bluuebunny/crossref_metadata_2025 huggingface dataset:
Customization design method for complex product systems based on a meta-model
Authors: Li Rong , Zhang Haizhu , Qin Shengfeng , Ding Guofu , Han Xin
Published: October 2019

In order to effectively reuse the design knowledge of product family life cycle development and support holistic and rapid individual product design, this article presents a new meta-model-based systemic customization design method for complex product systems within a product-pedigree. The proposed method aims to synthetically analyze the common and adaptive customer demands and product features of a product-pedigree of complex product systems and to quickly respond to the changing demands based on knowledge accumulation in the field of customization design. The key to implement such a method is (1) to construct a product-pedigree-oriented product meta-model with a four-layered architecture where it is possible to achieve a high degree of abstraction of product and (2) to develop a special technique for configuring the meta-model of the complex product systems. We have tested the proposed method with the rapid design of product-pedigree of a high-speed train’s bogies as an illustrative example. In this work, a rapid customization design prototype system has been developed and applied to the design of a high-speed train’s bogie to illustrate how to construct a product meta-model and how to conduct configuration design on different layers and variant design for generating new products.
Women’s behavior as implied consent: Male “reasonableness” in Australian rape law
Authors: Burgin Rachael , Flynn Asher
Published: October 2019

Defendants in rape trials rely on narratives of “implied consent,” situating women’s ordinary behavior as having indicated consent. Such narratives ignore women’s experiences, instead describing a male perpetrator’s subjective interpretation or inference of the woman’s actions. Implied consent narratives should have been eliminated by law reform introducing affirmative consent that redirected attention to steps that the perpetrator took to ensure the other party was consenting. Drawing from an Australian study, this article uses rape trial excerpts from the state of Victoria to argue that implied consent narratives endure in rape trials and form the key factor shaping a reasonable belief defense. Rape law allows men to interpret women’s behavior without restriction, providing evidence of the persisting influence of misogynistic views of women in law and legal practice. This article contributes to feminist jurisprudential and theoretical efforts to generate understandings of the ways rigid gender norms are enacted and performed in rape trials.
Counter-memory and and–and: Aesthetics and temporalities for living together
Authors: Tello Verónica
Published: October 2019

This essay traces a critical genealogy of counter-memory – spanning critical theory, film and contemporary art – bound to what Rosi Braidotti terms nomadic subjectivity. Engaging with the work of feminist and postcolonial theorists and artists, this essay charts the import of nomadic subjectivity as a method for staying with the many times and histories of global contemporaneity. It aims to move beyond thinking of counter-memory as simply a means to maintain or register erased and/or contested histories, or as a dialectical mnemonic system. It charts an alternative concept of counter-memory, one that is post-dialectical, not bound to the formulas of either/or, us/them or self/other, but which is instead committed to the endless accumulation and proximities of things – the and–and.
Attitude control of tiltwing aircraft using a wing-fixed coordinate system and incremental nonlinear dynamic inversion
Authors: Binz F , Islam T , Moormann D
Published: October 2019

In this paper, we present a novel concept for robustly controlling the attitude of tiltwing aircraft. Our main contribution is the introduction of a wing-fixed coordinate system for angular acceleration control, which forms the basis of a simple and robust attitude controller. Using the wing-fixed coordinate system allows us to describe the actuator effectivity using simple approximations based on the current operating conditions of the aircraft. Coupled with a robust angular rate control concept, which does not rely on an accurate aerodynamic model, we present a controller stabilizing the entire flight envelope of a tiltwing aircraft. The underlying angular acceleration controller uses the concept of Incremental Nonlinear Dynamic Inversion (INDI) to achieve robustness against aerodynamic uncertainties. The resulting controller is evaluated in both simulation studies and flight tests.
Recurrent Thymoma-Associated Paraneoplastic Encephalitis Resulting From Multiple Antibodies: A Case Report
Authors: Safadi Amy Li , Wang Tian , Maria Gianluca Di , Starr Amy , Delasobera Bronson E. , Mora Carlos Alberto , Tornatore Carlo
Published: October 2019

Few reports describe the clinical course and acute-care management of patients with recurrent multi-antibody paraneoplastic encephalitis. We describe a rare case of a patient having thymoma with multiple paraneoplastic syndromes who was found to have antibodies to α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) followed by N-methyl-d-aspartate (NMDA) receptor in the setting of residual thymic tissue. He initially presented to the hospital with severe, rapidly progressive encephalitis with simultaneous antibodies to AMPA and voltage-gated potassium channel complex receptor. Brain magnetic resonance imaging revealed scattered white matter hyperintensities and an enhancing lesion adjacent to the left caudate. Computerized tomography showed an anterior mediastinal mass that was resected and revealed to be a thymoma. He was refractory to treatment with intravenous immunoglobulin, high-dose steroids, and plasmapheresis. He was then started on monthly cyclophosphamide. After 3 cyclophosphamide infusions, he began to show improvement in his alertness, ability to speak, and capacity to follow commands. One month later, he was readmitted to the hospital for new and unusual behavioral outbursts and agitation. He was found to have new anti-NMDA receptor antibodies in his cerebrospinal fluid in the setting of residual hyperplastic thymic tissue that required another resection. He was treated with rituximab and then cyclophosphamide (due to an infusion reaction with rituximab) with positive outcomes. The presence of multiple antibodies may be associated with poor prognosis, requiring prompt recognition and aggressive immunosuppressive treatment. New neurological symptoms should prompt a search for residual pathologic tissue or tumor recurrence causing new autoantibodies and additional paraneoplastic syndromes.
Targetoid form of polymorphic eruption of pregnancy: A case report
Authors: Bohdanowicz Michal , Ghazarian Danny , Rosen Cheryl F
Published: October 2019

