bluuebunny/crossref_metadata_2025_split huggingface dataset:
Artesunate Attenuate Chronic Graft-Versus-Host Disease By Regulating Th17/Treg Balance
Authors: Xiaomei Chen , Weng Jianyu , Lai Peilong , Huang Xin , Yulian Wang , Geng Suxia , Guo Liyan , Huang Tian , Zeng Lingji , Du Xin 
Published: October 2019

Abstract
To investigate the effects of Artesunate treatment on chronic graft-versus-host disease (cGVHD). Recipient BALB/cJ mice received 8 × 106 bone marrow cells with 8 × 106 spleen cells from B10.D2 mice. Artesunate solubilized in acetone was injected intraperitoneally every day at the dose of 1 mg/kg at Day 28 after transplatation. The clinical scores, survival times and histopathological damage were analyzed. The frequency of Th17 and Tregs in PB and spleens from the mice was evaluated by flow cytometry. In addition, we cultured the CD4+ T cells from the spleens of mice in vitro, and stimulated these cells with artesunate, the frequency of Th17 and Tregs in these splenocytes was evaluated by flow cytometry. Artesunate administration diminishes clinical and histopathological damage and improved the survival of cGVHD mice. Artesunate contributed to Tregs development (4.45±0.04 vs 8.40±0.23; 6.62±0.24 vs 10.48±0.48; p<0.05) while decreased Th17 cells (4.45±0.04 vs 8.40±0.23; 6.62±0.24 vs 10.48±0.48; p<0.05)expressions both in PB and spleens, and decreased the Th17/Treg ratio (0.34±0.05 vs 0.09±0.03; 0.19±0.03 vs 0.06+0.02; P<0.01. Moreover, artesunate suppressed the Th17 cells expressions (0.82±0.37% vs 3.39±1.22% p<0.01) and contributed to Tregs development (34.63±1.29% vs 14.28±1.69% p<0.001), and also decreased the Th17/Treg ratio 0.24±0.09vs 0.02±0.01; p<0.05) in vitro. Artesunate suppressed the Th17 cells expressions and contributed to Tregs development. These studies provide nowsights into the artemisinin and identify artesunate as a novel drug for cGVHD.

Disclosures
No relevant conflicts of interest to declare.
Stroke in a Pediatric Immune Thrombocytopenia Case
Authors: Leblebisatan Goksel , Cil Metin , Sasmaz Hatice Ilgen , Kilinc Yurdanur , Soyupak Sureyya 
Published: October 2019

Abstract
Objective: Immune thrombocytopenia is a bleeding disorder related to deficiency of the main element in primary hemostasis, platelets. But it rarely results in an opposite clinical complication as thrombosis. The mechanism of this unexpected complication is not fully understood still and the treatment of the patients in this thrombocytopenic and thrombotic condition is critical and hard to manage. Here we present a case of immune thrombocytopenia with cerebral infarct with his clinical management details.
Case: A seven-year old boy diagnosed with Immune thrombocytopenia at the age of 1 in another clinic and treated with IVIG when the platelet levels fell and bleeding symptoms appeared. The patient refered to our clinic with severe refractory thrombocytopenia. After being diagnosed with ITP, IVIG treatment and HDDXM treatment (20 mg/m2/day for 4 days) was given 2 cure in a month then the patient follow up in the clinic. It is learned that the family continued with HDDXM and sometimes IVIG treatment when needed in the local hospital. When the child was 6 years old, because of inability to walk and talk he was referred and it was revealed that he had cerebral infarct in the ICA area with a platelet level of 11000/mm3. He was followed in pediatric ICU. He was treated with thrombocytopenia with replacement and steroids in addition to physical treatment for stroke. Anticoagulation was used as the clinical and laboratory picture allowed.
Result: The patient was discharged with the better clinical Picture and his chronic treatment planned mostly with steroids.
Conclusion: Stroke in the immune thrombocytopenic clinic is a rare but hard to manage condition. Although its mechanism not understand well, the clinician should be identify and manage it carefully.

