bluuebunny/crossref_metadata_2025_split huggingface dataset:
Polyethylene Glycol - Polyethylenimine Nanoparticle-Mediated Transfection of Mir-150 Into the Leukemia Cells and Determination of the Expression Pattern Changes in Apoptosis and Cell Cycle Genes
Authors: Balci Tugce , Avci Cigir Biray , Ozcan Ipek , Susluer Sunde Yilmaz , Kayabasi Cagla , Saydam Guray , Gunduz Cumhur 
Published: October 2019

Abstract
Abstract 4679
The aims of this study are transfection of tumor suppressor miR-150 that is down-regulated in leukemogenesis into leukemia cells mediated by Polyethylene Glycol - Polyethylenimine (PEG-PEI) nanoparticle and determine the changes in gene expression pattern in chronic myeloid leukemia model cell lines; K-562 and KU812 and non-leukemia cell lines NCI-BL2347 and NCI-BL2171.
Characterization studies and stability studies of PEG-PEI copolymers were performed and by using these copolymers 9 nanoparticle formulations were prepared. Three copolymers (named T1, T3 and T9) were eligible for further analysis and nanoparticle complex (named F1, F2 and F3) formulations, particles that their size less than 300 nM have been evaluated.
Nanoparticle-mediated substitution of miR-150 reduced the expressions of cell cycle control genes CDK2 and CCNG2, CDK6 7-fold and 2-fold, respectively in K562 leukemia cell model.
Nanoparticle-mediated substitution of miR-150 has been evaluated KU812 leukemia cell model and reduced the expression of cell cycle control genes CHEK2, CDK5RAP1 and CCNG2 and CDKN1A 6- fold 5 –fold and 2-fold, respectively.
Substitution of deregulated tumor suppressor miR-150 to leukemia cells with non-viral transfection yielded promising results for the treatment of leukemia. Taking into account these results, it should be supported by preclinical studies in animal models, which would add benefit to current treatment protocols in clinical application.

Disclosures:
No relevant conflicts of interest to declare.
Normal Values for High Affinity Fc Gamma Receptor I (CD64) From Neonate to Young Adults
Authors: Matsushita Shoko , Asakura Yoshiko , Hayashi Yuko , Endo Mikiya , Nakatsuji Yukie , Ito Shigeki 
Published: October 2019

Abstract
Abstract 4685
The high affinity Fc gamma receptor I (Fc gamma RI, cluster of differentiation 64 [CD64]) is normally expressed to a very low extent by neutrophils; during bacterial infections, the expression has been shown to increase in adults and newborn infants. However, the normal range in steady status remains unknown. To ultimately develop the novel measurement technique and clinical application of CD64 expression as an early diagnostic marker for bacterial infection, normal CD64 expression levels were determined in normal neonates, infants, young children, and adults who had no symptoms of infection, stable temperature, normal leukocyte counts and normal C-reactive protein levels, and who were not using steroids, immunosuppressive drugs, aspirin, or other non-steroid anti-inflammatory drugs in this study. To quantify the expression levels of neutrophil CD64, mean fluorescence intensity (MFI) on neutrophils was measured by flow cytometry in 420 samples. Subjects were assigned to 7 groups by age: neonates (group A, n = 76); 1 – 11 months (group B, n = 51); 1 – 5 years (group C, n = 87); 6 – 10 years (group D, n = 76); 11 – 15 years (group E, n = 77); 16 – 20 years (group F, n = 33); and over 21 years (group G, n = 20). Each expression level in groups A, B, and C was significantly higher than that in group G (P < 0.01); no differences were found among groups D, E, F, and G. These results suggest that normal cutoff levels of CD64 should be determined according to age to apply to the clinical setting because CD64 expression has a tendency to increase according to age, especially over 21 years.

