bluuebunny/crossref_metadata_2025_split huggingface dataset:
Restoring Cell Responsiveness to GM-CSF and IL-3 by PTEN Is Via Controlling the Regulation of Egr-1 In TF-1a Cell Line
Authors: Liu Y. Lucy , Malaviarachchi Priyangi , Emanuel Peter Dean 
Published: October 2019

Abstract
Abstract 2504
Cytokine independence is a common event in leukemia. The mechanism is often linked to deregulation of Ras/PI3K, Ras/MAP, and JAK/STAT pathways. PTEN, a major negative regulator of the Ras pathway, was initially isolated as a tumor suppressor in a variety of malignancies. We recently reported that PTEN is deficient in 67% of juvenile myelomonocytic leukemia (JMML) patients. Other groups have also found alterations of PTEN, PI3K, or AKT in 47.7% of T-cell acute lymphoblastic leukemia. Mice who are rendered pten-deficient develop myeloproliferative disorders and acute myeloid and lymphoid leukemias, and the leukemia-initiating stem cell in these mouse models could be distinguished from the normal hematopoietic stem cell by a differential response to rapamycin. The transcription factor, Egr-1, has been known to be a positive regulator of PTEN expression. It plays an important role in controlling proliferation as well as mobilization of hematopoietic stem cells. It is unknown whether PTEN had a feedback loop on regulating Egr-1 expression. Since poorly differentiated progenitors are often mobilized to the peripheral blood in leukemia patients, we hypothesized that PTEN deficiency alters the feedback loop of Egr-1 in response to cytokine stimulation. In this manner, PTEN deficiency might contribute to the cytokine independence observed in many types of leukemia. In order to evaluate this hypothesis we investigated the role of PTEN in cells responding to GM-CSF stimulation in the leukemia cell line, TF-1a. TF-1a is PTEN deficient as well as cytokine independent, and displays a constitutively high expression of Egr-1. We restored PTEN expression in TF-1a cells. This was accomplished by packaging the coding sequence of wild-type PTEN in a retrovirus vector, pBMN-GFP, that carried a gene encoding green fluorescent protein (GFP). TF-1a cells were transfected with pBMN-PTEN-GFP or pBMN-GFP by spinoculation. The pure population of transfected cells was sorted by FACS, followed by limiting dilution. The transfected single colonies were expanded for further evaluation of PTEN expression and biological function. Data from these experiments showed: 1) over-expression PTEN in TF-1a down-regulated the expression of Egr-1 and restored the responsiveness of Egr-1 to GM-CSF stimulation, 2) PTEN restored the responsiveness of TF-1a to GM-CSF and IL-3, and 3) these events are CREB-independent. In conclusion, our data demonstrates that PTEN contributes to the cell responsiveness to GM-CSF and IL-3, and has a negative feedback role to regulate Egr-1 expression in TF-1a cells. In addition to Egr-1 regulating PTEN in a positive manner, our data indicates the presence of a feedback loop from PTEN back to Egr-1. This appears to be an event independent of CREB.

Disclosures:
No relevant conflicts of interest to declare.
4-1BBL-Expressing aAPCs Attenuate IL-15-Induced NK Cell Expansion and Cytokine Production In Vitro but Induce NK Cell-Mediated Gvhd In Vivo
Authors: Capitini Christian M. , Meadors Joanna L. , Cho Monica M. , Orentas Rimas J. , Mackall Crystal L. , Fry Terry J. 
Published: October 2019

