bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rs1982809 is a functional biomarker for the prognosis of severe post-traumatic sepsis and MODs
Authors: Gan Lebin , Hu Chen , Deng Zhihong , Lu Hongxiang , Sun Jiali , Peng Guoxuan , Jiang Jianxin , Zeng Ling , Deng Jin 
Published: October 2019

BTLA is a useful biomarker to characterize the immune states of sepsis patients. We investigated the association between genetic variations of BTLA and morbidity of sepsis and MODS in severe traumatic patient. Three tag single nucleotide polymorphisms of BTLA were genotyped in 562 severe trauma patients. To further elucidate the mechanism, mRNA stability, BTLA 3ʹ-UTR activity, and its expression on T lymphocytes were measured. Only rs1982809 which located in 3ʹ-UTR of BTLA showed a significant clinical relevance with the incidence rate of sepsis and MOD scores. The sepsis incidence and MOD score of rs1982809 CC genotype carriers were higher than TT carriers. The percentage of circulating BTLA + CD4 + CD3 +  T lymphocytes was markedly lower in CC genotype carriers. Luciferase activity in plasmids containing C allele was lower than that of T allele. Thus, the differential expression of BTLA on T lymphocytes might be caused by the different 3ʹ-UTR activity induced by rs1982809 T/C. Therefore, rs1982809 is a useful clinical biomarker in the prognosis evaluating of sepsis and subsequent MODS. Moreover, it is also a functional single nucleotide polymorphism affecting the activity of BTLA 3ʹ-UTR and the expression of BTLA in peripheral blood T lymphocytes. Impact statement This work is useful in the field of genetic mechanism of severe post-traumatic complications, as it provides important evidence for the influence of BTLA gene polymorphism on sepsis and MODS susceptibility. The results are useful and of importance because rs1982809 is a useful clinical biomarker in the prognosis evaluating of sepsis and subsequent MODS. It is also a functional single nucleotide polymorphism affecting the activity of BTLA 3ʹ-UTR and the expression of BTLA in peripheral blood T lymphocytes.
Structure and development of single domain antibodies as modules for therapeutics and diagnostics
Authors: Hoey Robert J , Eom Hyeyoung , Horn James R 
Published: October 2019

Since their discovery just over 25 years ago, the single variable domain from heavy-chain-only antibodies plays a role in an increasing number of antibody-based applications. Structural and biophysical studies have revealed that the small, ∼15 kDa, single variable domain found in camelids displays versatility in target recognition. Such insight has served as the foundation to develop and engineer VHH domains with enhanced properties capable of targeting a range of therapeutically relevant protein antigens or low-molecular weight haptens. Furthermore, the modular nature of VHH domains allows them to be introduced into constructs that are simply not possible with conventional antibodies. Here, we review the structural and biophysical properties of VHH domains, highlight recent VHH-based therapeutics and diagnostics, and provide insight into VHH engineering that may pave the way to next-generation single domain antibody applications.Impact statementThe development of novel antibody formats, beyond conventional antibodies, opens new possibilities in medical therapies, diagnostics, and general life science applications requiring affinity reagents. The camelid VHH domain, from heavy-chain-only antibodies, has emerged as a jack-of-all-trades module for novel affinity reagents. Applications include targeted cancer therapies, novel antimicrobial agents, conformation specific reagents, and tailor-made molecular switch entities. The breadth of unique uses for the VHH will continue to grow, opening new opportunities to treat and understand disease.
Evaluation of a Novel Wellness Assessment Device (Preventiometer): A Feasibility Pilot Study
Authors: Nanda Sanjeev , Mahapatra Saswati , Lindeen Stephanie A , Bernau Joleen L , Cutshall Susanne M , Schierwater Bernd , Chon Tony Y , Wahner-Roedler Dietlind L , Bauer Brent A 
Published: October 2019

