The Interaction of Temozolomide with Blood Components Suggests the Potential Use of Human Serum Albumin as a Biomimetic Carrier for the Drug

The interaction of temozolomide (TMZ) (the main chemotherapeutic agent for brain tumors) with blood components has not been studied at the molecular level to date, even though such information is essential in the design of dosage forms for optimal therapy. This work explores the binding of TMZ to human serum albumin (HSA) and alpha-1-acid glycoprotein (AGP), as well as to blood cell-mimicking membrane systems. Absorption and fluorescence experiments with model membranes indicate that TMZ does not penetrate into the lipid bilayer, but binds to the membrane surface with very low affinity. Fluorescence experiments performed with the plasma proteins suggest that in human plasma, most of the bound TMZ is attached to HSA rather than to AGP. This interaction is moderate and likely mediated by hydrogen-bonding and hydrophobic forces, which increase the hydrolytic stability of the drug. These experiments are supported by docking and molecular dynamics simulations, which reveal that TMZ is mainly inserted in the subdomain IIA of HSA, establishing -stacking interactions with the tryptophan residue. Considering the overexpression of albumin receptors in tumor cells, our results propose that part of the administered TMZ may reach its target bound to plasma albumin and suggest that HSA-based nanocarriers are suitable candidates for designing biomimetic delivery systems that selectively transport TMZ to tumor cells

Silica/Proteoliposomal Nanocomposite as a Potential Platform for Ion Channel Studies

The nanostructuration of solid matrices with lipid nanoparticles containing membrane proteins is a promising tool for the development of high-throughput screening devices. Here, sol-gel silica-derived nanocomposites loaded with liposome-reconstituted KcsA, a prokaryotic potassium channel, have been synthesized. The conformational and functional stability of these lipid nanoparticles before and after sol-gel immobilization have been characterized by using dynamic light scattering, and steady-state and time-resolved fluorescence spectroscopy methods. The lipid-reconstituted KcsA channel entrapped in the sol-gel matrix retained the conformational and stability changes induced by the presence of blocking or permeant cations in the buffer (associated with the conformation of the selectivity filter) or by a drop in the pH (associated with the opening of the activation gate of the protein). Hence, these results indicate that this novel device has the potential to be used as a screening platform to test new modulating drugs of potassium channels

Numerical simulation and experiments of the multiphase flow in a liquid-liquid cylindrical cyclone separator

A Liquid-Liquid Cylindrical Cyclone separator (LLCC) is a device used in the petroleum industry to separate the oil-water mixture obtained from the well. The use of this device has not been widespread due to the lack of tools for predicting its separation capability. This paper presents a numerical and experimental study of the fluid dynamic performance of this type of cylindrical cyclone separators. The use of numerical simulations would reduce the time and cost necessary to obtain information for predicting the behavior of the equipment. The objective of this study is to determine if CFD (Computational Fluid Dynamics) techniques are able to reproduce the behavior of a LLCC separator. The CFD software examined was ANSYS-CFX 5.6TM and numerical simulations were carried out using the dispersed model with oil as the dispersed phase. The oil and water mixture entering the separator is divided due to centrifugal and buoyancy forces in an upper (oil rich) exit and a bottom (water rich) exit. The separation capability is determined as the maximum amount of water removed from the mixture with the minimum amount of oil content in the water rich exit. The experiments were conducted in a transparent LLCC separator that allows the visualization of the mixture and the measurement of the oil content. Experiments were conducted for three variables: mixture velocity and water content at the entrance, and the split ratio. The split ratio is defined as the bottom exit flow rate divided by the water flow rate at the entrance. The results showed that CFD tools are able to reproduce the oil content obtained from the experiments for all analyzed conditions. Additionally, the mixture distribution images from numerical and experimental data showed good agreement. This study confirms the capacity of CFD tools for the multiphase flow analysis of LLCC separators

Numerical simulation and experiments of the multiphase flow in a liquid-liquid cylindrical cyclone separator