Disclosures:
No relevant conflicts of interest to declare.
Disseminated Tuberculosis with Hemophagocytic Lymphohistiocytosis (HLH)
Authors: Rao Suparna Ajit , Dutta Tarun K. , Naik Rakesh V , Krishnamurthy Aishwarya , Viswanath Vinod K. 
Published: October 2019

Abstract
Abstract 4686
A 38 year old woman presented with high grade fever and jaundice for one month. Patient also had reduced appetite and loss of weight for the same period. On examination, patient had significant pallor, icterus and pedal edema. Ultrasonogram showed enlarged liver (20 cms) and enlarged spleen (17.2 cms). Patient was empirically treated for malaria. Her subsequent investigations revealed Hb 35g/dl, total leucocyte count 2×109/l, differential leucocyte count - neutro 82%, lympho 18%, platelet count − 59×109/l, and red cell indices were as follows: MCV 71.6 fL, MCH 21.6 pg/cell, MCHC 30.2 g/dl. Her reticulocyte count was 0.5%. Peripheral blood smear showed pancytopenia with moderate anisopoikilocytosis. Her total bilirubin was 4.2 mg/dl and serum ferritin was found to be 1720 μg/L. In view of pancytopenia and non-response to antimalarials, patient was treated in line of septicemia with piperacillin and tazobactam, and simultaneously a bone marrow biopsy was performed. Bone marrow biopsy subsequently revealed a hypercellular marrow with erythroid hyperplasia. Number of macrophages was increased with some showing ingested red cells (hemophagocytosis) within them. In view of fever, splenomegaly, pancytopenia, hemophagocytosis and hyperferritinemia, a diagnosis of hemophagocytic lymphohistiocytosis (HLH) was made as per HLH 2004 diagnostic criteria. Bone marrow also revealed multifocal epithelial granulomas with caseation, pointing the etiology to that of disseminated tuberculosis. Patient expired before any anti-tuberculous treatment could be instituted.
Causes of HLH are broadly malignancy, collagen diseases and infections. Though malignancy and collagen diseases are common causes in the Western countries, tuberculosis is an important cause in a tropical country like India.

Disclosures:
No relevant conflicts of interest to declare.
Comparison of a Rotary Powered Bone Marrow Aspiration and Biopsy Device to the Traditional Manual Device in an Adolescent
Authors: Falcon Maria G. , Assanasen Chatchawin , Thomas Paul , Saldivar Victor 
Published: October 2019

Abstract
Abstract 4718
Bone marrow examination is important for the diagnosis of hematological malignancies and nonmalignant diseases in children. In patients with newly diagnosed lymphoproliferative diseases and certain non-hematopoietic malignancies, bone marrow examination is also part of the staging process. Core biopsy length has been found to be critical in diagnosing, predicting relapse or identifying residual disease following chemotherapy in patients. The larger the amount of marrow obtained increases the chance of finding a focal lesion. Unfortunately, the current practice of obtaining trephine biopsies and bone marrow aspirates in children via the manual method has a poor success rate for obtaining adequate specimens. In 2007, an FDA-cleared battery powered bone marrow aspiration and biopsy system (OnControl™ by Vidacare) was developed. Multiple studies have evaluated the use of the powered device in adults and found decreased time of procedure, decreased pain, and improved core biopsy specimens.
Here we present a direct comparison of the rotary powered device versus the traditional manual device (e.g. Jamshidi) when obtaining bilateral bone marrow aspirates and biopsies in a 17 year-old female with relapsed alveolar rhabdomyosarcoma. This patient required bilateral bone marrow biopsies to stage her disease and evaluate for bone marrow involvement. One aspirate and biopsy was obtained using the powered device from the right posterior superior iliac crest, and specimens were obtained from the left iliac crest using the traditional manual device. The endpoints measured were quality of the biopsy, length and width of the biopsy, time to obtain aspirate and biopsy, number of attempts to obtain the biopsy, post-procedural pain, and operator satisfaction with the device (O.S.).
Device Quality Rating Length (mm) Width (mm) Aspirate Time (sec) Biopsy Time (sec) Attempts Pain Score (0–10) O.S. (0–10) Powered 2 9 1.5 20 108 1 0 10 Manual 1 14 2 25 225 1 0 9
In conclusion, the powered device was superior to the manual device in terms of time to obtain the aspirate and biopsy and operator satisfaction with the device. It was found to be equivalent to the manual device in regards to number of attempts to obtain the biopsy, and post-procedural pain score. The manual device produced a biopsy that was longer, wider, and of higher quality than the biopsy obtained via the powered method. A randomized controlled trial in the pediatric population comparing the rotary powered device to the traditional device is currently underway as further studies are needed to evaluate the use of the powered bone marrow aspiration and biopsy device in children.

