bluuebunny/crossref_metadata_2025 huggingface dataset:
Revision of the Neotropical species of Aphilodontinae (Geophilomorpha, Geophilidae), with eight new species and a first phylogenetic analysis of the subfamily
Authors: CALVANESE VICTOR C. , BRESCOVIT ANTONIO D. , BONATO LUCIO
Published: November 2019

Aphilodontinae are a well distinct but poorly known group of soil centipedes (Chilopoda: Geophilomorpha: Geophilidae) inhabiting two disjunct areas in South America and South Africa respectively. A comprehensive revision of the Neotropical species is presented based on the examination of >150 specimens, the phylogenetic analysis of the entire subfamily based on 47 morphological characters, and the description of eight new species from Southeastern Brazil. The Aphilodontinae were confirmed a monophyletic clade within Geophilidae and closely related to Geoperingueyia, with which they share synapomorphic characters in the labrum, second maxillae and forcipules. Particular groups of clypeal setae, inconspicuous lateral parts of labrum and coalescent forcipular throchanteroprefemur and femur were recovered as synapomorphies of the Aphilodontinae. Four monophyletic genera are recognized within the subfamily Aphilodontinae Silvestri, 1909 (1908) (= Brasilophilidae Verhoeff, 1908; = Mecistaucheninae Verhoeff, 1925 n. syn.), three exclusively Neotropical and one exclusively South African. The Neotropical genera are: Aphilodon Silvestri, 1898 (= Mecistauchenus Brölemann, 1907 n. syn.; = Brasilophilus Verhoeff, 1908 n. syn.), with ten species (including A. caboclos n. sp., A. indespectus n. sp., A. meganae n. sp., A. pereirai n. sp. and A. silvestrii n. sp.), characterized by a small terminal spine on the ultimate legs, in both males and females; Mecophilus Silvestri, 1909, with two species (including M. tupiniquim n. sp.), characterized by elongated forcipular segment and first leg-bearing segment; Mairata n. gen., with two species (M. butantan n. sp. and M. itatiaiensis n. sp.), characterized by the partial reduction of the distal article of the second maxillary telopodite and a particular shape of ultimate legs, in both males and females. The South African genus, Philacroterium Attems, 1926, comprises eleven known species (including P. weberi (Silvestri, 1909) n. comb., P. caffrarium (Verhoeff, 1937) n. comb., P. porosum (Verhoeff, 1937) n. comb., P. brevipes (Verhoeff, 1938) n. comb., P. bidentatum (Lawrence, 1955) n. comb., P. longipes (Lawrence, 1955) n. comb., P. macronyx (Lawrence, 1955) n. comb., P. maritimum (Lawrence, 1963) n. comb., P. pauciporum (Lawrence, 1963) n. comb. and P. transvaalicum (Lawrence, 1963) n. comb.) and is characterized by vestigial lateral parts of the labrum and the complete suppression of the distal article of the telopodites of the second maxillae. Identification keys are provided for the Aphilodontinae genera and for the species of Aphilodon, Mecophilus and Mairata n. gen. The known occurrence of Aphilodon in Brazil is extended from the states of São Paulo and Mato Grosso to the states of Minas Gerais and Rio de Janeiro, that of Mecophilus from the state of Paraná to the state of São Paulo.
Filtered prioritized and cluster-based optimization towards correction of antipatterns
Authors: Sreeji KS , Lakshmi C
Published: November 2019

When systems evolve, the maintainability and the complexity of the system increase, resulting in the poor antidesign patterns or antipatterns in software maintenance. Refactoring is an operation used to improve the structure of an existing system by changing its internal structure without affecting its external behavior. Many studies revealed that refactoring is underused by software development team in industry due to its increased effort and lack of knowledge about existing approaches. Thus, a prioritized correction of antipatterns based on relevance of classes, analyzed development history, risk to refactor, minimized code changes, and maintainers context and preferences is needed for good refactoring recommendations. We propose an approach based on filtered relevant classes with generic linkage-based clustering and adapted nondominated sorting genetic algorithm III to find good refactoring recommendations, which maximize the software quality and reduce the refactoring effort. Our approach uses our own node centrality-based sub-graph isomorphism algorithm to dynamically understand the system with good accuracy prior to optimization. Our approach is evaluated on six open-source systems and the results have shown the effectiveness of our approach compared to the other existing ones.
Dynamic impact behavior of syntactic foam core sandwich composites
Authors: Breunig P , Damodaran V , Shahapurkar K , Waddar S , Doddamani M , Jeyaraj P , Prabhakar P
Published: November 2019

Sandwich composites and syntactic foams independently have been used in many engineering applications. However, there has been minimal effort towards taking advantage of the weight saving ability of syntactic foams in the cores of sandwich composites, especially with respect to the impact response of structures. To that end, the goal of this study is to investigate the mechanical response and damage mechanisms associated with syntactic foam core sandwich composites subjected to dynamic impact loading. In particular, this study investigates the influence of varying cenosphere volume fraction in syntactic foam core sandwich composites subjected to varying dynamic impact loading and further elucidates the extent and diversity of corresponding damage mechanisms. The syntactic foam cores are first fabricated using epoxy resin as the matrix and cenospheres as the reinforcement with four cenosphere volume fractions of 0% (pure epoxy), 20%, 40%, and 60%. The sandwich composite panels are then manufactured using the vacuum assisted resin transfer molding process with carbon fiber/vinyl ester facesheets. Dynamic impact tests are performed on the sandwich composite specimens at two energy levels of 80 J and 160 J, upon which the data are post-processed to gain a quantitative understanding of the impact response and damage mechanisms incurred by the specimens. A qualitative understanding is obtained through micro-computed tomography scanning of the impacted specimens. In addition, a finite element model is developed to investigate the causes for different damage mechanisms observed in specimens with different volume fractions.
Creep in interlaminar shear of an Hi-Nicalon™/SiC–B4C composite at 1300℃ in air and in steam
Authors: Ruggles-Wrenn MB , Wallis TA
Published: November 2019