Disclosures
No relevant conflicts of interest to declare.
Protein Phosphorylation Mechanisms Underlying Chronic Pain in Sickle Cell Disease
Authors: He Ying , Wilkie Diana J. , Molokie Robert E. , DeSimone Joseph , Wang Zaijie J. 
Published: October 2019

Abstract
Pain is one of the most dreadful symptoms in sickle cell disease (SCD) and is often refractory to currently available analgesics. Besides acute painful vaso-occlusive crises, SCD is also accompanied by intractable chronic pain. This persistent, and often unrelieved, pain starts early in childhood and continues throughout life. The neurobiological mechanisms of chronic pain in SCD remain unclear, which markedly limits effective pain management and the quality of life in patients with SCD. Taking advantage of two humanized mouse models of SCD, this study aimed to investigate protein phosphorylation mechanisms for chronic pain in SCD.
We characterized pain in two transgenic mice models of SCD that exclusively express human alleles encoding normal α- and sickle β-globin. Berkeley SCD mice (BERK mice) and Townes' SCD mice (TOW mice) exhibited ongoing spontaneous pain behavior and increased sensitivity to evoked pain stimuli compared with littermate control mice expressing normal human hemoglobins. To investigate the underlying protein phosphorylation mechanisms of chronic pain in SCD, we examined PKC isoform mediated nociceptive signaling. Prominent activation of multiple PKC isoforms were observed in the superficial laminae of the spinal cord dorsal horn in BERK and TOW mice. Functional inhibition and silencing of specific PKC isoforms attenuated spontaneous pain, mechanical allodynia, and heat hyperalgesia in both transgenic SCD mice. Furthermore, employing hematopoietic stem cell transplantation approach, we were able to generate a sickle cell anemia model in PKC-null mice, allowing us to specifically target neuronal PKC in SCD. Neither spontaneous pain nor evoked pain was detected in the mice lacking specific PKC isoform despite the full establishment of SCD phenotypes.
In summary, this study is the first to identify the presence of ongoing spontaneous pain in preclinical sickle cell models, which closely mimic the most prevalent manifestation in patients with SCD. Moreover, we found that spinal PKC is a critical mechanism for the generation and maintenance of ongoing and evoked pain in SCD. These findings offer insights into sickle cell pain mechanisms, which may become a potential target for pharmacological interventions.

Disclosures
No relevant conflicts of interest to declare.
Natural Killer Cell Derived Exosomes As a Novel Therapeutic for Acute Myeloid Leukemia
Authors: Boyiadzis Michael , Hong Chang Sook , Whiteside Theresa L 
Published: October 2019

Abstract
Introduction: Natural killer (NK) cells play a critical role in the innate immune response through their capacity to lyse malignant cells without prior antigen-specific priming. NK cells have been evaluated for safety and efficacy in acute myeloid leukemia (AML), both in the transplant and non-transplant settings. Exosomes, small, 30-150 nm-sized extracellular vesicles originating from the endocytic compartment of parent cells, have recently emerged as a universal intercellular communication system. Exosomes are released by virtually all cells and carry proteins, lipids, and nucleic acids from the parent to recipient cells at short and long distances. The exosome molecular cargo reflects the content of the parent cell and is delivered to recipient cells in a membrane-protected vesicle. We hypothesize that exosomes produced by human activated NK cells carry the machinery necessary for the killing of leukemic cells.
Methods: Venous blood (20-50 mL) was obtained from healthy donors (n=10). NK cells were isolated using Ab-based immunomagnetic selection from the recovered peripheral blood mononuclear cells. NK cells were cultured in the presence of interleukin-2 and interleukin-15, and NK-cell supernatants were used for exosome isolation by size exclusion chromatography. Protein levels, numbers and size (qNano), and exosome morphology (transmission electron microscopy) were determined. Exosome cargos were studied by Western blots and/or flow cytometry for NK cell activating and inhibitory receptors, immune inhibitory molecules and for perforin and granzyme B. Cytotoxicity of the NK cell-derived exosomes for K562 targets, AML cell lines (Kasumi, MLL-1) and primary leukemia blasts was measured using flow cytometry-based assays.
Results: Activated human NK cells produced large quantities of exosomes. PKH-26-labeled NK cell-derived exosomes were avidly taken up by leukemic blasts. NK cell derived exosomes carried activating NK cell receptor NKG2D, natural cytotoxicity receptors, perforin, granzyme B, transforming growth factor beta (TGF-β), killer-cell immunoglobulin-like receptors and PD-1. NK cell derived exosomes mediated anti-leukemia activity against K562 targets, AML cell lines and primary leukemia blasts. Lysis of leukemic blasts by NK cell-derived exosomes was exosome concentration dependent.
Conclusion: We report that NK cell derived exosomes have anti leukemia activity in vitro. These data provide a foundation for the future development of new therapeutic strategies using NK cell-derived exosomes for the elimination of leukemia.