Disclosures:
Falcon: Vidacare Corporation: Research Funding. Assanasen:Vidacare Corporation: Research Funding.
Serum Fructosamine As an Alternative to Glycated Hemoglobin (HbA1c) for Glycemic Assessment in Diabetic Patients with Hemoglobin (Hb) Louisville
Authors: Lorenzo-Medina Mercedes , De-La-Iglesia Silvia , Quintana-Hidalgo Lucia , Ruiz-Garcia Lidia , Martin-Alfaro Ruth , Herrada Juan 
Published: October 2019

Abstract
Abstract 4771

Background:
HbA1c is widely used in the management of patients with diabetes mellitus (DM). Hb Louisville is an unstable hemoglobin that differs from HbA by the substitution of a phenylalanine residue for a leucine residue in position 42 of the b-chain, resulting in instability in the Hb molecule with increased erythrocytic destruction, clinically manifested as hemolytic anemia. The Hb Louisville heterozygotes suffer from a mild anemia, jaundice and hemolytic crisis. Abnormally low HbA1c concentrations are usually encountered in patients with high turnover rates of hemoglobin. Fructosamine is unaffected by disorders of red blood cells nor is influenced by hemoglobin variants, thus serving as an indicator of blood glucose concentrations of previous 2–3 weeks.

Methods:
Case report.

Results:
We report on three patients that belonged to the same family and had history of heterozygosity trait for Hb Louisville. All three patients had type 2 diabetes mellitus with falsely low HbA1c values (patient A: 2.8%, 2.9%, 3.2%; patient B: 4.4%, 3.8%, 3.7%; patient C: 4.2%, 4.6%, 3.7%). The patients had consistent elevation of fasting blood glucose values (patient A: 162, 171, 233 mg/dL; patient B: 262, 232, 235 mg/dL; patient C: 152, 130, 136 mg/dL). All three patients had evidence of hemolytic anemia, and their average hemoglobin levels were: patient A: 9.41 mg/dL, patient B: 10.9 mg/dL, patient C: 8.6 mg/dL. We therefore determined serum fructosamine levels, and the results were: patient A (patient deceased), patient B: 420 mcmol/L and patient C: 295 mcmol/L (Reference Range >285 mcmol/L).

Conclusion:
Fructosamine serum level determination is advocated for glycemic assessment in diabetic patients with Hb Louisville when discrepancies between glucose measurements and HbA1c values>are encountered.

Disclosures:
No relevant conflicts of interest to declare.
Changes in Regional Oxygenation At the Site of Sickle Cell Vaso-Occlusive Pain
Authors: Kato Roberta Miyeko , Nord Anne , McGuire Erin , Gardner Daniel , Bush Adam M , Detterich Jon , Khoo Michael , Meiselman Herbert J , Wood John C , Berdoukas Vasilios , Coates Thomas D. 
Published: October 2019

Abstract
Abstract 4773

Background:
The pathophysiology of sickle cell disease (SCD) is commonly thought to be in part a result of sickled red blood cells obstructing blood flow leading to hypoxic cellular injury. The most common SCD complication is painful vaso-occlusive crisis (VOC). However, the physiologic changes that occur at the site of VOC are poorly understood and there are no objective measurements of VOC. We hypothesized that regional oxygenation at the site of pain would be decreased from the patient's asymptomatic baseline and could be used as an objective measurement of the VOC process.

Methods:
Subjects were enrolled at the time of hospitalization for unilateral painful VOC. The subjects completed the adolescent pediatric pain tool (APPT) to assess the location, severity and quality of their pain. Regional oxygenation (RSO2) was measured by near infrared spectroscopy (NIRS) at the site that the subject identified as where they experienced the most severe VOC related pain. NIRS measurements were also made on the contralateral body site. NIRS measurements were made for 15 minutes with the subject in the supine position. Measurements were made in the first 24 hours of hospitalization. Blood pressure, heart rate, pulse oximetry, hematologic parameters and use of oxygen were determined. The subject returned 4 weeks after discharge for repeat testing to determine their physiologic post VOC RSO2 baseline. Mean values for the NIRS testing period were determined and the change in RSO2 during VOC from post VOC baseline were calculated.

