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In this study, we investigated the complexation of bioactive isoflavones, specifically biochanin A (BCA) and genistein (GEN), with hen egg white lysozyme (HEWL) and explored their inhibitory effects on HEWL modification using a combination of multi-spectroscopic and computational methods. The observed binding affinity was of a moderate nature (on the order of 104 M−1), and a static quenching mechanism was identified in the fluorescence quenching process. Notably, the binding constant (Kb) for GEN (4.449 ± 0.262 × 104 M−1) was found to be higher than that for BCA (3.707 ± 0.108 × 104 M−1) towards HEWL. Our spectroscopic measurements, complemented by molecular docking calculations, suggested the involvement of Trp62 in the binding site of the isoflavones within the geometry of HEWL. The micro-environment surrounding the Trp-residues exhibited an increase in hydrophilicity, as indicated by Synchronous fluorescence (SFS) and three-dimensional fluorescence (3D) studies. Interestingly, circular dichroism (CD) studies revealed no marked alteration in the secondary structure of HEWL upon binding with the isoflavones. Furthermore, our investigation into the interaction patterns, employing FTIR and molecular docking studies, revealed a predominance of hydrogen bonding and hydrophobic interactions. Beyond the binding study, the isoflavones demonstrated a promising inhibitory effect on the d-ribose-mediated glycation of HEWL, as well as on HEWL fibrillation, as evidenced by fluorescence emission studies. Our findings not only exhibited an excellent correlation with experimental observations but also provided precise insights into the location and dynamics of isoflavones within the binding site through detailed analyses of molecular docking and molecular dynamics simulation data.

The H1 receptor belongs to the family of rhodopsin-like G-protein-coupled receptors activated by the biogenic amine histamine. H1 receptor antagonists are widely used in the treatment of allergies. However, these drugs could have a much broader spectrum of activity, including hypoglycemic effects, which can broaden the spectrum of their use. The aim of the study was to evaluate the antiglycation potential of twelve H1 receptor antagonists (diphenhydramine, antazoline, promethazine, ketotifen, clemastine, pheniramine, cetirizine, levocetirizine, bilastine, fexofenadine, desloratadine, and loratadine). Bovine serum albumin (BSA) was glycated with sugars (glucose, fructose, galactose, and ribose) and aldehydes (glyoxal and methylglyoxal) in the presence of H1 blockers. The tested substances did not induce a significant decrease in the content of albumin glycation end-products, and the inhibition rate of glycoxidation was not influenced by the chemical structure or generation of H1 blockers. None of the tested H1 receptor antagonists exhibited strong antiglycation activity. Antiglycemic potential of H1 blockers could be attributed to their antioxidant and anti-inflammatory activity, as well as their effects on carbohydrate metabolism/metabolic balance at the systemic level.

The differences in maximum Maillard glycation extent of proteins incubated with saccharides of different size/charge might be explained by differences in saccharide reactivity towards each protein modification site. Mixtures of α-lactalbumin-glucose (AG), α-lactalbumin-maltotriose (AMTT) or α-lactalbumin-galacturonic acid (AGalA) were incubated for 10 h, reaching a glycation extent of 42, 19 and 30 %, respectively. Glycated protein samples were enzymatically hydrolyzed followed by UPLC-UV-MS determination and quantification of glycation in individual modification sites. On average, around 80 % of protein glycation was recovered as glycated peptides. For all protein-saccharide mixtures, the N-terminal amino group, K58, K62 and K114 were never glycated, whilst K5 and K122 were always the most glycated sites. The glycated sites R10, K79, K93, K94 and K108 found in AGalA, were absent from AG and AMtt. Differences in glycated/non-glycated sites could not explain the differences in total protein glycation between the various samples.

The objective of this study was to investigate a replacement for phosphate in meat products. Protein structural modification was employed in this study, and grafted myofibrillar protein (MP) with palatinose was added to meat emulsion without phosphate. Here, 0.15% of sodium polyphosphate (SPP) was replaced by the same (0.15%) concentration and double (0.3%) the concentration of grafted MP. Although the thermal stability was decreased, the addition of transglutaminase could increase stability. The rheological properties and pH also increased with the addition of grafted MP and transglutaminase. The addition of grafted protein could be perceived by the naked eye by observing a color difference before cooking, but it was not easy to detect after cooking. The cooking loss, emulsion stability, water holding capacity, lipid oxidation, and textural properties improved with the addition of grafted MP. However, the excessive addition of grafted MP and transglutaminase was not recommended to produce a high quality of phosphate replaced meat emulsion, and 0.15% was identified as a suitable addition ratio of grafted MP.

Diabetes and ulcer are the major health problems all over the world. The present study reports synthesis and bio-evaluation of 19 benzimidazole analogs in search of antiglycation, antioxidant and antiulcer agents. The synthetic analogs were characterized using 1H NMR, 13C NMR and HR-EIMS. All compounds were checked for their antiglycation, antiurease and antioxidant activities. The fluorophenyl benzimidazole analogs 12–14 strongly inhibited glycation with IC50 values ranging from 142 µM to 193 µM. The same fluorophenyl analogs (12–14) were also found to exhibit the highest antioxidant activity with IC50 values ranging from 1.2 µM to 6.6 µM which further highlights the significance of these bioactive analogs. The dihydroxyphenyl analogs 6–9 demonstrated the most potent enzyme inhibitory activity with IC50 values ranging from 3.10 µM to 5.90 µM. Molecular docking studies were performed on the active analogs to investigate their interactions with the urease enzyme and provide a plausible explanation for their observed urease inhibitory activity.

Protein glycation, hyper-inflammatory reactions, and oxidative stress play a crucial role in the pathophysiology of numerous diseases. The current work evaluated the protective ability of ethyl alcohol extract of leaves from holy basil (Ocimum sanctum Linn) against inflammation, oxidative stress, glycation and advanced glycation endproducts formation. Various in vitro assays assessed prementioned properties of holy basil. In addition, molecular docking was conducted. The highest hydrogen peroxide reduction activity (72.7 %) and maximum percentage of DPPH scavenging (71.3 %) depicted its vigorous antioxidant abilities. Furthermore, it showed the most excellent protection against proteinase activity (67.247 %), prevention of denaturation of egg albumin (65.29 %), and BSA (bovine serum albumin) (68.87 %) with 600 µg/ml. Percent aggregation index (57.528 %), browning intensity (56.61 %), and amyloid structure (57.0 %) were all reduced significantly using 600 μg/ml of extract. Additionally, the antimicrobial potential was also confirmed. According to a molecular docking study, active leaf extract ingredients were found to bind with superoxide dismutase, catalase, and carbonic anhydrase. As a conclusion, O. sanctum has a variety of health-promoting properties that may reduce the severity of many diseases in diabetic patients. However, in order to ascertain the mechanisms of action of the components of its leaves in disease prevention, more thorough research based on pharmacological aspects is needed.