Disclosures
No relevant conflicts of interest to declare.
The Hundred-Person Platelet Releasate Project (100PREPs): A Barcode for Human Health and Disease
Authors: Maguire Patricia , Szklanna Paulina , Parsons Martin , Wynne Kieran , O'Connor Hugh , O'Connell Karen , Egan Karl , McGuigan Christopher , Rickard Scott , Ni Ainle Fionnuala 
Published: October 2019

Abstract
Background:
Upon activation platelets release a host of soluble and vesicular signals, collectively termed the 'platelet releasate' (PR). The contents of this PR play a significant role in haemostasis, wound healing, tissue regeneration, inflammation, and pathologic sequelae. We recently established that the proteome of the platelet releasate was highly reproducible across 32 healthy adult donors, with low variation in secretion levels for individual PR proteins. As there is accumulating evidence that circulating platelets may sense or be 'educated' by their environment, we hypothesised that the PR may store a relevant and bespoke collection of molecules during pregnancy and in platelet-related disease.
Methods:
PR from 18 healthy pregnant women, 22 pregnant women diagnosed with early-onset preeclampsia (EOP) (onset <34 gestational weeks), 15 people with relapsing-remitting multiple sclerosis and 15 people with primary-progressive multiple sclerosis were obtained with informed consent. After trypsin/lys-C double digest, each platelet releasate was individually subjected to label-free quantitative proteomic analysis. A machine learning approach based on differential peptides was used as a training set and then to predict outcome.
Results:
69 PR proteins were found to be differentially released from platelets in healthy pregnancy in comparison to age-matched non-pregnant female controls, including several pregnancy-specific proteins. A further 36 PR proteins were statistically modified in EOP, a disease associated with life-threatening consequences for both mother and baby, and another 26 proteins were altered in multiple sclerosis. Based on a training set of differential PR peptides, we developed several diagnostic algorithms, including a proof-of-concept pregnancy algorithm that could predict outcome with 100% accuracy. Strikingly, our proprietary algorithm based on several platelet releasate peptides could predict the severity of maternal outcome in preeclampsia. Moreover, again based on specific platelet-derived peptides, we could differentiate between clinical subtypes of multiple sclerosis.
Conclusion:
We found that the PR proteome significantly changes in physiologic and pathologic conditions. We conclude that the PR is a barcode for the health status of an individual at a given time and provides an easily accessible discrete set of peptides and proteins that can differentiate disease.

Disclosures
Ni Ainle: Leo Pharma: Research Funding; Actelion: Research Funding; Bayer: Research Funding; Bayer: Membership on an entity's Board of Directors or advisory committees; BMS: Membership on an entity's Board of Directors or advisory committees; Daiichi Sankyo: Membership on an entity's Board of Directors or advisory committees; Boehringer: Membership on an entity's Board of Directors or advisory committees.
Diffuse Alveolar Hemorrhage Secondary to Direct Oral Anticoagulants: A Systematic Review of Reported Cases
Authors: Abu Rous Fawzi , al-Abcha Abdullah , El-khatib Layan , Shrotriya Shiva , Dhakal Prajwal 
Published: October 2019