Creep behavior in interlaminar shear of an advanced SiC/SiC composite with a self-healing matrix was investigated at 1300℃ in laboratory air and in steam. The composite was processed via chemical vapor infiltration (CVI). The composite has a self-healing oxidation-inhibited matrix comprising alternating layers of silicon carbide and boron carbide and is reinforced with laminated woven Hi-Nicalon™ fibers. Fiber preforms were coated with pyrolytic carbon followed by a boron carbon overlay. The interlaminar shear properties were evaluated at 1300℃. The creep behavior was examined for interlaminar shear stresses ranging from 13 to 20 MPa in air and in steam. Primary and secondary creep regimes were observed in all tests conducted in air and in steam. Creep run-out (defined as 100 h at creep stress) was achieved at 13 MPa in air and in steam. Presence of steam had little influence on creep strain rates and creep lifetimes. However, larger creep strains were accumulated in steam than in air. The retained properties of all specimens that achieved creep run-out were characterized. Composite microstructure and damage and failure mechanisms were investigated.
The Tactile Quartet: Comparing Ambiguous Apparent Motion in Tactile and Visual Stimuli
Authors: Haladjian Harry H. , Anstis Stuart , Wexler Mark , Cavanagh Patrick
Published: November 2019

In the visual quartet, alternating diagonal pairs of dots produce apparent motion horizontally or vertically, depending on proximity. Here, we studied a tactile quartet where vibrating tactors were attached to the thumbs and index fingers of both hands. Apparent motion was felt either within hands (from index finger to thumb) or between hands. Participants adjusted the distance between their hands to find the point where motion changed directions. Surprisingly, switchovers occurred when between-hand distances were as much as twice that of within-hand distances—a general bias that was also found for tactile judgments of static distances. This expansion of within-hand felt distances was again seen when lights were placed on the hands rather than vibrating tactors. Importantly, switchover points were similar when the hands were placed at different depths, indicating that representations governing tactile motion were in perceptual three-dimensional space, not retinal two-dimensional space. This was true whether the quartets were visual stimuli on the hands or were purely visual on a monitor, suggesting that proximity is generally determined in three-dimensional coordinates for motion perception. Finally, the similarity of visual and tactile results suggests a common computation for apparent motion, albeit with different built-in distance biases for separate modalities.
Extruded high-temperature thermoplastic tufting yarns for enhanced mechanical properties of composites
Authors: McGarrigle Cormac , Fernández Daniel , Middendorf Peter , Dooher Thomas , Kelly John , McIlhagger Alistair , Harkin-Jones Eileen , Archer Edward
Published: November 2019

Through-thickness stitching, in the form of tufting, has been shown to be a potentially successful method of improving resistance to delamination. Tufting is a single-sided stitching technique that involves the insertion of a yarn through a fabric, in the z-direction. However, further research into the development of a tailored tufting yarn could yield a greater improvement in the mechanical properties of the overall composite. Unlike other published works which rely on commercially available materials, for this study four thermoplastic yarns were produced from polyetheretherketone, polysulfone, polyethersulfone and polyphenylsulfone. Their ability to be tufted into a composite was examined along with their influence on the overall mechanical properties of the composite.
Adaptive data-driven nonlinear synchro squeezed transform with single class radial basis function kernel support vector machine applied to wind turbine planetary gearbox fault diagnostics
Authors: Uma Maheswari R , Umamaheswari R
Published: November 2019

Planetary stage gears operated at low rotational speed and varying wind speed result variation in load. Variable speed and variable load induce nonstationary operating conditions. Vibration signal measured from Wind power gear transmission systems are embedded with multiple sources of vibration and attenuated considerably as it travels from source of vibration to measuring point. Efficacious multi-component decomposition without mode mixing ensures the accurate fault signature recognition. Synchro squeezing transform is the promising tool that represents the ridges with high resolution in time as well as in frequency axis. An efficient vibration analysis technique, short windowed Fourier synchro squeezing transform with nonlinear radial basis function kernel support vector machine is proposed to detect the mechanical faults in low speed planetary stage of wind turbines. Raw vibration is modeled in time–frequency plane to extract fault pattern signatures effectively with high resolution by adapting an empirical nonlinear tool synchro squeezing transforms. Amplitude modulation and frequency modulation parameters are sculpted from instantaneous amplitude and instantaneous phase, frequency. Hybrid feature space with signal attributes, statistical moments, and randomness measures are extricated from amplitude modulation-frequency modulation components. Single class radial basis function support vector machine is trained with hybrid features. The fault detection accuracy of the proposed method is compared with the standard variants of empirical mode decomposition. The proposed short windowed Fourier synchro squeezing transform-radial basis function kernel support vector machine shows 98.2% accuracy, 98% sensitivity, and 98% specificity.
Bicalutamide-associated hallucinations in a metastatic prostate cancer patient: A case report
Authors: Turkkan Gorkem , Dogan Caner , Tek Burak
Published: November 2019

Introduction

Bicalutamide is widely used in the treatment of prostate cancer. Among its side effects, central nervous system disorders are relatively rare, and the information about bicalutamide-associated hallucinations is limited.

Case report

We report an uncommon case of a patient with metastatic prostate cancer, who had hallucinations due to the use of bicalutamide.

Management and outcome

The patient accepted to receive only hormonal therapy (bicalutamide and leuprolide acetate). But he developed hallucinations due to bicalutamide use. His hallucinations disappeared after discontinuation of bicalutamide. A good response was obtained with the use of luteinizing hormone-releasing hormone agonist monotherapy.