Polymorphic eruption of pregnancy, formerly known as pruritic and urticarial papules and plaques of pregnancy, is an uncommon cutaneous eruption that can affect women during their third trimester of pregnancy. As its name implies, it has a variety of morphologies and it is important to consider other diagnoses, such as pemphigoid gestationis, which polymorphic eruption of pregnancy can mimic. Sometimes, as in this case, polymorphic eruption of pregnancy can have a targetoid morphology reminiscent of erythema multiforme. A thorough workup and conservative management plan helps reassure the patient that the correct approach is being taken during the challenging period of a pregnancy nearing completion.
Impact of Age and Subscapularis Tendon Reparability on Return to Recreational Sports Activities and 2-Year Outcomes After Reverse Total Shoulder Arthroplasty
Authors: Godin Jonathan A. , Pogorzelski Jonas , Horan Marilee P. , Lacheta Lucca , Hussain Zaamin B. , Altintas Burak , Frangiamore Salvatore , Mannava Sandeep , Fritz Erik M. , Millett Peter J.
Published: October 2019

Background:

As the indications for reverse total shoulder arthroplasty (RTSA) have continued to expand, the average age of patients undergoing RTSA has decreased.

Purpose/Hypothesis:

The purpose of this study was to report the minimum 2-year outcomes after RTSA and to evaluate the impact of surgical variables on outcomes. We hypothesized that younger patients, patients with larger glenosphere, and patients with irreparable subscapularis tendons would experience worse subjective patient-reported outcome scores (PROS) and that younger patients and those with a reparable subscapularis would demonstrate a higher rate of return to recreational sports activities.

Study Design:

Cohort study; Level of evidence, 3.

Methods:

Patients who underwent RTSA by a single surgeon between November 2005 and September 2014 were considered for this study. Patient characteristics, surgical details, PROS, and rates of return to recreational sports activity were prospectively collected and retrospectively reviewed. We assessed correlations between PROS and both patient age and subscapularis reparability. PROS collected included the American Shoulder and Elbow Surgeons (ASES) score, Short Form 12 (SF-12), Quick Disabilities of the Arm, Shoulder and Hand (QuickDASH) score, Single Assessment Numeric Evaluation (SANE), and postoperative patient satisfaction.

Results:

A total of 110 patients with an average age of 68.0 years (range, 45-87 years) were included. Minimum 2-year follow-up was obtained in 94 patients (85.4%), with a mean follow-up of 3.6 years (range, 2.0-9.3 years). The subscapularis was reparable in 58 patients (57.4%). All PROS significantly improved from pre- to postoperatively ( P < .001). The median patient postoperative satisfaction was 9 out of 10 (range, 1-10). Increasing patient age was correlated with significant improvements in QuickDASH and ASES scores. Postoperatively, 76.1% (67/88) of patients were able to return to recreational sports activity. However, of those who did return, 68.2% indicated that they had to modify their activity in some way. Glenosphere size was not significantly associated with PROS or the rate of return to recreational sports activity, but patients with reparable subscapularis tendons demonstrated higher PROS and return to activity rates.

Conclusion:

Patients who had a reparable subscapularis tendon showed a significantly higher rate of return to recreational sports activity than patients who had a irreparable subscapularis tendon. Older age was correlated with greater improvement in several, although not all, patient-reported outcome measures. Glenosphere size was not significantly associated with higher PROS or rate of return to recreational sports activity.
Do More-Assimilated Latinxs Leave the Barrio and Move to “White” Neighborhoods? Latinxs, Young Adults, and Spatial Assimilation
Authors: Goldsmith Pat Rubio , Puga Martin
Published: October 2019

Studies of Latinx–white residential segregation and of Latinx residential attainment consistently report findings consistent with spatial assimilation. Nevertheless, most studies of this theory use statistical models that cannot account for multiple dimensions of neighborhoods that may influence residential attainment. In this article, we test predictions of the spatial assimilation model using discrete choice analyses, a multidimensional model. We use data from the National Educational Longitudinal Study on the residential attainment of 1,080 Latinx young adults, most of whom have recently left their parents’ homes. After accounting for the multiple dimensions of neighborhoods, we find little evidence that assimilation from income, generation, or barrio background influences young adult residential attainment. The consequences of language assimilation are modest. However, we find that Latinx young adults with a bachelor’s degree live in “whiter” and “less Latinx” neighborhoods than those without a BA net of these multiple dimensions. The findings suggest that increased assimilation among Latinx young adults is unlikely to improve their residential attainment unless it specifically includes greater education.
View from Helm: Corporate Governance Through HR Prism
Authors: Hota Deepak Kumar
Published: October 2019

The rigorous corporate governance frameworks in Company Law and Securities and Exchange Board of India regulations, have not prevented incidents of fraud in listed companies in India. The top management should therefore play a critical role, by fostering desirable organisational culture, clarifying clear boundaries of organisational behaviour and also enunciating value statements and requisite policies in the company.

