Polycomb repressor complex 2 regulates HOXA9 and HOXA10, activating ID2 in NK/T-cell lines

<p>Abstract</p> <p>Background</p> <p>NK- and T-cells are closely related lymphocytes, originating from the same early progenitor cells during hematopoiesis. In these differentiation processes deregulation of developmental genes may contribute to leukemogenesis. Here, we compared expression profiles of NK- and T-cell lines for identification of aberrantly expressed genes in T-cell acute lymphoblastic leukemia (T-ALL) which physiologically regulate the differentiation program of the NK-cell lineage.</p> <p>Results</p> <p>This analysis showed high expression levels of HOXA9, HOXA10 and ID2 in NK-cell lines in addition to T-cell line LOUCY, suggesting leukemic deregulation therein. Overexpression experiments, chromatin immuno-precipitation and promoter analysis demonstrated that HOXA9 and HOXA10 directly activated expression of ID2. Concomitantly elevated expression levels of HOXA9 and HOXA10 together with ID2 in cell lines containing MLL translocations confirmed this form of regulation in both ALL and acute myeloid leukemia. Overexpression of HOXA9, HOXA10 or ID2 resulted in repressed expression of apoptosis factor BIM. Furthermore, profiling data of genes coding for chromatin regulators of homeobox genes, including components of polycomb repressor complex 2 (PRC2), indicated lacking expression of EZH2 in LOUCY and exclusive expression of HOP in NK-cell lines. Subsequent treatment of T-cell lines JURKAT and LOUCY with DZNep, an inhibitor of EZH2/PRC2, resulted in elevated and unchanged HOXA9/10 expression levels, respectively. Moreover, siRNA-mediated knockdown of EZH2 in JURKAT enhanced HOXA10 expression, confirming HOXA10-repression by EZH2. Additionally, profiling data and overexpression analysis indicated that reduced expression of E2F cofactor TFDP1 contributed to the lack of EZH2 in LOUCY. Forced expression of HOP in JURKAT cells resulted in reduced HOXA10 and ID2 expression levels, suggesting enhancement of PRC2 repression.</p> <p>Conclusions</p> <p>Our results show that major differentiation factors of the NK-cell lineage, including HOXA9, HOXA10 and ID2, were (de)regulated via PRC2 which therefore contributes to T-cell leukemogenesis.</p

Validation of a new cardiac image fusion software for three-dimensional integration of myocardial perfusion SPECT and stand-alone 64-slice CT angiography

Purpose: Combining the functional information of SPECT myocardial perfusion imaging (SPECT-MPI) and the morphological information of coronary CT angiography (CTA) may allow easier evaluation of the spatial relationship between coronary stenoses and perfusion defects. The aim of the present study was the validation of a novel software solution for three-dimensional (3D) image fusion of SPECT-MPI and CTA. Methods: SPECT-MPI with adenosine stress/rest 99mTc-tetrofosmin was fused with 64-slice CTA in 15 consecutive patients with a single perfusion defect and a single significant coronary artery stenosis (≥50% diameter stenosis). 3D fused SPECT/CT images were analysed by two independent observers with regard to superposition of the stenosed vessel onto the myocardial perfusion defect. Interobserver variability was assessed by recording the X, Y, Z coordinates for the origin of the stenosed coronary artery and the centre of the perfusion defect and measuring the distance between the two landmarks. Results: SPECT-MPI revealed a fixed defect in seven patients, a reversible defect in five patients and a mixed defect in three patients and CTA documented a significant stenosis in the respective subtending coronary artery. 3D fused SPECT/CT images showed a match of coronary lesion and perfusion defect in each patient and the fusion process took less than 15min. Interobserver variability was excellent for landmark detection (r = 1.00 and r = 0.99, p < 0.0001) and very good for the 3D distance between the two landmarks (r = 0.94, p < 0.001). Conclusion: 3D SPECT/CT image fusion is feasible, reproducible and allows correct superposition of SPECT segments onto cardiac CT anatom

