Role of extracellular matrix components and structure in new renal models in vitro

The extracellular matrix (ECM), a complex set of fibrillar proteins and proteoglycans, supports the renal parenchyma and provides biomechanical and biochemical cues critical for spatial-temporal patterning of cell development and acquisition of specialized functions. As in vitro models progress towards biomimicry, more attention is paid to reproducing ECM-mediated stimuli. ECM’s role in in vitro models of renal function and disease used to investigate kidney injury and regeneration is discussed. Availability, affordability, and lot-to-lot consistency are the main factors determining the selection of materials to recreate ECM in vitro. While simpler components can be synthesized in vitro, others must be isolated from animal or human tissues, either as single isolated components or as complex mixtures, such as Matrigel or decellularized formulations. Synthetic polymeric materials with dynamic and instructive capacities are also being explored for cell mechanical support to overcome the issues with natural products. ECM components can be used as simple 2D coatings or complex 3D scaffolds combining natural and synthetic materials. The goal is to recreate the biochemical signals provided by glycosaminoglycans and other signaling molecules, together with the stiffness, elasticity, segmentation, and dimensionality of the original kidney tissue, to support the specialized functions of glomerular, tubular, and vascular compartments. ECM mimicking also plays a central role in recent developments aiming to reproduce renal tissue in vitro or even in therapeutical strategies to regenerate renal function. Bioprinting of renal tubules, recellularization of kidney ECM scaffolds, and development of kidney organoids are examples. Future solutions will probably combine these technologies

Human hydroxytyrosol's absorption and excretion from a nutraceutical

Among the various (poly)phenols that are being sold as such or as part of a more complex mixture, hydroxytyrosol (HT) is the only one that bears a European Food Safety Authority health claim. Therefore, several HT-based products are being developed and sold and it becomes necessary to evaluate its accessibility following ingestion. Twenty-one volunteers were recruited for a randomized, crossover, placebo-controlled, and double-blind intervention study.We performed a Latin square design: after one-week washout, i.e. olive-free diet, subjects were randomly assigned to the placebo (maltodextrin), 5, or 25 mg/day HT group. Twenty-four hour urine samples were collected after the intervention week, and baseline urines were collected the week before the study and during periods of washout. The results show that HT given as the foremost component of a nutraceutical preparation is bioavailable and is recovered in the urine chiefly as sulphate-3′

On the de Haas - van Alphen oscillations in quasi-two-dimensional metals: effect of the Fermi surface curvature

Here, we present the results of theoretical analysis of the de Haas-van Alphen oscillations in quasi-two-dimensional normal metals. We had been studying effects of the Fermi surface (FS) shape on these oscillations. It was shown that the effects could be revealed and well pronounced when the FS curvature becomes zero at cross-sections with extremal cross-sectional areas. In this case both shape and amplitude of the oscillations could be significantly changed. Also, we analyze the effect of the FS local geometry on the angular dependencies of the oscillation amplitudes when the magnetic field is tilted away from the FS symmetry axis by the angle $\theta.$ We show that a peak appears at $\theta \approx 0$ whose height could be of the same order as the maximum at the Yamaji angle. This peak emerges when the FS includes zero curvature cross-sections of extremal areas. Such maximum was observed in experiments on the $\alpha-(BETS)_4TIHg(SeCN)_4.$ The obtained results could be applied to organic metals and other quasi-two-dimensional compounds.Comment: 9 pages, 4 figures, text added, references adde

Endotoxin increase after fat overload is related to postprandial hypertriglyceridemia in morbidly obese patients.

The low-grade inflammation observed in obesity has been associated with a high-fat diet, though this relation is not fully understood. Bacterial endotoxin, produced by gut microbiota, may be the linking factor. However, this has not been confirmed in obese patients. To study the relationship between a high-fat diet and bacterial endotoxin, we analyzed postprandial endotoxemia in morbidly obese patients after a fat overload. The endotoxin levels were determined in serum and the chylomicron fraction at baseline and 3 h after a fat overload in 40 morbidly obese patients and their levels related with the degree of insulin resistance and postprandial hypertriglyceridemia. The morbidly obese patients with the highest postprandial hypertriglyceridemia showed a significant increase in lipopolysaccharide (LPS) levels in serum and the chylomicron fraction after the fat overload. Postprandial chylomicron LPS levels correlated positively with the difference between postprandial triglycerides and baseline triglycerides. There were no significant correlations between C-reactive protein (CRP) and LPS levels. The main variables contributing to serum LPS levels after fat overload were baseline and postprandial triglyceride levels but not glucose or insulin resistance. Additionally, superoxide dismutase activity decreased significantly after the fat overload. Postprandial LPS increase after a fat overload is related to postprandial hypertriglyceridemia but not to degree of insulin resistance in morbidly obese patients