Abstract
Introduction:
Direct oral anticoagulants (DOACs), also known as non-vitamin K oral anticoagulants, include rivaroxaban, dabigatran, apixiban, edoxaban, and betrixaban. Guidelines have recently identified DOACs as the first line treatment in venous thromboembolism and nonvalvular atrial fibrillation. As the use of these anticoagulants has gradually increased in last few years, the reports of bleeding-related adverse drug events with these agents has increased. There are very few data related to diffuse alveolar hemorrhage (DAH) with DOACs, a dreaded complication of any anticoagulant.
Objective:
To perform a systematic review of all reported cases of DAH secondary to DOAC use.
Methods:
Using several search terms, we reviewed all cases of DAH associated with DOACs which were published and indexed in English language in PubMed and EMBASE in the last 10 years. The bibliography of each relevant article was searched for additional related reports. Statistical analysis was performed using Statistical Packages for the Social Science (SPSS).
Results:
A total of 34 cases were included in the study. Mean age was 69±15 years; 68% were male, 32% were female. Rivaroxaban, dabigatran, apixaban, and edoxaban were used in 41%, 35%, 21%, 3% of patients respectively. Atrial fibrillation was the commonest indication for DOAC use, 68%. Hemoptysis was the commonest presenting symptom, 74%. DAH was diagnosed with Computed Tomography (CT), Bronchoalveolar Lavage (BAL), and both in 68%, 56%, and 35% respectively. A total of 18 (53%) cases required mechanical ventilation. There was a total of 7 (21%) deaths, while 20 (59%) survived.
Conclusion:
Despite reports of lower risk of major bleeding compared to other anticoagulants, DAH is still a possibility with DOAC use. DOACs' absorption and elimination relies largely on P-glycoprotein (P-gp) efflux transporters, coadministration of P-gp inhibitors such as Amiodarone, a common antiarrhythmic used in atrial fibrillation in conjunction with anticoagulation, may result in disturbance of DOAC's concentrations in the blood, increasing the risk of potential side effects. Autoimmune diseases can lead to DAH independently. Owing to the diseased lung tissue in patients with autoimmune disease, the use of DOAC's may enhance the already present risk of DAH. Renal function plays a role in excretion of DOAC's, AKI may cause accumulation of DOAC's in the body leading to major side effects. Newer cases reported the use of idracuzimab; a newly FDA-approved drug for dabigatran reversal. In conclusion; caution should be taken with the increasing use of DOAC's in management of complex care patients requiring anticoagulation, prompt diagnosis and management is key for survival and prevention of complications, and new reversal agents may play a future role in the management of such complications.
Table. Table.

Disclosures
No relevant conflicts of interest to declare.
Comparative Analysis of Proinflammatory Cytokine Levels in Peripheral Blood and Bone Marrow of Patients with Myeloproliferative Neoplasms
Authors: Chen Pu , Wu Boting , Shao Xia , Liu Chanjuan , Yu Zhenglin , Li Feng , Cheng Yunfeng 
Published: October 2019

Abstract
Background
Myeloproliferative neoplasms (MPNs) are characterized by marker somatic mutations involving JAK2, MPL, and CALR, which lead to constitutive activation of tyrosine kinase signaling cascades, subsequently dysregulated proliferative of myeloid linages, and eventually myelofibrosis or leukemic transformation. Recently, it has been argued that proinflammatory processes are crucial to the pathogenesis and progression of MPNs. A number of proinflammatory cytokines including lipocalin, IL-1, IL-2R, IL-6, IL-8, IL-12, IL-15, and IP-10 have been found elevated in the peripheral blood (PB) of MPN patients. However, there has been limited data on the levels of proinflammatory cytokines in the bone marrow (BM) of MPN patients. The present study determined and compared 40 proinflammatory cytokine levels in the PB and BM plasma of MPN patients with unequivocal molecular background, thus intending to illustrate the proinflammatory features of BM microenvironment as well as to evaluate the credibility of PB cytokine profiles.
Methods
Newly diagnosed MPN patients (n=12, 8 had JAK2 V617F mutation, 4 had CALR mutations) were included in the present study. PB samples were taken within 48 hours of BM samples. Paired PB and BM plasma cytokine profiles were measured as well as 10 health control PB plasma samples in a single procedure by Quantibody Human Inflammatory Array 3 (RayBiotech, Norcross, GA) which permitted detection of 40 inflammation-associated cytokines.
Results
Among 12 MPN patients, 8 had JAK2 V617 mutation (6 males, median age 61.5 years), and 4 had CALR mutations (3 males, median age 53.5 years). Positive linear correlations between PB and BM levels were found in 12 proinflammatory cytokines including BLC (r=0.613, p=0.034), I-309 (r=0.872, p<0.001), IL-1α (r=0.666, p=0.018), IL-1β (r=0.929, p<0.001), IL-12p40 (r=0.642, p=0.024), IL-15 (r=0.608, p=0.036), M-CSF (r=0.906, p<0.001), MIG (r=0.596, p=0.041), MIP-1α (r=0.787, p=0.002), MIP-1δ (r=0.648, p=0.023), sTNFRI (r=0.827, p=0.001), and sTNFRII (r=0.644, p=0.024). Compared to health controls, BM levels of G-CSF, IL-2, IL-4, IL-6R, IL-7, IL-8, IL-10, IL-13, MIP-1β, PDGF-BB, RANTES, and TIMP-1 were markedly elevated (all p<0.01), among which IL-2, IL-4, and IL-7 levels were even higher in patients with CALR mutations than those with classic JAK2 V617F mutation (all p<0.05).
Conclusions
Optimal linear correlation could only be found in limited species of proinflammatory cytokines, especially I-309, IL-1β, M-CSF, and sTNFRI, between PB and BM plasma of MPN patients. Therefore, caution should be recommended during attempts to illustrate the status of inflammation in MPN patients by circulating cytokine markers. A stronger inflammatory component might exist in MPN patients with CALR mutations than those with classic JAK2 V617F mutation.