Abstract
Abstract 2536
Methods to expand natural killer (NK) cells ex vivo for adoptive cell therapy are being explored to improve outcomes after allogeneic blood and marrow transplant (alloBMT). Artificial antigen presenting cells (aAPCs) can present cytokines and/or co-stimulatory molecules that can potentially improve expansion and activity. 4-1BBL (CD137L) has demonstrated mixed results on murine and human NK cells, but the impact on murine NK cell biology after alloBMT has not been explored. NK cells were harvested from either C57BL/6 (B6) or CB6F1 spleens and cultured ex vivo with a recombinant interleukin (IL)-15/IL-15 receptor alpha (Ra) complex in the presence or absence of a CD137L+ aAPC. Because IL-15 is typically presented in trans by IL-15Ra, the complex was utilized to potently increase agonist bioactivity. NK cells cultured with IL-15/IL-15Ra alone showed a peak of 20-fold expansion, but this expansion was decreased with the addition of CD137L+ aAPCs if the ratio of aAPC to NK cells was greater than 1:1. In the presence of IL-15/IL-15Ra, the impact of CD137L+ aAPCs on expression of the inhibitory receptors, Ly49C+I and activating receptor Ly49H was variable and strain dependent, with increased expression in B6 NK cells, but decreased expression in CB6F1 NK cells. The expression of major histocompatibility complex (MHC) class I was not affected in NK cells from either strain by the presence of CD137L+ aAPCs. The production of gamma interferon and tumor necrosis factor-a was robust in NK cells expanded by IL-15/IL-15Ra alone, but attenuated with the addition of CD137L+ aAPCs. Animal experiments showed that administration of NK cells expanded ex vivo with IL-15/IL-15Ra alone was well tolerated after T cell depleted MHC-mismatched alloBMT (CB6F1–>B6), but surprisingly the addition of CD137L+ aAPCs to cultures caused NK cells to induce GVHD-associated weight loss. In summary, IL-15/IL-15Ra expanded murine NK cells demonstrate increased cytokine production and do not cause toxicity when infused after alloBMT. The presence of CD137L+ aAPCs attenuated cytokine production and increased Ly49 receptor expression in NK cells from B6 mice. Remarkably, NK cells expanded by IL-15/IL-15Ra in the presence of CD137L+ aAPCs demonstrate increased propensity to cause GVHD. Ongoing studies are exploring the anti-tumor efficacy of IL-15/IL-15Ra expanded murine NK cells cultured in the presence and absence of CD137L.

Disclosures:
No relevant conflicts of interest to declare.
The Efficacy and Safety of Recombinant Human Thrombopoietin in Patients with Hematological Malgnancies After Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Miao Miao , Depei Wu , Sun Aining , Wang Ying , Yan Lingzhi , Wu Qian 
Published: October 2019

Abstract
Abstract 4565

OBJECTIVE:
To evaluate the efficacy and sefety of recombinant human thrombopoietin(rhTPO) prior to engraftment in adults with hematological malignancie who received allogeneic haematopoietil stem cell transplantation(Allo-HSCT).

METHODS:
This sutdy was a randomized, controlled clinical trial,38 patients were hematological malignancie, inclulding acute and chrinic myeloid leukemia, acute lymphoblastic leukemia, lymphoma.They received Allo-HSCT and were randomly divided into groups(group A 19 cases, group B 19 cases).The group A was no-rhTPO as control, the group B were received rhTPO 15000U/d from +1 day, and continued until the untransfused platelet count was >70×109/L for two consecutived days. Patients received platelete transfusion when they developed severe thrombocytopenia<20×109/L. Efficacy and sefety of rhTPO on the outcome of Allo-HSCT were evaluated.

RESULTS:
In both group A and B, platelet decrease after Allo-HSCT had no sognificant difference. The platelet engraftment duration of group A and B was 15.68±1.36(range 11–31) days and 13.47±0.72(range 9–21) days, respectively. The amount of platelet transfusion of group A and B was 4±0.55(range 20–130) units and 2.89±0.36(range 0–50) units, respectively. The effects of rhTPO on neutrophil engraftment, hepatic function, renal function, alloergic reations and acute GVHD were not found.

CONCLUSION:
The platelet engraftment duration of group B was shorter than that of group A(t=27.2, p<0.001), the amount of need platelet transfusion was significently less than those in the group A.There was a statistically significant difference in platelet engraftment and platelet transfusion needed(t=2.523, p<0.05). Administration of rhTPO prior to platelet engraftment after Allo-HSCT seem to be efficacy and safe.

Disclosures:
No relevant conflicts of interest to declare.
Immune Tolerance Therapy in Children with FVIII Inhibitors
Authors: Vdovin Vladimir , Svirin Pavel , Shiller Ekaterina , Schwartz Bruce A. 
Published: October 2019

