Determinants of HDL Cholesterol Efflux Capacity after Virgin Olive Oil Ingestion: Interrelationships with Fluidity of HDL Monolayer

Scope: Cholesterol efflux capacity of HDL (CEC) is inversely associated with cardiovascular risk. HDL composition, fluidity, oxidation, and size are related with CEC. We aimed to assess which HDL parameters were CEC determinants after virgin olive oil (VOO) ingestion. Methods and results: Post‐hoc analyses from the VOHF study, a crossover intervention with three types of VOO. We assessed the relationship of 3‐week changes in HDL‐related variables after intervention periods with independence of the type of VOO. After univariate analyses, mixed linear models were fitted with variables related with CEC and fluidity. Fluidity and Apolipoprotein (Apo)A‐I content in HDL was directly associated, and HDL oxidative status inversely, with CEC. A reduction in free cholesterol, an increase in triglycerides in HDL, and a decrease in small HDL particle number or an increase in HDL mean size, were associated to HDL fluidity. Conclusions: HDL fluidity, ApoA‐I concentration, and oxidative status are major determinants for CEC after VOO. The impact on CEC of changes in free cholesterol and triglycerides in HDL, and those of small HDL or HDL mean size, could be mechanistically linked through HDL fluidity. Our work points out novel therapeutic targets to improve HDL functionality in humans through nutritional or pharmacological interventions.Fil: Fernández Castillejo, Sara. Universitat Rovira I Virgili; EspañaFil: Rubió, Laura. Universitat Rovira I Virgili; España. Universidad de Lleida; EspañaFil: Hernáez, Álvaro. Centro de Investigación Biomédica en Red Fisiopatología de la Obesidad y Nutrición; EspañaFil: Catalán, Úrsula. Universitat Rovira I Virgili; EspañaFil: Pedret, Anna. Universitat Rovira I Virgili; EspañaFil: Valls, Rosa M.. Universitat Rovira I Virgili; EspañaFil: Mosse, Juana Inés. Universidad de Lleida; España. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Covas, Maria Isabel. Centro de Investigación Biomédica en Red Fisiopatología de la Obesidad y Nutrición; EspañaFil: Remaley, Alan T.. National Institutes of Health; Estados UnidosFil: Castañer, Olga. Centro de Investigación Biomédica en Red Fisiopatología de la Obesidad y Nutrición; EspañaFil: Motilva, Maria José. Universidad de Lleida; EspañaFil: Solá, Rosa. Universitat Rovira I Virgili; Españ

Transverse instability for non-normal parameters

We consider the behaviour of attractors near invariant subspaces on varying a parameter that does not preserve the dynamics in the invariant subspace but is otherwise generic, in a smooth dynamical system. We refer to such a parameter as ``non-normal''. If there is chaos in the invariant subspace that is not structurally stable, this has the effect of ``blurring out'' blowout bifurcations over a range of parameter values that we show can have positive measure in parameter space. Associated with such blowout bifurcations are bifurcations to attractors displaying a new type of intermittency that is phenomenologically similar to on-off intermittency, but where the intersection of the attractor by the invariant subspace is larger than a minimal attractor. The presence of distinct repelling and attracting invariant sets leads us to refer to this as ``in-out'' intermittency. Such behaviour cannot appear in systems where the transverse dynamics is a skew product over the system on the invariant subspace. We characterise in-out intermittency in terms of its structure in phase space and in terms of invariants of the dynamics obtained from a Markov model of the attractor. This model predicts a scaling of the length of laminar phases that is similar to that for on-off intermittency but which has some differences.Comment: 15 figures, submitted to Nonlinearity, the full paper available at http://www.maths.qmw.ac.uk/~eo

Optimal neural network feature selection for spatial-temporal forecasting

In this paper, we show empirical evidence on how to construct the optimal feature selection or input representation used by the input layer of a feedforward neural network for the propose of forecasting spatial-temporal signals. The approach is based on results from dynamical systems theory, namely the non-linear embedding theorems. We demonstrate it for a variety of spatial-temporal signals, with one spatial and one temporal dimensions, and show that the optimal input layer representation consists of a grid, with spatial/temporal lags determined by the minimum of the mutual information of the spatial/temporal signals and the number of points taken in space/time decided by the embedding dimension of the signal. We present evidence of this proposal by running a Monte Carlo simulation of several combinations of input layer feature designs and show that the one predicted by the non-linear embedding theorems seems to be optimal or close of optimal. In total we show evidence in four unrelated systems: a series of coupled Henon maps; a series of couple Ordinary Differential Equations (Lorenz-96) phenomenologically modelling atmospheric dynamics; the Kuramoto-Sivashinsky equation, a partial differential equation used in studies of instabilities in laminar flame fronts and finally real physical data from sunspot areas in the Sun (in latitude and time) from 1874 to 2015.Comment: 11 page