Discussion

The pathophysiology of bicalutamide-induced hallucinations is unclear. We hypothesize that antiandrogens can indirectly cause hallucinations through changes in plasma testosterone and cerebral reelin expression. Additionally, luteinizing hormone-releasing hormone agonist monotherapy is a good option in metastatic prostate cancer patients who have intolerable side effects due to the use of antiandrogens.
Social work interventions with survivors of acts of political violence
Authors: Schiff Miriam , Lesser Lior , Levine Tova , Savo Yaeliy , Dashti Tzlil , Rosenne Hadas
Published: November 2019

Social workers and students in a large teaching hospital explored the characteristics of psychosocial interventions during acts of political violence. The associations between hospital length of stay, Acute Stress Reaction, and the characteristics of the implemented psychosocial interventions were also examined. One hundred and forty patients (61.4% males) treated during the so-called stabbing intifada/uprising in the years 2014–2015 in Israel in one large hospital were included. Data collection was based on clinical data mining.

Findings

The most frequent patient interventions were trauma-focused, while the most frequent family interventions were needs assessment and support-system building. Most of the interventions with the family (but not with the patient) were associated with longer hospital stays. Greater severity of Acute Stress Reaction was associated with greater use of trauma-focused interventions with the patient (but not with the family).

Applications

The findings suggest that social workers hold implicit trauma-focused intervention theories that should be written up in order to develop practice-based intervention models in the context of political violence.
Identification and structural characterization of four novel degradation products and a process impurity of isavuconazonium sulfate for injection formulation bulk by LC-ESI-QTOF-MS/MS
Authors: Regella Venkata Rama Prabhakara Sastry , Subramanian Venkatesan Chidambaram , Bhetanabhotla Sarveswara Sastry
Published: November 2019

Isavuconazonium sulfate is a highly water-soluble pro-drug of active triazole isavuconazole. The degradation profile of isavuconazonium sulfate for injection formulation bulk under acid hydrolysis, alkaline hydrolysis, oxidation, heat, and light conditions was studied using LC-PDA/MS technique. Four degradation products (DP1 to DP4) and one process impurity (PI1) were identified in isavuconazonium sulfate formulation bulk and characterized the identified impurities by liquid chromatography-electron spray ionization-quadruple-time of flight tandem mass spectrometry. Isavuconazole (DP1) was found to be one of the major degradation products of isavuconazonium sulfate.
Customization in mobile health apps: explaining effects on physical activity intentions by the need for autonomy
Authors: Bol Nadine , Høie Nina Margareta , Nguyen Minh Hao , Smit Eline Suzanne
Published: November 2019

Given the widespread adoption and technical possibilities of mobile technology, mobile health apps could be potentially effective tools to intervene in people’s daily routines and stimulate physical activity. Self-determination theory and the motivational technology model both suggest that mobile technology can promote health behaviour change by allowing users to customize their online experience when using mobile health apps. However, we know very little about why and for whom customization is most effective. Using a between-subjects experimental design, we tested the effects of customization in mobile health apps among a convenience sample ( N = 203). We assessed the effects of customization on perceived active control over mobile health apps, autonomous motivation to use mobile health apps, and intention to engage in physical activity, and tested the moderating role of need for autonomy. Structural equation modelling showed that customization in mobile health apps does not increase perceived active control, autonomous motivation, or the intention to engage in physical activity. However, an interaction effect between customization and need for autonomy showed that customization in mobile health apps leads to higher intentions to engage in physical activity for those with a greater need for autonomy, but not for those with a lesser need for autonomy. The implications for theory and practice are discussed.
The Protean Environmental State in Dongguan: Reconceptualising the local state and ecological development in China
Authors: Flynn Andrew , Yu Li
Published: November 2019

How China copes with the deep-rooted challenges of improving its environmental quality raises questions about how economic and environmental tensions arise, whether they can be reconciled, and if so, how that might be achieved. In this paper, we critically examine evolving environmental governance challenges at the city level since it is here that much public policy delivery takes place. In major urban areas, problems of poor waste management, and air and water pollution have been experienced by citizens for some time. What is novel is that municipal governments now have some capacity to act to improve local environmental conditions. Moreover, municipal governments are being assisted, to some extent, in their rethinking of environment–economy relations by a central government policy agenda that is moving from an overwhelming preoccupation with development to taking on board welfare and environmental concerns. This is demanding a reconceptualisation of the local state from being one that is almost wholly development oriented to one that recognises environmental imperatives. The paper outlines key perspectives on the local state, particularly the ideas of a developmental, entrepreneurial and green state. It draws on these accounts to produce a more nuanced analysis of the multi-faceted nature of environmental governance. We call this the Protean Environmental State. To put these debates into a local context we analyse how one early industrialising city, Dongguan, is facing up to its polluted environment and seeking to steer its way towards a new, more ecologically friendly development pathway.
Tropomyosin 1 Genetically Constrains in Vitro Megakaryopoiesis
Authors: Thom Christopher , Jobaliya Chintan , Lorenz Kimberly , Maguire Jean Ann , Gagne Alyssa , Gadue Paul , French Deborah L , Voight Benjamin F
Published: November 2019

Platelet size and count are heritable, quantifiable outcomes of hematopoiesis. A better understanding of genetic mechanisms that regulate hematopoiesis could yield insights into blood disorders and augment clinical translational efforts to generate platelets in vitro. For example, induced pluripotent stem cell-derived blood cells could someday ameliorate adverse effects and increase transfusable platelet supplies. However, these methods remain inefficient.