Bharat Earth Movers Limited (BEML) (see https://www.bemlindia.in/ ) which is in a tough and competitive business environment, has codified its values statement ‘BEML FIRST’ (Focus on Customer, Innovation & Technology, Reliability & Quality, Speed & Responsiveness and Trust & Teamwork). Taking support of its committed workforce and leveraging technology, the company has set high standards in corporate governance, and has zero tolerance to unethical behaviour. BEML has a well-established Enterprise Resource Planning (ERP), an online file management system and a committee of functional directors which plays a major role in taking critical decisions, bringing in transparency in business processes and decision-making in the company.
Modeling Fanconi Anemia Using Human Induced Pluripotent Stem Cells By Reversible Complementation
Authors: Rowe R. Grant , Marion William , Ruiz-Torres Sonya , Lummertz da Rocha Edroaldo , Girvan Yosra , Schlaeger Thorsten , North Trista E. , Wells Susanne , Daley George Q.
Published: October 2019

Modeling of Fanconi anemia (FA) using human induced pluripotent stem cells (iPSCs) has been hindered by the requirement for an intact FA DNA repair pathway for effective reprogramming and maintenance of pluripotency in patient-derived iPSCs. Temporary complementation of FA pathway defects could permit effective reprogramming, with removal of complementation upon directed differentiation to hematopoietic stem and progenitor cells (HSPCs) to model the hematopoietic phenotypes of FA. In this study, we used three FA patient-derived iPSC lines engineered with doxycycline inducible complementation of disease-causing FANCA mutations. We maintained complementation with doxycycline exposure in the pluripotent state and during morphogen directed differentiation of hemogenic endothelium (HE) within embryoid bodies, which possesses the capacity to differentiate to HSPCs. Upon initiation of the endothelial-to-hematopoietic transition (EHT) in purified iPSC-derived HE, doxycycline was either removed or maintained in culture to either inactivate or sustain FANCA expression. Morphologic EHT and emergence of CD34+CD45+ immunophenotypic human HSPCs were not affected by the status of FANCA expression, which allowed for the isolation of otherwise isogenic FANCA-expressing and FANCA-deficient HSPCs for disease modeling. FANCA-deficient HSPCs were unable to form FANCD2/γH2AX foci relative to FANCA-expressing HSPCs in response to genotoxic stress, confirming impairment of the function of the FA pathway. We found that uncomplimented, FANCA-deficient HSPCs showed impaired cell cycle progression, increased apoptosis, and markedly decreased colony forming activity in methylcellulose compared to complemented, FANCA-expressing HSPCs. Unexpectedly, we also found that FANCA-deficient HSPCs showed accelerated erythroid differentiation compared to cells with an intact FA pathway. RNA sequencing analysis showed enrichment of signatures of normal HSPCs in complemented HSPCs, while uncomplimented HSPCs showed signatures of more mature hematopoietic cells and cells undergoing erythroid differentiation. Together, these findings demonstrate the utility of reversible complementation in modeling FA with patient-derived iPSCs, and provide an inexhaustible source of otherwise isongenic complimented and uncomplimented human FA HSPCs for use in the investigation of new therapies.

Disclosures

No relevant conflicts of interest to declare.
Lipoprotein(a) a Risk Factor for Venous Thrombosis and Pulmonary Embolism in Patients Younger Than 50 Years of Age
Authors: Nguyen Sonha , Ilano Lynette , Oluoha Nneka , Pakbaz Zahra
Published: October 2019

Introduction: Despite convincing evidence toward causal role of Lipoprotein(a) in occlusive arterial disease, the data is conflicting when it comes to venous thromboembolism (VTE) and pulmonary embolism (PE). Also it is not known if intervention to normalize Lipoprotein(a) will decrease risk of recurrent VTE and PE eliminating the need for life long anticoagulation. To our knowledge this isfirst data set report focusing on evaluating association between Lipoprotein(a) and VTE in patients younger than 50 years old. Methods: Inthis retrospective study, chart review was completed for twenty-six consecutive patients referred to hematology clinic with diagnosis of deep vein thrombosis (DVT) or PE in year 2017-2018 . Four patients older than 50 years old at the time of acute events were excluded. Lipoprotein(a) had only been measured in patients who had negative hypercoagulable work up with normal lipid panel but had obesity . Protein C, S and anti-thrombin were not measured if patients were already on anticoagulation. Serum level of Lipoprotein(a) greater than 75 nmol/L was considered to be elevated. Results: Total of 22 patients (18 females) were included in the data analysis. Nine patients had DVT, 5 patients had PE, and 8 patients had both DVT and PE. Median age was 34 (12-47). Lipoprotein(a) level was not checked on eight patients who had SLE (n=1), surgery (n=1), Factor V Leiden mutation (n=1), protein S deficiency (n=1), anti-phospholipid syndrome (n=2), prothrombin gene mutation (n=1), and one patient who had lost follow up. The median Lipoprotein(a) level was 107 (8-276). Serum Lipoprotein(a) was elevated in 8 out of the 14 screened patients (57%) with the median of 135 (107-276). Out of 8 patients with elevated Lipoprotein(a), only 1 patient had additional clinical risk factors for thrombosis (history of smoking, alcohol abuse, hypertriglyceridemia and elevated LDL). In an attempt to normalize Lipoprotein(a) level, 3 patients were started on niacin but only one tolerated maximum 1000 mg niacin daily which resulted in decrease in level but did not achieve normalization. Conclusion: This data suggests that elevated serum Lipoprotein(a) in females younger than 50 years old is associated with DVT/PE events. Further investigation is required to confirm this finding. At this time it is not known if attempt to normalize Lipoprotein(a) will prevent recurrent PE/DVT and eliminate need for long life anticoagulation in patients with unprovoked VTE who have elevated lipoprotein(a).

