bluuebunny/crossref_metadata_2025_split huggingface dataset:
RhoH Downregulates Rac Through RhoGAP Family Protein
Authors: Katsumi Akira , Kobayashi Miki , Matsushita Tadashi , Kiyoi Hitoshi , Kojima Tetsuhito , Maruyama Mitsuo , Kaibuchi Kozo , Naoe Tomoki 
Published: October 2019

Abstract
Abstract 4792
Rho GTPases have been widely studied because they are the critical regulators of cellular signaling mediated by G-protein-coupled receptors, tyrosine kinase receptors and integrins. Among more than 20 members of the Rho GTPases, Rac1, Cdc42 and RhoA have been extensively characterized. Rac plays essential roles in the control of actin cytoskeleton organization, migration, phagocytosis, metastasis, transformation, gene expression and cell-cycle progression. Moreover, it has been found that Rac plays as a key molecular switch of hematopoietic stem/progenitor cells engraftment and marrow retention. RhoH, a member of the Rho family, is specifically expressed in the hematopoietic cells, including bone marrow progenitor, differentiated myeloid and lymphoid cells. RhoH antagonizes Rac, however, the molecules that inactivate Rac downstream of RhoH have not been identified. Here we perform GTP-RhoH affinity column chromatography by using platelet lysates. Using triple quadrupole liquid chromatography tandem mass spectrometer, 83 different proteins associated with active RhoH were identified. They include Rho GTPase activating proteins (RhoGAP), serine/threonine kinases, phosphatidylinositol kinases, ERM family proteins and others. Among them, we focused on RhoGAP family. RhoGAP binds to RhoH through its GAP domain. Expression of both full length and GAP domain of RhoGAP effectively deactivates Rac, whereas GAP domain deleted RhoGAP did not. These results suggest that RhoH negatively regulates Rac through RhoGAP.

Disclosures:
Naoe: Kyowa-Hakko Kirin.: Research Funding; Dainipponn-Sumitomo Pharma.: Research Funding; Chugai Pharma.: Research Funding; Novartis Pharma.: Honoraria, Speakers Bureau; Zenyaku-Kogyo: Research Funding; Otsuka Pharma.: Research Funding.
Cord Blood Hematopoietic Progenitor Cell in Co-Culture with Bone Marrow Mesenchymal Stromal Cells: A Synergic Effect
Authors: Almici Camillo , Neva Arabella , Verardi Rosanna , Braga Simona , Di Palma Andrea , Bianchetti Andrea , Russo Domenico , Marini Mirella 
Published: October 2019

Abstract
Abstract 4811
The number of hematopoietic stem and progenitor cells (HPCs) in cord blood units are limited and this can result in delayed engraftment. In vitro expansion of HPCs provides a perspective to overcome these limitations. Different combinations of cytokines as well as mesenchymal stromal cells (MSC) have been shown to separately support HPCs ex vivo expansion, but the combining effects are under evaluation. Data derived from ex vivo co-culture systems using MSC as a feeder layer suggest that cellular contacts could have a significant impact on expansion. We have evaluated the expansion rate of thawed cord blood samples (n=6) in a medium containing SCF (100 ng/ml) and G-CSF (100 ng/ml) plated over a pre-established bone marrow derived MSC layer in comparison to the absence of either MSC layer or cytokines. After 7 days cultures were demi-depopulated. At 14 days of culture adherent and non-adherent cells were harvested, counted and evaluated for antigens expression and clonogenic capacity. Immunophenotypic analysis was performed using CD34-PE, CD38-FITC, CD45-PE-Cy7, CD133-APC. Clonogenic assay was performed in semisolid methylcellulose culture medium (MethoCult, Stem Cell Technologies), CFU frequencies and total CFU numbers per cord blood sample were determined. After 14 days of culture, in the presence of MSC layer, an 11.2-fold increase (range 4.4–18.4) in total number of cells was observed, in comparison to a 4.8-fold increase (range1.1-10.35) in the absence of MSC layer. The presence of MSC layer generated a 4.3-fold increase (range 1.5–7.2) in the number of CD34 positive cells, compared to a 3.3-fold increase (range 0.9–5.7) in the absence of MSC; when considering the more immature CD34+/CD38− subpopulation the corresponding increase were 26.9-fold vs 2.85-fold, respectively. Moreover, the percentage of the CD34+/CD38− subpopulation was higher in the adherent compared to the non-adherent fraction (76% vs 15%). The selection effect given by the MSC layer was confirmed by the presence of hematopoiesis foci growing onto the MSC layer. Our data show that cord blood HPCs can be expanded in vitro, moreover the co-culture on a MSC layer shows a synergistic effect on TCN, CD34+ cells and on more primitive CD34+/CD38− cells. Therefore, a clinical protocol of cord blood HPCs and MSC co-culture could represent a promising approach for improving engraftment kinetics in cord blood transplant recipients.