Glycosylation is a crucial post-translational modification process where sugar molecules (glycans) are covalently linked to proteins, lipids, or other biomolecules. In this highly regulated and complex process, a series of enzymes are involved in adding, modifying, or removing sugar residues. This process plays a pivotal role in various biological functions, influencing the structure, stability, and functionality of the modified molecules. Glycosylation is essential in numerous biological processes, including cell adhesion, signal transduction, immune response, and biomolecular recognition. Dysregulation of glycosylation is associated with various diseases. Glycation, a post-translational modification characterized by the non-enzymatic attachment of sugar molecules to proteins, has also emerged as a crucial factor in various diseases. This review comprehensively explores the multifaceted role of glycation in disease pathogenesis, with a specific focus on its implications in osteoarthritis (OA). Glycosylation and glycation alterations wield a profound influence on OA pathogenesis, intertwining with disease onset and progression. Diverse studies underscore the multifaceted role of aberrant glycosylation in OA, particularly emphasizing its intricate relationship with joint tissue degradation and inflammatory cascades. Distinct glycosylation patterns, including N-glycans and O-glycans, showcase correlations with inflammatory cytokines, matrix metalloproteinases, and cellular senescence pathways, amplifying the degenerative processes within cartilage. Furthermore, the impact of advanced glycation end-products (AGEs) formation in OA pathophysiology unveils critical insights into glycosylation-driven chondrocyte behavior and extracellular matrix remodeling. These findings illuminate potential therapeutic targets and diagnostic markers, signaling a promising avenue for targeted interventions in OA management. In this comprehensive review, we aim to thoroughly examine the significant impact of glycosylation or AGEs in OA and explore its varied effects on other related conditions, such as liver-related diseases, immune system disorders, and cancers, among others. By emphasizing glycosylation's role beyond OA and its implications in other diseases, we uncover insights that extend beyond the immediate focus on OA, potentially revealing novel perspectives for diagnosing and treating OA.

In this study, we systematically investigated the emulsifying capabilities of myofibrillar protein (MP)- and MP peptide (MPP)-based conjugates synthesized through intensification techniques: water bath (WB), microwave, ultrasound, and the combined ultrasound-microwave (UM) methods. Compared with WB, microwave, and ultrasound treatments, the combined UM treatment greatly promoted the glycation reaction because ultrasound and microwave mutually reinforced modification effects. The resultant conjugate structure tended to unfold with more flexible conformation and homogeneous morphology. Moreover, the emulsifying properties of conjugates developed with single and combined ultrasound-assisted glycation displayed substantial improvement, and pre-hydrolysis further enhanced these performances, as observed in the Principal Component Analysis as well. Remarkably, MPP grafted by maltodextrin with the assistance of a combined UM field produced the smallest and most uniform emulsion system, positioning it as the most efficient emulsifier among all the fabricated glycoconjugates. Our study highlighted the potential of synergistically applying ultrasound and microwave techniques to develop a well-performance glycation with an ideal conjugate structure, in which they would be associated into a strong film that provided the robust physical barrier, creaming stability, heat retention, and oxidation resistance. These findings offered a basis for better utilizing complex ultrasonic technology to develop novel and improved MP-based food products.

Glycated proteins are effective materials for improving the environmental stability and bioavailability of biologically active chemicals. The whey protein concentrate (WPC)-xylose (D) conjugates produced by the Maillard process were used to load and protect purple sweet potato anthocyanins in this study. Response surface optimization was used to determine the best parameters for preparing WPC-D conjugates. Glycation altered the secondary structure of proteins, according to structural studies, which also had an impact on their functional characteristics, which showed better antioxidant activity before and after simulated gastrointestinal (GI) digestion (p

Poor emulsification of myofibrillar proteins (MPs) limits the production of meat protein emulsion-type products, and it is related to the myosin self-assembles in low-salt settings. The effect of high-intensity ultrasound (HIU) pretreatment combined with non-enzymatic glycation on MP-stabilized emulsions in low-salt settings was investigated in this study, and the potential mechanism was revealed. The results indicated that, compared to using either HIU or glycation treatment alone, HIU pretreatment in combination with glycation significantly improves the physical stability of emulsions while increasing the distribution uniformity and reducing the droplet particle size from 18.05 μm to 2.54 μm (P

This study determined the effect of polyherbal tea blend (PTB) from Roselle (Hibiscus sabdariffa) Calyx, Soursop (Annona. Muricata), and Moringa (Moringa oleifera) leave on fasting blood glucose level (FBGL), glycated hemoglobin, insulin, and enzymes [arginase, angiotensin-I converting enzyme, thiobarbituric acid reactive species, phosphodiesterase-5 (PDE-5), and purinergic and antioxidant enzymes] related to erectile dysfunction (ED) in the penile homogenate of diabetes mellitus (DM) rats’ model. The samples were prepared and blended in the ratio of 2:2:6 and bagged to contain 1 gram. The sweetening/sweetening (S/US- PTB) teabags, were infused with 20 ml of boiled water for 5 (S/US-PTB-5) and 10 (S/US-PTB-10) min, respectively. The animals were made diabetic [except the normal control (NC) group] with streptozotocin intraperitoneally, and grouped into NC, DM, DM+Acarbose (ACA), DM+S-PTB-5, DM+S-PTB-10, DM+US-PTB-5, and DM+US-PTB-10. The result revealed that the PTB decreased FBGL and related enzymes, but improved insulin level and antioxidant status in the diabetes-induced ED (DMED), while the S/US-PTB-10 appeared better. Since the formulated PTB is in line with the therapeutic effect, the tea can be used as a therapeutic agent for managing not only diabetes but also DMED. However, sexual behavior, clinical trials, and phytochemical screening of the PTB should be carried out.

ABSTRACT: The Maillard reaction (MR), under proper environmental conditions, has been used to improve protein functionality. In the present work, 2 high temperatures (50–80°C) and water activity (Aw; 0.45–0.67) were used to promote exogenous glycosylation of glycomacropeptide (GMP) while minimizing processing times (0, 8, 24, 48, and 96 h at 50°C; 0, 2, 4, 8, and 24 h at 80°C). Maltodextrin, a polysaccharide commonly used in the food industry as a functional ingredient, was used as a reducing sugar, and compared with lactose, a native milk sugar. The progression of MR was evaluated by tracking changes in molecular weight using SDS-PAGE, the formation of Amadori compounds, and browning. Aqueous glycosylated GMP solutions (5 to 20% wt/vol) were tested for solubility, rheological properties, and foam formation. As expected, MR progression was faster with Aw = 0.67 and 80°C. Glycosylated GMP powders showed no change in their solubility after MR. However, the apparent viscosity (γ=30s-1) of the 20% wt/vol suspensions exhibited a slight increase when GMP was glycosylated with maltodextrin for 24 h at 80°C, and a 2-log increase when GMP was glycosylated with lactose, with a high browning development in both cases. The foam expansion index of the resuspended glycosylated powders was increased by between 25 and 66% compared with the nonglycosylated powders. Better foam stability (approximately 2 h) and no browning development were observed for GMP glycosylated with maltodextrin for 2 h at Aw = 0.67 and 80°C. The results show that GMP has undergone further glycosylation by means of controlled MR, which improves viscosity and foaming index without negatively affecting solubility. These preliminary studies provide a basis for the future creation of a new ingredient with GMP and reducing sugars.