Background Periodic wellness assessments can provide an estimate of a person’s relative risks for major diseases, but wellness visits are underused. Our suggestion is to use a comprehensive device during a single visit. Objective The goal of this pilot study was to evaluate the feasibility of a novel one-stop wellness device (Preventiometer; iPEx5 GmbH, Greifswald, Germany) for performing multiple tests and providing a comprehensive wellness assessment in a short period. Methods A Preventiometer was used to provide wellness assessments for 10 healthy volunteers who then answered a 25-question survey to rate their satisfaction with the testing and their overall impression. Results All volunteers agreed or strongly agreed with the following: The assessment reports were easy to understand, the Preventiometer met their satisfaction, the participants were comfortable during the assessment, and all measurements and testing were well coordinated. Participants liked the instant test result feature. Most (90%) agreed that the machine was useful for a quick health assessment for busy people, and 70% felt that it was time efficient. Conclusion In this feasibility pilot study, the Preventiometer performed multiple tasks and provided a comprehensive wellness assessment in a short period. Participants reported remarkably high satisfaction with the tests. A larger study is needed to prove that this is a pragmatic approach to help individuals improve their health.
Removal of Normal Competition Increases Proliferation of Pre-Leukemic Cells in a Mouse Model of Pre-B Acute Lymphoblastic Leukemia.
Authors: Reid Gregor SD , Barrett David , Seif Alix Eden , Brown Valerie I. , Grupp Stephan A. 
Published: October 2019

Abstract
Abstract 1430
Poster Board I-453
B cell precursor (BCP) acute lymphoblastic leukemia (ALL) is the most common childhood malignancy. While the peak age of onset for this disease is 4 years, abnormal cells that will eventually give rise to disease can be detected in blood taken from patients at birth. Interestingly, similar abnormal cell populations can also be detected in blood taken at birth from many children who do not go onto develop ALL. Understanding the factors that influence the survival and expansion or elimination of these abnormal pre-leukemic cells will shed light on the process of leukemogenesis and may suggest strategies for the prevention of BCP-ALL progression. We have used a mouse model of BCP-ALL to investigate the influence of normal immune cells on the pre-leukemic population. An abnormal BCP population arises in Eμ-RET (RFP) transgenic mice during fetal development, with overt leukemia developing between 3 and 9 months of age. We show that purified BCP from bone marrow of 4-week old pre-leukemic RFP mice undergo rapid expansion when adoptively transferred into immune-deficient mice, in many cases reaching white blood cell counts of >105 (with >90% showing the abnormal BCP phenotype). Secondary and tertiary transfer of expanded BCP into immune-deficient mice again produced a severe BCP lymphoproliferative disease. Such an expansion was not observed when purified BCP were transferred into wild-type mice or when unsorted pre-leukemic RFP bone marrow was transferred into immune-deficient mice. In addition, outgrowth of the abnormal cells was significantly curtailed if the purified RFP cells were mixed with a tenfold excess of wild-type bone marrow cells prior to adoptive transfer. This inhibition of BCP outgrowth was not achieved with Rag1-deficient bone marrow cells, suggesting that competition with normal adaptive immune cells for limited growth factors may impair the expansion of RFP pre-leukemic cells. Consistent with this hypothesis, blockade of the IL-7 receptor on pre-leukemic cells abrogated their expansion following adoptive transfer. As production of IL-7 is elevated in lymphopenic settings, we evaluated the impact of T cell-depletion on pre-leukemic cells in RFP mice. Pre-leukemic cell populations were significantly elevated in the spleen (31.3% +/- 15.4 vs 9% +/- 9, p<0.01) and blood (17% +/-19.2 vs 3% +/- 4.8, p<0.05) of T cell-depleted mice compared to controls. Taken together, these results suggest that competition for growth factors such as IL-7 may be a limiting step in the early expansion of pre-leukemic cell populations and may provide a mechanistic link between immune responses and pediatric ALL through the cytokine-mediated expansion of the pre-leukemic cell pool.

Disclosures
No relevant conflicts of interest to declare.
Purinergic Signaling Modulates Human Bone Marrow-Derived Mesenchymal Stem Cells Function.
Authors: Salvestrini Valentina , Ferrari Davide , Gulinelli Sara , Caione Luisa , Piacibello Wanda , Zini Roberta , Ciciarello Marilena , Manfredini Rossella , Lucchetti Giovanna , Ferrari Sergio , Virgilio Francesco Di , Lemoli Roberto M 
Published: October 2019