Disclosures:
No relevant conflicts of interest to declare.
Ex Vivo Expansion of CD34+ Cells From Cord Blood, Bone Marrow, and Mobilized Peripheral Blood Using NANEX Nanofiber Scaffold
Authors: Fischer Stephen L , Fonseca Jacqueline M , Zhao Yukang , Kelley Linda L. , Sakthivel Ramasamy 
Published: October 2019

Abstract
Abstract 4816
Hematopoietic stem cell (HSC) transplantation has become the standard of care for patients with hematologic cancers, anemia, and a variety of other malignant and non-malignant disorders, with greater than 50,000 such procedures being performed globally each year, according to the Worldwide Network for Blood and Marrow Transplantation. Although mobilized peripheral blood (MPB) has become a preferred source of HSCs for transplants, bone marrow (BM) and umbilical cord blood (UCB) are also frequently utilized. Regardless of source, several groups have reported that grafts containing lower total nucleated cell (TNC) and CD34+ cell doses contribute to delayed engraftment and higher graft failure rate. Therefore, methods to increase the total cell number while maintaining the progenitor phenotype, especially the CD34+ progenitor cells, from individual grafts would have a significant clinical impact.
Ex vivo expansion of HSCs prior to transplantation is one approach that offers tremendous promise for increasing cell doses and improving clinical outcomes. In many ex vivo culture systems, HSCs are cultured as a suspension cells and cultured in the presence of various media additives that act to enhance cell proliferation while reducing differentiation. An often-overlooked factor influencing fate decisions is the interaction of HSCs with a substrate. In the natural bone marrow microenvironment, HSCs maintain close contact with a complex network of stromal cells and extracellular matrix, likely indicating that cell-cell and cell-matrix interactions play an important role in maintaining their stem cell phenotype. With the goal of mimicking the bone marrow stem cell niche, Arteriocyte, Inc. has developed a 3-D NANEX nanofiber based cell culture substrate. The functionalized NANEX substrate is designed to provide topographical and substrate-immobilized biochemical cues that act in synergy with media additives to enhance HSC proliferation while maintain the progenitors stem cell phenotype.
Here, we present our recent work with the NANEX platform towards comparing and achieving a high yield ex vivo expansion of CD34+ cells from MPB, BM, and UCB. Additionally, through the use of flow cytometry and CFU assays, we quantify and characterize NANEX-expanded cells from each source. Furthermore, we compared NANEX to a variety of commercially available products and demonstrate that NANEX significantly improves expansion and reduces phenotype loss during ex vivo culture. Our data indicates that NANEX technology provides a robust ex vivo expansion of HSCs and, with further GMP and clinical development, offers great potential for clinical applications.