Genome wide association studies (GWAS) have linked hundreds of DNA loci with altered human platelet traits, but little is known about how these GWAS loci mechanistically impact hematopoiesis, megakaryocyte or platelet biology. Using publicly available genome-wide association and epigenetic data sets, we applied a machine-learning framework to identify epigenetic features enriched at established platelet trait association sites. From these results, we derived a quantitative prediction model that identified hematopoiesis-, megakaryocyte-, and platelet-relevant genomic loci and related genes more accurately than any prior computational method. In addition to specifying exact genetic variants known to regulate platelet traits and function, our model highlighted several novel variants in established platelet trait variation GWAS loci that were predicted to disrupt key transcription factor binding sites.

Among loci nominated by our statistical model was a variant (rs11071720) that alters Tropomyosin 1 (TPM1) gene expression. TPM1 regulates cytoskeletal biology in many cell types, and cytoskeletal functions critically impact hematopoiesis and terminal blood cell biology. We used CRISPR/Cas9-mediated genome editing to create TPM1-knockout human induced pluripotent stem cells. TPM1KO cells were healthy and showed normal developmental progression through primitive streak, mesoderm, and early primitive hematopoietic differentiation programs. Unexpectedly, TPM1KO cells demonstrated enhanced formation of hemogenic endothelium and hematopoietic progenitor cells, increasing total megakaryocyte yield by more than 2-fold. TPM1KO megakaryocytes demonstrated normal morphology, gene expression patterns and functional responses to platelet agonists.

Our findings help explain human genetics associations and identify a novel strategy to enhance in vitro hematopoiesis. More broadly, our now-validated machine learning-based approach might specify additional functional loci underlying hematopoiesis, megakaryopoiesis, or thrombopoiesis.

Disclosures

No relevant conflicts of interest to declare.
Interim Serum Soluble Interleukin-2 Receptor Levels Are Strongly Associated with Prognosis in Newly Diagnosis DLBCL Patients
Authors: Okamoto Haruya , Miyashita Akihiro , Nagata Hiroaki , Tsutsumi Yasuhiko , Kamitsuji Yuri , Kobayashi Yutaka , Uoshima Nobuhiko
Published: November 2019

 Serum soluble interleukin-2 receptor (sIL2R) levels are often measured to evaluate the state of lymphoma. The serum sIL2R level at diagnosis has been reported to be correlated with the prognosis of diffuse large B cell lymphoma (DLBCL) patients treated with the R-CHOP regimen. However, it is unclear whether interim sIL2R levels are associated with prognosis in DLBCL. Here, we analyzed the prognostic impact of interim serum sIL2R levels in DLBCL.

 We retrospectively examined data for DLBCL patients who started receiving chemotherapy at the Japanese Red Cross Society Kyoto Daini Hospital between January 2012 and December 2018. All of the patients received R-CHOP-like regimens (rituximab plus pirarubicin or adriamycin, cyclophosphamide, vincristine, and prednisolone). The interim sIL2R level (I-IL2R) was defined as the value measured after the third chemotherapy cycle. I-IL2R levels of >700 U/ml were regarded as positive. The primary endpoints of this study were progression-free survival (PFS) and overall survival (OS). The unadjusted probabilities of PFS and OS were estimated using the Kaplan-Meier method. The log-rank test and multivariate Cox regression analysis were used to assess the prognostic value of each clinical variable.

 In total, 102 patients were enrolled. The patients' median age was 73.5 years (range, 35-88), 58 patients (56.9%) were male, and 52 (51.0%) had poor revised International Prognostic Index scores. The median follow-up time was 25.2 months (range, 3.7-88.6). Twenty-three patients (22.5%) were I-IL2R-positive (>700 U/ml). Univariate analysis revealed that I-IL2R-positivity was associated with a poor prognosis. The 3-y PFS rates of the I-IL2R-negative (<700 U/ml) and I- IL2R-positive (>700 U/ml) patients were 60.4% (95% confidence interval [95%CI], 46.2-71.9) and 37.5% (95%CI, 15.7-59.4; p<0.001, log-rank test), respectively, and their 3-y OS rates were 82.2% (95%CI, 69.7-89.9) and 37.4% (95%CI, 13.8-61.4; p<0.001, log-rank test), respectively. Multivariate analysis confirmed that the I-IL2R level is independently associated with prognosis.

 The I-IL2R level of >700 U/ml patients had poor prognosis. The I-IL2R level can be used to predict the outcomes of DLBCL patients. IL2R levels should be measured during chemotherapy, and I-IL2R-positive patients could be targeted with high-dose or novel therapies. As this study was based on a retrospective analysis and involved a small cohort and a limited follow-up period, further studies are needed to confirm the prognostic impact of I-IL2R.

Figure

Disclosures

No relevant conflicts of interest to declare.
Endothelial JAK3 Expression Enhances Pro-Hematopoietic Angiocrine Function of Sinusoidal Endothelial Cells
Authors: Barcia Durán José Gabriel
Published: November 2019

Unlike Jak1, Jak2, and Tyk2, Jak3 is the only member of the Jak family of secondary messengers that signals exclusively by binding the common gamma chain of interleukin receptors IL2, IL4, IL7, IL9, IL15, and IL21. Jak3-null mice display defective T and NK cell development, which results in a mild SCID phenotype. Still, functional Jak3 expression outside the hematopoietic system remains unreported. Our data show that Jak3 is expressed in endothelial cells across hematopoietic and non-hematopoietic organs, with heightened expression in the bone marrow and spleen. Increased arterial zonation in the bone marrow of Jak3-null mice further suggests that Jak3 is a marker of sinusoidal endothelium, which is confirmed by fluorescent microscopy staining and single-cell RNA-sequencing. We also show that the Jak3-null niche is deleterious for the maintenance of long-term repopulating hematopoietic stem and progenitor cells (LT-HSCs) and that Jak3-overexpressing endothelial cells have increased potential to expand LT-HSCs in vitro. In addition, we identify the soluble factors downstream of Jak3 that provide endothelial cells with this functional advantage and show their localization to the bone marrow sinusoids in vivo. Our work serves to identify a novel function for a non-promiscuous tyrosine kinase in the bone marrow vascular niche and further characterize the hematopoietic stem cell niche of sinusoidal endothelium.

