilized healthy volunteer donors has been implemented.
Patients and Methods
After a median follow up of 94 months (7.8 years) we reviewed and analyzed safety data of our HDs database according to HDs age: HDs-1 (162), patients younger than 50 years old, HDs-2 (62), patients 50-59 years old and HDs-3 (23), patients 60 or more years old. Long-term follow-up included monitoring of neoplastic, cardiac or autoimmune diseases.
Results
Two hundred and forty-seven successive donors were evaluated and their characteristics are well balanced among age-groups and no statistical differences have been detected: most of them were male (55.9% ), sibling (97.2%) and HLA matching (93.1%). Mobilization failure rate has been 4.3% (7/162), 1.6% (1/62) and 13% (3/23) in HDs-1, HDs-2 and HDs-3, respectively (p=0.075).
Short- and long-term safety was not different among age-groups. Bone pain was reported as the most frequent short-term adverse event (76.5%). Other commonly observed short-term symptoms included headache (31.6%), fatigue (28.7%), insomnia (21.5%), nausea (13.0%) and fever (6.1%). For long-term safety surveillance no haematological malignancies were observed. The observed rate of solid tumors, cardiovascular and autoimmune events, was the expected incidence for these diseases in the western countries.
Conclusion
In our study, mobilization with lenograstim in elderly HDs seems to be as safe as for young HDs safe. This data contribute to the growing body of evidence of the long-term safety of G-CSF for allogenic donor stem cell mobilization also for elderly HDs.

Disclosures
No relevant conflicts of interest to declare.
Hypoferremia of Fasting in Mice Is Associated with Increased Hepcidin and Decreased Erythroferrone Expression
Authors: Li Yihang , Booth Gregory R , Feng Qi , Fleming Robert E 
Published: October 2019

Abstract
Background: Protein-calorie restriction in humans is associated with changes in iron metabolism that differ from dietary iron-deficiency. Dietary iron absorption and tissue iron distribution and are regulated by the hepatocellular peptide hormone hepcidin, which mediates a decrease in circulating iron concentrations and increase in cellular iron stores. Hepcidin is upregulated in response to increased serum and tissue iron, upregulated in response to ER-stress via the transcription factor CREB-H, and down-regulated in response to erythroid iron demand via the circulating signaling molecule erythroferrone/myonectin (ERFE). An overnight fast was reported to decrease expression of skeletal muscle ERFE in mice.
Objective: The aim of this study was to investigate the regulation of hepcidin in the hypoferremia of fasting.
Design/Methods: We analyzed parameters of iron homeostasis in five 5 week old male AKR mice fasted for 18 hours overnight (and allowed access to water) compared with mice with access to chow ad libitum. Serum and tissue iron concentrations were measured. Liver hepcidin mRNA was quantified by real-time RT-PCR and verified by Northern blot analysis. Marrow ERFE and liver CREB-H mRNA expression was quantified by RT-PCR and normalized to beta actin.
Results: As anticipated, fasted mice had significantly lower serum iron concentrations (203 vs 321 mcg/dL, P<0.001) and transferrin saturations (64.4 vs 79.6%, P<0.05). Duodenal iron concentrations were increased in fasted mice, 2.3-fold (P<0.01). Histochemical staining demonstrated iron retention in the absorptive enterocytes. Liver (916 vs 735 mcg/g, P=0.1) and splenic (480 vs 414 mcg/g, P=0.001) iron concentrations were modestly higher in the fasted mice. Despite hypoferremia, the fasted mice demonstrated increased liver hepcidin expression (1.9 fold, P < 0.05). Liver CREB-H mRNA expression was increased ~10 fold, P<0.01; however there was no change in liver XBP-1 processing (marker of ER stress). Marrow ERFE expression was downregulated by approximately one third (P<0.01).
Conclusions: The fasting state in mice is associated with increased hepcidin expression, iron retention in absorptive enterocytes, and decreased circulating iron. These observations suggest that the hypoferremia of fasting is not consequent to limited iron availability, but rather to hepcidin-mediated retention of iron in the duodenum and storage tissues. They moreover support a role for nutritional state, independent of iron status, as a regulator of hepcidin and mediated by changes in expression of ERFE and CREB-H.