Disclosures:
No relevant conflicts of interest to declare.
Gender Difference in Periodic Limb Movements in Adults with Sickle Cell Disease
Authors: Roizenblatt Suely , Roizenblatt Marina , Filho Frederico Pollack , Matsuda Sandra S , Mecabo Grazielle , Arruda Martha M A S , Tufik Sergio , Figueiredo Maria Stella 
Published: October 2019

Abstract
Abstract 4843
Periodic limb movements during sleep (PLMS) are commonly described in children with Sickle Cell Disease (SCD), however the relevance of such events in sleep disturbance or complaint of daytime fatigue remain questionable. It has been known that the dopaminergic neurotransmission has a modulator role in pain perception. Since PLMS have been suggested to be a potential biological marker of dopaminergic mechanisms, this study aimed to assess PLMS in adults with SCD and its correlations with clinical and sleep parameters.

Methods:
Seventy adults with SCD (50% females, matched for age and body mass index), underwent Brief Pain Inventory, overnight polysomnogram and laboratorial tests for hemoglobin, reticulocytes, ferritin, transferin saturation, haptoglobin, fetal hemoglobin, lactate dehydrogenase (LDH), and bilirrubins.

Results:
The mean PLMS index was higher in females (16.5±10.7/h vs. 8.7±8.2/h, p< 0.05), with 88.6% of the females having increased PLMS index (≥ 15/h) in comparison to males (22.6), p<0.01. Total arousal index and PLMS arousal index were increased in females with PLMS ≥ 15/h compared to those with PLMS < 5/h (p<0.05, both), but not in males. Females also exhibited correlation of increased PLMS index and clinical parameters such as pain score (rS =0.71), indirect bilirrubin (rS =0.42) and LDH (rS =0.38), p<0.01 in all. An interaction effect of the non-use of hydroxyurea treatment in PLMS abnormality was also observed (p<0.05).

Conclusions:
Elevated PLMS were more common in females with SCD than in males and correlated with pain symptoms and hematological parameters of hemolysis. Hydroxyurea treatment showed a protective effect against the PLMS index increase.
Supported by AFIP, CEPID-FAPESP 98/14303-3, and CAPES/SUS.

Disclosures:
No relevant conflicts of interest to declare.
A Study of Chronic Hydroxyurea Administration in Sickle Cell Patients
Authors: Koutsouka Freideriki , Makis Alexandros , Lagos Konstantinos , Pliakou Xanthi , Bourantas Dimitrios , Kapsali Eleni , Briasoulis Evangelos 
Published: October 2019

Abstract
Abstract 4857

Background.
The hemoglobin F (HbF) promoting agent hydroxyurea (HU) has been established as an effective therapy for adults with sickle cell disease (SCD) who develop frequent pain episodes, acute chest syndrome, severe vaso-occlusive events, or severe symptomatic anemia. However, concerns have been raised for long-term safety issues of HU in relation to its cytotoxic effects, which are attributed to ribonucleotide reductase inhibition. We analyzed the clinical effects of chronic use of HU in adult SCD patients treated in a referral Academic center.

Methods.
Medical files of 29 adult patients (10 male, median age 42 years, range 29–68 years) followed at our Thalassemia Unit for the last 17 years were retrieved, and detailed data were recorded and analyzed. Four patients had homozygous SCD and 25, sickle cell/beta thalassemia.

Results.
The median follow up time was 10 years (range 3–17 years), the total follow-up time was 289 patient-years and the total HU exposure period was 308 patient-years. The mean dose of HU administered was 29.3 mg/kg/day. The main clinical outcomes of HU therapy are shown in Table 1. In terms of efficacy, HU induced marked increase of HbF and substantial reduction of painful crises and transfusions needed, all outcomes designated statistically significant. HU therapy was also associated with significant increase of hemoglobin (Hb) levels, as well as with reduction of serum lactate dehydrogenase levels (LDH) and white blood cell count (WBC). In terms of toxicity, one poor HU compliant female patient died of pulmonary embolism. Disease complications observed during the long follow up time were: pulmonary hypertension in 2 patients, leg ulcers in one patient and renal impairment in one patient. Seven patients discontinued HU therapy (mean treatment duration 8.4 years) because of scheduled pregnancy (3 patients), severe neutropenia (2 patients) and non-compliance (2 patients). No patient developed cancer.

