Inhibition of Class I Histone Deacetylase Activity Blocks the Induction of TNFAIP3 Both Directly and Indirectly via the Suppression of Endogenous TNF-α

Histone deacetylase inhibitors (HDIs) are promising drugs for the treatment of inflammatory diseases. However, their therapeutical exploitation is slowed down by severe adverse manifestations that can hardly be foreseen, mainly due to incomplete knowledge of how HDIs impact the delicate balance of inflammatory mediators. In this work, we characterized the effects of the HDI trichostatin A (TSA) on the expression of TNFAIP3, which is a crucial inhibitor of the classical NF-kB pathway and an LPS-induced negative feedback regulator. The accumulation of TNFAIP3 mRNA after LPS stimulation showed biphasic behavior, with one wave within the first hour of stimulation and a second wave several hours later, which were both reduced by TSA. By using inhibition and knockdown approaches, we identified two temporally and mechanistically distinct modes of action. The first wave of TNAIP3 accumulation was directly blunted by the histone deacetylase (HDAC) blockade. By contrast, the second wave was decreased mainly because of the lack of endogenous TNF-α induction, which, in turn, depended on the intact HDAC activity. In both cases, class I HDACs appeared to play a nonredundant role, with HDAC3 required, but not sufficient, for TNF-α and TNFAIP3 induction. In addition to TNFAIP3, TNF-α is known to induce many response genes that orchestrate the inflammatory cascade. Thus, suppression of TNF-α may represent a general mechanism through which HDIs regulate a selected set of target genes.     

Magnetic-Based Human Tissue 3D Cell Culture: A Systematic Review

Cell-based assays, conducted on monolayer (2D) cultured cells, are an unquestionably valuable tool for biomedical research. However, three-dimensional (3D) cell culture models have gained relevance over the last few years due to the advantages of better mimicking the microenvironment and tissue microarchitecture in vivo. Recent magnetic-based 3D (m3D) cell culture systems can be used for this purpose. These systems are based on exposing magnetized cells to magnetic fields by levitation, bioprinting, or ring formation to promote cell aggregation into 3D structures. However, the successful development of these structures is dependent on several methodological characteristics and can be applied to mimic different human tissues. Thus, a systematic review was performed using Medline (via Pubmed), Scopus, and Web of Science (until February 2022) databases to aggregate studies using m3D culture in which human tissues were mimicked. The search generated 3784 records, of which 25 met the inclusion criteria. The usability of these m3D systems for the development of homotypic or heterotypic spheroids with or without scaffolds was explored in these studies. We also explore methodological differences specifically related to the magnetic method. Generally, the development of m3D cultures has been increasing, with bioprinting and levitation systems being the most used to generate homotypic or heterotypic cultures, mainly to mimic the physiology of human tissues, but also to perform therapeutic screening. This systematic review showed that there are areas of research where the application of this method remains barely explored, such as cancer research.     

Extracellular Vesicles in Diffuse Large B Cell Lymphoma: Characterization and Diagnostic Potential

Diffuse large B cell lymphoma (DLBCL) is an aggressive B cell lymphoma characterized by a heterogeneous behavior and in need of more accurate biological characterization monitoring and prognostic tools. Extracellular vesicles are secreted by all cell types and are currently established to some extent as representatives of the cell of origin. The present study characterized and evaluated the diagnostic and prognostic potential of plasma extracellular vesicles (EVs) proteome in DLBCL by using state-of-the-art mass spectrometry. The EV proteome is strongly affected by DLBCL status, with multiple proteins uniquely identified in the plasma of DLBCL. A proof-of-concept classifier resulted in highly accurate classification with a sensitivity and specificity of 1 when tested on the holdout test data set. On the other hand, no proteins were identified to correlate with non-germinal center B-cell like (non-GCB) or GCB subtypes to a significant degree after correction for multiple testing. However, functional analysis suggested that antigen binding is regulated when comparing non-GCB and GCB. Survival analysis based on protein quantitative values and clinical parameters identified multiple EV proteins as significantly correlated to survival. In conclusion, the plasma extracellular vesicle proteome identifies DLBCL cancer patients from healthy donors and contains potential EV protein markers for prediction of survival.     

Inverse analysis with integral transformed temperature fields: Identification of thermophysical properties in heterogeneous media

The objective of this work is to introduce the use of integral transformed temperature measured data for the solution of inverse heat transfer problems, instead of the common local transient temperature measurements. The proposed approach is capable of significantly compressing the measured data through the integral transformation, without losing the information contained in the measurements and required for the solution of the inverse problem. The data compression is of special interest for modern measurement techniques, such as the infrared thermography, that allows for fine spatial resolutions and large frequencies, possibly resulting on a very large amount of measured data. In order to critically address the use of integral transformed measurements, we examine in this paper the simultaneous estimation of spatially variable thermal conductivity and thermal diffusivity in one-dimensional heat conduction within heterogeneous media. The direct problem solution is analytically obtained via integral transforms and the related eigenvalue problem is solved by the Generalized Integral Transform Technique (GITT). The inverse problem is handled with Bayesian inference by employing a Markov Chain Monte Carlo (MCMC) method. The unknown functions appearing in the formulation are expanded in terms of eigenfunctions as well, so that the unknown parameters become the corresponding series coefficients. Such projection of the functions in an infinite dimensional space onto a parametric space of finite dimension also permits that several quantities appearing in the solution of the direct problem be analytically computed. Simulated measurements are used in the inverse analysis; they are assumed to be additive, uncorrelated, normally distributed, with zero means and known covariances. Both Gaussian and non-informative uniform distributions are used as priors for demonstrating the robustness of the estimation procedure.     