Metabolic fingerprint after acute and under sustained consumption of a functional beverage based on grape skin extract in healthy human subjects

Grape-derived polyphenols are considered to be one of the most promising ingredients for functional foods due to their health-promoting activities. We applied a HPLC-MS-based untargeted metabolomic approach in order to evaluate the impact of a functional food based on grape skin polyphenols on the urinary metabolome of healthy subjects. Thirty-one volunteers participated in two dietary crossover randomized intervention studies: with a single-dose intake (187 mL) and with a 15-day sustained consumption (twice per day, 187 mL per day in total) of a functional beverage (FB). Postprandial (4-hour) and 24-hour urine samples collected after acute consumption and on the last day of sustained FB consumption, respectively, were analysed using an untargeted HPLC-qTOF-MS approach. Multivariate modelling with subsequent application of an S-plot revealed differential mass features related to acute and prolonged consumption of FB. More than half of the mass features were shared between the two types of samples, among which several phase II metabolites of grape-derived polyphenols were identified at confidence level II. Prolonged consumption of FB was specifically reflected in urine metabolome by the presence of first-stage microbial metabolites of flavanols: hydroxyvaleric acid and hydroxyvalerolactone derivatives. Overall, several epicatechin and phenolic acid metabolites both of tissular and microbiota origin were the most representative markers of FB consumption. To our knowledge, this is one of the first studies where an untargeted LC-MS metabolomic approach has been applied in nutrition research on a grape-derived FB

Effect of Theobroma cacao flavonoids on immune activation of a lymphoid cell line

We analysed the effect of (-)-epicatechin and cocoa extract on the activation of a lymphoid cell line. Particularly the expression of IL-2 receptor alpha (IL-2Ralpha or CD25) and, the secretion of IL-2 and IL-4 were established after flavonoid treatment. Two media culture conditions (1 and 10 % of fetal calf serum supplementation) and the different moments of flavonoid addition (simultaneously or 2 h before cell-activation) were compared. IL-2Ralpha (CD25) expression on activated cells was significantly reduced by epicatechin and cocoa extract in a dose-dependent manner, achieving the highest inhibition of about 50 % when flavonoids were added 2 h before stimulation. IL-2 secretion was also inhibited by the presence of both epicatechin and cocoa extract, displaying 60 and 75 % of inhibition, respectively. Cocoa flavonoids were also able to enhance 3-4.5-fold IL-4 release. In summary, cocoa extract down-modulated T lymphocyte activation and therefore the acquired immune response. This fact could be important in some states of the immune system hyperactivity such as autoimmune or chronic inflammatory diseases

Clinical Phenotype Clustering in Cardiovascular risk patients for the identification of Responsive Metabotypes after red Wine Polyphenol intake

This study aims to evaluate the robustness of clinical and metabolic phenotyping through, for the first time, the identification of differential responsiveness to dietary strategies in the improvement of cardiometabolic risk conditions. Clinical phenotyping of 57 volunteers with cardiovascular risk factors was achieved using k-means cluster analysis based on 69 biochemical and anthropometric parameters. Cluster validation based on Dunn and FOM analysis for internal coherence and external homogeneity were employed. k-means produced four clusters with particular clinical profiles. Differences on urine metabolomic profiles among clinical phenotypes were explored and validated by multivariate OSC-PLS-DA models. OSC-PLS-DA of 1H-NMR data revealed that model comparing 'obese and diabetic cluster' (OD-c) against 'healthier cluster' (H-c) showed the best predictability and robustness in terms of explaining the pairwise differences between clusters. Considering these two clusters, distinct groups of metabolites were observed following an intervention with wine polyphenol intake (WPI, 733 equivalents of gallic acid/day) per 28 days. Glucose was significantly linked to OD-c metabotype (p<0.01), and lactate, betaine and dimethylamine showed a significant trend. Whereas, associated to wine polyphenol intervention (OD-c_WPI and H-c_WPI) was tartrate (p<0.001), and mannitol, threonine methanol, fucose and 3-hydroxyphenylacetate showed a significant trend. Interestingly, 4-hydroxyphenylacetate significantly increased in H-c_WPI (p<0.05) compared to OD-c_WPI and to basal groups (gut microbial derived metabolite after polyphenol intake), thereby exhibiting a clear metabotypic intervention effect. Results revealed gut microbiota responsive phenotypes to wine polyphenols intervention. Overall, this study illustrates a novel metabolomic strategy for characterizing inter-individual responsiveness to dietary intervention and identification of health benefits