Conclusions.
This retrospective study, which analyzed data of most prolonged administration of HU, provides data supporting the efficacy and safety of chronic administration of HU in SCD adults.

Disclosures:
No relevant conflicts of interest to declare.
Paraneoplastic Pemphigus (PNP) As a Presenting Finding in a Patient with Hodgkin's Lymphoma
Authors: Marjon Philip , Kim Byung , Lazic Tamara , Cernik Christina , Rathore Ritesh 
Published: October 2019

Abstract
Abstract 4864

Introduction:
We report a case of PNP in a patient who was subsequently diagnosed with classical Hodgkins Lymphoma mixed cellularity type. PNP is an autoimmune mediated blistering disorder that presents with skin lesions demonstrating characteristics of erythema multiforme and pemphigus vulgaris.

Case:
A 76 year-old Hispanic male presented to the Emergency department with intermittent low fevers of 4 months duration associated with progressive weakness and fatigue; he now had mouth ulcers of 1-week duration along with anorexia. Physical exam was significant for a temperature of 102.4F, bilateral conjunctivitis with mild white discharge. Labs included WBC 5,000/mm3, hemoglobin 9.5 gm/dL, platelets 99,000/mm3; differential showed lymphocytes 22%, monocytes 11%, eosinophils 3%, and myelocytes 9%. Oral examination revealed dry mucosa with bleeding perioral ulcers. An ulcer biopsy was obtained and demonstrated necrotic keratinocytes and necrotic neutrophilic inflammation consistent with PNP. Chest CT showed a multinodular goiter, enlarged mediastinal and hilar nodes with calcified granulomas bilaterally as well as a new noncalcified nodule in the right lung. PET scan revealed FDG avid lymph nodes in the left supraclavicular, hilar and left axilla areas as well as multiple regions in the spleen and retroperitoneal nodes. A left axillary node biopsy showed classical Hodgkins lymphoma of mixed cellularity type. The patient was started on prednisone 75 mg per day in divided doses and his oral ulcerations and clinical status improved after 2 days of therapy. Subsequently the patient was started on gemcitabine, vinorelbine and liposomal doxorubicin (GVD) chemotherapy for his malignancy.

Discussion:
Although PNP has been described as being associated with hematologic malignancies such as Non-Hodgkins Lymphoma, Chronic Lymphocytic Leukemia and Castleman's disease, it has rarely been demonstrated in association with classical Hodgkins disease. This case report highlights the need for a higher index of suspicion of underlying malignancy including classical Hodgkins disease when confronted with a patient presenting PNP.

Disclosures:
No relevant conflicts of interest to declare.
Hodgkin's Lymphoma (HL) Presenting As An Abnormal Mammogram: A Case Report
Authors: Le Kim , Lorenzo-Medina Mercedes , Friedman Paul , Dane Langsjoen , Noble Luis , Richards Matthew , Arango Carmen , Gupta Anuradha , Campos-Gines Angel F , Landeros Mark M , Herrada Juan 
Published: October 2019

Abstract
Abstract 4865

Background:
Although not a common presentation, Hodgkin's lymphoma (HL) can first manifest as an enlarged axillary lymph node. We report a case of a patient diagnosed with nodular sclerosing HL, presenting as an abnormal axillary lymph node incidentally noted during routine mammogram.

Case report:
A 79 year-old asymptomatic female was referred to our center after being discovered a 3 cm left axillary lymph node detected during a routine screening mammogram. Except from the enlarged left axillary lymphadenopathy, the remaining physical exam was within normal limits. An ultrasound-guided fine needle biopsy revealed histologic features consistent with Hodgkin's lymphoma, nodular sclerosing type. Immunostains confirmed the presence of Reed Sternberg cells that were positive for CD 30, focally positive for CD 15, and negative for CD 20 and EMA. Bilateral bone marrow showed no evidence of HL involvement. A positron-emission tomography detected abnormal uptake in multiple enlarged lymph nodes above and below the diaphragm, and in the thoracic spine, at T9, T11 and T12 vertebral body levels. The patient was successfully treated with doxorubicin-based combination chemotherapy regimen.