Forecasting electricity prices and their volatilities using Unobserved Components

The liberalization of electricity markets more than ten years ago in the vast majority of developed countries has introduced the need of modelling and forecasting electricity prices and volatilities, both in the short and long term.Thus, there is a need of providing methodology that is able to deal with the most important features of electricity price series, which are well known for presenting not only structure in conditional mean but also time-varying conditional variances.In this work we propose a new model, which allows to extract conditionally heteroskedastic common factors from the vector of electricity prices. These common factors are jointly estimated as well as their relationship with the original vector of series, and the dynamics affecting both their conditional mean and variance. The estimation of the model is carried out under the state-space formulation.The new model proposed is applied to extract seasonal common dynamic factors as well as common volatility factors for electricity prices and the estimation results are used to forecast electricity prices and their volatilities in the Spanish zone of the Iberian Market.Several simplified/alternative models are also considered as benchmarks to illustrate that the proposed approach is superior to all of them in terms of explanatory and predictive power.     

Prediction of fatty acid composition using a NIRS fibre optics probe at two different locations of ham subcutaneous fat

NIR technology was used to analyze the fatty acid composition of fat tissue from conjugated linoleic acid fed pigs by direct application of a fibre optics probe on two different locations of subcutaneous fat. One hundred and fifty-three pigs were fed three different diets and the fatty acid profile of subcutaneous fat from Gluteus medius was analyzed using gas chromatography and FT-NIR in a longitudinal and transversal cut. Spectra were acquired using a Bruker Optics Matrix-F duplex spectrometer equipped with a fibre optics probe (IN-268-2). Total saturated, monounsaturated and polyunsaturated together as well as oleic and stearic fatty acids were predicted accurately (R² > 0.70), myristic, and linoleic fatty acids were predicted with lower accuracy (0.60 < R² < 0.70), while palmitic, asclepic and α-linolenic fatty acids were poorly predicted (R² < 0.60). Results indicated that NIR technology has potential as a rapid tool to discriminate carcasses from animals fed diets with different fatty acid composition by a direct measurement on the fat from the ham.     

Changes on the levels of serotonin precursors – tryptophan and 5-hydroxytryptophan – during roasting of Arabica and Robusta coffee

The levels of free and total tryptophan and of 5-hydroxytryptophan (5-HTP) were investigated in green and roasted grains and beverages of Coffea arabica L. (Arabica) and Coffea canephora Pierre var. robusta (Robusta). Grains were light, medium and dark roasted. Free and protein tryptophan were extracted before and after hydrolysis. The levels of tryptophan and 5-HTP were quantified simultaneously by ion-pair HPLC and fluorimetric detection after derivatisation with o-phthalaldehyde. Robusta green coffee had higher total and protein tryptophan, whereas Arabica had higher free tryptophan levels. 5-HTP was not detected in the samples before and after roasting. Free tryptophan was completely degraded during roasting. Roasting significantly affected protein tryptophan. The rate of loss was smaller in Arabica compared to Robusta at every roasting degree. A beverage prepared the Brazilian way with a medium-roasted coffee provided 1.4–2.5 mg tryptophan/50 ml cup.     

Determination of naturally occurring progestogens in bovine milk as their oxime derivatives using high performance liquid chromatography–electrospray ionization–tandem mass spectrometry

BACKGROUND: Hormones and hormone‐like substances which are present in the environment have been repeatedly accused of being the cause of most endocrine disruption. However, the possible role of endogenous hormones in food of animal origin deserves to be discussed as well. The relation between steroid hormones and several human health problems has been previously reported, such as prostate and breast cancer, perturbation of human reproduction and endocrine disruption on humans and wildlife. This research is particularly concerned with cow's milk, which contains a considerable amount of sex hormones. RESULTS: A liquid chromatography–tandem mass spectrometry method has been developed for the simultaneous detection and quantification of four naturally occurring steroid hormones in commercial bovine milk (pregnenolone (P₅), progesterone (P₄), 17‐hydroxypregnenolone (17‐OHP₅), 17‐hydroxyprogesterone (P₄)). Oxime derivatives of steroids were analyzed in positive ionization and multiple reaction monitoring mode. Methodology has been validated according to Decision 2002/657/EC criteria. CONCLUSION: This method has been successfully used in real samples. It is fast and easy‐handling and provides a useful tool for the assessment of progestogens in bovine milk. Copyright © 2010 Society of Chemical Industry     

Enhancement of sweetness intensity in gels by inhomogeneous distribution of sucrose

Model systems consisting of layers of mixed agar/gelatin gel were used to investigate the effect of the spatial distribution of sucrose on perceived sweetness intensity. 2-Alternative forced choice tests were performed with consumers to compare the sweetness of layered samples with an inhomogeneous distribution of sucrose to the sweetness of a reference sample with a homogeneous distribution. All samples had the same overall sucrose concentration (10%) and similar mechanical and rheological properties. Inhomogeneous samples that had large concentration gradients of sucrose between layers were perceived sweeter than the homogeneous reference. No differences in sweetness were observed between the reference and samples with small concentration gradients of sucrose. Additionally, the position of the layers containing sucrose did not affect sweetness. Results showed that an inhomogeneous distribution of sucrose can be used to reduce sucrose content by 20% without a decrease in sweetness intensity.