High-content screen in human pluripotent cells identifies miRNA-regulated pathways controlling pluripotency and differentiation

Background: By post-transcriptionally regulating multiple target transcripts, microRNAs (miRNAs or miR) play important biological functions. H1 embryonic stem cells (hESCs) and NTera-2 embryonal carcinoma cells (ECCs) are two of the most widely used human pluripotent model cell lines, sharing several characteristics, including the expression of miRNAs associated to the pluripotent state or with differentiation. However, how each of these miRNAs functionally impacts the biological properties of these cells has not been systematically evaluated. Methods: We investigated the effects of 31 miRNAs on NTera-2 and H1 hESCs, by transfecting miRNA mimics. Following 3-4 days of culture, cells were stained for the pluripotency marker OCT4 and the G2 cell-cycle marker Cyclin B1, and nuclei and cytoplasm were co-stained with Hoechst and Cell Mask Blue, respectively. By using automated quantitative fluorescence microscopy (i.e., high-content screening (HCS)), we obtained several morphological and marker intensity measurements, in both cell compartments, allowing the generation of a multiparametric miR-induced phenotypic profile describing changes related to proliferation, cell cycle, pluripotency, and differentiation. Results: Despite the overall similarities between both cell types, some miRNAs elicited cell-specific effects, while some related miRNAs induced contrasting effects in the same cell. By identifying transcripts predicted to be commonly targeted by miRNAs inducing similar effects (profiles grouped by hierarchical clustering), we were able to uncover potentially modulated signaling pathways and biological processes, likely mediating the effects of the microRNAs on the distinct groups identified. Specifically, we show that miR-363 contributes to pluripotency maintenance, at least in part, by targeting NOTCH1 and PSEN1 and inhibiting Notch-induced differentiation, a mechanism that could be implicated in na\uefve and primed pluripotent states. Conclusions: We present the first multiparametric high-content microRNA functional screening in human pluripotent cells. Integration of this type of data with similar data obtained from siRNA screenings (using the same HCS assay) could provide a large-scale functional approach to identify and validate microRNA-mediated regulatory mechanisms controlling pluripotency and differentiation

Production of human factor VIII-FL in 293T cells using the bicistronic MGMT(P140K)-retroviral vector

Hemophilia A is the most common X-linked bleeding disorder; it is caused by deficiency of coagulation factor VIII (FVIII). Replacement therapy with rFVIII produced from human cell line is a major goal for treating hemophilia patients. We prepared a full-length recombinant FVIII (FVIII-FL), using the pMFG-P140K retroviral vector. The IRES DNA fragment was cloned upstream to the P140K gene, providing a 9.34-kb bicistronic vector. FVIII-FL cDNA was then cloned upstream to IRES, resulting in a 16.6-kb construct. In parallel, an eGFP control vector was generated, resulting in a 10.1-kb construct. The 293T cells were transfected with these constructs, generating the 293T-FVIII-FL/P140K and 293T-eGFP/P140K cell lines. In 293T-FVIII-FL/P140K cells, FVIII and P140K mRNAs levels were 4,410 (+/- 931.7)- and 295,400 (+/- 75,769)-fold higher than in virgin cells. In 293T-eGFP/P140K cells, the eGFP and P140K mRNAs levels were 1,501,000 (+/- 493,700)- and 308,000 (+/- 139,300)-fold higher than in virgin cells. The amount of FVIII-FL was 0.2 IU/mL and 45 ng/mL FVIII cells or 4.4 IU/mu g protein. These data demonstrate the efficacy of the bicistronic retroviral vector expressing FVIII-FL and MGMT(P140K), showing that it could be used for producing the FVIII-FL protein in a human cell line.FINEP [01.07.0652.00]FINEPFAPESPFAPESP [1998/14247-6]CNPq [314458/2009-3, 2008/57877-3]CNP

Development of a molecular platform for HTLV confirmatory diagnosis: importance of the internal amplification control

Finnancial Support: CTC/INCTC, FAPESP, FUNDHERP, FINEP