Senescence is a natural phenomenon of growing old. It accelerates under certain conditions like diabetes mellitus resulting in early decline of bodily functions, which can be avoided by many claimed functional foods. The present study aims to investigate the anti-aging ability of Fenugreek seeds (Trigonellafoenum-graecum); a common ingredient of Indo-Pak cuisines. Briefly, the Fenugreek seeds extract (FgSE) in concentrationsof0.1, 0.5 and 1 mg/ml inhibited the formation of Advanced Glycation End products (AGEs) and fructosamine adducts in Bovine serum albumin (BSA)/fructose model in vitro. The BSA conformational analysis via Circular Dichorism and Congo red assays showed that it preserves secondary structure of BSA in aforementioned model. Although mechanistic studies revealed insignificant lysine blocking ability of Fenugreek by OPA assay, however carbonyl entrapping was found to be 24%, 34% and 42% at 0.1, 0.5 and 1 mg/ml, respectively. In vivo model of High Fructose diet (HFD) induced glycation, FgSE treatment in doses of 10, 25 & 50 mg/kg markedly improved Escape latency (p

Presently, many studies assess allergenicity via IgE immunoblotting and IgE binding tests; however, IgE detection does not always signal the manifestation of a clinical allergy. However, the capacity of food allergens to trigger basophils makes it possible to use the in vitro functional basophil activation test (BAT) to assess allergenicity. The effect of the Maillard reaction (MR) on the allergenic potential of processed soy proteins was evaluated by two IgE binding tests (Competitive ELISA and Inhibition ImmunoCAP), a Western Blot and a functional BAT; with the aim to analyze whether the sIgE binding results correspond to the functional assay results. The results between the IgE binding tests and the functional assay were in-line for 2 of the 6 studied patients. For one patient there was no correlation between any of the results. For the raw soy protein extract heated with glucose for 10 and 30 min (SH SPE + Glu and LH SPE + Glu, respectively), the results were in-line for only 3 out of the 6 patients. Thus, the present study shows a discrepancy between IgE binding tests and basophil stimulation when assessing the effect of soy processing on its allergenicity. Since IgE-binding capacity does not always correlate to IgE cross-linking capacity, the conclusions of the allergenic potential based on the IgE binding tests alone should be drawn with care and further studies on this matter would benefit from the inclusion of a functional assay such as the BAT.

Objectives: Diabetes mellitus (DM), cancer and cardiovascular diseases (CVD) are major contributors to human miseries and death. Additionally, previous findings stated that cancer is also known to be associated with diabetes. Therefore, in the given study, facile synthesis of gold nanoparticles (AuNPs) using glycation reaction was achieved and further, their bioconjugation with herbal drug garcinol (G) was performed (G-AuNPs) in order to enhance the antiglycative, antidiabetic and anticancer efficacy of the garcinol. Methods: The antiglycation and antidiabetic potential of garcinol bioconjugated AuNPs (G-AuNPs) were assessed through different experiments. Further, anticancer potential against adenocarcinoma cells were also assessed. Results: The study confirmed 78.6 % bioconjugation of G over the surface of AuNPs. Furthermore, the antiglycation study depicted that G-AuNPs are found to be more effective inhibitor of the glycation reaction in contrast to pure G. The findings represented that G-AuNPs significantly blocks the formation of early glycation adducts and AGEs, also they mask the glycation prone free arginine and lysine residues from participating in the glycation process. Additionally, our α-amylase inhibition assay results demonstrated that pure G (IC50 − 8.9 µM) has significantly higher α-amylase inhibition activity compared to standard inhibitor, acarbose (IC50 − 0.118 mM). Also, the anticancer study described that G and G-AuNPs treatment resulted in death of the cells via apoptosis, which resulted in elevated permeability, condensed chromatin, deep blue fluorescent, and condensed nucleus of A549 cells. Our findings also revealed that the potential of mitochondrial membrane (ΔΨm) of A549 cells were also disrupted at much lower concentration of G-AuNPs (13.3 µM) as compared to pure G (28.7 µM). Conclusion: The key findings of the investigation suggested that G-AuNPs acted as a potent antiglycation, antidiabetic, and anticancer agent.

Low-density lipoprotein (LDL) transports cholesterol to various tissues via the blood. Glycation of LDL occurs during hyperglycemic condition which is characterised by persistently high blood glucose level. Circulating erythrocytes can come in direct contact with glycated LDL (G-LDL). The objective of this study was to investigate the effect of G-LDL on human erythrocytes, specifically on hemoglobin, intracellular generation of reactive species and the antioxidant defence system. Isolated erythrocytes were incubated with G-LDL (3 and 6 mg/ml) and native LDL (6 mg/ml) at 37 °C for 24 h. Native LDL and G-LDL untreated erythrocytes were similarly incubated at 37 °C and served as control. G-LDL treatment increased hemolysis compared to control and native LDL-treated erythrocytes. Incubation of erythrocytes with G-LDL led to an increase in protein oxidation and lipid peroxidation while greatly decreasing the total sulfhydryl content. It also significantly enhanced hemoglobin oxidation, heme degradation, and the release of free iron moiety. Treatment with G-LDL led to an appreciable increase in the production of reactive oxygen and nitrogen species. The antioxidant power and activities of major antioxidant enzymes were drastically reduced, while critical membrane-bound enzymes were inhibited. The surface morphology of G-LDL-treated erythrocytes was altered leading to the formation of echinocytes. Importantly, treatment of erythrocytes with native LDL did not significantly affect the above-mentioned parameters and values were similar to the corresponding controls. Thus, G-LDL is cytotoxic to human erythrocytes and causes oxidative damage to cell components. This can reduce the oxygen-transporting ability of blood and also result in red cell senescence and anemia., Competing Interests: Declaration of competing interest The authors of this paper declare no social or financial conflict of interest in this research., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: The non-enzymatic glycation of proteins and their advanced glycation end products (AGEs) are associated with protein transformations such as in the development of diseases and biopharmaceutical storage. The characterization of heavily glycated proteins at the intact level is of high interest as it allows to describe co-occurring protein modifications. However, the high heterogeneity of glycated protein makes this process challenging, and novel methods are required to accomplish this., Results: In this study, we investigated two novel LC-HRMS methods to study glycated reference proteins at the intact protein level: low-flow hydrophilic-interaction liquid chromatography (HILIC) and native size-exclusion chromatography (SEC). Model proteins were exposed to conditions that favored extensive glycation and the formation of AGEs. After glycation, complicated MS spectra were observed, along with a sharply reduced signal response, possibly due to protein denaturation and the formation of aggregates. When using HILIC-MS, the glycated forms of the proteins could be resolved based on the number of reducing monosaccharides. Moreover, some positional glycated isomers were separated. The SEC-MS method under non-denaturing conditions provided insights into glycated aggregates but offered only a limited separation of glycated species based on molar mass. Overall, more than 25 different types of species were observed in both methods, differing in molar mass by 14-162 Da. 19 of these species have not been previously reported., Significance: The proposed strategies show great potential to characterize highly glycated intact proteins from native and denaturing perspectives and provide new opportunities for fast clinical diagnoses and investigating glycation-related diseases., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ziran Zhai reports financial support was provided by Chinese Scholarship Council. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

We aimed to characterize the metabolomic profiles in preterm small-for-gestational age (SGA) infants using cord blood. We conducted a gestational age (GA)-matched case-control study that included 30 preterm infants who were categorized into two groups: SGA infants, with a birth weight (BW) < 10th percentile for GA (n = 15) and non-SGA infants, with BW ≥ 10th percentile for GA (n = 15). SGA infants with chromosomal or genetic abnormalities were excluded. At birth, the umbilicus was double-clamped, and the cord blood was sampled from the umbilical vein. Metabolomic analyses were performed using capillary electrophoresis time-of-flight mass spectrometry. The median GA at birth was not significantly different between the two groups [SGA, 32 (26-36) weeks; non-SGA, 32 (25-35) weeks; p = 0.661)]. Of the 255 metabolites analyzed, 19 (7.5%) showed significant differences between SGA and non-SGA infants. There were significant reductions in the carnosine, hypotaurine, and S-methylcysteine levels in SGA infants as compared to non-SGA infants (p < 0.05). Carnosine was correlated with gestational age, BMI before pregnancy, body weight gain during pregnancy (p = 0.002, p = 0.023, and p = 0.020, respectively). In conclusion, preterm SGA infants have low levels of cord blood antioxidative- and antiglycation-related metabolites, making them vulnerable to oxidative stress., (Copyright © 2024 Taiwan Pediatric Association. Published by Elsevier B.V. All rights reserved.)