Abstract
Abstract 1441
Poster Board I-464
Nucleotides triphosphates are extracellular messengers binding to specific plasma membrane receptors (P2Rs) modulating a wide variety of biological responses in several cell types. In this study, we show that bone marrow-derived human mesenchymal stem cells (hMSCs) express several functional P2R subtypes at the molecular and protein level. hMSCs are very resistant to the cytotoxic effects of high concentrations of extracellular ATP, as demonstrated by the lack of morphological and mitochondrial changes or release of intracellular markers of cell death. Gene expression profiling revealed that hMSCs stimulated with ATP underwent a down-regulation of genes involved in cell proliferation, whereas those involved in cell migration were strongly up-regulated. Functional studies confirmed the inhibitory activity of ATP on proliferation of hMSCs and clonogenic stromal progenitors. Furthermore, ATP potentiated the chemotactic response of hMSCs to the chemokine CXCL12, and increased their spontaneous migration. In vivo, xenotransplant experiments demonstrated that the homing capacity of hMSCs to murine bone marrow was significantly increased by pre-treatment with ATP. Moreover ATP increased the production of the pro-inflammatory cytokines IL-2, IFN-gamma, and IL-12p70, while decreasing the anti-inflammatory cytokine IL-10. Thus, our data show that purinergic signaling modulates hMSCs functions and highlight a role for extracellular nucleotides in hMSCs biology.

Disclosures
Di Virgilio: F. Di Virgilio serves as a consultant for Cordex Pharma Inc. (USA) involved in the development of “P2 receptor-based drugs”. : Consultancy; F. Di Virgilio serves as a consultant for Affectis Pharmaceuticals AG (Germany), involved in the development of “P2 receptor-based drugs”. : Consultancy.
Stress Response Gadd45a Gene as Tumor Suppressor in MYC Expressing Myeloid Cells Is Cytokine Specific.
Authors: Mohamed-Hadley Alisha A , Liebermann Dan A. , Hoffman Barbara 
Published: October 2019

Abstract
Abstract 1481
Poster Board I-504
MYC, which regulates proliferation, apoptosis/survival and differentiation, is implicated in the etiology of a wide variety of hematologic malignancies. Multiple cooperating molecular pathways of cell survival and apoptosis determine if a cell lives or dies, and understanding how c-MYC interfaces with these pathways to influence the survival of cells is important to understand tumor initiation and progression, and response of tumors to different treatment regimens. Previously, this laboratory has shown that deregulated c-MYC blocks terminal myeloid differentiation and prematurely recruits both the Type I and Type II CD95/Fas apoptotic pathways, promoting an incompletely penetrant apoptotic response (Amanullah et al., Oncogene 19:2967-77, 2000; Amanullah et al., Oncogene 21;1600-10, 2002). Here we provide data to show that the response of myeloid cells to deregulated MYC expression depends on the status of the Gadd45 family of stress response genes. The gadd45 gene family plays pivotal roles as stress sensors that modulate signaling in response to physiological and environmental stressors, also modulating susceptibility of cells for transformation in vitro and tumor development in vivo. Gadd45 behaves as either tumor suppressor or oncogene depending upon the transforming oncogene and the cell type (Tront et al., Cancer Research 66:8448-54, 2006; Tront et al., manuscript in press). To elucidate the role Gadd45a plays in response to the proto-oncogene c-MYC in myeloid cells, bone marrow (BM) cells from wild type (WT) and Gadd45a null mice were retrovirally infected to constitutively express c-MYC. We showed that Gadd45a null BM expressing constitutive c-MYC exhibited less apoptosis than its WT counterpart in expansion media (IL-3, IL-6, SCF), demonstrating that Gadd45a is required for optimal MYC mediated apoptosis. In addition, enhancement of cell cycle progression was observed. Therefore, loss of gadd45a in conjunction with constitutive MYC expression results in enhanced proliferation. Furthermore, in GM-CSF treated cells the MYC–mediated block/delay in differentiation was more extensive in the Gadd45a null cells compared to similarly treated WT cells. Interestingly, the percent of apoptosis was higher in the Gadd54a null cells expressing constitutive MYC as compared to the WT counterpart. This observation was in contrast to the results seen in expansion media, suggesting that the role Gadd45a plays in the presence of deregulated MYC may be cytokine specific. Data will be presented to explain how gadd45 regulates both the apoptotic response, depending upon the specific cytokine, and differentiation of MYC-expressing myeloid cells. Furthermore, experiments to determine how loss of gadd45a influences MYC-mediated leukemia, including assessing the effect of manipulating the cytokine milieu, are currently underway.