Abstract
Abstract 4656
In up to 40% of cases, Hemophilia A treatment is complicated by the development of FVIII inhibitors, rendering the patient resistant to replacement therapy and increasing the risk of unmanageable bleeding, particularly arthropathy and disability. Presence of an inhibitor dramatically increases overall treatment costs 3–5 fold. Bleeding episodes in inhibitor patients require hospitalisation and bypassing agents such as rFVIIa. Immune tolerance (ITI) therapy, involving repetitive and persistent treatment with FVIII replacement is the only proven strategy for inhibitor eradication.
Since 2006, 30 children with inhibitors and poor-prognosis for ITI success have been treated with octanate® for ITI at seven Russian haemophilia treatment centers in the frame of the ongoing Observational Immune Tolerance Induction (ObsITI) study. The median age was 5 years.
The objective of this study was to evaluate the progress of prospective ITI with octanate® in children with inhibitors and a poor-prognosis for ITI success. In addition, joint status using a new musculoskeletal assessment tool before, during or after ITI was evaluated. A cost-of-care analysis to assess the consumption of octanate® in ITI in comparison with rFVIIa and to quantify the average direct medical costs for the treatment of young patients with FVIII inhibitors was also performed.
Despite the stringent success criteria, octanate®, along with the high-dose Bonn protocol, has led to a complete success rate of about 80% in children with inhibitors and poor prognostic factor for ITI success.
Successful tolerisation with octanate® allows resumption of fully effective replacement treatment with a consequent improvement in the patients’ musculoskeletal outcome and estimated savings over 5 years of € 600000. In addition, breakpoint analyses estimated that the number of years after which ITI with octanate® pays off compared with on-demand treatment with rFVIIa is 1.4 years.
In summary, octanate® is safe, well tolerated and effective in the induction of immune tolerance, even in poor-prognosis patients. The comparative cost-of-care analysis will help to optimize the therapy for the inhibitor patient and to ensure an effective allocation of financial resources.

Disclosures:
Schwartz: Octapharma: Employment. Off Label Use: This product Octanate is licensed in a number of countries world wide but not yet licensed in the US.
Frequency of Bleeding Disorders in Women Presenting Menorrhagia in the North of Iran
Authors: Janbabaei Ghasem , Borhani Samaneh , Rashidi Masoumeh , Farazmandfar Touraj , Shekarriz Ramin , Khademloo Mohammad , Ghavamzadeh Ardeshir 
Published: October 2019

Abstract
Abstract 4660

Objectives:
Menorrhagia is a common presentation of bleeding disorders, especially VWD in women. We decided to determine the frequency of these disorders in women with menstruation problems.

Materials and Methods:
208 patients in reproductive age with menorrhagia were investigated for bleeding disorders in two steps. Step one includes CBC, PT, PTT and BT, which performed for all patients, and for patients who had an abnormality in step one, step two was considered. Step two includes VWAg, RCO, factors level, RIPA and platelet aggregometry.

Results:
Among 208 patients who investigated for bleeding disorders 53 patients (25%) had abnormalities in coagulation tests or platelet count. In our survey frequencies of bleeding disorder was VWD=14(6.73%), thrombocytopenia=13(6.25%), deficiency of factor II= 2(0.96%), factor V=1(0.48%), factor VII=3(1.44%), factor VIII=2(0.96%), Factor XI =4(1.92%), factor XII=4(1.92%), Bernard Soulier=2(0.96%). Furthermore, we found 18 patients (8.65%) who had abnormal PT, PTT or BT with no definite diagnosis.

Conclusion:
In this study, the most common bleeding disorder was VWD and thrombocytopenia ranked second disorders. Although other bleeding disorders are rare, in our study a number of them were found. So, we recomment the above coagulation test for women with menorrhagia.

Disclosures:
No relevant conflicts of interest to declare.
Plasma VEGF Levels and Etiology of Thrombocytopenia, a Biomarker Study
Authors: Xiao Ying-Yi , Liu Yang , Zeng Thanthanaye , Khiani Komal , Burton Jack , Huang Yiwu , Xu Yiqing 
Published: October 2019

Abstract
Abstract 4684

Background:
Thrombocytopenia is a major medical problem associated with bleeding. Etiology of thrombocytopenia includes myelosuppression, splenic sequestration and increased peripheral consumption and/or destruction. Vascular Endothelial Growth Factor (VEGF) is produced in both endothelial cells and platelets and is stored in alpha granules in the platelets. It plays multiple roles in the regulation of vascular endothelial cell proliferation and vascular integrity.

Hypothesis:
VEGF release from the platelets in vivo due to platelet activation or destruction may result in changes in plasma and serum VEGF levels, and the level could be associated with various etiologies of thrombocytopenia.

Methods:
This was a prospective IRB approved on-going study. Eligibility includes platelet count less than 120,000/ul, no current diagnoses of solid tumor and thrombocytopenia is not due to chemotherapy. Blood was collected at the time of diagnosis, and VEGF was measured in serum and plasma specimens using a commercial ELISA kit from Ray Biotech (Norcross, GA). Plasma adjusted VEGF levels are defined as plasma VEGF value divided by the platelet count.