Results:
5 female subjects with SCD were studied, mean age 15.8±3.0years. There was an average pain score of 7.4/10+1.9. Pain occurred in the back of 2 subjects and the lower extremities in the others. The RSO2% in the painful and not painful sites was increased from baseline by an average of 10.6±4.0% and 16.6±5.6%, respectively. The painful site had a decreased RSO2% in comparison to the contralateral side in 2 of 5 subjects (see figure). The RSO2% VOC baseline at the painful site post was higher in 3 of 5 subjects. There was no difference from baseline in blood pressure, heart rate, or arterial oxygen saturations during VOC. There were changes in the hematologic parameters consistent with VOC and active hemolysis.

Conclusions:
The SCD subjects demonstrated increased regional oxygenation at both the painful and contralateral not painful sites during VOC compared to the post VOC baseline. During VOC there was no predictable difference in regional oxygenation at the site of VOC pain contrary to our original hypothesis.

Disclosures:
Wood: Ferrokin Biosciences: Consultancy; Shire: Consultancy; Apotex: Consultancy, Honoraria; Novartis: Honoraria, Research Funding. Coates:Novartis: Speakers Bureau; Apopharma: Consultancy.
Second Cancers After Treatment with Stanford V Regimen in Eastern Cooperative Oncology Group (ECOG) Pilot Study E1492 At a Median Follow up of 17 Years
Authors: Advani Ranjana H. , Li Hailun , Hoppe Richard T. , Bartlett Nancy L. , Bennett John M. , Neuberg Donna S. , Cassileth Peter A , Kahl Brad S. , Horning Sandra J. 
Published: October 2019

Abstract
Abstract 4779

Purpose:
The Stanford V regimen is a combined modality approach for treatment of Hodgkin lymphoma (HL). E1492, an ECOG pilot study, consisted of 12 weeks of Stanford V chemotherapy followed by 36 Gy radiation therapy (RT) to sites > 5 cm or macroscropic splenic disease at diagnosis. Efficacy results were reported previously (Horning et al. J Clin Onc 18, 2000). The study now has a median follow up of 17 years and patients have been followed for overall survival (OS) and development of second cancers.

Methods:
47 eligible patients with stage bulky mediastinal (mass > one third of the maximum intrathoracic diameter) stage I-II or stage III-IV HL were enrolled between March 1992 and February 1995. Patients were followed every 3 months during the first year off therapy, every 6 months during years 2–5 and annually thereafter. The ECOG database was reviewed for OS and reported second cancers. Patient characteristics at baseline, type of second cancer and time to development of second cancer were assessed. RT summary forms were reviewed for patients with second cancers.

Results:
41 patients were treated with combined modality therapy and 6 with chemotherapy alone. The median age was 32 years (range 20–56 years). The 5 and 10 year OS are 96%, and 89% respectively. Seven second cancers were reported, as shown in the Table. The cumulative incidence for second cancers accounting for death as a competing risk is 0.02 (95% CI, 0–0.06) at 5 years, 0.07 (95% CI, 0–0.14) at 10 years, and 0.15 (95% CI, 0.04–0.27) at 15 years. Complete details of the exact location, histologic subtype and subsequent management of second cancers were not available for review.
Five of the 7 cancers [2 skin, 1 prostate, 2 acute myeloid leukemia (AML)] were not radiation-related. One patient developed AML after primary therapy as reported in the initial publication. A second patient developed AML 5 years after salvage therapy followed by autologous stem cell transplant for relapsed HL. It is likely the 2 cases of breast cancer were treatment related.

Conclusion:
Within the caveats of a retrospective analysis from a small cooperative group phase 2 trial, the mature 10 year OS of 89% and low frequency of secondary cancers are encouraging in comparison to historic treatment with combined modality treatment. Longer follow up of other Stanford V regimen data sets (i.e. United Kingdom National Cancer Research Institute Lymphoma Group Study ISRCTN 64141244 and the Eastern Cooperative Oncology Group E2496) are required to confirm these findings.

Disclosures:
No relevant conflicts of interest to declare.
A New Algorithm and Panel Construction for Pediatric Leukemia Immunophenotyping Using 10-Color Flow Cytometry
Authors: Keller Bettina , Radsak Markus P , Faber Joerg , Russo Alexandra 
Published: October 2019

Abstract
Abstract 4799

Background:
Rapid identification and quantification of abnormal cell populations in minimal specimen are crucial for diagnosis and longitudinal minimal residual disease (MRD) testing of childhood leukemia. So far, most standard immunophenotypic analyses are performed using antibody panels with up to five-colors and require high cell numbers. For infant and pediatric specimen, high-level multicolor analyses is highly desirable to gather sufficient data for initial diagnostic and follow up monitoring of pathologic populations.

