Protein–phenolic interactions and inhibition of glycation – combining a systematic review and experimental models for enhanced physiological relevance

Background: While antiglycative capacity has been attributed to (poly)phenols, the exact mechanism of action remains unclear. Studies so far are often relying on supra-physiological concentrations and use of non-bioavailable compounds.<p></p> Methods: To inform the design of a physiologically relevant in-vitro study, we carried out a systematic literature review of dietary interventions reporting plasma concentrations polyphenol metabolites. Bovine Serum Albumin (BSA) was pre-treated prior to in vitro glycation: either no treatment (native), pre-oxidised (incubated with 10nM H2O2, for 8 hours) or incubated with a mixture of phenolic acids at physiologically relevant concentrations, for 8 hours). In-vitro glycation was carried out in presence of i) glucose only (0, 5 or 10mM), ii) glucose (0, 5 or 10mM) plus H2O2 (10nM), or iii) glucose (0, 5 or 10mM) plus phenolic acids (10-160nM). Fructosamine was measured using the nitroblue tetrazolium method.<p></p> Results: Following (high) dietary polyphenol intake, 3-hydroxyphenylacetic acid is the most abundant phenolic acid in peripheral blood (up to 338μM) with concentrations for other phenolic acids ranging from 13nM-200μM. Presence of six phenolic acids with BSA during in-vitro glycation did not lower fructosamine formation. However, when BSA was pre-incubated with phenolic acids, significantly lower concentration of fructosamine was detected under glycoxidative conditions (glucose 5 or 10mM plus H2O2 10nM) (p<0.001 vs. native BSA).<p></p> Conclusion: Protein pre-treatment, either with oxidants or phenolic acids, is an important regulator of subsequent glycation in a physiologically relevant system. High quality in-vitro studies under conditions closer to physiology are feasible and should be employed more frequently.<p></p&gt

Changing distributions of body size and adiposity with age

Background: Adiposity and health risks are better indicated by waist circumference than body mass index (BMI). Patterns of change with age are incompletely documented.<p></p> Methods: Adults aged 18–92 years in the Scottish and English Health Surveys of 1994–96 and 2008–10 were divided into fifteen 5-year age bands. Sex-specific prevalences of overweight/obesity and of increased/high waist circumference against age were compared using analysis of covariance. <p></p> Results: Data available for 7932 Scottish and 55 925 English subjects in 1994–96, and for 27 391 Scottish and 30 929 English in 2008–10, showed generally similar patterns of change in the two countries. Prevalences of both elevated BMI and waist circumference rose with age for longer in 2008–10 than in 1994–96, reaching higher peaks at greater ages, particularly among men. Between 1994–96 and 2008–10, maximum prevalences of BMI >30 increased from 25 to 38% (larger increases in men than women), reaching a peak at age 60–70 years in both sexes. This peak prevalence was 5–10 years later than in 1994–96 for men and remained unchanged for women. Between 1994–96 and 2008–10, maximum prevalences of high waist circumference (men>>102 cm, women>88 cm) increased from 30 to –70% in both sexes, peaking in 2008–10 at ages 80–85 years (men) and 65–70 years (women). In 2008–10, proportions of adults with ‘normal’ BMI (18.5–25) fell with age to 15–20% at age 60–70 years (men) and 75 years (women). Among all those with BMI=18.5–25, aged>65 years, the proportions with unhealthily elevated waist circumference were 30 (men>94 cm) and 55% (women>80 cm). <p></p> Conclusions: Almost 40% of men and women are now becoming obese. People are growing fatter later in life, with waist circumference rising more persistently than BMI, which may indicate increased loss of muscle mass and sarcopenia in old age. Among older people, few now have ‘normal’ BMI, and of these up to half have elevated waist circumference, raising questions for the suitability of BMI as a measure of adiposity in this age group. <p></p&gt

Role of oxidative stress in physiological albumin glycation: a neglected interaction