Results:
Forty seven patients were enrolled, the clinical diagnoses included AML or MDS (designated as A/M, n=6), hypersplenism (n=13), ITP (n=14), sepsis (n=11) and confirmed TTP (n=3). Blood was also collected from 4 healthy volunteers as controls. Plasma VEGF levels and plasma adjusted VEGF levels (defined as plasma VEGF value divided by the number of platelets) were selected as the main measurements of outcome as shown below

Conclusion:
Plasma VEGF adjusted with platelet count should be the measurement for in vivo release of VEGF from endothelial or platelet sources. Plasma adjusted VEGF level has a wide range of variation in patients with thrombocytopenia, about 50% of our test subjects had undetectable VEGF plasma levels. Although not statistically significant, patients with thrombocytopenia due to AML or MDS had lowest plasma VEGF values and least variation, while patients with sepsis had highest plasma VEGF readings and variations, probably due to increased endothelial cell production. The study is ongoing in large scales, and we are also interested in following the trend of serial plasma adjusted VEGF levels along with platelet recovery.

Disclosures:
No relevant conflicts of interest to declare.
Neutropenia in Glycogen Storage Disease 1b (GSD1b)
Authors: Dale David C. , Bolyard Audrey Anna , Marrero Tracy M. , Phan Lan , Boxer Laurence A. , Kishnani Priya S. , Kurtzberg Joanne , Weinstein David A. 
Published: October 2019

Abstract
Abstract 4791
Neutropenia and recurrent infections are common manifestations of glycogen storage disease 1b (GSD1b), a disease caused by mutations in G6PT1, the glucose 6 phosphate transporter 1 gene. Patients with GSD1b have complex defects in neutrophil deployment and function. GSD1b causes recurrent infections – gingivitis, mouth ulcers, respiratory infections, abscesses and enterocolitis. Granulocyte colony-stimulating factor (G-CSF) has been used to treat these patients for more than 2 decades.
We reviewed the presentations, manifestations, complications and G-CSF treatment for 54 GSD1b patients (31 adults and 23 children) at Duke University, the University of Florida and through the Severe Chronic Neutropenia International Registry at the University of Washington. Observational data were available on most patients for at least 3 years (mean = 12 years, median = 13 years, range = 1 to 23 years). Available data showed that before treatment with G-CSF, all of these patients had mild to moderate neutropenia; the mean of the median absolute neutrophil count (ANC) was 0.839 × 109/L ± 0.165 SEM (median = 0.503 × 109/L, range = 0.038 to 7.665 × 109/L). All of the patients also had recurrent infections; the most common events were mouth ulcers, otitis and skin abscesses. There were 7 patients who had enterocolitis prior to G-CSF treatment.
G-CSF therapy was initiated at various ages ranging from birth to age 30 (mean = 6 years, median = 4 years). Initial daily doses of G-CSF ranged from 0.50 to 63.28 mcg/kg/day (mean = 5.86, median = 3.65) and were generally titrated upward to achieve a normal neutrophil count. The mean ANC on treatment was 2.598 × 109/L ± 0.136 SEM (median = 1.856 × 109/L, range = 0.0 to 12.645 × 109/L). G-CSF treatment was uniformly associated with an increase in blood neutrophils and reduced patterns of infection. In most patients the symptoms of enterocolitis improved within a few days to a few weeks after starting G-CSF therapy. An increase in spleen size also commonly occurred. In 630 patient-years of observation, there have been 3 deaths attributable to the following causes: 1 subject died of sepsis, 1 subject died of complications from hepatomegaly and neutropenia, and 1 subject died 6 years after a liver transplant. The subject developed AML 5 years after the liver transplant while receiving a combination of immunosuppressant drugs.
Patients with GSD1b associated neutropenia respond well to G-CSF with reduced infections and symptoms of enterocolitis. Long-term complications are infrequent and G-CSF is well tolerated in most patients.

