bluuebunny/crossref_metadata_2025_split huggingface dataset:
Physicochemical Properties of Circulating Red Blood Cells of Lethally X-irradiated Mice Treated with Rat Bone Marrow
Authors: MAKINODAN TAKASHI , ANDERSON NORMAN G. 
Published: October 2019

Abstract
1. Two months after injection of rat bone marrow into lethally X-irradiated mice (950 r-RBM mice), 100% of the circulating RBC were serologically of the rat type, indicating that the surface molecular configuration of RBC from these experimental mice are of the rat type.
2. The hemoglobin was found to be also very much like the rat type in its ease in crystallization, its alkali denaturation property, its electrophoretic property, and its tendency to form a paracrystalline state at low temperature.
3. These cells possessed dual osmotic properties; the relative hemoglobin concentration released when cells were lysed in water was more comparable to the rat type, but its temperature dependency was more comparable to the mouse type.
Desoxyribonucleic Acid Synthesis by Bone Marrow Cells in Vitro
Authors: THOMAS E. DONNALL , LOCHTE HARRY L. 
Published: October 2019

Abstract
The incorporation of C14-formate into thymine is described as a method for measuring desoxyribonucleic acid synthesis by bone marrow cells in vitro. The method is satisfactorily reproducible and permits measurement of DNA synthesis over short periods of time with small amounts of bone marrow.
Normal serum was found to be essential for rapid rates of DNA synthesis and could not be replaced by supplemented media. The rate of DNA synthesis was almost constant during the first 4-6 hours of incubation, but fell off rapidly thereafter. The ability to promote DNA synthesis was found to differ from one normal serum to another. The optimal pH for DNA synthesis was approximately 7.8.
Because DNA synthesis is vitally concerned with cell duplication, measurement of the rate of its synthesis should be a valuable tool for measuring viability of those marrow cells young enough to proliferate.
The Mode of Action and Nature of a Heat Stable Plasma Erythropoietic Factor
Authors: RAMBACH WALTER A. , ALT HOWARD L. , COOPER JOHN A. D. 
Published: October 2019

Abstract
Boiled filtrates of "anemic" rabbit plasma have been studied by physicochemical means, and the response to their injection into the rat has been followed utilizing tracer and morphologic technics. It has been demonstrated that the erythropoietic activity of these filtrates is associated with an alpha-2 globulin, most probably a mucoprotein; and that this material produces, in the rat, an increase in the rate of red cell division and maturation, and an acceleration of release of these cells from the bone marrow and accessory organs of hemopoiesis.
The Pseudo-L.E. Cell Phenomenon, with Report of a Case
Authors: KURNICK N. B. 
Published: October 2019

Abstract
An example of leuko-phagocytosis secondary to metastic bronchogenic carcinoma is presented. The transitory development of leukoagglutination and leukophagocytosis (pseudo-L.E. cell phenomenon) led to the erroneous diagnosis of systemic lupus erythematosus. The differentiation between the L.E. cell phenomenon and the pseudo-phenomenon is discussed.
Electron Microscopy of Formed Elements of Normal Human Blood
Authors: GOODMAN JOSEPH R. , REILLY EMMETT B. , MOORE ROGER E. 
Published: October 2019

Abstract
The history of the development of practicable electron microscope technics for examination of the formed elements of the blood was reviewed. A method found to yield good reproductions of ultrathin sections of blood cells was presented in considerable detail. The various phases of the technic were critically evaluated.
The morphologic characteristics of the various formed elements of normal peripheral blood were defined in association with sample electron micrographs. Ultramicroscopic detail of neutrophils, eosinophils, basophils, lymphocytes, monocytes, platelets and erythrocytes was discussed.
The Separate Existence of the Pituitary Erythropoietic Hormone
Authors: VAN DYKE DONALD C. , SIMPSON M. E. , CONTOPOULOS A. N. , EVANS H. M. 
Published: October 2019