Diabetes-mediated development of micro and macro-vascular complications is a global concern. One of the factors is hyperglycemia induced the non-enzymatic formation of advanced glycation end products (AGEs). Accumulated AGEs bind with receptor of AGEs (RAGE) causing inflammation, oxidative stress and extracellular matrix proteins (ECM) modifications responsible for fibrosis, cell damage and tissue remodeling. Moreover, during hyperglycemia, aldosterone (Aldo) secretion increases, and its interaction with mineralocorticoid receptor (MR) through genomic and non-genomic pathways leads to inflammation and fibrosis. Extensive research on individual involvement of AGEs-RAGE and Aldo-MR pathways in the development of diabetic nephropathy (DN), cardiovascular diseases (CVDs), and impaired immune system has led to the discovery of therapeutic drugs. Despite mutual repercussions, the cross-talk between AGEs-RAGE and Aldo-MR pathways remains unresolved. Hence, this review focuses on the possible interaction of Aldo and glycation in DN and CVDs, considering the clinical significance of mutual molecular targets., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Human serum albumin (HSA) is known to undergo modifications by glucose during diabetes. This process produces glycated HSA that can have altered binding to some drugs. In this study, high-performance affinity microcolumns and competition studies were used to see how glycation affects the binding by two thiazolidinedione-class drugs (i.e., pioglitazone and rosiglitazone) at specific regions of HSA. These regions included Sudlow sites I and II, the tamoxifen and digitoxin sites, and a drug-binding site located in subdomain IB. At Sudlow site II, the association equilibrium constants (or binding constants) for pioglitazone and rosiglitazone with normal HSA were 1.7 × 10 5 M -1 and 2.0 × 10 5 M -1 at pH 7.4 and 37 °C, with values that changed by up to 5.7-fold for glycated HSA. Sudlow site I of normal HSA had binding constants for pioglitazone and rosiglitazone of 3.4 × 10 5 M -1 and 4.6 × 10 5 M -1 , with these values changing by up to 1.5-fold for glycated HSA. Rosiglitazone was found to also bind a second region that had a positive allosteric effect on Sudlow site I for all the tested preparations of HSA (binding affinity, 1.1-3.2 × 10 5 M -1 ; coupling constant for Sudlow site I, 1.20-1.34). Both drugs had a strong positive allosteric effect on the tamoxifen site of HSA (coupling constants, 13.7-19.9 for pioglitazone and 3.7-11.5 for rosiglitazone). Rosiglitazone also had weak interactions at a site in subdomain IB, with a binding constant of 1.4 × 10 3 M -1 for normal HSA and a value that was altered by up to 6.8-fold with glycated HSA. Neither of the tested drugs had any significant binding at the digitoxin site. The results were used to produce affinity maps that described binding by these thiazolidinediones with HSA and the effects of glycation on these interactions during diabetes., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: David S Hage reports financial support was provided by National Institutes of Health. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In hyperglycemia, accelerated glycation and oxidative stress give rise to many diabetic complications, such as diabetic cardiomyopathy (DCM). Glycated human serum albumin (GHSA) has disturbed structural integrity and hampered functional capabilities. When GHSA accumulates around cardiac cells, Nrf-2 is dysregulated, aiding oxidative stress. L-Arginine (L-Arg) is prescribed to patients with diabetes and cardiovascular diseases. This research contributes to the mechanistic insights on antiglycation and antioxidant potential of L-Arg in alleviating DCM. HSA was glycated with methylglyoxal in the presence of L-Arg (20-640 mM). Structural and functional modifications of HSA were studied. L-Arg and HSA, GHSA interactions, and thermodynamics were determined by steady-state fluorescence. H9c2 cardiomyocytes were given treatments of GHSA-L-Arg along with the inhibitor of the receptor of AGEs. Cellular antioxidant levels, detoxification enzyme activities were measured. Gene, protein expressions, and immunofluorescence data examined the activation and nuclear translocation of Nrf-2 during glycation and oxidative stress. L-Arg protected HSA from glycation-induced structural and functional modifications. The binding affinity of L-Arg was more towards HSA (10 4  M -1 ). L-Arg, specifically at lower concentration (20 mM), upregulated Nrf-2 gene, protein expressions and facilitated its nuclear translocation by activating Nrf-2 signaling. The study concluded that L-Arg can be of therapeutic advantage in glycation-induced DCM and associated oxidative stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The Reactive carbonyl species (RCS) like, 3-deoxyglucosone (3-DG) is a highly reactive intermediate of the glycation reaction which may result into the formation of protein advanced glycation end-products (AGEs), like myoglobin-AGEs (Mb-AGEs) which further lead to the structural perturbations. In the present work, we studied the glycation of myoglobin (Mb) by physico-chemical techniques and the resulting immunogenicity by ELISA techniques. The level of free lysine and arginine determined was found to be significantly decreased indicating the establishment of the glycation reaction. The glycated myoglobin (G-Mb) shows the increased absorbance at 280 nm and enhanced fluorescence emission at 350 nm excitation wavelengths. The levels of ketoamine and hydroxymethylfurfural (HMF) were recorded to be enhanced in G-Mb when compared to its native conformer. Furthermore, the New-Zealand white female rabbits immunized with native and its glycated analogue ‘G-Mb’ induced high titer antibodies as compared to the native one.

The present study simulates the fracture behavior of diabetic cortical bone with high levels of advanced glycation end-products (AGEs) under dynamic loading. We consider that the increased AGEs in diabetic cortical bone degrade the materials heterogeneity of cortical bone through a reduction in critical energy release rates of the microstructural features. To simulate the initiation and propagation of cracks, we implement a phase field fracture framework on 2D models of human tibia cortical microstructure. The simulations show that the mismatch between the fracture properties (e.g., critical energy release rate) of osteons and interstitial tissue due to high AGEs contents can change crack growth trajectories. The results show crack branching in the cortical microstructure under dynamic loading is affected by the mismatches related to AGEs. In addition, we observe cortical features such as osteons and cement lines can prevent multiple cracking under dynamic loading even with changing the mismatches due to high AGEs. Furthermore, under dynamic loading, some toughening mechanisms can be activated and deactivated with different AGEs contents. In conclusion, the current findings present that the combination of the loading type and materials heterogeneity of microstructural features can change the fracture response of diabetic cortical bone and its fragility., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Natural products exist in various natural foods such as plants, herbs, fruits, and vegetables. Furthermore, marine life offers potential natural products with significant biological activity. The biochemical reaction is known as advanced glycation end products (AGEs) occurs in the human body. On the other hand, foods are capable of a wide range of processing conditions resulting in the generation of exogenous AGEs adducts. Protein glycation and the formation of advanced glycation end products both contribute to the pathogenesis of hyperglycemic complications. AGEs also play a pivotal role in microvascular and macrovascular complications progression by receptors for advanced glycation end products (RAGE). RAGE activate by AGEs leads to up-regulation of transcriptional factor NF-kB and inflammatory genes. Around the globe, researchers are working in various approaches for therapeutical implications on controlling AGEs mediated disease complications. In this regard, one of the potential promising agents observed with a wide range of AGEs inhibition by food-derived natural products. Current biotechnological tools have been turned to natural products or phytochemicals to manufacture the molecules without compromising their functionality. Metabolic engineering and bioinformatics perspectives have recently enabled the generation of a few potent metabolites with anti-diabetic activity. As the primary focus, this review article will also discuss multidisciplinary approaches that emphasize current advances in anti-diabetic therapeutic action and future perspectives of natural products.