A Liquid-Liquid Cylindrical Cyclone separator (LLCC) is a device used in the petroleum industry to separate the oil-water mixture obtained from the well. The use of this device has not been widespread due to the lack of tools for predicting its separation capability. This paper presents a numerical and experimental study of the fluid dynamic performance of this type of cylindrical cyclone separators. The use of numerical simulations would reduce the time and cost necessary to obtain information for predicting the behavior of the equipment. The objective of this study is to determine if CFD (Computational Fluid Dynamics) techniques are able to reproduce the behavior of a LLCC separator. The CFD software examined was ANSYS-CFX 5.6TM and numerical simulations were carried out using the dispersed model with oil as the dispersed phase. The oil and water mixture entering the separator is divided due to centrifugal and buoyancy forces in an upper (oil rich) exit and a bottom (water rich) exit. The separation capability is determined as the maximum amount of water removed from the mixture with the minimum amount of oil content in the water rich exit. The experiments were conducted in a transparent LLCC separator that allows the visualization of the mixture and the measurement of the oil content. Experiments were conducted for three variables: mixture velocity and water content at the entrance, and the split ratio. The split ratio is defined as the bottom exit flow rate divided by the water flow rate at the entrance. The results showed that CFD tools are able to reproduce the oil content obtained from the experiments for all analyzed conditions. Additionally, the mixture distribution images from numerical and experimental data showed good agreement. This study confirms the capacity of CFD tools for the multiphase flow analysis of LLCC separators

The chromatin remodeller CHD8 is required for E2F-dependent transcription activation of S-phase genes

The precise regulation of S-phase-specific genes is critical for cell proliferation. How the repressive chromatin configuration mediated by the retinoblastoma protein and repressor E2F factors changes at the G1/S transition to allow transcription activation is unclear. Here we show ChIP-on-chip studies that reveal that the chromatin remodeller CHD8 binds ∼2000 transcriptionally active promoters. The spectrum of CHD8 target genes was enriched in E2F-dependent genes. We found that CHD8 binds E2F-dependent promoters at the G1/S transition but not in quiescent cells. Consistently, CHD8 was required for G1/S-specific expression of these genes and for cell cycle re-entry on serum stimulation of quiescent cells. We also show that CHD8 interacts with E2F1 and, importantly, loading of E2F1 and E2F3, but not E2F4, onto S-specific promoters, requires CHD8. However, CHD8 recruiting is independent of these factors. Recruiting of MLL histone methyltransferase complexes to S-specific promoters was also severely impaired in the absence of CHD8. Furthermore, depletion of CHD8 abolished E2F1 overexpression-dependent S-phase stimulation of serum-starved cells, highlighting the essential role of CHD8 in E2F-dependent transcription activation

Genome-wide study of chromatin remodeling factor CHD8 role in transcription

1 página. Cold Spring Harbor Laboratory (CSHL) Meeting on Mechanisms of Eukaryotic Trasncription 2011. August 30 - september 3, 2011.CHD8 (Chromodomain-Helicase-DNA binding protein 8) is a member of the chromodomain helicase DNA-binding (CHD) subfamily of enzymes, which also belongs to the SNF2 family of ATP-dependent chromatin remodelers.Peer reviewe

Disposable Electrochemical Biosensor Based on the Inhibition of Alkaline Phosphatase Encapsulated in Acrylamide Hydrogels