Abstract
Although it is recognized that a number of factors are involved in the mechanism controlling red blood cell production, the anterior pituitary furnishes one factor of importance. This pituitary factor is apparently a hormone distinct from other trophic hormones. It is allied chemically only to ACTH and yet both biological and chemical evidence indicates that the pituitary erythropoietic hormone and ACTH are not the same. The major problem in further elucidating the role of the pituitary in the control of erythropoiesis resolves itself into developing a satisfactory method of preparation of erythropoietic hormone which will separate it from ACTH. It is true that no ACTH preparation has been made which does not possess some erythropoietic activity, and no pituitary erythropoietic hormone has been prepared which is entirely devoid of ACTH. Nevertheless, the proportion of these two activities in different preparations has varied so greatly as to assure their separate existence. Autolytic digestion consistently reduces the ACTH activity, while erythropoietic activity is retained. It has been found that oxycellulose is a better adsorbent of ACTH than of erythropoietic hormone. These observations add chemical evidence to the accumulated biological evidence establishing the separate existence of these two pituitary principles.
TYK2-STAT1 Pathway Positively Regulates BCL2 Gene Expression in T-Cell Acute Lymphoblastic Leukemia
Authors: Sanda Takaomi , Tyner Jeffrey W , Gutierrez Alejandro , Ngo Vu N , Glover Jason M , Chang Bill H , Yost Arla J , Weng Andrew P , Ma Wenxue , Tatarek Jessica , Fleischman Angela G. , Ahn Yebin , Yang Yandan , Zhou Wenjun , Neuberg Donna S. , Moriggl Richard , Müller Mathias , Kelliher Michelle , Jamieson Catriona , Gray Nathanael S , Staudt Louis M. , Druker Brian J. , Look Thomas 
Published: October 2019

Abstract
Abstract 1470
To discover oncogenic pathways that are characteristically deregulated in T-cell acute lymphoblastic leukemia (T-ALL), we performed RNA interference screens both in T-ALL cell lines and primary specimens. We found that the JAK tyrosine kinase family member, TYK2, and its downstream effector, STAT1, are each required for the survival of T-ALL cells. To identify the effector molecules downstream of the TYK2-STAT1 pathway in T-ALL, we analyzed global gene expression profiles in TYK2-dependent T-ALL cell lines after silencing of TYK2 or STAT1. As expected, gene set enrichment analysis revealed that genes downregulated by TYK2 knockdown were generally also downregulated by knockdown of STAT1. Importantly, we found that expression of the anti-apoptotic gene BCL2 was significantly downregulated after silencing of both TYK2 and STAT1. Analysis by quantitative PCR of additional T-ALL cell lines revealed that silencing of TYK2 resulted in significant reductions of BCL2 mRNA expression in multiple TYK2-dependent cell lines. Expression of the wild-type but not the kinase-dead TYK2 protein was sufficient to rescue BCL2 protein expression and to prevent apoptosis after knockdown of endogenous TYK2, indicating that the tyrosine kinase activity of TYK2 is required for BCL2 upregulation. Similarly, expression of the shRNA-resistant wild-type STAT1A protein partially rescued BCL2 protein expression and prevented apoptosis, while a variant of STAT1A (Y701F) that is incapable of becoming phosphorylated on a requisite tyrosine residue did not rescue BCL2 levels. Taken together, our findings indicate that aberrant activation of a TYK2-STAT1 pathway upregulates BCL2 expression in T-ALL cells, and that the T-ALL cells develop pathway dependence, in that they require these sustained high levels BCL2 expression for survival.

Disclosures:
No relevant conflicts of interest to declare.
NCI Common Toxicity Criteria and Mortality After Chemotherapy for Acute Myeloid Leukemia (AML)
Authors: Veatch Joshua R. , Sandhu Ravinder K , Shannon-Dorcy Kathleen , Pagel John M. , Becker Pamela S. , Petersdorf Stephen , Appelbaum Frederick R. , Estey Elihu H. 
Published: October 2019