Diabetic nephropathy (DN) is a diabetic complication characterized by disruption of renal microvasculature, reactive oxygen species accumulation and increased inflammation, all of which contribute to renal injury. Phenethyl isothiocyanate (PEITC) is a naturally occurring isothiocyanate well known for its antioxidant and anti-inflammatory effects, yet its reno-preventive effects against DN has not been investigated. The current study looked into the in vivo reno-protective effects of PEITC in STZ-induced DN in rats. PEITC (3, 10 and 30 mg/kg) was administered orally for 8 weeks post DM establishment. PEITC treatment significantly improved kidney and liver functions, renal histopathological features, tissue fibrosis, macrophage infiltration and blood glucose levels compared to DN control. Mechanistically, PEITC treatment alleviated DN-induced renal damage via modulating glycation and oxidative stresses and inflammatory response. As such, PEITC activated glyoxalase 1 (GLO1) that induced a retraction in renal tissue expression of advanced glycation end products (AGEs) and its receptor (RAGE). PEITC activated nuclear erythroid 2-related factor 2 (Nrf2) and increased expression of its downstream targets, hemeoxygenase-1 (HO-1) and gamma glutamate–cysteine (γ-GCS). Additionally, PEITC treatment decreased the expression of Nrf2 repressor protein, keap1. The anti-inflammatory effect of PEITC was driven, at least in part, via reducing the NLRP3 inflammasome activation as indicated by down regulation of NLRP3, TXNIP, capsase-1 and IL-1β, TNF-alpha and IL-6. In conclusion; PEITC attenuated DN progression in a dose dependent manner mainly via interruption of AGE/RAGE and NLPR3/TXNIP/NrF2 crosstalk.

Low-density lipoprotein (LDL) modification such as oxidation and glycation has been known to play a significant role in the initiation and progression of atherosclerosis. The inhibitors on LDL modification have been emerged as a new strategy to be a supplement for prevention of atherosclerosis. In an effort to discover inhibitors on LDL modification from natural products, phytochemical investigation on Cynanchum wilfordii roots were performed, leading to isolation of acetophenones (1–6) including a new one (1) and nine pregnane-type steroids (7–15). The structure of the compound 1 was determined by analysis of 1D and 2D NMR spectral data analysis. All the isolated compounds were evaluated for their inhibitory activities against LDL modification. Among them, compounds 2 and 5 strongly inhibited LDL oxidation and glycation. These results suggest that compounds 2 and 5 from C. wilfordii may provide potential protective effects against atherosclerosis via inhibition of LDL oxidation and glycation.

Paraoxonase 2 (PON2) is an intracellular anti-oxidant protein ubiquitously expressed in all cells and reduces reactive oxygen species, endoplasmic reticulum (ER) stress, further improves mitochondrial function and thereby shows anti-apoptotic function. In diabetes and its complications this PON gets glycated and becomes in effective. The PON activity is reported to be reduced in diabetic retinopathy and we have earlier showed Carboxy methyl lysine (AGE) decreased PON2 expression and activity in Human retinal endothelial cells (HREC) . In this study, we have designed and developed a mutated PON2 by in silico and in vitro approach which can resist glycation. Where in glycation-prone residues in PON2 was predicted using in silico analyses and a mutated PON2 was developed using in vitro site directed mutagenesis (SDM) assay mPON2 (mutant PON2-PON2-K70A) and its efficacy was compared with wPON2 (wild type PON2). CML glycated wPON2 and reduced its activity when compared with mPON2 in HREC confirmed by immunoprecipitation and in vitro experiments. Additionally, mPON2 interaction efficiency with its substrates was higher than wPON2 by insilico assay and demonstrated enhanced inhibition against CML-induced oxidative stress, ER stress, pro-inflammation, and mitochondrial fission than wPON2 by invitro assay. Further mPON2 showed increased inhibition of phosphorylation of NFĸB induced by CML. Our investigation establishes that the over expression of mPON2 in HREC can defy glycation and therefore mitigate ER stress and inflammation against CML than endogenous wPON2. These findings imply that mPON2 can be a beneficial therapeutic target against diabetic retinopathy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The process of glycation, characterized by the non-enzymatic reaction between sugars and free amino groups on biomolecules, is a key contributor to the development and progression of both microvascular and macrovascular complications associated with diabetes, particularly due to persistent hyperglycemia. This glycation process gives rise to advanced glycation end products (AGEs), which play a central role in the pathophysiology of diabetes complications, including nephropathy. The d-ribose-mediated glycation of fibrinogen plays a central role in the pathogenesis of diabetes nephropathy (DN) and retinopathy (DR) by the generation and accumulation of advanced glycation end products (AGEs). Glycated fibrinogen with d-ribose (Rb-gly-Fb) induces structural changes that trigger an autoimmune response by generating and exposing neoepitopes on fibrinogen molecules. The present research is designed to investigate the prevalence of autoantibodies against Rb-gly-Fb in individuals with type 2 diabetes mellitus (T2DM), DN & DR. Direct binding ELISA was used to test the binding affinity of autoantibodies from patients' sera against Rb-gly-Fb and competitive ELISA was used to confirm the direct binding findings by checking the bindings of isolated IgG against Rb-gly-Fb and its native conformer. In comparison to healthy subjects, 32% of T2DM, 67% of DN and 57.85% of DR patients' samples demonstrated a strong binding affinity towards Rb-gly-Fb. Both native and Rb-gly-Fb binding by healthy subjects (HS) sera were non-significant (p > 0.05). Furthermore, the early, intermediate, and end products of glycation have been assessed through biochemical and physicochemical analysis. The biochemical markers in the patient groups were also significant (p < 0.05) in comparison to the HS group. This study not only establishes the prevalence of autoantibodies against d-ribose glycated fibrinogen in DN but also highlights the potential of glycated fibrinogen as a biomarker for the detection of DN and/or DR. These insights may open new avenues for research into novel therapeutic strategies and the prevention of diabetes-related nephropathy and retinopathy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Advanced glycation end products are the most important species of glycation pathway, and cause disorders such as oxidative stress and diabetes. Sulfonamide compounds, which are generally known as widespread inhibitors, are potential agents used in different drug products, which can readily enter biological matrices. In the present work, the structure and activity of human carbonic anhydrase II studied in the presence of glucose as well as four sulfonamide agents from different views. These included enzyme kinetics, free lysine content, fluorescence spectroscopy, circular dichroism, and ROS measurement. Our results indicated that upon glycation, the structure of HCA II collapses and 8 to 13 lysine residues will be more available based on ligand incubation. Secondary and tertiary structural changes were also observed in the presence and absence of sulfonamide agents using fluorescence and circular dichroism methods, respectively. These spectroscopic data also showed a remarkable increase in hydrophobicity and decrease in α-helix contents during glycation, especially after 35 days of incubation. ROS assay was studied in the presence of glucose and sulfonamide compounds, that demonstrated the role of sulfonamide compounds in ROS formation in the presence of glucose in a synergistic manner. Overall, our data indicated that sulfonamides act as a stimulant factor upon prolonged interaction with HCA II and may intensify the complications of some disorders, such as diabetes and other conformational diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Black soldier fly (Hermetia illucens) has received considerable interest as an alternative protein source. Aqueous solutions of black soldier fly larvae (BSFL) protein and glucose (2:1 w.w−1, pH 9) were heated at 50, 70 and 90 °C, for 2–10 h at 2 h intervals, respectively. The zeta-potential (ζ) of BSFL-Glu conjugates heat-treated at 70 °C ranged from -10.25 to -25.25 mV while the native BSFL protein ranged from -12.84 to -16.70 mV. The ζ-potential analysis revealed that the glycation reaction modified the surface charge density of the BSFL protein as a function of reaction time and temperature. In addition, an increase in thermal stability of the BSFL-Glu conjugates was observed by means of Thermo-gravimetric analysis (TGA) and differential scanning calorimetry (DSC). Fourier transform infrared spectroscopy (FT-IR) analysis indicated that the most apparent structural changes in the BSFL protein were in the amide I and amide II region. Well-separated clusters permitting differentiation between native BSFL and BSFL-Glu conjugates were observed by using principal component analysis (PCA) on FT-IR spectra. At 50, 70 and 90 °C the first two principal components (PC1 and PC2) showed an accumulated total variance of 91, 96 and 95%, respectively. A classification efficiency of 91% was obtained when using soft independent modelling of class analogy (SIMCA). Infrared spectroscopy combined with SIMCA is a powerful tool to monitor the formation of edible insect protein–sugar conjugates by Maillard reaction. As a result, combining FT-IR spectroscopy with multivariate techniques (PCA and SIMCA) exhibited a strong potential to differentiate between native and glycated protein samples from black soldier fly larvae.