Disclosures:
Dale: Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding. Boxer:Amgen: Equity Ownership.
Autoimmune Hemolytic Anemia
Authors: CROSBY WILLIAM H. , RAPPAPORT HENRY 
Published: October 2019

Abstract
1. The records of 57 patients with autoimmune hemolytic anemia have been reviewed to learn if the hematologic data were of any prognostic importance. It appeared that higher mortality was associated with severe anemia, reticulocytopenia, thrombocytopenia and leukopenia. The presence of spherocytosis and erythroblastosis seemed to have no prognostic significance with regard to mortality.
2. Thrombocytopenia in several patients was associated with thromboembolic disease.
3. There was a definite seasonal incidence of hemolytic crises in AHA, more crises occurring during the winter months.
Syndrome of Hypogammaglobulinemia, Splenomegaly and Hypersplenism
Authors: PRASAD ANANDA S. , REINER E. , WATSON C. J. 
Published: October 2019

Abstract
Two cases of acquired idiopathic hypogammaglobulinemia associated with splenomegaly and hemolytic anemia due to hypersplenism have been reported. Splenomegaly and hypersplenism appears to be the result of reticulum cell hyperplasia caused by the lack of gamma globulin and resultant repeated infections. Both the cases had splenectomy with marked hematologic improvement.
A review of the literature indicates that this syndrome is relatively common, in comparison to the incidence of total number of acquired hypogammaglobulinemia that has been reported in the literature so far. It also seems very important to recognize it because of the obvious therapeutic implications. Serum gamma globulin level should be determined in cases with unexplained hepatosplenomegaly, hypersplenism and hemolytic anemia.
The Hemolytic Effect of Ionizing Radiations and Its Relationship to the Hemorrhagic Phase of Radiation Injury
Authors: STOHLMAN FREDERICK , BRECHER GEORGE , SCHNEIDERMAN MARVIN , CRONKITE EUGENE P. 
Published: October 2019

Abstract
Post-irradiation anemia is primarily the consequence of red cell aplasia and of hemorrhage secondary to thrombocytopenia. Data have been presented which indicate that in addition there is intravascular red cell damage. Of particular interest was the observation herein presented that the damage to the red cells produced by ionizing radiations is of an indirect nature. Using the chromium technic it was possible to demonstrate red cell damage as early as the 1st-3rd post-irradiation day. In these studies tagged cells were transfused 15-55 days prior to irradiation in order to establish the elution rate of Cr51. On the 1st-3rd day following irradiation there was an abrupt increase in the rate of Cr51 loss. When normal cells were transfused immediately after irradiation there was also a striking increase in the rate of Cr51 loss above that observed in normal animals, indicating that radiation damage to the red cell was indirect. When cells were transfused from irradiated donors into normal recipients, it was not possible to demonstrate red cell damage prior to the 10th post-irradiation day, indicating that the damage was progressive.
Since there was a selective loss of Cr51 it is suggested that chromium produced an additive damage to the cell. The possible implication of this observation in the use and interpretation of Cr51 survival curves is discussed.
It was also determined that red cells which were recirculated following extravasation during the thrombocytopenic phase of radiation injury had a shortened survival, whereas normal cells, tagged and injected intramuscularly into normal recipients survived normally on return to the general circulation. Cells collected from dogs one day after irradiation, tagged with Cr51 and then injected intramuscularly into normal recipients, showed a striking increase in the rate of destruction. Cells collected on the 4th post-irradiation day were destroyed even more rapidly. These studies confirmed the early onset of radiation injury to the cells, its progressive nature, and demonstrated the damage incurred as a result of passage through the extravascular cycle. As might be expected there was also a shortened survival of red cells collected from the thoracic lymph of thrombopenic irradiated animals.
Conclusions: 1. Ionizing radiations produce intravascular red cell damage in addition to the known loss incident to hemorrhage. This damage is evident within 24-72 hrs. of irradiation.
2. The red cell damage is an indirect and progressive effect of irradiation.
3. Those cells which are returned to the general circulation, following passage through an extravascular cycle, are damaged. The extent of injury to normal cells is such that it is not detectable by current methods. However, the addition of a second injury, such as that following irradiation, enables the detection of this damage.
4. Chromium "elution" is altered by irradiation injury.
Preservation of Radiation Recovery Factor in Frozen Marrow
Authors: FERREBEE JOSEPH W. , BILLEN DANIEL , URSO IRENE M. , LU WAN CHING , THOMAS E. DONNALL , CONGDON CHARLES C. 
Published: October 2019

Abstract
Injection of mouse marrow preserved in glycerol at -80 C. for six weeks has been shown to be quantitatively similar to injections of fresh marrow in inducing recovery in mice after lethal radiation. The effect of the preservation procedure on subsequent rates of DNA synthesis has been determined in vitro for both preserved human marrow and preserved mouse marrow.
Since DNA synthesis is a satisfactory measure of rates of cell multiplication and since preservation is shown to affect DNA synthesis in human marrow to a lesser degree than in mouse marrow, it may be concluded that the procedure used (-80 C. plus 15% glycerol) is satisfactory for the maintenance of viability in human marrow.