Disclosures

No relevant conflicts of interest to declare.
ETH-155008, a Novel Selective Dual Inhibitor of FLT3 and CDK4/6 in Preclinical Treatment of Acute Myeloid Leukemia
Authors: Liu Changnian , Liu Bin , Xu Chunping , Zhang Pan , Li Bin , Ji Bin , Zhang Bin , Liu Qingjun , Zhang James , Yu Chengzhi
Published: November 2019

Fms-like tyrosine kinase 3 (FLT3) is part of a family of receptor tyrosine kinases (RTKs), and a molecular therapeutic target of acute myeloid leukemia (AML). There are three FTL3 inhibitors, midostaurin, gilteritinib, and quizartinib have been approved for clinical treatment of AML. We have evaluated the inhibition of cell proliferation in vitro and antitumor activity in vivo of ETH-155008, a novel triple inhibitor of FLT3, Pim-3 and CDK4/6. ETH-155008 was found to have a strong inhibitory activity against key target enzymes (FLT3 IC50 at 0.5nM; Flt3 -Itd IC50 at 0.6 nM; CDK4/D1 IC50 at 0.8nM, CDK6/D1 IC50 at 1.4nM), which were more potent than the tested positive controls gilteritinib, quizartinib, and AMG-925. ETH-155008 showed to have highly potent antiproliferative activities against cultured human acute myeloid leukemia cells (MV4-11 and Molm-13) with IC50 at 4.1nM and 12 nM, respectively, and showed remarkable antitumor efficacy in vivo.

The in vivo antitumor activity of ETH-155008 was evaluated and compared with midostaurin and gilteritinib in the treatment of the two models with mutant forms in TKD and gatekeeper region, namely BaF3 FLT3-ITD-F691L and BaF3 FLT3-ITD-D835Y, respectively. Treatment with ETH-155008 (40 mg/kg/daily x 18, oral) significantly delayed tumor growth and resulted in a smaller mean tumor size (tumor growth inhibition (TGI) of 86% (p<0.001 compared with the control group) in the BaF3 FLT3-ITD-F691L model, and cured all of the tumor-bearing mice in the BaF3 FLT3-ITD-D835Y model. The antitumor activity of ETH-155008 was comparable to gilteritinib, but much better than midostaurin, the latter showed only mild antitumor activity with ~25% of TGI in the two FLT3 mutant models. We also tested the therapeutic efficacy of ETH-155008 in the human AML xenograft model MV4-11, ETH-155008 cured all of the tumor-bearing mice (mean tumor size at time of treatment 126 mm3), at a dose of 40mg/kg/daily for 14 days, that showed very little toxicity (only 2% body weight loss) which were well below the maximum tolerated dose.

ETH-155008 represents a novel dual inhibitor that may yet fulfill the promise of effective AML therapy through dual targeting of FLT3 and CDK4/6. These preclinical results supported the safety and efficacy observed in the preclinical studies and worthy of further clinical evaluation.

Disclosures

Liu: Euthare LLC: Other: founder. Zhang:shengke therapeutics: Other: cofounder. Yu:shengke Therapeutics: Other: co-founder.
Combination of BCL-2 Inhibition and Triptolide Causes Synthetic Lethality in Acute Myeloid Leukemia through Preventing Reciprocal Resistance
Authors: Xu Bing , Shi Yuanfei , Liu Long , Carter Bing Z
Published: November 2019

BCL-2 inhibition exerts effective pro-apoptotic activities in acute myeloid leukemia (AML) but clinical efficacy as a monotherapy was limited in part due to the treatment-induced MCL-1 increase. Triptolide (TPL) exhibits anti-tumor activities in part by upregulating pro-apoptotic BCL-2 proteins and decreasing MCL-1 expression in various malignant cells. We hypothesized that combined BCL-2 inhibition and TPL exert synergistic anti-leukemia activities and prevent the resistance to BCL-2 inhibition in AML. We here report that TPL combined with BCL-2 inhibitor ABT-199 synergistically induced apoptosis in leukemic cells regardless of p53 status through activating the intrinsic mitochondrial apoptotic pathway in vitro. Although ABT-199 or TPL alone inhibited AML growth in vivo, the combination therapy demonstrated a significantly stronger anti-leukemic effect. Mechanistically, TPL significantly upregulated BH3 only proteins including PUMA, NOXA, BID and BIM and decreased MCL-1 but upregulated BCL-2 expression in both p53 wild type and p53 mutant AML cell lines, while the combination decreased both BCL-2 and MCL-1 and further increased BH3 only BCL-2 proteins. MCL-1 and BCL-2 increases associated with respective ABT-199 and TPL treatment and resistance were also observed in vivo. Significantly downregulating MCL-1 and elevating BH3 only proteins by TPL could not only potentially block MCL-1-mediated resistance but also enhance anti-leukemic efficacy of ABT-199. Conversely, BCL-2 inhibition counteracted the potential resistance of TPL mediated by upregulation of BCL-2. The combination further amplified the effect, which likely contributed to the synthetic lethality. This mutual blockade of potential resistance provides a rational basis for the promising clinical application of TPL and BCL-2 inhibition in AML independent of p53 status.