This study aims to optimize the ultrasound treatment conditions for enhancing the degree of glycation (DG) of ovalbumin (OVA)-xylose conjugates through Maillard reaction and investigate the correlation between DG and functional properties affected by structural changes. The structural and functional properties of classical heating OVA, glycated OVA, ultrasonic treated OVA, and ultrasound-assisted glycated OVA were investigated to explore the interaction mechanism of ultrasound treatment on foaming and emulsifying properties improvement. Results indicated that the ultrasound assistance increased free sulfhydryl content, surface hydrophobicity and particle size of OVA-xylose conjugates, and thus enhancing the surface properties, which were strongly linear correlated with DG under different glycation parameters (pH, xylose/OVA ratio, heating time). Additionally, circular dichroism spectroscopy analysis revealed that ultrasound promoted the conversion of α-helices to β-sheets and unfolded structures, which was consistent with the formation of short amyloid-like aggregates that observed by atomic force microscopy phenomenon. Overall, our study provides new insights into the effects of ultrasound treatment on Maillard-induced protein functional properties enhancement, which may be a new strategy to tune the DG and functionality of protein-saccharide grafts during ultrasound processing.

Background: Kappaphycus alvarezii (Doty) Doty ex P.C.Silva is a red algae with antioxidant and antiglycation activities. Algae still have not been widely used for treating diabetes, especially to prevent complications. The purpose of this study was to examine the effect of active fractions from Kappaphycus alvarezii on plasma glucose level, glycation process and renal RAGE gene expression. Methods: This study used bioassay-guided fractionation, consisting of three stages: extraction, partition, and fractionation. These processes were monitored with Thin Layer Chromatography and the BSA-Glucose method to select the best extract with antiglycation activity (calculated as the percentage of inhibition and IC50). The selected active fraction from four fractions was further used for in vivo study, which was conducted with hyperglycemic Wistar male rats. Plasma glucose level was measured using GOD-PAP methods, while plasma glycated albumin (GA) and Nε- (carboxymethyl) lysine (CML) levels were measured using ELISA. Renal RAGE gene expression was analyzed using qPCR. Results: Fraction II was selected as the active fraction of Kappaphycus alvarezii showing antiglycation activity with the highest percentage of inhibition and the lowest IC50. This fraction significantly reduced plasma GA and CML levels, but it did not significantly reduce plasma glucose level. Furthermore, renal RAGE gene expression was lower in the diabetic rat group treated with this active fraction compared to the untreated group. Conclusions: This study successfully identified an active fraction of Kappaphycus alvarezii with antiglycation activity to reduce plasma GA and CML levels as well as renal RAGE gene expression. Therefore, this fraction could be developed as a potential candidate for treating diabetes.

Our previous studies suggested that early glycation products (EGPs) generated in the first step of Maillard reaction/glycation were anti-inflammatory. The objectives of the present study were to determine the effects of EGPs derived from the whey protein isolate-glucose system on type 1 diabetes (T1D), and the underlying immunological mechanisms. In non-obese diabetic (NOD) mice, EGPs at the physiological dose of 600 mg/kg/day increased glucose metabolism, decreased non-fasting blood glucose levels and T1D incidence, decreased insulin resistance, and decreased the pancreatic immune infiltration. The protective effects were accompanied with decreases in CD4−CD8+ thymocytes, CD8+ T cells and serum insulin autoantibody levels, and increases in splenic CD4+CD25+ T cells, macrophage M2/M1 ratio and serum IL-10 level. However, similar treatment with EGPs produced minimal effect on the multiple low-dose streptozotocin-induced hyperglycemia. In conclusion, EGPs protected NOD mice against T1D via increasing anti-inflammatory immune responses and decreasing autoreactivity to self-antigens.

We investigated the effect of cyanidin-3-rutinoside (C3R) against methylglyoxal (MG)-advanced glycation end products (AGEs) mediated cardio-vascular abnormalities in rats pre-treated with MG. In the rats, plasma MG level rose following oral administration of MG (60–120 mg/rat/day) for 8 weeks and was accompanied by a buildup of MG-derived AGE, Nɛ-(carboxymethyl) lysine, in aortic tissue both of which were decreased by co-treatment with C3R (30 and 100 mg/kg/day). Similarly, MG-impaired vascular contraction/relaxation responses were normalized by C3R with concomitant upregulation of endothelial nitric oxide synthase mRNA expression. In the whole animal subjected to coronary artery ligation, C3R (100 mg/kg/day) reduced ventricular extra beats and ventricular tachycardia precipitated during acute ischemic episode. We suggest that C3R reduces the formation and accumulation of AGEs via direct trapping of the MG and upregulation of mRNA expression of glyoxalase I. The findings implicate C3R as a candidate compound for preventing MG-AGE mediated cardiovascular abnormalities in diabetes.