Disclosures

Pakbaz: Novartis: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau.
The Clinical Characteristics and Efficacy of Chronic Myeloid Leukemia Patients with T315I Mutation
Authors: Chen Chen , Xu Na , WanEr Wu , Liang Liu , Zhou Xuan , JiXian Huang , Changxin Yin , Cao Rui , Liu Qifa , Xiaoli Liu
Published: October 2019

Background: The application of tyrosine kinase inhibitor (TKIs) has greatly improved the overall survival (OS) and quality of life of chronic myeloid leukemia (CML) patients.However, in the TKIs era, 10% to 25% of CML patients still develop TKIs resistance, and ABL kinase point mutations are the most common reason.Most of the ABL kinase region mutations resistant to imatinib could be alleviated by second generation TKIs, but the T315I mutation resistance to the first and second generation TKIs. Ponatinib is a multi-target tyrosine kinase inhibitor, which belongs to the third generation of TKI inhibitors,and is sensitive for CML or Ph postitive ALL patients with T315I mutation. But,how to apply ponatinib bridging graft or whether ponatinib preventive therapy is needed after transplantation is uncertainty.

Methods: 18 CML patients with T315I mutation detected by ABL1 kinase region mutation in Southern Hospital from March 2013 to April 2018 were retrospectively analyzed.G-banding method was used for chromosome analysis and real-time quantitative PCR method was used to detect mutations in ABL1 kinase region by BCR-ABL1 fusion gene Sanger sequencer.

Result:18 CML patients with T315I mutation :13 cases chronic phase (CP) ,2 cases in accelerated phase,3 cases in blastcrisis phase(BP); 9 cases in high risk group, 6 cases in middle risk group and 3 cases in low risk grou by Sokal score score system.15 patients by imatinib ,3 patients first-line treatment with dasatinib .In imatinib group, 13 cases conversed to dasatinib because of drug resistance or intolerance, and 5 cases (5 / 13) were converted to ponatinib because of T315I mutation.One case in dasatinib group converted to ponatinib because of T 315 I mutation.A total of 6 patients (6 / 18) were treated with ponatinib. 6 patients (6 / 18) treated by allogeneic hematopoietic stem cell transplantation (Allo-SCT).The median stage of T315I mutation was 12.5 m from the beginning of treatment to the detection of T 315I mutation in 18 patients.At the end of the follow-up, 8 cases died of recurrence and 10 survived: (CMR 2 cases, CHR 1 cases, PR 3 cases, NR 3 cases, 1 cases not regularly followed up, unable to evaluate the disease state), including 6 patients with PO treatment.

Conclusion:The point of T315I mutation was detected in patients with CML resistance after long-term sequential therapy frequently. The recurrence rate was still high even if these patients experience allogeneic hematopoietic stem cell transplantation.However,these patients treatment with ponatinib before and after transplantation maybe reduce the recurrence rate and improve prognosis.

Key words:Chronic myeloid leukemia;BCR-ABL;T315I;ponatinib.

Disclosures

No relevant conflicts of interest to declare.
CHD1 - a Novel Epigenetic Regulator in Myeloid Malignancies with a Role in DNA Repair
Authors: Sinha Arpan , De La Garza Adriana , Verma Amit , Frazer J. Kimble , Bowman Teresa V.
Published: October 2019

Myelodysplastic syndromes (MDS) and acute myeloid leukemias (AML) are clonal hematopoietic disorders resulting from genetic alterations in hematopoietic stem cells. These myeloid disorders are clinically heterogeneous and biologically complex. Despite major advances in understanding the genetic and molecular landscape of MDS/AML, along with the introduction of newer and targeted therapies, the cure rates in AML are still only about 60% in children, and much lower in adults. Exploiting the genetic tractability of the zebrafish (Danio rerio) vertebrate model, we are investigating the role of a novel epigenetic regulator, Chromodomain helicase DNA binding protein-1 (CHD1) in hematopoiesis and its misregulation leading to MDS and AML. CHD1 is located at chromosome 5q21, which lies within the most frequent breakpoints seen with the deletion of the long arm of chromosome 5 [del (5q)] in patients with MDS and AML. In addition to del (5q), we found that CHD1 levels are significantly lower in bone marrow cells of patients with other forms of MDS, relative to normal controls. Using CRISPR/Cas9-mediated targeted mutagenesis in zebrafish, we created chd1 homozygous mutant fish. We confirmed a marked decrease in chd1 gene expression in these mutant fish. Chd1 homozygous mutants are viable and fertile as adults, with no significant developmental or hematopoietic phenotypes observed during embryogenesis. As CHD1 can act as a tumor suppressor and is linked to the DNA damage response, we hypothesized that chd1 mutant zebrafish would be more sensitive to DNA damaging agents. Indeed, we found that chd1 mutants have increased sensitivity to ionizing radiation as evidenced by elevated brain cell death measured by whole mount imaging of live embryos and immunofluorescence for activated Caspase 3, a marker of apoptosis. We also generated chd1het; tp53het zebrafish to test whether chd1 haploinsufficiency could accelerate tumor rates in tp53 mutant fish. Single heterozygotes chd1het or tp53het usually do not form tumors at one year of age, but chd1het; tp53het double heterozygous zebrafish showed substantial tumor growth by one year of age. Taken together, our data suggest that CHD1 may play a key role in protecting genomic integrity, explaining why diminished CHD1 levels could contribute to the pathogenesis of MDS and AML. This genetic interaction may be especially crucial in patients with combined del (5q) and TP53 alterations, and could contribute to the increased severity seen in this group. Our findings suggest these and other CHD1-deficient patients may be resistant to standard therapies due to attenuated DNA damage responses, allowing their AML to survive DNA damage caused by conventional anti-cancer treatments.

