Gating-induced large aqueous volumetric remodeling and aspartate tolerance in the voltage sensor domain of Shaker K+ channels

Indexación: Scopus.ACKNOWLEDGMENTS. We thank Chris Lingle and Yu Zhou (Washington University) for critical reading of the manuscript and Victoria Prado for Xenopus care and oocyte preparation. We also thank Millennium Scientific Initiative P029-022-F. This work was supported by Fondecyt Postdoctoral Grants 3170599 (to I.D.-F.) and 3160321 (to H.M.).Neurons encode electrical signals with critically tuned voltage-gated ion channels and enzymes. Dedicated voltage sensor domains (VSDs) in these membrane proteins activate coordinately with an unresolved structural change. Such change conveys the transmembrane translocation of four positively charged arginine side chains, the voltage-sensing residues (VSRs; R1–R4). Countercharges and lipid phosphohead groups likely stabilize these VSRs within the low-dielectric core of the protein. However, the role of hydration, a sign-independent charge stabilizer, remains unclear. We replaced all VSRs and their neighboring residues with negatively charged aspartates in a voltage-gated potassium channel. The ensuing mild functional effects indicate that hydration is also important in VSR stabilization. The voltage dependency of the VSR aspartate variants approached the expected arithmetic summation of charges at VSR positions, as if negative and positive side chains faced similar pathways. In contrast, aspartates introduced between R2 and R3 did not affect voltage dependence as if the side chains moved outside the electric field or together with it, undergoing a large displacement and volumetric remodeling. Accordingly, VSR performed osmotic work at both internal and external aqueous interfaces. Individual VSR contributions to volumetric works approached arithmetical additivity but were largely dissimilar. While R1 and R4 displaced small volumes, R2 and R3 volumetric works were massive and vectorially opposed, favoring large aqueous remodeling during VSD activation. These diverse volumetric works are, at least for R2 and R3, not compatible with VSR translocation across a unique stationary charge transfer center. Instead, VSRs may follow separated pathways across a fluctuating low-dielectric septum. © National Academy of Sciences. All rights reserved.https://www.pnas.org/content/115/32/820

Development of a three-dimensional bone-like construct in a soft self-assembling Peptide matrix

This work describes the development of a three-dimensional (3D) model of osteogenesis using mouse preosteoblastic MC3T3-E1 cells and a soft synthetic matrix made out of self-assembling peptide nanofibers. By adjusting the matrix stiffness to very low values (around 120 Pa), cells were found to migrate within the matrix, interact forming a cell-cell network, and create a contracted and stiffer structure. Interestingly, during this process, cells spontaneously upregulate the expression of bone-related proteins such as collagen type I, bone sialoprotein, and osteocalcin, indicating that the 3D environment enhances their osteogenic potential. However, unlike MC3T3-E1 cultures in 2D, the addition of dexamethasone is required to acquire a final mature phenotype characterized by features such as matrix mineralization. Moreover, a slight increase in the hydrogel stiffness (threefold) or the addition of a cell contractility inhibitor (Rho kinase inhibitor) abrogates cell elongation, migration, and 3D culture contraction. However, this mechanical inhibition does not seem to noticeably affect the osteogenic process, at least at early culture times. This 3D bone model intends to emphasize cell-cell interactions, which have a critical role during tissue formation, by using a compliant unrestricted synthetic matrix

Modificación hidrotérmica del almidón de yuca para su empleo como estabilizador de helados