Disclosures

Carter: Amgen: Research Funding; AstraZeneca: Research Funding; Ascentage: Research Funding.
SF3B1/DNMT3A Double Mutations Are Associated with Better Prognosis Than DNMT3A Mutation Alone in Myeloid Neopasms
Authors: Song Jinming , Hussaini Mohammad , Moscinski Lynn C. , Zhang Ling , Zhang Xiaohui , Shao Haipeng , Zhang Hailing
Published: November 2019

Background: DNMT3A and SF3B1 are two of the most commonly mutated genes in myeloid neoplasms. DNMT3A mutations have been found to be associated poor prognosis in acute myeloid leukemia (AML), while SF3B1 mutations have been reported to be associated with favorable prognosis in myelodysplastic syndrome (MDS). Studies exploring the clinical significance of DNMT3A and SF3B1 co-mutation are limited and conflicting. Furthermore, the previous studies did not control for the presence of other co-mutated genes, which could be confounding factors.

Materials and Methods: 4390 in-house patients with confirmed or suspected myeloid neoplasms were tested by Next Generation Sequencing (NGS) (up to 54 gene panel). From these, we identified 68 patients with only DNMT3A mutation, 61 patients with only SF3B1 mutation, and 23 patients with only SF3B1/DNMT3A co-mutation, for a total of 152 patients. Patient demographics, diagnosis, cytogenetic abnormalities, and overall survival time were collated and analyzed.

Results: Patients with SF3B1 mutation alone or SF3B1/DNMT3A double mutations were found to be significantly older (median age of 76 and 75, respectively) than patients with isolated DNMT3A mutation (median age of 66, p values = 0.00016 and 0.00174 respectively). Patients with SF3B1/DNMT3A double mutations or isolated SF3B1 mutations occurred more commonly in MDS. They occurred less frequently in acute myeloid leukemia (AML) (1.63% and 21.74% respectively) than isolated DNMT3A mutation (41.76%, p values < 0.00001 and p = 0.00529 respectively). Patients with double mutations demonstrated increased ring sideroblasts (100%) at a rate similar to patients with isolated SF3B1 mutation (92.16%), which are unsurprisingly higher than patents with DNMT3A mutation alone (8%, p value < 0.000001). The patients with SF3B1 mutation alone or SF3B1/DNMT3A co-mutation were less likely to have poor cytogenetics and more likely to have intermediate cytogenetics than patients with DNMT3A mutation alone. Finally, patients with sole SF3B1 mutation or SF3B1/DNMT3A co-mutation showed significantly longer median survival time (26.60 months and 19.93 months respectively) and better overall survival curve (Figure 1) than patients with DNMT3A mutation alone (8.32 months, p value = 0.001 and 0.01078 respectively), for all the patients in this study as well as for patients with only myelodysplastic syndrome.

Conclusion: Patients with SF3B1/DNMT3A double mutations showed a prognosis similar to patients with isolated SF3B1 mutation and better clinical outcomes than patients with isolated DNMT3A mutation. This suggests that the favorable prognosis of SF3B1 mutation in MDS or AML patients is not abrogated by the concurrent presence of a DNMT3A mutation.

Figure 1

Disclosures

No relevant conflicts of interest to declare.
Predictive Significance of Selected Gene Mutations Identified Using Next Generation Sequencing in Relapsed and Refractory Chronic Lymphocytic Leukemia Patients Treated with Ibrutinib
Authors: Machnicki Marcin M , Górniak Patryk , Pepek Monika , Szymczyk Agnieszka , Iskierka-Jażdżewska Elżbieta , Steckiewicz Pawel , Bluszcz Aleksandra , Rydzanicz Malgorzata , Hus Marek , Ploski Rafal , Juszczynski Przemyslaw , Stoklosa Tomasz , Jamroziak Krzysztof , Pula Bartosz
Published: November 2019

Ibrutinib, a selective inhibitor of the Bruton's kinase (BTK), is characterized by high efficacy in the therapy of treatment-naïve and relapsed and refractory chronic lymphocytic leukemia patients (RR-CLL). Numerous clinical trials and "real-world evidence" confirmed its clinical efficacy also in high-risk patients, including those with TP53 aberrations. Predictive factors to ibrutinib therapy present at time of treatment initiation are not well recognized, therefore we aimed at analyzing the potential significance of mutational status of selected 22 genes in the disease outcome. For this purpose, targeted sequencing of approx. 2 Mb of exonic regions was carried out in 45 RR-CLL patients using custom-made gene panel and sequencing on Illumina MiSeq FGx platform (median 73x mean coverage, range 21.23-149x). Median patient age was 65 years, 28 (62.2%) patients were male and 25 (55.5%) patients had Rai 3-4 stage at ibrutinib initiation. Mean observation time was 34.5 (±19.9 SD) months, while ibrutinib was administered for the mean time of 33.8 (±20.8 SD) months. Median progression-free survival (PFS) and overall survival (OS) was not reached. The 36-month estimated rate of PFS and OS was 79.1% and 68.2%, respectively. The overall response rate (ORR) was noted in 23 patients (51.1%). Complete remission was noted in one (2.2%) patient, while partial remission and partial remission with lymphocytosis was noted in 16 (35.6%) and 6 (13.3%) patients, respectively. Twenty (44.4%) patients had stable disease, whereas progression following ibrutinib initiation was noted in 2 (4.5%) cases. The highest rate of pathogenic gene mutations was observed in TP53 (n=18; 40.0%), NOTCH1 (n=13; 28.8%), SF3B1 (n=11; 24.4%), ATM (n=7; 15.6%), MED12 (n=6, 13.3%), CHD2 (n=5; 11.1%), XPO1 (n=5; 11.1%), NFKBIE (n=5; 11.1%), BIRC3 (n=4; 8.9%), SPEN (n=4; 8.9%), POT1 (n=4; 8.9%), EGR2 (n=3; 6.7%), RPS15 (n=3; 6.7%). Single cases of mutation in ZMYM3, PIM2, PIM3, BRAF, RB1, TNFAIP3 were detected. No mutations in MYD88, BTK or BCL2 were detected. Univariate survival analysis revealed that none of the analyzed mutations had predictive significance in the analyzed group. Interestingly, ibrutinib treatment was characterized by stable PFS and OS despite the low ORR levels with a significant number of patients achieving disease stability.