Disclosures

No relevant conflicts of interest to declare.
A Restructured Approach to Diagnosis of Heparin Induced Thrombocytopenia in a Large Tertiary Hospital
Authors: Swarup Sriman , Ball Somedeb , Adhikari Nimesh , Welch Courtney Alice , Fackrell Jaden , Thein Kyaw Zin , Sultan Anita , Swarup Khatrina , Awasthi Sanjay , Tijani Lukman
Published: October 2019

Background

The standard practice for diagnosis of heparin-induced thrombocytopenia (HIT) involves a combination of 4T score and laboratory tests, such as enzyme immunoassay for detection of antibodies. We noted a lack of widespread use of 4T score in our practice setting. We also found that our laboratory utilized Particle Immunofiltration Assay (PIFA) for HIT screening, which has been shown to have questionable diagnostic utility in HIT diagnosis (Warkentin et al., 2007). The study aims to improve the rate of 4T score usage in conjunction with an improved laboratory diagnostic test for patients with suspected HIT in a cost-efficient manner.

Method

We initiated a quality improvement project involving the review of all patients with laboratory orders for PIFA testing between March 2017 to March 2018, explicitly assessing for documented 4T scores before the ordering of PIFA. Three of the investigators also calculated 4T scores for these patients at the time of laboratory testing and noted the results of the serotonin release assay (SRA), if ordered. We further collected data on any alternative anti-coagulation used in such patients for a cost-efficacy analysis later.

Results

A total of 170 PIFA tests were ordered during the period of investigation. Only five (0.02%) of these patients had a documented 4T score at the time of testing. One hundred thirteen patients (66.4%) had a low 4T score per investigator-calculation. Forty-seven patients (27.6%) were noted to have intermediate 4T scores. Lastly, ten patients (0.05%) were observed to have high 4T scores. A total of 32 SRAs were ordered; five of which were positive (four with an intermediate 4T score and one with high 4T score). PIFA was false-negative in two confirmed cases of positive SRA and false-positive in 13 instances of negative SRA. Thus, in this study, the sensitivity of PIFA was noted to be 60%, and specificity was observed to be 50%. Nineteen patients also received alternative parenteral anti-coagulation (fondaparinux or argatroban); seven of these were with low, eight with intermediate, and four with high 4T scores.

Conclusion

The study highlights the need for improving 4T score usage rates in our hospital as well as a need for switching to an alternative HIT screening test to promote patient safety and cost efficacy. Hence, we have begun the integration of 4T score with laboratory testing into the electronic medical record, alongside a shift in our HIT screening test from PIFA to the recently FDA-approved automated latex immunoturbidimetric assay. We will be continuing analysis of patients with suspected HIT for another six months to assess the effects of the above interventions.

Figure. Figure.

Disclosures

No relevant conflicts of interest to declare.
Adenosine Regulation of cAMP through Phosphodiesterases
Authors: Teng Bunyan , Darlington Daniel N , Cap Andrew P
Published: October 2019

Introduction: Adenosine, an autacoid and metabolite of ATP, has been known to have anti-platelet properties. Of the 4 adenosine receptors (ARs), only A2A AR have been implicated in adenosines anti-platelet properties in human. A2A AR is a G-Protein Coupled Receptors associated with a stimulatory G-Protein (Gs) that can activate adenylyl cyclase (AC) and increase intracellular cAMP. An elevation of cAMP has been shown to inhibit platelet aggregation to natural stimuli. Regulation of intracellular cAMP is balanced between synthesis by adenylate cyclase and degradation by phosphdiesterases (PDE). There are 3 PDE subtypes found in platelets: PDE2, PDE3, and PDE5. However, it is not know which subtype(s) is (are) responsible for regulating cAMP level in human platelets after adenosine stimulation.

Materials and Methods: Platelet-rich plasma (PRP) was isolated from whole blood of human volunteers, and centrifuged at 200g for 10min. Light transmission aggregometry was performed after stimulation of platelets with 100uM ADP, with or without NECA (non-specific AR agonist), DPCPX (A1 AR antagonist), and Sch 58261 (A2A AR antagonist). PRP treated with NECA, DPCPX, Sch 58261, and PDE inhibitors (EHNA, E in figures, for PDE2, Trequinsin, T in figures, for PDE3, and 4-{[3'4'-(methylenedioxy) benzyl]amino}-6-methoxyqunazolin, 4 in figures, for PDE 5). Cyclic AMP was measured in platelets after treatment by liquid chromatography/ Tandem Mass Spectroscopy (Quantiva, ThrermoFisher) after treated with these drugs.

Results: ADP-induced platelet aggregation was inhibited in a dose dependent manner by the non-specific adenosine agonist, NECA (Figure 1) and the effect was blocked by A2A specific antagonist Sch 58261, not by the A1 AR antagonist, DPCPX (Figure 2). NECA inhibition of platelet aggregation was likely due to an elevation of intracellular cAMP (1 uM, 5min incubation, Figure 3). Inhibition of PDE3 alone, significantly increased intracellular cAMP, suggesting that basal PDE3 activity is present. PDE 3 inhibition combined with NECA elevated cAMP even higher than PDE inhibition or NECA alone (Figure 3), suggesting that NECA (A2A stimulation) effects PDE activity. Inhibition of PDE2 or 5 had no effect on basal or NECA stimulated cAMP (Figure 3). Inhibition of all 3 PDE (2,3,5) combined with NECA elevated cAMP to levels higher then NECA+ PDE3 inhibition, again suggesting that NECA maybe effecting the activity of the PDEs (Figure 3). The potentiation of cAMP by PDE3 inhibition + NECA was block by A2A, but not A1 antagonist (Figure 4) suggesting that the nonspecific adenosine agonist is elevating cAMP through A2A.