Abstract
Abstract 1479
Abnormal values for creatinine, bilirubin, AST, and ALT following chemotherapy, in particular in phase 1 trials, are interpreted using NCI common toxicity criteria. By convention, grade 3 or 4 toxicity leads to dose reduction while grade 1 or 2 toxicity does not. However, grade 3 – 4 toxicity is typically asymptomatic, calling into question the relevance of these criteria. In an attempt to provide a more empirical basis for decisions we analyzed the relation between (a) grades of NCI toxicity for bilirubin, creatinine, ALT, and AST following chemotherapy for AML and (b) death. Specifically we examined 752 courses of chemotherapy in 533 patients delivered at the University of Washington or Fred Hutchinson Cancer Research Center for newly- diagnosed or relapsed/refractory AML between 2002 and 2012. Patients had at least 2 sets of renal and liver function test values but were excluded if either pretreatment bilirubin or creatinine was greater than 2.0 to better approximate a population in an early phase trial. We compared initial and maximal lab values in the first 28 days in the 40 courses that resulted in death before 28 days, and in the 712 courses that did not (6% death rate; the death rate per patient was 7%). Maximum lab values for creatinine were strongly related to mortality, with grade O having a 1.0% mortality rate and with each increasing grade having a statistically significant increase in mortality over the immediately preceding level: grade I: 5%, grade II: 22%, grade III or IV: 50%. Bilirubin had a significant increase in mortality at grade II (10.0%, p=.009 relative to grade O) and grade III-IV (15.4%, p=.0007 relative to grade O). AST was also associated with increased mortality at grade II (9.5%, p=.04 relative to grade O) and grade III-IV (15.6%, p =.0006 relative to grade O). ALT was not correlated to mortality, even at grade III-IV (9.5% versus 5.3% at grade 0, p=0.23). Although these correlations obviously do not imply causation, it might be argued that grade II toxicity for creatinine, associated with a 4-fold increase in mortality, should result in dose reduction. In contrast, it might be argued that dose reduction should not occur for grade III/IV ALT toxicity, which correlated poorly with mortality. In any event our data suggest that if the principal purpose of toxicity grading is to guide subsequent dose reduction to prevent death, the NCI toxicity criteria should be revisited.

Disclosures:
No relevant conflicts of interest to declare.
CNS Relapse in Acute Promyelocytic Leukemia: Incidence, Management and Outcome
Authors: Kohrt Holbrook E , Coutre Steven 
Published: October 2019

Abstract
Abstract 1489

Background:
A standardized approach to the diagnosis, prognosis and therapy of patients with APL and CNS involvement is limited by the rare nature of the complication. We report the incidence, risk factors, management and outcome of APL with CNS involvement at Stanford University and review the published cases in the literature during the prior three decades.

Methods:
Between 1983 and Jan 2012, 7 cases of APL with CNS involvement were identified at Stanford University and 65 in the literature during the same interval. All pts in the Stanford series received ATRA and anthracycline containing induction regimens followed by consolidation with anthracycline and cytarabine (7 pts), plus ATRA (5 pts), ATO (1 pt), or IT prophylaxis (1 pt). Five pts received maintenance with ATRA containing regimens in all cases (2 pts relapsed prior to starting maintenance). Median follow-up was 6 years.

Results:
Seventy-two relapses involving the CNS were seen after 1 to 50 months (median 8 months) and 61 (84.7%) occurred in CR1 with 8 (11.1%) in CR2, 2 (2.8%) in CR3 and 1 (1.4%) in CR4. Forty-seven (66.2%) were accompanied by bone marrow involvement, 14 (29.8%) molecular and 33 (70.2%) hematological. Median age was 35y with 9 pediatric pts (range 1.3 to 20y) and 63 adult pts (range 22 to 69y). Median WBC count was 28,100 cells/mm3 (range 1,100 to 162,000 cells/mm3) with 61.4% >10 and 6.8% >100 cells/mm3. PML/RARα isoform was reported in 29 pts with 37.9% bcr1, 3.4% bcr2, and 58.6% bcr3. Five pts presented with concurrent CNS-localized hemorrhage. Therapy included methotrexate (88.9%), cytarabine (80.6%), arsenic trioxide (ATO, 47.2%), and anthracycline (11.1%) containing regimens with 88.9% receiving intrathecal therapy. Eighty-six percent received cranial radiation with dose ranging 18 to 30 Gy. Autologous or allogeneic transplantation followed salvage therapy in 30.8% and 5.6% of pts, respectively. Fifty-four (75.0%) pts were salvaged successfully, including 88.2% of pts receiving ATO, however 18 (33.3%) pts later relapsed, 4 (22.2%) of which were salvaged with additional treatment.