Disclosures

Juszczynski: Selvita S.A.: Consultancy, Membership on an entity's Board of Directors or advisory committees. Jamroziak:Janssen: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding; Celgene: Consultancy, Honoraria, Research Funding.
Bile Acids Induce Platelet Activation Leading to Degranulation and a Prothrombotic Phenotype
Authors: Zobel Joachim , Strini Tanja , Tischitz Martin , Pohl Sina , Greimel Theresa , Jahnel Jörg , Schlagenhauf Axel
Published: November 2019

Background: Previous articles have identified the farnesoid X receptor (FXR) as an integral part in the formation of coated platelets. Coated platelets are preactivated platelets featuring degranulation, increased fibrinogen binding, and increased serine protease activity leading to fibrin generation. Furthermore, phosphatidylserine exposure is increased and integrin α2bβIII is inhibited - leading to a prothrombotic phenotype despite decreased platelet aggregation. We hypothesize that bile acids, as natural ligands of FXR, lead to a change of platelet phenotype and therefore play a pivotal role in the formation of coated platelets, especially in presence of cholestasis.

Methods: Based on previous findings, we incubated human washed platelets of healthy adult volunteers with the synthetic FXR ligand GW4064 in various concentrations (0, 10, 20, 50, 100µM) and used flow cytometry to detect a shift in p-selectin expression, PAC-1 binding and annexin-V-binding. Moreover, we used different concentrations (0, 100, 200, 400, 600µM) of three bile acids (ursodeoxycholic acid, UDCA; chenodeoxycholic acid, CDCA; glycochenodeoxycholic acid, GCDCA) to see if natural FXR ligands induce an effect on the platelet phenotype.

Results: We observed a dose dependent shift in annexin-V-binding when treating washed platelets with GW4064 as well as CDCA and GCDCA. Similarly, GW4064 led to increased p-selectin expression while increased PAC-1-binding was only detected at the highest concentration. In contrast, CDCA and GCDCA showed merely slight changes in p-selectin expression whereas PAC-1-binding seemed to be unaffected. However, none of these effects were seen when using UDCA.

Conclusion: We conclude that pretreatment of washed platelets with CDCA and GCDCA initiate a dose-dependent shift towards a prothrombotic platelet phenotype. Therefore, we assume that increased levels of certain bile acids drive thrombosis in patients with cholestatic liver injury. Furthermore, a recent mouse model study suggested that platelet derived growth factor β (PDGFβ), a component of α-granula, drives liver fibrosis. Hence, in addition to their prothrombotic effects, coated platelets might exacerbate liver fibrosis.

Disclosures

No relevant conflicts of interest to declare.
Novel KMT2A Partner Gene NUTM2A Revealed By Anchored Multiplex PCR in ALL
Authors: Lebedeva Svetlana , Zerkalenkova Elena , Soldatkina Olga , Maschan Michael , Maschan Alexey A. , Novichkova Galina , Olshanskaya Yulia
Published: November 2019

Objectives

Chromosomal rearrangements of the human MLL/KMT2A gene are associated with infant, pediatric and adult acute leukemias. Such rearrangements are more frequent in infants, in whom they are found in 60% - 80% of acute lymphoblastic leukemias (ALL). KMT2A-r group itself is genetically heterogeneous. KMT2A-r can occur with at least 94 partner genes previously described in «THE MLL RECOMBINOME <…> IN 2017». Detection of KMT2A-r is an essential part of AL initial diagnostics. Also, the accurate detection of all KMT2A-r types is crucial in order to perform minimal residual disease (MRD) monitoring, as it is clear now that MRD-based therapy adjustment has a very strong impact on outcome. Here we report a case of novel KMT2A partner in pediatric ALL - NUT family member 2A (NUTM2A).

Methods

The patient is 8 y.o. girl with T-cell acute lymphoblastic leukemia with pleural effusion. Bone marrow and pleural fluid aspirates were analyzed by G-banded karyotyping and FISH with KMT2A breakapart probe. Pleural fluid aspirates were also analyzed by real-time RT-PCR for 8 most common KMT2A rearrangements screening, long-distance inversed PCR and targeted RNA-seq with FusionPlex Myeloid kit (ArcherDX, CO, USA). Sanger sequencing was used for validation.

Results

Conventional cytogenetics and FISH showed 47,XX,t(10;11)(q22;q23),+mar[5] karyotype with 100% KMT2A-rearranged nuclei. Gene fusion analysis identified novel fusion KMT2A-NUTM2A with exon 11 - exon 1 breakpoint junction.

NUTM2A is a gene at 10q23.2. This gene is not fully described in the literature. Rearrangements of this gene were identified in endometrial stromal sarcomas (ESS) (Cheng-Han Lee et al. 2012) and small round cell sarcoma (SRCS) (Sugita et al. 2017). In case of ESS, this rearrangement results in an in-frame fusion between YWHAE and NUTM2A (or highly homologous gene NUTM2B), and in case of SRCS it results in an in-frame fusion between NUTM2A and CIC. To our knowledge, KMT2A-NUTM2A fusion in our study is the first case demonstrating NUTM2A rearrangement in hematological malignancies.

Conclusions

Here we for the first time show the novel KMT2A-NUTM2A fusion transcript, which was found in pediatric T-cell acute lymphoblastic leukemia. Anchored multiplex PCR is one of the most sensitive way to detect rare variants of KMT2A rearrangements. It also allows selecting patient-specific primers for further PCR detection of MRD.