Conclusion:
There are multiple clinical presentations for Hodgkin's lymphoma. Routine mammograms can aid in the detection of cancer types other than breast cancer.

Disclosures:
No relevant conflicts of interest to declare.
Metabolism of Human Leukemia Cells in NOD/Scid Mice: Effectiveness of 2-Deoxy-D-Glucose
Authors: Tsunekawa Norikazu , Miwa Hiroshi , Mizuno Shohei , Gotoh Mineaki , Takahashi Miyuki , Goto Mayuko , Suganuma Kazuto , Hanamura Ichiro , Shikami Masato , Nitta Masakazu 
Published: October 2019

Abstract
Abstract 4881
We have recently shown that NB4,an APL cell line, is dependent on glycolysis, while THP-1, a monocytic cell line, is relatively dependent on oxidative phosphorylation in mitochondria, where oxidation of fatty acid is important. (Leukemia and Lymphoma 51:2112,2010)
Here, we examined the energy metabolism of these leukemia cell lines in immunodeficient mice. 1.0×107 cells of each cell line were inoculated subcutaneously in NOD/scid mice with the treatment of anti-asialo GM1 antibody. Mice were divided into two groups: normal diet (carbohydrate;65.3%, fat;6.3%) or carbohydrate-restricted (high fat) diet (carbohydrate;19.6%, fat;62.2%). At day 42, tumor volume (TV) of NB4 in three normal diet-fed mice became 694±358mm3, which was greater than that in three high fat diet-fed mice:173±216mm3 (P= 0.043). On the other hand, tumor volume of THP-1 in three normal diet-fed mice became 1130 ± 600mm3, which was smaller than that in three high fat diet-fed mice: 3300 ± 1053mm3 (p = 0.050).
Then, since 14 days after inoculation of leukemia cells in NOD/scid mice fed with normal diet, glycolysis inhibitor 2-deoxy-D-glucose (2-DG) (35mg/mouse) was administered intraperitoneally once a week. Tumor size at day 41 of both leukemia cells were suppressed by 2-DG treatment. (THP-1: 2-DG (-): TV = 3099 ± 193mm3, 2-DG (+): TV = 513 ± 550mm3) (THP-1: p = 0.040). (NB4: 2-DG (-): TV = 495 ± 417mm3, 2-DG (+): TV = 87 ± 89mm3) (NB4: p = 0.313).
Pathological examination of tumors have shown that 2-DG treated tumor had larger areas of dead cells in both cell lines.
Our study demonstrated that NB4 is dependent largely on glucose and THP-1 on fat. In addition, 2-DG is effective in growth suppression of leukemia cells in vivo. These findings will help the future therapy for leukemia targeting energy metabolism.

Disclosures:
No relevant conflicts of interest to declare.
Analysis of the Clinical Features and the Prognosis of Acute Lymphoblastic Leukemia with Complex Chromosomal Aberrations
Authors: Ye Xiujin , Zhu Lixia , Li Li , Zhang Jie , He Jingsong , Zhu Jingjing , Huang He , Cai Zhen 
Published: October 2019

Abstract
Abstract 4914

Background:
Acute lympoblastic leukemia (ALL) with complex chromosomal aberrations (CCAs) which patients harboring three or more acquired chromosomal aberrations always have a poor outcome. Philadelphia chromosome–positive (Ph+) ALL is the largest genetically defined subtype in adult ALL with the most unfavorable prognosis. It is necessary to investigate the clinical and laboratory features of ALL patients with CCAs, analyze the distribution of Ph+ in these patients and assess the prognosis.

Methods:
In a retrospective follow-up study from 2005.1∼2011.6, 38 ALL patients with CCAs were investigated. All cases were diagnosed with morphological,immunological,cytogenetic classification and molecular biology. The karyotypes were interpreted according to the International System for Human Cytogenetic Nomenclature (ISCN). Clinical characteristics, laboratory features, treatment response and prognosis of ALL patients with CCAs were analyzed.