Mortalidade por câncer em trabalhadores da indústria da borracha de São Paulo

OBJECTIVE: To investigate cancer deaths among rubber industry workers according to company size. METHODS: This historical cohort study included 9,188 male workers, engaged as members of a workers' union in São Paulo, Southeastern Brazil, between 1975 and 1985, alive in January 1990 and followed until the end of 2000. Overall and specific cancer deaths were compared within the cohort, with subjects stratified according to company size and economic activity, sector of work, and worker qualification. Risk set assessment and conditional logistic regression were used to estimate relative risks, adjusted for age, time since first job, and duration of employment in the rubber industry. RESULTS: When compared to employees of larger companies, workers employed in small companies showed greater risk of death due to any type of cancer (RR=2.33, 95% CI: 0.90-6.03), stomach tumors (RR=3.47; 95% CI: 2.57-4.67), and upper aero-digestive tract tumors (RR=2.49; 95% CI: 1.78-3.48). Risk of death was greater among workers employed in maintenance (RR=2.23; 95% CI: 0.73-6.76); expedition, storage and transportation (RR=2.97; 95% CI: 0.77-11.38); and production (RR=3.51; 95% CI: 1.07-11.57), when compared to workers engaged in clerical work and support activities. CONCLUSIONS: Our results provide further evidence for an increased risk of stomach and upper aero-digestive tract tumors - and possibly of all types of cancer - among rubber industry workers employed in small companies when compared to employees of larger companies.OBJETIVO: Avaliar as mortes por câncer em trabalhadores da indústria da borracha de acordo com o tamanho da empresa. MÉTODOS: Coorte histórica com 9.188 trabalhadores masculinos, matriculados em sindicato de trabalhadores da indústria de borracha do Estado de São Paulo entre 1975 e 1985, sobreviventes em janeiro de 1990 e acompanhados até o final de 2000. As mortes por câncer foram comparadas internamente à coorte, cujos membros foram subdivididos em estratos por porte e sub-ramo da empresa, setor de trabalho e qualificação profissional. A abordagem por conjuntos de risco e regressão logística condicional foi utilizada para o cálculo dos riscos relativos, ajustados por idade, tempo desde o primeiro emprego e tempo de trabalho na indústria da borracha. RESULTADOS: Quando comparados aos empregados de grandes empresas, trabalhadores de pequenas empresas mostraram excesso de mortes por todos tipos de câncer (RR=2,33; IC 95%: 0,90-6,03); tumores de estômago (RR=3,47; IC 95%: 2,57-4,67) e do trato aero-digestivo superior (RR=2,49; IC 95%: 1,78-3,48). Identificou-se excesso de mortes por câncer nos trabalhadores dos setores de manutenção (RR=2,23; IC 95%: 0,73-6,76); expedição, armazenamento e transporte (RR=2,97; IC 95%: 0,77-11,38); e produção (RR=3,51; IC 95%: 1,07-11,57), comparados aos trabalhadores dos setores administrativos. CONCLUSÕES: Os resultados mostram evidências de aumento do risco de tumores de estômago e do trato aero-digestivo superior e, possivelmente, por câncer em geral em trabalhadores de empresas de pequeno porte da indústria da borracha quando comparados aos trabalhadores das grandes empresas

Towards the growth of Cu2ZnSn1 xGexS4 thin films by a single-stage process: Effect of substrate temperature and composition

9 págs.; 7 figs.; 2 tabs.Cu2ZnSn1-xGexS4 (CZTGS) thin films prepared by flash evaporation of a Zn-rich Cu2ZnSn0.5Ge0.5S4 bulk compound in powder form, and a subsequent thermal annealing in S containing Ar atmosphere are studied. The effect of the substrate temperature during evaporation and the initial composition of the precursor powder on the growth mechanism and properties of the final CZTGS thin film are investigated. The microstructure of the films and elemental depth profiles depend strongly on the growth conditions used. Incorporation of Ge into the Cu2ZnSnS4 lattice is demonstrated by the shift of the relevant X-ray diffraction peaks and Raman vibrational modes towards higher diffraction angles and frequencies respectively. A Raman mode at around 348-351 cm-1 is identified as characteristic of CZTGS alloys for x = [Ge]/([Sn]+[Ge]) = 0.14-0.30. The supply of Ge enables the reduction of the Sn loss via a saccrifical Ge loss. This fact allows increasing the substrate temperature up to 350º C during the evaporation, forming a high quality kesterite material and therefore, reducing the deposition process to one single stage & 2015 Elsevier B.V. All rights reserved.RC acknowledges financial support from Spanish MINECO within the Ramón y Cajal programme (RYC-2011-08521) and VIR for the Juan de la Cierva fellowship (JCI-2011-10782). GB also acknowledges the CSIC-JAE Pre-doctoral Program, co-funded by the European Social Fund. This work was supported by the Marie Curie-IRSES Project (PVICOKEST, GA: 269167), Marie Curie-ITN project (KESTCELL, GA: 316488), DAAD project (INTERKEST, Ref: 57050358), and MINECO projects (SUNBEAM, ENE2013-49136-C4-3-R) (TEC2012- 38901-C02-01). A. Scheu is acknowledged for GDOES measurements.Peer Reviewe