Total urinary polyphenols and longitudinal changes of bone properties. The InCHIANTI study

SummaryThe aim of this study was to evaluate the association of levels of urinary total polyphenols considered as a proxymeasure of polyphenol intake, with longitudinal changes of bone properties, in the InCHIANTI study. Dietary intake ofpolyphenols appears to be associated with future accelerated deterioration of bone health.IntroductionPolyphenols, micronutrients ingested through plant-based foods, have antioxidant and anti-inflammatory propertiesand may contribute to osteoporosis prevention. We evaluated associations of high levels of urinary total polyphenols (UTP), aproxy measure of polyphenol intake, with longitudinal changes of bone properties in a representative cohort of free-livingparticipants of the InCHIANTI study.MethodsThe InCHIANTI study enrolled representative samples from the registry list of two towns in Tuscany, Italy. Baselinedata were collected in 1998 and follow-up visits in 2001 and 2004. Of the 1453 participants enrolled, 956 consented to donate a24-h urine sample used to assess UTP, had dietary assessment, a physical examination, and underwent a quantitative comput-erized tomography (pQCT) of the tibia. From pQCT images, we estimated markers of bone mass (BM), diaphyseal design (DD),and material quality (MQ). Mixed models were used to study the relationship between baseline tertiles of UTP with changes ofthe bone characteristics over the follow-up.ResultsAt baseline, higher levels of UTP were positively correlated with markers of BM, DD, and MQ. Compared with lowertertile of UTP, participants in the intermediate and highest tertiles had higher cortical bone area, cortical mineral content, andcortical thickness. However, participants in the intermediate and highest UTP tertiles experienced accelerated deterioration ofthese same parameters over the follow-up compared with those in the lowest UTP tertile.ConclusionsDietary intake of polyphenols estimated by UTP and dietary questionnaire was associated with long-term acceler-ated deterioration of bone health. Our study does not support the recommendation of increasing polyphenol intake for osteopo-rosis prevention

Development and characterization of a microfluidic model of the tumour microenvironment

The physical microenvironment of tumours is characterized by heterotypic cell interactions and physiological gradients of nutrients, waste products and oxygen. This tumour microenvironment has a major impact on the biology of cancer cells and their response to chemotherapeutic agents. Despite this, most in vitro cancer research still relies primarily on cells grown in 2D and in isolation in nutrient- and oxygen-rich conditions. Here, a microfluidic device is presented that is easy to use and enables modelling and study of the tumour microenvironment in real-time. The versatility of this microfluidic platform allows for different aspects of the microenvironment to be monitored and dissected. This is exemplified here by real-time profiling of oxygen and glucose concentrations inside the device as well as effects on cell proliferation and growth, ROS generation and apoptosis. Heterotypic cell interactions were also studied. The device provides a live ‘window’ into the microenvironment and could be used to study cancer cells for which it is difficult to generate tumour spheroids. Another major application of the device is the study of effects of the microenvironment on cellular drug responses. Some data is presented for this indicating the device’s potential to enable more physiological in vitro drug screening

Metabolomics-guided insights on bariatric surgery versus behavioral interventions for weight loss.

Despite evidence to support their utility, lifestyle-based strategies for weight loss and treatment of obesity (i.e., based on diet and physical activity) have met so far with little success in the long term in terms of permanent weight loss (1). Bariatric surgery is the only current treatment for obesity leading to sustained weight loss (2) and to improvements in glucose regulation, up to a complet