Disclosures
No relevant conflicts of interest to declare.
Thrombospondin-1 Stimulates Calcium Influx and Echinocytosis in Sickle Cell-Derived Red Blood Cells
Authors: Kanias Tamir , Isenberg Jeffrey , Bullock Grant C , Schrott Valerie M , Stolz Donna B , Novelli Enrico M 
Published: October 2019

Abstract
Thrombospondin-1 (TSP1) is a secreted matricellular protein found preformed in platelet α-granules and upregulated in the plasma of patients with sickle cell disease (SCD). In preclinical models TSP1 enhances sickle red blood cell (RBC) binding to endothelial cells, while in murine sickle mice TSP1 promotes RBC echinocytosis and vaso-occlusion. Recently we reported elevated plasma TSP1 was associated with vaso-occlusive complications in SCD patients. Herein we tested the hypothesis that TSP1 compromises RBC membrane integrity via enhanced calcium influx leading to morphological changes, such as echinocytosis. RBC were collected via centrifugation of heparinized blood from human HbAA donors (n=3), SCD patients with homozygous HbSS SCD (n=4), and wild type C57BL/6J mice (n=2). Cells were then re-suspended in glucose and calcium-supplemented PBS, and treated with several concentrations of exogenous TSP1 (2.75 nM, 11 nM, 22 nM and 44 nM) for 2 hours at 37°C. RBC morphology was assessed by both light and electron microscopy. TSP1 modulation of RBC calcium influx was studied using the calcium fluorophore Fluo-3, AM. TSP1 treatment, in a dose-dependent manner, stimulated echinocytosis in human SCD and non-SCD RBC and in murine cells. In some instances electron microscopy showed echinocyte membrane projections with a string-of-beads appearance, suggestive of imminent microparticle shedding. TSP1 treatment, again in a dose-dependent manner, also significantly increased calcium influx in SCD (Figure, right panel) and non SCD RBC (Figure, left panel) and murine RBC (not shown). Interestingly, SCD RBC displayed more sensitivity to TSP1-mediated increases in cellular calcium as compared to normal cells. Our results show for the first time that TSP1, at doses found in patient plasma, stimulates significant echinocytosis and calcium influx in both normal and SCD RBC. These data identify a novel role for plasma TSP1 in promoting RBC pathology. Future studies are underway to elucidate this mechanism and its pathogenicity in humans with and without SCD.
Figure 1 Figure 1.

Disclosures
Isenberg: Vasculox: Membership on an entity's Board of Directors or advisory committees; Radiation Control Technologies: Membership on an entity's Board of Directors or advisory committees.
Leukemic Reticuloendotheliosis
Authors: BOURONCLE BERTHA A. , WISEMAN BRUCE K. , DOAN CHARLES A. 
Published: October 2019

Abstract
The clinical, hematologic, and pathologic findings in 26 cases of leukemic reticuloendotheliosis are presented.
A histopathologic correlation of the various forms of leukemia is suggested. Clinically the disease may follow an acute, subacute, or chronic course There is no idiopathic clinical sign or symptom, but the hematologic picture is pathognomonic and is characterized by the presence of reticulum cells in the peripheral blood and bone marrow. These cells are also present in the organs where the reticuloendothelial system predominates: spleen, liver, and lymph nodes.
The study of these cases supports the concept that leukemic reticuloendotheliosis is an independent hematologic and pathologic entity.
Paroxysmal Nocturnal Hemoglobinuria: The Role of Properdin in the Diagnostic Thrombin Test
Authors: CROSBY WILLIAM H. , BENJAMIN NAOMI R. 
Published: October 2019

Abstract
1. The "thrombin test" for PNH consists of adding thrombin to the standard acid-hemolysis test, thereby increasing hemolysis of PNH red cells. It has been suggested that the effect of the thrombin solution is not due to involvement of the coagulation system but to contamination of the thrombin by properdin, a globulin which is essential in the acid-hemolysis reaction of PNH.
2. In the present experiments the thrombin test was positive using preparations of thrombin which were free of properdin activity.
3. Thromboplastic materials devoid of properdin activity may be substituted for thrombin in the thrombin test.
4. It is concluded that the properdin which contaminates the commercially available thrombin preparations has little to do with the results of the thrombin test for PNH.