The present work describes the development of an easy-to-use portable electrochemical biosensor based on alkaline phosphatase (ALP) as a recognition element, which has been immobilized in acrylamide-based hydrogels prepared through a green protocol over disposable screen-printed electrodes. To carry out the electrochemical transduction, an electroinactive substrate (hydroquinone diphosphate) was used in the presence of the enzyme and then it was hydrolyzed to an electroactive species (hydroquinone). The activity of the protein within the matrix was determined voltammetrically. Due to the adhesive properties of the hydrogel, this was easily deposited on the surface of the electrodes, greatly increasing the sensitivity of the biosensor. The device was optimized to allow the determination of phosphate ion, a competitive inhibitor of ALP, in aqueous media. Our study provides a proof-of-concept demonstrating the potential use of the developed biosensor for in situ, real-time measurement of water pollutants that act as ALP inhibitors.This research was funded by Ministerio de Ciencia e Innovación (projects PID2019-105923RB-I00, PDC2021-120884-I00, European Union NextGenerationEU PRTR-C17.I1) and Generalitat Valenciana (project GVA-THINKINAZUL/2021/015 and PROMETEO/2018/087)

CHD3 Proteins and Polycomb Group Proteins Antagonistically Determine Cell Identity in Arabidopsis

Dynamic regulation of chromatin structure is of fundamental importance for modulating genomic activities in higher eukaryotes. The opposing activities of Polycomb group (PcG) and trithorax group (trxG) proteins are part of a chromatin-based cellular memory system ensuring the correct expression of specific transcriptional programs at defined developmental stages. The default silencing activity of PcG proteins is counteracted by trxG proteins that activate PcG target genes and prevent PcG mediated silencing activities. Therefore, the timely expression and regulation of PcG proteins and counteracting trxG proteins is likely to be of fundamental importance for establishing cell identity. Here, we report that the chromodomain/helicase/DNA–binding domain CHD3 proteins PICKLE (PKL) and PICKLE RELATED2 (PKR2) have trxG-like functions in plants and are required for the expression of many genes that are repressed by PcG proteins. The pkl mutant could partly suppress the leaf and flower phenotype of the PcG mutant curly leaf, supporting the idea that CHD3 proteins and PcG proteins antagonistically determine cell identity in plants. The direct targets of PKL in roots include the PcG genes SWINGER and EMBRYONIC FLOWER2 that encode subunits of Polycomb repressive complexes responsible for trimethylating histone H3 at lysine 27 (H3K27me3). Similar to mutants lacking PcG proteins, lack of PKL and PKR2 caused reduced H3K27me3 levels and, therefore, increased expression of a set of PcG protein target genes in roots. Thus, PKL and PKR2 are directly required for activation of PcG protein target genes and in roots are also indirectly required for repression of PcG protein target genes. Reduced PcG protein activity can lead to cell de-differentiation and callus-like tissue formation in pkl pkr2 mutants. Thus, in contrast to mammals, where PcG proteins are required to maintain pluripotency and to prevent cell differentiation, in plants PcG proteins are required to promote cell differentiation by suppressing embryonic development

Physiological Responses of Species to Microclimate Help explain Population Dynamics along Succession in a Tropical Dry Forest of Yucatan, Mexico

We investigated relationships between population dynamics and microclimate, physiology, and the degree of mycorrhizal colonization, for three species (Piscidia piscipula L.(Sarg.)) (Fabaceae), Bunchosia swartzianaGriseb. (Malpighiaceae) and Psidium sartorianum (Bergius) Nied. (Myrtaceae)) of a tropical sub deciduous forest in Yucatan, Mexico that were growing in plots of different successional ages. We hypothesized that abundance and persistence were related to increased plasticity in CO2assimilation. We found that Piscidia piscipula had greater abundance in intermediate plots (18 to 21 years), presented higher levels of plasticity in CO2 assimilation (greater variability among individuals, plots, and seasons), presented the highest CO2 assimilation rates, and presented greater drought resistance (higher water potentials and capacitance). Conversely, Psidium sartorianum had greater abundance in older plots (more than 50 years of secondary succession), lower assimilation rates, and low levels of plasticity in CO2 assimilation. Bunchosia had intermediate values. Locally, the degree of mycorrhizal colonization was consistent with abundance across plots. Regionally (but not locally), plasticity in CO2 assimilation was consistent with abundance. We found differences in microclimates among plots and within plots among species. Physiological adjustments appeared to play an important role in the capacity to regenerate and in the successional persistence of these species in this tropical dry forest