Conclusion:
CNS relapse is a rare complications of APL with the majority of cases occurring in pts with WBC >10 cells/mm3, and in first CR. CNS relapse was frequently associated with concurrent bone marrow relapse and approximately one-third occurred with only molecular involvement. A preventative role of IT prophylaxis could not be determined due to only 1 pt receiving prophylaxis. A high percentage of pts salvaged with ATO achieved CR, supporting its therapeutic efficacy and ability to cross the blood brain barrier. Incorporation of ATO in first line therapy for APL may reduce the risk of CNS relapse.

Disclosures:
No relevant conflicts of interest to declare.
Reactivating PP2A by FTY720-Loaded Lipid Nanoparticles, a Novel Therapeutic Strategy in Acute Myeloid Leukemia
Authors: de Mendoza Ander Estella-Hermoso , Castello-Cros Remedios , Cirauqui Cristina , Pippa Raffaella , Moreno Isabel , Blanco-Prieto Maria J , Odero Maria D 
Published: October 2019

Abstract
Abstract 1511
Protein phosphatase 2A (PP2A) is a human tumor suppressor that inhibits cellular transformation by regulating the activity of several signaling proteins critical for malignant cell behavior. Our group has shown that PP2A activity is reduced in acute myeloid leukemia (AML), and that restoration of PP2A activity in AML cells blocks proliferation, induces caspase-dependent apoptosis, and affects AKT and ERK1/2 activity. It has been proposed that FTY720 suppress PP2A activity by directly interacting with PP2AA-C dimers, and possibly interfering with the SET-PP2A interplay. Reactivation of PP2A by FTY720 in vivo restricts leukemogenesis in different types of hematological malignancies; however, its antitumor effects in AML remain underexplored. Here, we developed lipid nanoparticles (LN) encapsulating FTY720, and evaluated its pharmacokinetic profile and biodistribution in mice, and its antileukemic effects in AML in vitro and in vivo. FTY720-LN show suitable size (around 210 nm) and encapsulation efficiency (nearly 75%) for the oral and intravenous administration of the compound. Blood concentration of FTY720 decreased rapidly 2 hours after the intravenous administration of FTY720-LN; however, orally administered LN provided both a sustained release of the drug over four days and a high distribution in tissues, significantly increasing the accumulation of FTY720 in spleen, compared to the administration of the same dose of FTY720 solution and intravenous administration of FTY720-LN. In vitro studies revealed that FTY720 and FTY720-LN restore PP2A phosphatase activity in AML cells, leading to cell growth arrest, induction of caspase-dependent apoptosis and decrease in phosphorylation of PP2A targets AKT and ERK. Moreover, in vivo daily intraperitoneal administration of FTY720 in RAG2−/− γc−/− mice bearing an AML xenograft tumor reduced significant tumor growth. Of note, oral administration of FTY720-LN every three days was as efficient in reducing AML xenograft tumor growth as a daily oral administration of FTY720. These results provide the first evidence for the potential use of lipid nanoparticles encapsulating FTY720 as a therapeutic agent in AML.

Disclosures:
No relevant conflicts of interest to declare.
Functional Characterization of HLA SNPs Associated with Risk of Follicular Lymphoma
Authors: Skibola Christine F. , Conde Lucia , Smith Martyn T. , Sille Fenna C.M. 
Published: October 2019

Abstract
Abstract 1551

Background:
Genome-wide association studies (GWAS) suggest an important role of genetic variation in the etiology of follicular lymphoma (FL). Multiple susceptibility alleles in the human leukocyte antigen (HLA) Class I and II regions on chromosome 6p21.33–32 have been identified for FL, including rs10484561 (OR = 1.95, P = 1.12×10−29) and rs2647012 SNP (OR = 0.64, P = 2×10−21) (Conde et al., Nat Gen, 2010; Smedby et al. Plos Gen, 2011). Moreover, high-resolution HLA typing using next-generation sequencing has identified a number of HLA alleles associated with FL including the extended haplotype DRB1*01:01—DQA1*01:01—DQB1*05:01 that is in complete linkage disequilibrium with rs10484561 (D'=1), and DRB1*15-DQA1*01-DQB1*06, which is highly correlated with rs2647012 (D'=1) (Skibola et al., Tissue Antigens, 2012). We hypothesized that these SNPs or protein variants may alter immune responses to specific antigens and influence tumor development through effects on gene expression or protein structure. To gain a further understanding of the mechanisms involved, here we explored the effects of HLA alleles on expression at the transcriptional and protein level.