CLEC-2 and Syk in the megakaryocytic/platelet lineage are essential for development

The C-type lectin receptor CLEC-2 signals through a pathway that is critically dependent on the tyrosine kinase Syk. We show that homozygous loss of either protein results in defects in brain vascular and lymphatic development, lung inflation and perinatal lethality. Furthermore, we find that conditional deletion of Syk in the haematopoietic lineage, or conditional deletion of CLEC-2 or Syk in the megakaryocyte/platelet lineage, also causes defects in brain vascular and lymphatic development, although the mice are viable. In contrast, conditional deletion of Syk in other haematopoietic lineages had no effect on viability or brain vasculature and lymphatic development. We show that platelets, but not platelet releasate, modulate the migration and intercellular adhesion of lymphatic endothelial cells through a pathway that is dependent on CLEC-2 and Syk. These studies demonstrate that megakaryocyte/platelet expression of CLEC-2 and Syk is required for normal brain vasculature and lymphatic development and that platelet CLEC-2 and Syk directly modulate lymphatic endothelial cell behaviour in vitro

Towards the growth of Cu2ZnSn1-xGexS4 thin films by a single-stage process : effect of substrate temperatura and composition

Cu2ZnSn1-xGexS4 (CZTGS) thin films prepared by flash evaporation of a Zn-rich Cu2ZnSn0.5Ge0.5S4 bulk compound in powder form, and a subsequent thermal annealing in S containing Ar atmosphere are studied. The effect of the substrate temperature during evaporation and the initial composition of the precursor powder on the growth mechanism and properties of the final CZTGS thin film are investigated. The microstructure of the films and elemental depth profiles depend strongly on the growth conditions used. Incorporation of Ge into the Cu2ZnSnS4 lattice is demonstrated by the shift of the relevant X-ray diffraction peaks and Raman vibrational modes towards higher diffraction angles and frequencies respectively. A Raman mode at around 348-351 cm-1 is identified as characteristic of CZTGS alloys for x = [Ge]/([Sn]+[Ge]) = 0.14-0.30. The supply of Ge enables the reduction of the Sn loss via a saccrifical Ge loss. This fact allows increasing the substrate temperature up to 350º C during the evaporation, forming a high quality kesterite material and therefore, reducing the deposition process to one single stag

A transcriptomic snapshot of early molecular communication between Pasteuria penetrans and Meloidogyne incognita

© The Author(s). 2018Background: Southern root-knot nematode Meloidogyne incognita (Kofoid and White, 1919), Chitwood, 1949 is a key pest of agricultural crops. Pasteuria penetrans is a hyperparasitic bacterium capable of suppressing the nematode reproduction, and represents a typical coevolved pathogen-hyperparasite system. Attachment of Pasteuria endospores to the cuticle of second-stage nematode juveniles is the first and pivotal step in the bacterial infection. RNA-Seq was used to understand the early transcriptional response of the root-knot nematode at 8 h post Pasteuria endospore attachment. Results: A total of 52,485 transcripts were assembled from the high quality (HQ) reads, out of which 582 transcripts were found differentially expressed in the Pasteuria endospore encumbered J2 s, of which 229 were up-regulated and 353 were down-regulated. Pasteuria infection caused a suppression of the protein synthesis machinery of the nematode. Several of the differentially expressed transcripts were putatively involved in nematode innate immunity, signaling, stress responses, endospore attachment process and post-attachment behavioral modification of the juveniles. The expression profiles of fifteen selected transcripts were validated to be true by the qRT PCR. RNAi based silencing of transcripts coding for fructose bisphosphate aldolase and glucosyl transferase caused a reduction in endospore attachment as compared to the controls, whereas, silencing of aspartic protease and ubiquitin coding transcripts resulted in higher incidence of endospore attachment on the nematode cuticle. Conclusions: Here we provide evidence of an early transcriptional response by the nematode upon infection by Pasteuria prior to root invasion. We found that adhesion of Pasteuria endospores to the cuticle induced a down-regulated protein response in the nematode. In addition, we show that fructose bisphosphate aldolase, glucosyl transferase, aspartic protease and ubiquitin coding transcripts are involved in modulating the endospore attachment on the nematode cuticle. Our results add new and significant information to the existing knowledge on early molecular interaction between M. incognita and P. penetrans.Peer reviewedFinal Published versio