Conclusion: 1. In human platelets, NECA stimulates cAMP through A2A receptors and this elevation is likely due to an elevation in adenylate cyclase via Gs coupled to A2A. PDE3 is basally active and likely regulated by adenosine receptors.

Disclosures

No relevant conflicts of interest to declare.
Protective Effect of Thrombopoietin on CoCl2-Induced Apoptosis of HUVEC Cells through PI3-k/Akt Pathways
Authors: Ye Jieyu , Wang Jun-yan , Liang En-yu , Yang Mo
Published: October 2019

Endothelial damage is a serious syndrome following by stem cell transplantation. However, a useful treatment for attenuating the periods of this disorder was still under explored. Our previous studies have demonstrated that in patients with acute inflammation states, serum thrombopoietin (TPO) levels were significant higher compared with healthy subjects (181.11 ± 35.38 vs 96.13 ± 9.7 pg/ml, p<0.001), which may act as an acute response protein to protect the body. TPO is now recognized as a potent proliferative factor for outer hematopoietic system, such as cardiomyocytes and neurocytes. This study is aim to investigate the protective effect of TPO on human endothelial cells, and to explore its potential mechanism. Four experimental groups were set up using human umbilical vein endothelial (HUVEC) cells, which are: Normal control, CoCl2 alone (800 μmol/L), TPO(100 μg/L) + CoCl2 and TPO alone groups. CoCl2 alone decreased the viability of HUVEC cells in a dose dependent manner (r=-0.997), while addition of TPO significantly rescued this effect (n=5, p<0.01). We further tested the anti-apoptotic effect of TPO on HUVEC cells. CoCl2 alone was able to induced apoptosis as demonstrated by Annexin V (n=4, p<0.001), active-caspase-3 (n=4, p<0.01) and Mitochondrial Membrane potential (JC-1) assays (n=4, p<0.01). Consistently, TPO plus CoCl2 showed a significant anti-apoptotic effect compared with CoCl2 alone group in Annexin V (n=4, p<0.01), active-caspase-3 (n=4, p=0.039) and Mitochondrial Membrane potential (JC-1) assays (n=4, p=0.038). In order to find out the underlying mechanism, we further tested the expression of phospho-Akt (p-Akt). Our results demonstrated that CoCl2 alone suppressed the PI-3k/Akt pathway. TPO plus CoCl2 was shown to activate the expression of p-Akt by western blot. In summary, our findings showed that TPO had a protective effect on human endothelial cells. This effect was likely mediated by activation of PI-3k/Akt pathway, which leads to anti-apoptosis on these cells. This may provide us a new clue to identify novel strategy in treating endothelial syndromes.

Disclosures

No relevant conflicts of interest to declare.
Bendamustine with Total Body Irradiation Limits Murine Graft-Versus-Host Disease in Part Via Myeloid-Derived Suppressor Cells
Authors: Hoffman Emely , Stokes Jessica , Molina Megan Stanley , Katsanis Emmanuel
Published: October 2019

Background

GvHD remains a significant challenge in allogeneic hematopoietic cell transplantation (HCT). An under-investigated strategy to reduce GvHD is the modification of the preparative conditioning regimen. Cyclophosphamide (CY) and total body irradiation (TBI) is a commonly utilized myeloablative conditioning regimen, but is associated with GvHD. Bendamustine (BEN) is an alkylating agent that has been applied recently in nonmyeloablative chemotherapy-based conditioning regimens, demonstrating reduced myelosuppression, transplant related mortality, and acute graft versus host disease (GvHD). We have recently published that BEN can effectively replace CY when given following murine haploidentical bone marrow transplantation. We therefore hypothesized that BEN can also have advantages over CY when given in conjunction with TBI as a myeloablative preparative regimen.

Methods

We used a major histocompatibility mismatched murine BMT model to evaluate GvHD following BEN-TBI and CY-TBI conditioning. BALB/c (H-2d) mice received equivalent doses of BEN or CY (based on maximum tolerated dose), followed by TBI (400 cGy) and infusion of bone marrow (BM) cells with splenocytes (SC) or purified T-cells from C57BL/6 (H-2b) mice. GvHD morbidity and mortality and engraftment were monitored and MDSC frequencies were assessed by flow cytometry and immunofluorescence.

Results

We demonstrate that BEN-TBI conditioning, while facilitating complete donor chimerism, results in significantly less GvHD and improved survival compared to CY-TBI. In BEN-TBI conditioned mice, suppressive CD11b+Gr-1high myeloid cells are increased in the blood, bone marrow (BM), and spleen, with no evident differences in suppressive function. The ratio of Gr-1+ cells to T-cells is significantly increased in BEN-TBI mice in the intestines. When Gr-1high cells are depleted prior to transplant, the beneficial effects of BEN-TBI are partially lost. Alternatively, administration of G-CSF, which promotes CD11b+Gr-1+ myeloid cell expansion, trends towards an increase in survival in BEN-TBI mice.

Conclusions

These findings indicate a potential role of granulocytic myeloid-derived suppressor cells (MDSCs) in the mechanism by which BEN allows engraftment with reduced GvHD. BEN-TBI conditioning reduces GvHD morbidity and mortality compared to CY-TBI and may present a safer alternative to CY-TBI as a myeloablative conditioning regimen in allogeneic HCT.