Methods:
HLA-DRB1, HLA-DQB1 and HLA-DQA1 mRNA expression and protein levels were measured by RT-qPCR and flow cytometry, respectively, in 40 lymphoblastoid cell lines (Coriell) where wild type and variant rs10484561 and rs2647012 genotypes were compared.

Results:
HLA-DQB1 gene expression changes were significantly associated with rs2647012 genotypes, where the presence of the variant (protective) allele correlated with higher HLA-DQB1 transcript levels. Flow cytometry with HLA-specific antibodies and pan-HLA antibodies (against DR/DQ/DP) also confirmed that the variant rs2647012 allele was associated with higher HLA-DQB1 protein levels. No significant changes were observed in HLA-DRB1, HLA-DQB1 or HLA-DQA1 transcript or protein levels when comparing rs10484561 wild type versus variant alleles.

Conclusions:
We did not detect significant differences in transcript or surface protein levels based on rs10484561 genotypes. These findings suggest that the increased risk of FL associated with rs10484561 that is in high LD with the HLA-DRB1*0101—DQA1*0101—DQB1*0501 extended haplotype is not due to transcriptional modulation, but may be due to effects on protein structure. On the other hand, the protective rs2647012 variant was associated with higher HLA-DQB1 mRNA and protein expression, suggesting that rs2647012 or a SNP in LD influences FL risk through effects on gene expression. We are currently testing the influence of additional FL risk alleles in the 6p21.3 region on HLA gene and protein expression, as well as investigating whether FL risk alleles alter HLA expression in other cell types.

Disclosures:
No relevant conflicts of interest to declare.
Prognositc Inpact of Tumor-Infiltrating FOXP3 + Regulatory T Cells in DLBCL Treated with R-CHOP
Authors: Goto Naoe , Tsurumi Hisashi , Shibata Yuhei , Mabuchi Ryoko , Nakamura Nobuhiko , Nakamura Hiroshi , Kitagawa Junichi , Kanemura Nobuhiro , Hara Takeshi , Moriwaki Hisataka 
Published: October 2019

Abstract
Abstract 1555

Background:
Tumor-infiltrating immune cells perform important functions in host immune reaction against tumor cells including diffuse large B cell lymphoma (DLBCL). Recently, variable tumor-infiltrating cells were reported to give a influence for prognosis, for example, regulatory T cell (T reg), cytotoxic T cell, macrophage and mast cell etc. Among these microenvironmental cells, we focused on Treg cell, and we assessed the distribution and prognostic significance of these cells in DLBCL. The forkhead/winged helix transcription factor 3 (FOXP3) is a transcriptional factorshown to be the key control gene in the development and function of Tregs both in mice and humans.

Patients and Methods:
We examined samples from 94 patients (54 men and 40 women; median age, 70 years) at diagnosis who were prospectively enrolled between 2002 and 2008. All patients treated with R-CHOP(rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone). The pattern of FOXP3 protein expression was evaluated using immunehistochemistry in paraffin-embeded tissue samples. In addition, these samples were stained with antibodies for CD10, bcl-6 and MUM-1 via tissue microarray to classify into subgroups.

Results:
The median percentage of FOXP3+ cells was 91/mm2 (range 4–2100 /mm2). Patients with poor performance status (PS), and high serum lactate dehydrogenase (LDH) showed lower numbers of FOXP3+ cells. (PS; p= 0.014, LDH; p=0.0048) Patients with high counts of FOXP3+ cells (>90/mm2) have better prognosis than those of low counts (5 year (5-y) overall survival (OS); 72.1%, 49.7% p=0.024, respectively). Although no prognostic difference was observed between GCB type and non-GCB type (5-y OS: GCB 71.2%, non GCB 53.1%, p=0.12), low counts of FOXP3+ cell and non-GCB type patient was poorer prognosis than high counts and non GCB type. (low 5-y OS 31.2%, high 5-y OS 69.8% p=0.02).

Conclusion:
Increased count of FOXP3+ tumor-infiltrating cell might predict better prognosis of DLBCL.

Disclosures:
No relevant conflicts of interest to declare.