En el trabajo se desarrolla un procedimiento de modificación física de almidón nativo de yuca a través de un tratamiento hidrotérmico en la zona previa a la gelatinización, de la cual no existen reportes de experimentación. El objetivo es determinar la influencia de las principales variables del proceso sobre el comportamiento reológico, con el propósito de ser empleado como estabilizador en la Industria heladera. Se alcanzaron buenos resultados reológicos con una concentración de 6% p/p de almidón nativo a 56 0C. El material obtenido se usó en formulaciones de helados, sustituyendo parte de los productos convencionales con efectos muy positivos en cuanto a la evaluación sensorial y el índice de aireación. Los resultados alcanzados permiten establecer perspectivas para el empleo de almidones modificados de yuca cuyo tratamiento hidrotérmico en la zona previa a la gelatinización ofrece efectos positivos, como estabilizante de suspensiones acuosas, similares a los que se alcanzan con almidones pregelatinizados, además con la ventaja de obtener el producto modificado con mayor facilidad

Behavior patterns in hormonal treatments using fuzzy logic models

Assisted reproductive technologies are a combination of medical strategies designed to treat infertility patients. Ideal stimulation treatment has to be individualized, but one of the main challenges which clinicians face in the everyday clinic is how to select the best medical protocol for a patient. This work aims to look for behavior patterns in this kind of treatments, using fuzzy logic models with the objective of helping gynecologists and embryologists to make decisions that could improve the process of in vitro fertilization. For this purpose, a real-world dataset composed of one hundred and twenty-three (123) patients and five hundred and fifty-nine (559) treatments applied in relation to such patients provided by an assisted reproduction clinic, has been used to obtain the fuzzy models. As conclusion, this work corroborates some known clinic experiences, provides some new ones and proposes a set of questions to be solved in future experiments.Ministerio de Economía y Competitividad TIN2013-46928-C3-3-RMinisterio de Economía y Competitividad TIN2016-76956- C3-2-RMinisterio de Economía y Competitividad TIN2015-71938-RED

Influence of radiative forcing factors on ground–air temperature coupling during the last millennium: implications for borehole climatology

Past climate variations may be uncovered via reconstruction methods that use proxy data as predictors. Among them, borehole reconstruction is a well-established technique to recover the long-term past surface air temperature (SAT) evolution. It is based on the assumption that SAT changes are strongly coupled to ground surface temperature (GST) changes and transferred to the subsurface by thermal conduction. We evaluate the SAT–GST coupling during the last millennium (LM) using simulations from the Community Earth System Model LM Ensemble (CESM-LME). The validity of such a premise is explored by analyzing the structure of the SAT–GST covariance during the LM and also by investigating the evolution of the long-term SAT–GST relationship. The multiple and single-forcing simulations in the CESM-LME are used to analyze the SAT–GST relationship within different regions and spatial scales and to derive the influence of the different forcing factors on producing feedback mechanisms that alter the energy balance at the surface. The results indicate that SAT–GST coupling is strong at global and above multi-decadal timescales in CESM-LME, although a relatively small variation in the long-term SAT–GST relationship is also represented. However, at a global scale such variation does not significantly impact the SAT–GST coupling, at local to regional scales this relationship experiences considerable long-term changes mostly after the end of the 19th century. Land use land cover changes are the main driver for locally and regionally decoupling SAT and GST, as they modify the land surface properties such as albedo, surface roughness and hydrology, which in turn modifies the energy fluxes at the surface. Snow cover feedbacks due to the influence of other external forcing are also important for corrupting the long-term SAT–GST coupling. Our findings suggest that such local and regional SAT–GST decoupling processes may represent a source of bias for SAT reconstructions from borehole measurement, since the thermal signature imprinted in the subsurface over the affected regions is not fully representative of the long-term SAT variations.Ministerio de Industria, Comercio y Competitividad (FPI grant no. BES-2015-075019)Versión del editor3,50