Disclosures

No relevant conflicts of interest to declare.
Subtyping Diffuse Large B-Cell Lymphoma with Cell-of-Origin Using 32-Gene Expression Assay in Formalin-Fixed Paraffin-Embedded Tissue
Authors: Jiang Xiangnan , Yan Wanhui , Sun Yifeng , Xu Qinghua , Zhou Xiaoyan , Cao Junning , Hong Xiaonan , Li Xiaoqiu
Published: November 2019

Introduction

Diffuse large B-cell lymphoma (DLBCL) is a group of heterogeneous disease with distinct molecular subtypes. The most established subtyping algorithm, the Cell-of-Origin (COO) model, categorizes DLBCL into activated B-cell (ABC) and germinal center B-cell (GCB)-like subgroups through gene expression profiling. COO subtyping is mandatory for every newly diagnosed DLBCL patients, as it is critical for determining the therapeutic and surveillance strategies. We evaluated a newly developed assay using 32-gene expression profiling to determine the COO of DLBCL with formalin-fixed paraffin-embedded (FFPE) tissue.

Methods

The DLBCL-COO Test is a qPCR-based 32-gene expression assay for COO determination in FFPE samples. Biopsy of DLBCL patients with paired FFPE and fresh tissue were identified to assign COO, based on the immunohistochemistry (IHC) algorithm (Han's algorithm), DLBCL-COO qPCR assay and global gene expression profiling with RNA-seq, respectively. The global gene expression profiling with RNA-seq was taken as the "gold standard" for reference. Clinical information including the survival data were collected.

Results

160 cases of DLBCL with evaluable COO assignments with IHC, DLBCL-COO 32-gene assay and global gene expression profiling with RNA-seq were identified. Comparing with the 77.5% concordance between IHC algorithm and gold standard, there is 91.9% concordance between DLBCL-COO 32-gene assay and gold standard (P =0.005). 72 patients assigned as ABC subtype and 14 patients assigned as Type-3 subtype demonstrated a significantly inferior overall survival than 42 patients assigned as GCB subtype using DLBCL-COO assay (P =0.023). However, COO based the IHC algorithm failed to provide the predictive value regarding overall survival (P =0.09).

Conclusions

DLBCL-COO assay provides flexibility and accuracy in DLBCL subtype characterization. These subtype distinctions should help guide disease prognosis and treatment options within DLBCL clinical practice.

Disclosures

Sun: Canhelp Genomics: Employment. Xu:Canhelp Genomics: Employment.
LIM Domain Protein 1 (Ldb1) Is Required for Lmo2 Oncogene-Induced Thymocyte Self-Renewal and T-Cell Leukemia in a Mouse Model of Human T-ALL
Authors: Li Liqi , Mitra Apratim , Zhao Bin , Choi Seeyoung , Lee Jan , Stamos Daniel , Warzecha Claude , Pfeifer Karl , Hardwick Joyce , Venters Bryan , Love Paul , Dave Utpal
Published: November 2019

LIM domain Only-2 (LMO2) is one of the most frequently deregulated oncogenes in T-cell acute lymphoblastic leukemia (T-ALL) and is generally expressed in the clinically aggressive Early Thymocyte Precursor ALL. LMO2 encodes a small protein with 2 LIM domains that is part of a large multiprotein complex in hematopoietic stem and progenitor cells (HSPC), where it is required for HSC specification and maintenance. Many of LMO2's protein partners in HSPCs are expressed in T-ALL implying that protein complexes like those scaffolded by LMO2 in HSPCs also play a role in leukemia. LDB1 is concordantly expressed with LMO2 in human T-ALL although its expression is more widespread than LMO2. In this study, we analyzed a critical component of the Lmo2 associated complex, LIM domain binding 1 (Ldb1), in the CD2-Lmo2 transgenic mouse model of human T-ALL. To further define Ldb1's role in leukemia, we induced its conditional knockout in CD2-Lmo2 transgenic mice with the use of Lck-Cre, Rag1-Cre, and Il7r-Cre transgenic mice. CD2-Lmo2 transgenic mice develop T-ALL with high penetrance and closely model the human disease.

We discovered that the penetrance and latency of Lmo2-induced T-ALL were markedly attenuated in the Lck-Cre model and T-ALL onset was completely abrogated in the Rag1-Cre and Il7r-Cre models. The latter two models induced more efficient deletions of Ldb1, earlier in the T-cell differentiation program compared to Lck-Cre. Interestingly, Lck-Cre deletion was efficient in thymocytes without the Lmo2 transgene. In striking contrast, Ldb1 deletion could not be induced in CD2-Lmo2 transgenic T-cell progenitors. Consistent with this finding, T-ALLs that developed in CD2-Lmo2/floxed-Ldb1/Lck-Cre mice had incomplete deletion of Ldb1. These results imply that Ldb1 is a required factor for Lmo2 to induce T-ALL. To further probe the pathogenesis of Lmo2-induced T-ALL, we analyzed preleukemic phenotypes in the Rag1-Cre (or Il7r-Cre) conditional knockout models. Our results showed that Ldb1 is required for the induction of thymocyte self-renewal and radioresistance. Ldb1 was also required for the acquisition of the pre-leukemic ETP gene expression signature observed in immature CD2-Lmo2 transgenic thymocytes. Detailed biochemical experiments show that LMO2 protein is directly stabilized by LDB1 in leukemia cells perhaps on chromatin.

In conclusion, these results support a model where Lmo2-induced T-ALL is caused by a failure to downregulate Ldb1/Lmo2 nucleated transcription complexes that normally function to enforce self-renewal in bone marrow hematopoietic progenitor cells.

Disclosures

No relevant conflicts of interest to declare.