Results:
The study included 17 Ph+ ALL patients with an age range of 12 to 81 years (mean, 34.5 years) and a male to female ratio of 1:1. Ph+ ALL patients with CCAs were older than Ph- cases (P=0.096). The median survival time was 9.5 months (range, 1–44 months). The complete remission (CR) rate after two cycles of systemic chemotherapy was 53.6% (15/28) and the relapse rate was 53.3% (8/15). Ph+ ALL patients with CCAs showed a lower CR rate than Ph- cases. Eleven patients died during the follow-up period, six of them died within 2 months from the initial ALL diagnosis, and nine patients were failure to achieve CR. Three patients underwent allogeneic bone marrow transplantation (BMT): two of them presented Ph+ relapsed after 2 and 10 months, respectively. Using Kaplan-Meier survival analysis, we found the survival outcome of Ph+ ALL patients with CCAs is inferior to Ph- cases.

Conclusion:
Treatment outcomes of ALL patients with CCAs receiving chemotherapy or BMT are very poor. Ph+ as an unfavorable parameter influences on the CR rate and survival outcome. It is necessary to accumulate more clinical data to find innovative treatments that can improve the prognosis of this refractory leukemia.

Disclosures:
No relevant conflicts of interest to declare.
A Multicenter, Randomized, Open-Label Study to Evaluate Safety and Efficacy of Posaconazole Vs Fluconazole in the Prophylaxis of Invasive Fungal Infection in a Chinese Population
Authors: Yang Shen , Huang Xiaojun , Wang Jianxiang , Jin Jie , Hu Jianda , Yu Kang , Wu Depei , Wang Shujie , Yu Li , Chen Xiequn , Liu Ting , Liang Yingmin , Fangping Chen , Li Yan , Shen Zhixiang 
Published: October 2019

Abstract
Abstract 4916

Background
Invasive fungal infection (IFI) is a common and fatal complication in neutropenic patients with acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS).

Methods
In this multicenter, randomized study, we enrolled patients with persistent neutropenia resulting from chemotherapy for the treatment of AML and MDS. Prophylaxis with either posaconazole or fluconazole was given to the patients to compare the efficacy and safety of the 2 drugs. The treatment was given with each cycle of chemotherapy until the patient recovered from neutropenia and had complete remission, an IFI occurred, or until the patient had received a maximum of 12 weeks of treatment. The primary end point was the incidence of possible, proven, or probable diagnosis of IFI during the treatment phase in *the 2 groups; clinical failure rate, all-cause mortality, and time to first systemic antifungal treatment were secondary end points.

Results
A total of 252 patients were included in this study, and 7 patients were excluded due to withdrawal of informed consent or deviation of eligible criteria. Finally, 124 patients from the posaconazole group and 121 patients from the fluconazole group were included; and 234 patients (117 in each group) entered into statistical analysis, respectively. The incidence of proven, probable, and possible IFI was 9.4% (11/117) and 22.2% (26/117) in the posaconazole and fluconazole groups, respectively (P = 0.0114). The clinical failure rate was lower in the posaconazole group (31.6% [95% CI: 23.3–40.9], 37/117) than in the fluconazole group (41.88% [95% CI: 32.8–51.4], 49/117); however, statistical significance was not reached (P = 0.168). The patients receiving posaconazole had a later onset of first systematic antifungal therapy than those receiving fluconazole (P = 0.0139). The most common clinical adverse reactions were liver function abnormalities, with 11 cases (9.4%) in the posaconazole group and 6 cases (5.1%) in the fluconazole group (P = 0.221).

Conclusions
Posaconazole demonstrates efficacy as prophylaxis against IFI in high-risk neutropenic patients with AML and MDS undergoing chemotherapy and is well tolerated during long-term use. (ClinicalTrials.gov number, NCT00811928)

Disclosures:
No relevant conflicts of interest to declare.