Disclosures

No relevant conflicts of interest to declare.
Molecular and Clinical Aspects of Aα Dysfibrinogenemia in 25 Patients from the Czech Republic
Authors: Ceznerova Eliska , Stikarova Jana , Roselova Pavla , Kaufmanova Jirina , Suttnar Jiri , Louzil Jan , Kotlin Roman , Salaj Peter , Dyr Jan E.
Published: October 2019

Fibrinogen is a 340 kDA glycoprotein that plays a vital role in the hemostasis. The three pairs of polypeptide chains (Aα, Bβ and γ) form a heterodimer structure of fibrinogen, each chain is encoded by a distinct gene (FGA, FGB and FGG). Congenital dysfibrinogenemia is a rare qualitative disorder characterized by structural alterations in fibrinogen molecule resulting in its defective function. The clinical manifestation of majority cases is asymptomatic (50%); about 30% are associated with bleeding tendencies and only some cases are thrombotic (20%).

In this study, we evaluated 25 patients with heterozygous missense mutations in exon 2 of FGA gene from 13 unrelated families, all with low levels of functional fibrinogen and prolonged clotting time. All mutations were located around thrombin cleavage site at N-terminus of a fibrinogen Aα chain. Mutation Aα G13E was found in 6 families (13 patients), Aα R16H in 4 families (6 patients), Aα R16C in 1 family (4 patients), and Aα R19G in 1 family (2 patients). Correlations between the genotype and phenotype were studied by fibrin polymerization, fibrinolysis, kinetics of fibrinopeptides release, and electron and confocal microscopy. The viscoelastic properties of fibrin clots were measured. All mutations caused a delay or absence of fibrinopeptide A release and prolonged fibrin polymerization. Clinical phenotypes of mutations were, unexpectedly and importantly, highly heterogeneous. Out of 25 patients 17 were asymptomatic, 7 had the bleeding tendencies and 1 patient had thrombosis. One woman with Aα G13E had 2 spontaneous abortions and 3 pregnant women with Aα G13E, Aα R16C and Aα R19G mutations suffered for hemorrhages or postpartum bleeding. Eventually, all had successful delivery under provided care with a hematologist. Small surgeries were performed to patients with Aα R16H supplied with fibrinogen concentrates and antifibrinolytics. Our observations show that the patients should be carefully treated based both on the results of the laboratory analyzes and personal and family history.

This work was supported by the project of the Ministry of Health, Czech Republic, for conceptual development of research organization 00023736, by Grant from the Academy of Sciences, Czech Republic nr. P205/12/G118, and by ERDF OPPK CZ.2.16/3.1.00/24001.

Disclosures

No relevant conflicts of interest to declare.
Therapeutic Potential of AO-176, a Next Generation Humanized CD47 Antibody, for Hematologic Malignancies
Authors: Wilson W. Casey , Bouchlaka Myriam N , Capoccia Ben J , Hiebsch Ronald R , Donio Michael J , Puro Robyn J , Carter Alun J , Chakraborty Prabir , Manning Pamela T , Karr Robert W , Sung Vicki , Pereira Daniel S
Published: October 2019

Inhibitors of adaptive immune checkpoints have shown promise as cancer treatments. CD47 is an innate immune checkpoint receptor broadly expressed on normal tissues and over-expressed on several tumors. Binding of tumor CD47 to signal regulatory protein alpha (SIRPalpha) on macrophages and dendritic cells triggers a "don't eat me" signal that inhibits phagocytosis enabling escape of innate immune surveillance. Blocking CD47/SIRPα interaction promotes phagocytosis reducing tumor burden in numerous xenograft and syngeneic animal models.

We have developed a next generation humanized anti-CD47 antibody, AO-176, that not only blocks the CD47/SIRPalpha interaction and induces phagocytosis of hematologic and solid tumor cells, but also exhibits several unique functional properties. The first property is the ability of AO-176 to induce direct tumor cytotoxic cell death in hematologic (ex. Jurkat, Raji and Molt-4) as well as solid human tumor cell lines by a cell autonomous mechanism (not ADCC). Secondly, AO-176 exhibits preferential binding to tumor versus normal cells, including red blood cells (RBCs), T cells, endothelial cells, skeletal muscle cells and epithelial cells. A0-176 also does not affect the function of any of these primary cells when assayed ex vivo. The negligible binding of AO-176 to RBCs versus hematologic (ex. Jurkat, Raji or Molt-4) or solid tumor cells is particularly profound and different from other reported anti-CD47 antibodies. AO-176 also does not induce hemagglutination of RBCs. These properties are expected not only to decrease the antigen sink, but also to minimize on-target clinical adverse effects observed following treatment with other reported RBC-binding anti-CD47 antibodies. Consistent with this attribute, AO-176 was well tolerated in cynomolgus monkeys with no adverse effects in general nor with respect to RBCs which was consistent with ex vivo results. A third novel property of AO-176 is its enhanced binding to tumor cells at acidic pH. Because the microenvironment of leukemic bone marrow and solid tumors has an acidic pH, this enhanced binding of AO-176 at low pH has the potential added advantage of tumor-specific targeting. Lastly, we show that AO-176 demonstrates dose-dependent anti-tumor activity in hematologic and solid tumor xenograft models.

Taken together, the unique properties and anti-tumor activity of our next generation anti-CD47 antibody, AO-176, distinguishes it from other CD47/SIRPalpha axis targeting agents as it progresses to clinical development.