Objective:
In this study, we aimed to establish a newly defined pediatric multicolor flow cytometric panel algorithm with high reliability yet minimal specimen requirement.

Results:
We defined a 10-color flow cytometric panel using the new violet laser dye “KromeOrange (KO)”. Applying CD45-KO/Side Scatter gating, combined with 2 additional backbone markers the panel is designed in two consecutive steps. In the first step, a single standardized 10-color-“screening tube” (FITC-HLA-DR, PE-CD15/CD56, ECD-CD5, PC5.5-CD33, PC7-CD13, APC-CD117, APC A700-CD34, APC A750-CD19, PB-CD3, KrO-CD45) is applied for initial orientation of specific lineage assignment. Based on results obtained with the screening tube, a specific multi-tube “classification panel” is used to complete detailed characterization of lineage specific malignancy and maturation stage. Suitable specimens include fresh blood, bone marrow and all body fluids. All samples are stained directly with monoclonal antibodies, followed by the lyses of erythrocytes and a short wash. Compared to standard five color panel previously used the application of greater numbers of informative antibodies in the screening tube and in the 2ndstep muti-tube classification panel is cost and time efficient and results in a more precise characterization of any single event.

Conclusion:
Our panel construction and algorithm definition for infant and pediatric leukemia immunophenotyping is one of the first 10-color flow cytometry panels described for this application. Advantages are the possibility to obtain highly specific information from minimal specimens with significantly improved laboratory efficiency. The overall performance is currently tested in a routine clinical setting.

Disclosures:
No relevant conflicts of interest to declare.
Src Family Kinase Inhibitor PP2 Has Different Effects On ATRA or Arsenic Trioxide-Induced Differentiation of Acute Promyelocytic Leukemia Cell Line
Authors: Kim Kyoung Ha , Cheong Hee-Jeong , Kim Sook-Ja , Jeon Young Woo , Yoon Jina , Kim Han Jo , Kim Se Hyung , Kim Hyun Jung , Lee Sang-Cheol , Bae Sang Byung , Kim Chan-Kyu , Lee Namsu , Lee Kyu Taeg , Park Sung Kyu , Hong Dae Sik , Hee-Sook Park , Won Jong-Ho 
Published: October 2019

Abstract
Abstract 4800
Acute promyelocytic leukemia (APL) is a subtype of acute myeloid leukemia (AML) with distinctive biologic and clinical features that is now highly curable. Leukemic promyelocytes have the unique ability to undergo differentiation after exposure to retinoic acid and both differentiation and apoptosis after exposure to arsenic trioxide (As2O3, ATO). Emerging evidence has implicated Src family kinases (SFKs) as regulators of proliferation and survival of myeloid lineage cells. Recent studies showed that inhibition of SFKs resulted in enhancement of retinoic acid-induced myeloid differentiation. In this study, we demonstrated that the SFK inhibitor PP2 enhanced the differentiation of NB4 cells when combined with ATO as well when combined with as all-trans-retinoic acid (ATRA) and the difference in the retinoic acid-induced gene expression between the cells treated with PP2 in combination with ATRA and the cells treated with PP2 in combination with ATO.
SFK inhibitor PP2 significantly enhanced ATRA- or ATO-induced differentiation of NB4 cells. The synergistic effect was significantly stronger when PP2 was combined with ATRA than when PP2 was combined with ATO. Flow cytometric analysis demonstrated a significant increase in CD11b-positive granulocytes up to 60.73% and 31.58%, respectively. These results were confirmed by morphologic analysis using Wright stain and NBT staining. These effects were not related to apoptosis. Annexin-V-fluorescein staining revealed that PP2 combined with ATRA or PP2 combined with ATO did not induce apoptosis in NB4 cells. Retinoic acid-induced gene expression was different in both groups. ICAM-1 expression was significantly increased in the cells treated with PP2 in combination with ATRA whereas cathepsin D expression was significantly increased in the cells treated with PP2 in combination with ATO. These findings suggested that the synergistic effect of SFK inhibitor PP2 in combination with ATRA was significantly stronger than that of PP2 in combination with ATO on NB4 myeloid leukemia cell differentiation and this difference was related to the disparate activation of retinoic acid-induced genes.

Disclosures:
No relevant conflicts of interest to declare.