Disclosures:
No relevant conflicts of interest to declare.
Antioxidant Defense to Hemolysis Is Organ-Specific and Reflects a Heterogeneity in Vascular Permeability in Sickle Cell Disease.
Authors: Ghosh Samit , Adisa Olufolake , Tan Fang , Ofori-Acquah Solomon F 
Published: October 2019

Abstract
Abstract 1537
Poster Board I-560
Sickle cell disease (SCD) is characterized by intravascular hemolysis generating cell-free hemoglobin at concentrations that exceed the scavenging capabilities of heme-binding plasma proteins. Heme is a major source of oxidative stress, which is widely known to increase vascular permeability and cause tissue edema. Barrier disruptive effects of oxidative stress can however be counterbalanced by adaptive antioxidant defenses. Hitherto, the identity of specific antioxidant enzymes and cognate mechanisms protecting individual organs from oxidative stress in SCD remains poorly defined. In this study, microarray analysis was performed using non-toxic concentrations of hemin to analyze mechanisms of antioxidant defense by the endothelium. Multiple candidate antioxidant enzymes were identified each differentially elevated in several major organs in anemic SCD mice compared to non-anemic heterozygote and hemizygote littermates. Remarkably, none of the antioxidants was elevated in the brain of sickle mice. Moreover, the antioxidant phenotype in the kidney, spleen and liver of sickle mice were predominantly acute while the sickle lung was characterized by a predominantly chronic antioxidant phenotype. This latter finding was confirmed in SCD patients with chronic lung disease. Antioxidant enzyme activity was significantly higher in the lungs of adult sickle mice aged 3-6 months than in younger mice aged 4-6 weeks (p=0.004). However, this enhanced antioxidant activity declined significantly in middle-age mice 11-13 months old (p=0.005). Vascular permeability assessed by extravsasation of Evans blue dye was normal in the brain of sickle mice of all ages in agreement with our data indicating absence of oxidative stress in this organ. On the contrary, vascular permeability in the lung, kidney and heart of adult sickle mice was abnormally high. This abnormality deteriorated significantly exclusively in the lung (p= 0.04) but not in the heart or kidney in middle-age mice. Increased lung permeability in middle-age sickle mice was confirmed by overt tissue edema determined by lung wet/dry weight ratios. Our study has shown for the first time that the antioxidant response to the systemic chronic hemolysis of SCD is organ-specific. Furthermore, we have identified decline of antioxidant reserve concomitant with tissue edema as potential age-dependant risk factors for fatal lung complications in SCD. Finally, our data provide a framework to develop targeted antioxidant therapies to preserve organ function in SCD.

Disclosures
No relevant conflicts of interest to declare.
Lipoprotein Cholesterol and Triglyceride in Children with Steady-State Sickle Cell Disease.
Authors: Seixas Magda Oliveira , Rocha Larissa , Carvalho Mauricio , Menezes Joelma , Lyra Isa , Nascimento Valma , Couto Ricardo , Atta Ajax , Reis Mitermayer Galvão , Goncalves Marilda Souza 
Published: October 2019

Abstract
Abstract 1547
Poster Board I-570

Introduction
Levels of high-density lipoprotein cholesterol have been correlated with anti-inflammatory, anti-oxidative, anti-aggregation, anti-coagulant and pro-fibrinolytic activities. We hypothesized that lipoprotein cholesterol and triglycerides have important roles in sickle cell disease pathogenesis.

Patients and Methods
A prospective study of biochemical and hematological analyses of 152 steady-state children with sickle cell disease and 132 healthy subjects using immunochemistry, immunoassay and electronic cell counter respectively. Clinical data were collected from patient medical records. Data analyses were performed using Prism 5.01 (Graphpad Software, San Diego, CA), EPIinfo 6.04 (CDC, Atlanta, Georgia) and STATA SE 10 software (StataCorp, Texas, USA).