Protein glycation is a key mechanism involved in chronic disease development in both diabetic and nondiabetic individuals. About 12–18% of circulating proteins are glycated in vivo in normoglycemic blood, but in vitro studies have hitherto failed to demonstrate glucose-driven glycation below a concentration of 30 mM. Bovine serum albumin (BSA), reduced BSA (mercaptalbumin) (both 40 g/L), and human plasma were incubated with glucose concentrations of 0–30 mM for 4 weeks at 37 °C. All were tested preoxidized for 8 h before glycation with 10 nM H2O2 or continuously exposed to 10 nM H2O2 throughout the incubation period. Fructosamine was measured (nitroblue tetrazolium method) at 2 and 4 weeks. Oxidized BSA (both preoxidized and continuously exposed to H2O2) was more readily glycated than native BSA at all glucose concentrations (p = 0.03). Moreover, only oxidized BSA was glycated at physiological glucose concentration (5 mM) compared to glucose-free control (glycation increased by 35% compared to native albumin, p < 0.05). Both 5 and 10 mM glucose led to higher glycation when mercaptalbumin was oxidized than when unoxidized (p < 0.05). Fructosamine concentration in human plasma was also significantly higher when oxidized and exposed to 5 mM glucose, compared to unoxidized plasma (p = 0.03). The interaction between glucose concentration and oxidation was significant in all protein models (p < 0.05). This study has for the first time demonstrated albumin glycation in vitro, using physiological concentrations of albumin, glucose, and hydrogen peroxide, identifying low-grade oxidative stress as a key element early in the glycation process

Instabilities in freely expanding sheets of associating viscoelastic fluids

We use the impact of drops on a small solid target as a tool to investigate the behavior of viscoelastic fluids under extreme deformation rates. We study two classes of transient networks: semidilute solutions of supramolecular polymers and suspensions of spherical oil droplets reversibly linked by polymers. The two types of samples display very similar linear viscoelastic properties, which can be described with a Maxwell fluid model, but contrasting nonlinear properties due to different network structure. Upon impact, weakly viscoelastic samples exhibit a behavior qualitatively similar to that of Newtonian fluids: A smooth and regular sheet forms, expands, and then retracts. By contrast, for highly viscoelastic fluids, the thickness of the sheet is found to be very irregular, leading to instabilities and eventually formation of holes. We find that material rheological properties rule the onset of instabilities. We first provide a simple image analysis of the expanding sheets to determine the onset of instabilities. We then demonstrate that a Deborah number related to the shortest relaxation time associated to the sample structure following a high shear is the relevant parameter that controls the heterogeneities in the thickness of the sheet, eventually leading to the formation of holes. When the sheet tears-up, data suggest by contrast that the opening dynamics depends also on the expansion rate of the sheet.Comment: accepted for publication in Soft Matte

Serum levels of advanced glycation end-products (AGEs) and the decoy soluble receptor for AGEs (sRAGE) can discriminate non-alcoholic fatty liver disease in age-, sex- and BMI-matched normo-glycemic adults

Background: Non-alcoholic fatty liver disease (NAFLD) is a serious health problem affecting ~25% of the global population. While NAFLD pathogenesis is still unclear, multiple NAFLD parameters, including reduced insulin sensitivity, impaired glucose metabolism and increased oxidative stress are hypothesised to foster the formation of advance glycation end-products (AGEs). Given the link of AGEs with end organ damage, there is scope to examine the role of the AGE/RAGE axis activation in liver injury and NAFLD. Methods: Age, sex and body mass index matched normo-glycemic NAFLD adults (n = 58) and healthy controls (n = 58) were enrolled in the study. AGEs were analysed by liquid chromatography-mass spectrometry (CML, CEL), fluorescence (pentosidine, AGE fluorescence), colorimetry (fructosamine) and ELISA (sRAGE). Their association with liver function, inflammation, fibrosis and stage of NAFLD was examined. Results: Early and advanced glycation end-products, except Nε-carboxymethyl-L-lysine (CML), were 10–30% higher, sRAGE levels 1.7-fold lower, and glycation/sRAGE ratios 4-fold higher in the NAFLD cases compared to controls. While AGEs presented weak to moderate correlations with indices of liver function and damage (AST/ALT, HOMA-IR, TNF-α and TGF-β1), including sRAGE to characterize the AGEs/sRAGE axis strengthened the associations observed. High glycation/sRAGE ratios were associated with 1.3 to 14-fold likelihood of lower AST/ALT ratios. The sum of AGEs/sRAGE ratios accurately distinguished between healthy controls and NAFLD patients (area under the curve of 0.85). Elevated AGEs/sRAGE (>7.8 mmol/pmol) was associated with a 12-fold likelihood of the presence of NAFLD. Conclusion: These findings strengthen the involvement of AGEs-RAGE axis in liver injury and the pathogenesis of NAFLD