Glycation of amino or guanidino groups of proteins with glucose and glucose-derived reactive aldehydes, such as α-hydroxyaldehydes, leads to accumulation of advanced glycation end-products (AGEs) in the body, resulting in diabetic complications and age-related pathology. Although molecular structures of glycolaldehyde- and glyceraldehyde-derived AGEs have been described in previous studies, little is known about lactaldehyde-derived AGEs of α-hydroxyaldehydes. Here, we report a novel crosslinked type of AGE, named as lactaldehyde-derived lysine dimer (LAK2), which is produced due to non-enzymatic glycation of Nα-acetyl-L-lysine with lactaldehyde under physiological conditions. We have identified the molecular structure of LAK2 by extensive mass spectrometry and nuclear magnetic resonance analyses. Furthermore, we propose a reaction pathway to produce LAK2, in which it is formed through an intermediate common with the recently reported lactaldehyde-derived pyridinium-type lysine adduct (LAPL). Since lactaldehyde is known to be produced from L-threonine in a myeloperoxidase (MPO)-mediated reaction at sites of inflammation, LAK2 has the potential to be an oxidative stress marker of MPO-mediated reactions induced in inflammation.

Sunless (chemical) tanning is widely regarded as a safe alternative to solar UV-induced skin tanning known to be associated with epidermal genotoxic stress, but the cutaneous biology impacted by chemical tanning remains largely unexplored. Chemical tanning is based on the formation of melanin-mimetic cutaneous pigments (‘melanoidins’) from spontaneous amino-carbonyl (‘glycation’) reactions between epidermal amino acid/protein components and reactive sugars including the glycolytic ketose dihydroxyacetone (DHA). Here, we have examined the cutaneous effects of acute DHA-exposure on cultured human HaCaT keratinocytes and epidermal reconstructs, profiled by gene expression array analysis and immunodetection. In keratinocytes, DHA-exposure performed at low millimolar concentrations did not impair viability while causing a pronounced cellular stress response as obvious from rapid activation of phospho-protein signal transduction [p-p38, p-Hsp27(S15/S78), p-eIF2α] and gene expression changes (HSPA6, HMOX1, CRYAB, CCL3), not observable upon exposure to the non-ketose, tanning-inactive DHA-control glycerol. Formation of advanced glycation end products (AGEs) from posttranslational protein-adduction was confirmed by quantitative mass spectrometric detection of N-ε-(carboxyethyl)-l-lysine (CEL) and N7-carboxyethyl-l-arginine, and skin cells with CRISPR-Cas9-based elimination of the carbonyl stress response gene GLO1 (encoding glyoxalase 1) displayed hypersensitivity to DHA-cytotoxicity. In human epidermal reconstructs a topical use-relevant DHA-dose regimen elicited a comparable stress response as revealed by gene expression array (HSPA1A, HSPA6, HSPD1, IL6, DDIT3, EGR1) and immunohistochemical analysis (CEL, HO-1, p-Hsp27-S78). In DHA-treated SKH-1 hairless mouse skin IHC-detection revealed epidermal occurrence of CEL- and p-Hsp27-epitopes. For comparison, stress response gene expression array analysis was performed in epidermis exposed to a supra-erythemal dose of solar simulated UV (2 MEDs), identifying genes equally or differentially sensitive to either one of these cutaneous stimuli [DHA (‘sunless tanning’) versus solar UV (‘sun-induced tanning’)]. Given the worldwide use of chemical tanners in consumer products, these prototype data documenting a DHA-induced specific cutaneous stress response deserve further molecular exploration in living human skin.

Background and purpose: The formation and accumulation of advanced glycation end products (AGEs) and rat lens aldose reductase (RLAR) generated in the glycation process play an outstanding role in the complications of diabetes. Owing to the adverse effects of AGEs on diabetic patients, the search for new anti-AGE agents from plants without side effects has had significant interest from the researchers in the last decades for the development of a therapy that improves diabetic complications. Spinach could reverse the formation of AGEs and RLAR. This study aimed to investigate the ability of 10 known glucopyranosides flavonoids isolated from Spinacia oleracea on the formation of AGEs and RLAR in vitro and in vivo experiments. Materials and methods: Methanol extract of leaves of spinach was subjected to bioassay-guided fractionation using to silica gel column chromatographic followed by gel filtration by Sephadex LH-20. BSA glucose system and in vitro bioassays using rat lens aldose reductase (RLAR) were employed to evaluated inhibitory activity on the formation of AGEs. The induced diabetes in zebrafish by immersing in a 111 mM glucose solution for 14 days, revealed increased glycation of proteins in the eyes. Measurements of glycated hemoglobin and fructosamine were used to verify the anti-AGEs effect of the isolated flavonoids. Key results: Through bioassay-guided fractionation of methanol extract of leaves spinach, ten known glucopyranoside flavonoids (1-10) have been isolated, and spectroscopic studies established their structures. Among the isolated compounds are: patuletin-3-O-(2”-coumaroylglucosyl)-(1→6)-[apiosyl-(1→2)]- β-d-glucopyranoside (7), patuletin 3-O-(2”-feruloyl glucosyl)-(1→6)-[apiosyl-(1→2)]- β-d-glucopyranoside (8), they have shown potent inhibition on AGEs formation, stronger than the positive controls used in the different experiments. Conclusion and implications: The findings indicated that glucopyranoside flavonoids found in Spinacia oleracea might have therapeutic potential for decreasing protein glycation, and might ameliorate AGE-related diabetic complications.

Bidens gardneri Baker, popularly known as “picão-vermelho”, has been used, traditionally, as a medicinal plant for the treatment of Diabetes mellitus. This study aimed to evaluate antidiabetic effect of leaves from B. gardneri aqueous extract (BGAE). We also evaluated in vitro anti-glycation potential. Chemical composition was analyzed based on a colorimetrics and HPLC methods. The oral glucose tolerance test (OGTT), was performed in rats with different doses (30, 100 and 300 mg/kg). Alloxan-induced diabetic and hypercaloric diet-fed rats were treated with 300 mg/kg of BGAE, orally, for 10 days and then 10 weeks, respectively. The activity of intestinal disaccharidases (maltase, sucrase and lactase) and quantification of muscle and liver glycogen content were also evaluated. On chemical analysis, the extract showed high phenolics content and the chromatogram showed 4-O-caffeoylquinic acid as the major component. The extract presented inhibition in the formation of advanced glycation end products (AGEs) and disaccharidases activity. In OGTT the dose of 300 mg/kg significantly decreased the blood glucose. In diabetic animals, BGAE significantly decreased blood glucose levels, preventing weight loss. In addition, in hypercaloric diet-fed rats, the extract prevented hyperglycemia. Our results suggest that, aqueous extract of B. gardneri has a potential for therapeutic intervention of diabetes.