Spectropolarimetric multi line analysis of stellar magnetic fields

In this paper we study the feasibility of inferring the magnetic field from polarized multi-line spectra using two methods: The pseudo line approach and The PCA-ZDI approach. We use multi-line techniques, meaning that all the lines of a stellar spectrum contribute to obtain a polarization signature. The use of multiple lines dramatically increases the signal to noise ratio of these polarizations signatures. Using one technique, the pseudo-line approach, we construct the pseudo-line as the mean profile of all the individual lines. The other technique, the PCA-ZDI approach proposed recently by Semel et al. (2006) for the detection of polarized signals, combines Principle Components Analysis (PCA) and the Zeeman Do ppler Imaging technique (ZDI). This new method has a main advantage: the polarized signature is extracted using cross correlations between the stellar spectra nd functions containing the polarization properties of each line. These functions are the principal components of a database of synthetic spectra. The synthesis of the spectra of the database are obtained using the radiative transfer equations in LTE. The profiles built with the PCA-ZDI technique are denominated Multi-Zeeman-Signatures. The construction of the pseudo line as well as the Multi-Zeeman-Signatures is a powerful tool in the study of stellar and solar magnetic fields. The information of the physical parameters that governs the line formation is contained in the final polarized profiles. In particular, using inversion codes, we have shown that the magnetic field vector can be properly inferred with both approaches despite the magnetic field regime.Comment: Accepted for publication in Astronomy and Astrophysic

Microwave background anisotropies and non-linear structures II. Numerical computations

A new method for modelling spherically symmetric inhomogeneities is applied to the formation of clusters in an expanding Universe. We impose simple initial velocity and density perturbations of finite extent and we investigate the subsequent evolution of the density field. Photon paths are also calculated, allowing a detailed consideration of gravitational lensing effects and microwave background anisotropies induced by the cluster. We apply the method to modelling high-redshift clusters and, in particular, we consider the reported microwave decrement observed towards the quasar pair PC1643+4631 A&B. We also consider the effect on the primordial microwave background power spectrum due to gravitational lensing by a population of massive high-redshift clusters.Comment: 15 pages, 23 figures; Monthly Notices of the Royal Astronomical Society (MNRAS), in pres

Microwave Background Anisotropies and Nonlinear Structures I. Improved Theoretical Models

A new method is proposed for modelling spherically symmetric inhomogeneities in the Universe. The inhomogeneities have finite size and are compensated, so they do not exert any measurable gravitational force beyond their boundary. The region exterior to the perturbation is represented by a Friedmann-Robertson-Walker (FRW) Universe, which we use to study the anisotropy in the cosmic microwave background (CMB) induced by the cluster. All calculations are performed in a single, global coordinate system, with nonlinear gravitational effects fully incorporated. An advantage of the gauge choices employed here is that the resultant equations are essentially Newtonian in form. Examination of the problem of specifying initial data shows that the new model presented here has many advantages over `Swiss cheese' and other models. Numerical implementation of the equations derived here is described in a subsequent paper.Comment: 10 pages, 4 figures; Monthly Notices of the Royal Astronomical Society (MNRAS), in pres

Tracking Plum pox virus in Chile throughout the year by three different methods and molecular characterization of Chilean isolates

During 2007, a survey was performed to detect and identify Plum pox virus (PPV) in Chilean stone fruit commercial orchards. A total of 1396 trees were analyzed and 45 (3.22 %) of them resulted positive. A fragment of 467 bp, corresponding to the replicase-coat protein (Nib-CP) region from the virus genome, was amplified and the the sequences obtained permitted the characterization of all isolates as PPV-D type, confirming that, so far, this is the only serotype present in Chile. To optimize virus detection, 27 PPV-positive trees were selected and sampled monthly from December 2006 until December 2007, collecting plant tissues available at the time of sampling (leaves, cuttings, buds and flowers). Each sample was analyzed by three different techniques: DASI-ELISA, RT-PCR and non-isotopic molecular hybridization (MH). The results showed that RT-PCR was more sensitive for detection in all months excepting January 2007, when the three techniques showed the same sensitivity. In general, MH showed a better sensitivity compared with DASI-ELISA. The best plant materials for analysis were: leaves, in February, March, September and October; phloem from cuttings in June; buds in July, and flowers in August. Keywords: PPV, detection, phylogeny, samplin