Radial Migration in Disk Galaxies I: Transient Spiral Structure and Dynamics

We seek to understand the origin of radial migration in spiral galaxies by analyzing in detail the structure and evolution of an idealized, isolated galactic disk. To understand the redistribution of stars, we characterize the time-evolution of properties of spirals that spontaneously form in the disk. Our models unambiguously show that in such disks, single spirals are unlikely, but that a number of transient patterns may coexist in the disk. However, we also show that while spirals are transient in amplitude, at any given time the disk favors patterns of certain pattern speeds. Using several runs with different numerical parameters we show that the properties of spirals that occur spontaneously in the disk do not sensitively depend on resolution. The existence of multiple transient patterns has large implications for the orbits of stars in the disk, and we therefore examine the resonant scattering mechanisms that profoundly alter angular momenta of individual stars. We confirm that the corotation scattering mechanism described by Sellwood & Binney (2002) is responsible for the largest angular momentum changes in our simulations.Comment: accepted to MNRAS; substantial additions to Section 4 dealing with the radial mixing proces

All-cause mortality benefit of coronary revascularization vs. medical therapy in patients without known coronary artery disease undergoing coronary computed tomographic angiography: results from CONFIRM (COronary CT Angiography EvaluatioN For Clinical Outcomes: An InteRnational Multicenter Registry)

Aims To date, the therapeutic benefit of revascularization vs. medical therapy for stable individuals undergoing invasive coronary angiography (ICA) based upon coronary computed tomographic angiography (CCTA) findings has not been examined. Methods and results We examined 15 223 patients without known coronary artery disease (CAD) undergoing CCTA from eight sites and six countries who were followed for median 2.1 years (interquartile range 1.4-3.3 years) for an endpoint of all-cause mortality. Obstructive CAD by CCTA was defined as a ≥50% luminal diameter stenosis in a major coronary artery. Patients were categorized as having high-risk CAD vs. non-high-risk CAD, with the former including patients with at least obstructive two-vessel CAD with proximal left anterior descending artery involvement, three-vessel CAD, and left main CAD. Death occurred in 185 (1.2%) patients. Patients were categorized into two treatment groups: revascularization (n = 1103; 2.2% mortality) and medical therapy (n = 14 120, 1.1% mortality). To account for non-randomized referral to revascularization, we created a propensity score developed by logistic regression to identify variables that influenced the decision to refer to revascularization. Within this model (C index 0.92, χ2 = 1248, P < 0.0001), obstructive CAD was the most influential factor for referral, followed by an interaction of obstructive CAD with pre-test likelihood of CAD (P = 0.0344). Within CCTA CAD groups, rates of revascularization increased from 3.8% for non-high-risk CAD to 51.2% high-risk CAD. In multivariable models, when compared with medical therapy, revascularization was associated with a survival advantage for patients with high-risk CAD [hazards ratio (HR) 0.38, 95% confidence interval 0.18-0.83], with no difference in survival for patients with non-high-risk CAD (HR 3.24, 95% CI 0.76-13.89) (P-value for interaction = 0.03). Conclusion In an intermediate-term follow-up, coronary revascularization is associated with a survival benefit in patients with high-risk CAD by CCTA, with no apparent benefit of revascularization in patients with lesser forms of CA