Results
There was a significant positive association of high-density lipoprotein cholesterol with hemoglobin (p<0.001), hematocrit (p<0.001) and total cholesterol (p<0.001) and a negative association with reticulocytes (p=0.046), leukocytes (p=0.015), monocytes (p=0.004) and platelets (p=0.005), bilirubins [total bilirubin (p<0.001), direct bilirubin (p<0.001) and indirect bilirubin (p<0.001], iron (p<0.001), aminotransferases [aspartate aminotransferase (p=0.004), alanine aminotransferase (p=0.035)], lactate dehydrogenase (p<0.001), urea (p=0.030), alpha 1-antitrypsin (p<0.001), very low-density lipoprotein cholesterol (p=0.003), triglycerides (p=0.005) and hemoglobin S (p=0.002). Low high-density lipoprotein cholesterol concentration was associated with cardiac abnormalities (p<0.025), pneumonia history (p=0.033) and blood transfusion use (p=0.025). Triglycerides (p=0.047), very low-density lipoprotein cholesterol (p=0.044), low-density lipoprotein cholesterol (p=0.033), total cholesterol (p=0.007), alpha 1-antitrypsin (p=0.040) and ferritin (p=0.008) levels were associated with cholelithiasis.

Conclusion
We hypothesize that some SCD patients can have a specific dyslipidemic subphenotype characterized by hypertriglyceridemia, high VLDL-C and low plasma LDL-C and HDL-C in association with other biomarkers, including those related to inflammation. This represents an important step toward a more reliable clinical prognosis. We suggest further studies and continued research into new mechanisms involving this complex network of markers in order to establish their role in SCD pathogenesis.

Disclosures
No relevant conflicts of interest to declare.
FLT3, NPM1 and MLL Mutations Help Risk Stratification in Pediatric Acute Myeloid Leukemia.
Authors: Shen Shuhong , Liu Yin , Tang JingYan , Gu Long-Jun 
Published: October 2019

Abstract
Abstract 1574
Poster Board I-600

Introduction
Acute myeloid leukemia (AML) is a heterogeneous disease which harbors various genetic alterations. Among theses genetic events, Mutations of FLT3, NPM1, MLL and other genes often predict prognosis, particularly in cases cytogenetic normal (CN-AML). Could these be criteria for risk stratification in Pediatric AML ?

Patients and Methods
155 cases of de novo AML were diagnosed routinely according to morphology, immunology, cytogenetics, and molecular biology examination on bone marrow (BM) aspirates between Jan. 2002 and Dec. 2008. All patients received chemotherapy according to the AML-XH-99 protocol, which consist of Daunorubicin, Cytosine arabinoside, Etoposide, Homoharringtonine. For acute promyelocytic leukemia, all-trans retinoic acid and Arsenic trioxide were also included. Meanwhile, total RNA of leukemic cells form all diagnostic BM samples were extracted, and then reverse transcribed. MLL partial tandem duplication (MLL/PTD) fusion transcripts were screened by real-time quantitative polymerase chain reaction. FLT3 internal tandem duplication (FLT3/ITD), FLT3 tyrosine kinase domain mutation (FLT3/TKD) and NPM1 mutation were examined by High resolution melting analysis.

Results
Of the 155 children with de novo AML, 121(78.1%) had received chemotherapy for more than one week with data available for analysis. Among them, 55(45.5%) was cytogenetically normal (CN-AML). In this total cohort of patients 49(27.09%) had FLT3/ITD (32.70% in CN-AML), 14 (9.03%) had FLT3/TKD (7.30% in CN-AML), 62 (40%) had NPM1 mutation (49% in CN-AML), and additional 8 (5.16%) had MLL/PTD (5.50% in CN-AML). In this cohort of patients 98 (63.22%) had at least one mutation. The clinical outcomes were listed in table 1. Generally, patients with FLT3 mutation (ITD or TKD mutation) usually have worse results after chemotherapy, as reported previously by other researchers. Meanwhile, NPM1 mutations usually predict better prognosis in our cohort of AML patients. MLL/PTD always predicts the worst outcome in AML as other MLL rearrangements in leukemia. Among CN-AML patients, 5-year EFS and OS were similar to whole cohort of patients according to those mutations. Cox regression analysis in a univariate model revealed that the presence of FLT3/ITD and NPM1 was significant prognostic factor of EFS, (P<0.05). We therefore proposed a molecular-risk classification of pediatric AML patients based on the data we got in this study. For the newly classified groups of low, medium and high risk groups, EFS rate was 62.03%±8.42%, 45.42%±4.52%, and 14.85%±2.99%, respectively, P=0.00. CRD for the 3 groups was 27.69±21.34 months, 22.62±19.64 months, 13.26±11.95 months, respectively, p=.022. Our results indicate that combinations of these couple of molecular events may be the useful tool for further classify AML in children.

Disclosures
No relevant conflicts of interest to declare.