Disclosures

No relevant conflicts of interest to declare.
Phase II of Single-Agent Ibrutinib in Recurrent/Refractory Primary (PCNSL) and Secondary CNS Lymphoma (SCNSL)
Authors: Grommes Christian , Wolfe Julia , Gavrilovic Igor , Kaley Thomas , Stone Jacqueline , Daras Mariza , Nolan Craig , Pentsova Elena , Hatzoglou Vaios , Mellinghoff Ingo , DeAngelis Lisa
Published: October 2019

BACKGROUND: PCNSL is an aggressive primary brain tumor with median progression free survival (PFS) after upfront methotrexate-based chemotherapy of 2-3 years. Outcome and treatment options are poor for recurrent/refractory (r/r) disease. Ibrutinib has shown promising clinical response in various B-cell malignancies. This trial investigates Ibrutinib in patients with r/r PCNSL and SCNSL.

METHODS: Eligible patients had r/r PCNSL or SCNSL, age≥18, ECOG≤2, normal end-organ function, and unrestricted number of CNS directed prior therapies. In patients with SCNSL, systemic disease needed to be absent. All patients needed to have at least one prior CNS directed therapy.

RESULTS: Forty-four patients were enrolled; 3 received ibrutinib at 560mg and 41 at 840mg. Median age was 68 years (range 21-90); 20 were women. Median ECOG was 1 (0: 11, 1: 22, 2: 11); 29 had PCNSL and 15 SCNSL; 61% had recurrent disease. Twenty-four had isolated parenchymal disease, 4 isolated cerebrospinal fluid (CSF) involvement and 16 both. No grade 5 event has been observed. Eleven patients experienced 12 grade 4 adverse events: neutropenia (in 5 patients), lymphopenia (2), ALT elevation (1), combined ALT/AST elevation (1), sepsis (1), and pneumonitis (1). Seventeen patients experienced 35 grade 3 adverse events (most common: lymphopenia in 4, ALT elevation (3), hyperglycemia (3), urinary tract infection (3), decreased WBC (2), lung infection (2), and thrombocytopenia (2)). Treatment was stopped due to adverse events in 3 patients (grade 4 pulmonitis, grade 4 liver toxicity, grade 3 infectious encephalitis). Twenty-four patients did not experience ≥3 grade adverse events. The most common toxicities at any grade were thrombocytopenia (73%), hyperglycemia (55%), anemia (43%), hypercholesterolemia (43%) and hypertriglyceridemia (43%) of which most were grade 1/2. Only 1 Aspergillus infection has been observed in a patient with chronic steroid use. After a median follow-up of 22 months (range 1.5-39), 40/44 patients were evaluated for response (4 did not complete at least 15 days of drug treatment due to deteriorating clinical status or withdrawal from study). Overall response was 78% (31/40; 81% (22/27) in PCNSL; 69% (9/13) in SCNSL) with 17 CR, 14 PR, 4 SD and 5 PD as best response. The median PFS is 4 months (5.4 months in patients that completed at least 15 days of drug treatment; longest: 16.5 months). The median overall survival is 19.5 months.

CONCLUSION: Patients with CNS lymphoma tolerate Ibrutinib with manageable adverse events. Clinical response was seen in 78% of CNS lymphoma patients.

Disclosures

No relevant conflicts of interest to declare.
Effectiveness of End of Therapy PET Based Deauville Scoring in Predicting Outcomes in Hodgkin's Lymphoma: A Real-World Experience
Authors: Mehra Nikita , Ganesan Prasanth , Radhakrishnan Venkatraman , Ganesan Trivadi S , Dhanushkodi Manikandan , Sundersingh Shirley , Shankar S Krupa , P K Jayachandran , Kannan Krishnarathnam , S M Carthikeyan , Sagar Tenali Gnana
Published: October 2019

Background: The use of PET CT has become instrumental in the management of Hodgkin's lymphoma (HL), with multiple studies favoring PET adapted therapies. Current guidelines recommend using the Deauville score (DS) for assessment of end therapy PET-CT scans. We looked at the predictive ability of end therapy PET CT DS in a cohort of HL.

Methods: Adult patients diagnosed with, and treated for, Hodgkin's lymphoma between 2007 and 2015 with end-of-treatment PET CT scans were included in this retrospective analysis.

Results: One hundred and ninety-eight patients [median age- 35 (18-74) years, advanced disease: 147 (74%); bulky disease: 54 (27%)] underwent end-of-therapy PET CT scanning with Deauville scoring. The chemotherapy regimens used were as follows: ABVD in 167 patients (84%); COPP/AVD in 20 patients (10%) and other regimens in 11 patients (6%). The end of therapy Deauville score (DS) was 1 or 2 in 160 patients (81%), 3 in 9 patients (5%) and 4 or 5 in 29 patients (14%). The median follow-up was 62 months. The 5-year DFS for patients with DS 1 or 2 was 85%, DS 3 was 65% and DS 4 or 5 was 40% (p<0.05). The 5-year overall survival for DS 1 and 2 combined was 91%, for DS 3 was 76% and for DS 4 and 5 combined was 70% (p=0.001).

Based on this analysis, the positive and negative predictive values for predicting treatment outcome on the basis of end of therapy PET CT by Deauville scoring were 0.48 and 0.85 respectively.

Conclusions: DS 1 and 2 were highly predictive of excellent survival while DS 4 and 5 were predictive for treatment failure. However, the positive predictive value was overall lower than reported from other studies. DS score of 3 was seen in very few patients at end therapy. Further modification of this is required for clear interpretation.

Figure. Figure.

Disclosures

No relevant conflicts of interest to declare.