Disclosures
No relevant conflicts of interest to declare.
Targeted Delivery of CpG-Mir-146a Mimic Oligonucleotides As a Therapeutic Strategy to Reduce NF-Κb-Mediated Pathogenic Inflammation and Myeloid Leukemia Progression
Authors: SU Yu-Lin , Zhang Zhuoran , Mann Mati , Wang Xiuli , Nguyen Le Xuan Truong , Zhang Bin , Li Ling , Swiderski Piotr , Boldin Mark , Forman Stephen J , Marcucci Guido , Kortylewski Marcin 
Published: October 2019

Abstract
NF-kB signaling plays central role in the regulation of immune cell activity. The microRNA-146a-5p (miR-146a) provides negative feedback inhibition of the NF-kB pathway to prevent either excessive immunity, such as cytokine release syndrome. Low expression of miR-146a is also implicated in certain types of leukemia, especially in del(5q)-syndrome myelodysplastic and acute myeloid leukemia (MDS/AML). While miR-146a is a potential therapeutic target, the lack of efficient miRNA delivery methods limits clinical translation. We previously developed a strategy for targeted delivery of oligonucleotide therapeutics, such as siRNA, into human and mouse myeloid cells and B cells, using partly or completely phophorothioated (PS) single-stranded oligodeoxynucleotides containing CpG motif (CpG ODN). Here, we demonstrate that similar strategy, using CpG ODN optimized for targeting human monocytes, can be employed for the delivery of functional miR-146a mimic. The CpG-miR146a mimic conjugate, but not unconjugated miR-146a, was quickly internalized by target human and mouse myeloid cells, such as dendritic cells, macrophages and leukemic cells. In vitro CpG-miR146a mimic reduced protein levels of downstream targets, Irak1 and Traf6, as well as the inhibition of DNA binding and transcriptional activity of NF-kB in myeloid cells. We further verified functional activity of CpG-miR146a mimic against several models of acute inflammation. The intravenous injections of this oligonucleotide reduced myeloproliferation and excessive cytokine response to bacterial endotoxin or to live Listeria challenge in miR-146a-/- mice. Furthermore, CpG-miR146a mimic had similar anti-inflammatory effect also in a human model of cytokine release syndrome induced by CD19 chimeric antigen receptor (CAR) T-cell activity. CpG-miR146a effectively abrogated CAR T-cell induced IL-6 production by human monocytes in vitro. Finally, our studies indicated that CpG-miR146a mimic can target del(5q) MDS and AML cells in vitro and in vivo resulting in growth inhibition of MDSL and HL-60 acute myeloid leukemia. Our results suggest that CpG-miR146a strategy can be provide a new-in-class immunomodulatory therapeutics for treatment of acute inflammatory disorders, myeloproliferative diseases and myeloid leukemia.

Disclosures
Wang: Mustang Therapeutics: Other: Licensing Agreement, Patents & Royalties, Research Funding. Forman:Mustang Therapeutics: Other: Licensing Agreement, Patents & Royalties, Research Funding. Kortylewski:N/A: Patents & Royalties: I am an inventor on the US patent 9,976,147.