In this study, the fracture behavior of ribosylated bovine cortical bone is investigated under loading conditions simulating a fall event. Single edge notched specimens, separated into a control group (n = 11) and a ribosylated group (n = 8), were extracted from the mid-diaphysis of a single bovine femur harvested from a mature cow. A seven-day ribosylation process results in the accumulation of Advanced-Glycation End Products (AGEs) cross-links and AGE adducts. Specimens were subjected to symmetric three point bending (opening mode) and an impact velocity of 1.6 m/s using a drop tower. Near-crack displacement fields up to fracture initiation are determined from high-speed images post-processed using digital image correlation. A constrained over-deterministic least squares regression and orthotropic material linear elastic fracture mechanics theory are used to extract the in-plane critical stress intensity factors at fracture initiation (i.e., fracture initiation toughness values). Statistically significant differences were not observed when comparing the in-plane fracture initiation toughness values (p≥0.96) or energy release rate (p=0.90) between the control and seven-day ribosylated groups. The intrinsic variability of bone may require high sample numbers in order to achieve an adequately powered experiment when assessing dynamic fracture behavior. While there are no detectable differences due to the ribosylation treatment investigated, this is likely due to the limited sample sizes utilized., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Different SOD1 proteoforms are implicated## in both familial and sporadic cases of Amyotrophic Lateral Sclerosis (ALS), an aging-associated disease that affects motor neurons. SOD1 is crucial to neuronal metabolism and health, regulating the oxidative stress response and the shift between oxidative-fermentative metabolism, which is important for astrocyte-neuron metabolic cooperation. Neurons have a limited capacity to metabolize methylglyoxal (MGO), a potentially toxic side product of glycolysis. MGO is highly reactive and can readily posttranslationally modify proteins, in a reaction known as glycation, impacting their normal biology. Here, we aimed to investigate the effect of glycation on the aggregation and toxicity of human SOD1WT (hSOD1WT). Cells with deficiency in MGO metabolism showed increased levels of hSOD1WT inclusions, displaying also reduced hSOD1WT activity and viability. Strikingly, we also found that the presence of hSOD1WT in stress granules increased upon MGO treatment. The treatment of recombinant hSOD1WT with MGO resulted in the formation of SDS-stable oligomers, specially trimers, and thioflavin-T positive aggregates, which can promote cell toxicity and TDP-43 pathology. Together, our results suggest that glycation may play a still underappreciated role on hSOD1WT and TDP-43 pathologies in sporadic ALS, which could open novel perspectives for therapeutic intervention., Competing Interests: Declaration of competing interest We have no conflicts of interest to disclose. All authors declare that there are notcompeting financial interests in relation to this work. Elis Cristina Araujo Eleutherio reports financial support was provided by Carlos Chagas Filho Foundation for Research Support of Rio de Janeiro State. Tiago Fleming Outeiro reports financial support was provided by German Research Foundation. Cristian Follmer reports financial support was provided by Carlos Chagas Filho Foundation for Research Support of Rio de Janeiro State. Elis Cristina Araujo Eleutherio reports financial support was provided by Coordination of Higher Education Personnel Improvement., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The low thermal stability of myofibrillar proteins (MPs) is a technological barrier to them being applied in beverage formulas. In this study, we investigated the effect of high-intensity ultrasound (HIU) pretreatment combined with glycation on the thermal stability, structural characteristics, and in vitro digestion behavior of MPs in water. The results indicated that HIU pretreatment combined with glycation significantly inhibited thermal aggregation and reduced the particle size of MPs compared to using either HIU or glycation treatments individually. The grafting of dextran (DX) shielded the sulfhydryl (-SH) and hydrophobic groups and inhibited disulfide bond cross-linking and hydrophobic association. Moreover, HIU pretreatment facilitated the shielding effect of glycation by destroying the filamentous myosin structure and exposing the internal -SH and hydrophobic groups as well as the grafting sites, maximally inhibiting thermal aggregation. In addition, the smaller protein particles and more flexible structure caused by HIU pretreatment combined with glycation increased their binding affinity toward protease. Overall, these findings can promote the technological development of modulating the MP structure-digestion for formulating novel meat protein-based products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Glycation is a non-enzymatic reaction wherein sugars or dicarbonyls such as methylglyoxal (MGO) and glyoxal (GO) react with proteins, leading to protein inactivation. The hydrolysing enzyme human deglycase-1 (hDJ-1) is reported to decrease glycative stress by deglycating the modified proteins, specifically at cysteine, lysine, and arginine sites. This specificity of hDJ-1 is thought to be regulated by its active site cysteine residue (Cys106). Structural analysis of hDJ-1 by molecular docking and simulation studies, however, indicates a possible role of glutamate (Glu18) in determining its substrate specificity. To elucidate this, Glu18 present at the catalytic site of hDJ-1 was modified to aspartate (Asp18) by SDM, and the resultant mutant was termed mutant DJ-1 (mDJ-1). Both hDJ-1 and mDJ-1 were heterologously expressed in Escherichia coli BL21 (DE3) strain and purified to homogeneity. The hDJ-1 showed k cat values of 1.45 × 10 3 s -1 , 3.6 × 10 2 s -1 , and 3.1 × 10 2 s -1 , and K m values 0.181 mM, 18.18 mM, and 12.5 mM for N-acetylcysteine (NacCys), N-acetyllysine (NacLys), and N-acetylarginine (NacArg), respectively. The mDJ-1 showed altered k cat values (8 × 10 2 s -1 , 3.8 × 10 2 s -1 , 4.9 × 10 2 s -1 ) and K m values of 0.14 mM, 6.25 mM, 5.88 mM for NacCys, NacLys and NacArg, respectively. A single amino acid change (Glu18 to Asp18) improved the substrate specificity of mDJ-1 toward NacLys and NacArg. Understanding hDJ-1's structure and enhanced functionality will facilitate further exploration of its therapeutic potential for the treatment of glycation-induced diabetic complications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

The larches, the Larix genus of plants are known as a natural source of taxifolin (dihydroquercetin), and extracts of its taxifolin rich xylem are used in dietary supplements to maintain health. In the present study, to assess biological activities of a methanol extract of the Japanese larch, Larix kaempferi (LK-ME), the effects of LK-ME on cell viability, inflammatory cytokine expression, and glycation were investigated. The effects of taxifolin which is known to be a main compound of LK-ME, and its related flavonoids, quercetin and luteolin were also examined. The results show that taxifolin exhibits lower growth inhibition activity and lesser induction activity of inflammatory cytokines in a human monocyte derived cell line, THP-1 cells, while in vitro anti-glycation activities of taxifolin were inhibiting at comparable levels to those of quercetin and luteolin. The growth inhibition and the cytokine induction activities, and the anti-glycation effects of LK-ME are assumed to have properties similar to taxifolin. The results of high performance liquid chromatography (HPLC) analysis indicated that taxifolin was detected as the main peak of LK-ME at the absorbance of 280 nm, and the concentration of taxifolin was measured as 3.12 mg/ml. The actual concentration of taxifolin in LK-ME is lower than the concentration estimated from the IC50 values calculated by the results of glycation assays, suggesting that other compounds contained in LK-ME are involved in the anti-glycation activity.

Human serum albumin (HSA) is a major cargo protein, which undergoes glycation in hyperglycaemic conditions and results in impaired function. In physiological conditions, HSA plays a crucial role in pharmacological activities such as drug transport or delivery through its binding capacity and also by its enzymatic activity, which enables the translation of pro-drugs into active drugs. In this study, the impact of the methylglyoxal-mediated glycation on dynamic behaviour of inter-domain motion, Cys34 reactivity, binding site residual interaction and secondary structure transition were investigated through molecular dynamics simulation. The alteration in inter-domain motion reflects the effect of glycation-mediated changes on the structural conformation of albumin. The binding site residue interactions and volume analysis revealed the impact of glycation on the geometry of the binding site. We also found the correlation of Cys34 reactivity with increase of turns in the region between Ia-h4 and Ia-h5. The rise in turn formation in that region keeps Tyr84 farther away from Cys34 which could lead to higher Cys34 reactivity. In parallel, significant alterations in alpha helical content of helices in the binding sites were observed. These structural and conformational changes in glycated albumin could be the causative agents for functional impairment which leads to diabetic complications., Competing Interests: Declaration of competing interest The author(s) declare(s) that there is no conflict of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Protein glycation and aggregation have a pivotal role in many diseases including diabetes and neurodegenerative disorders. N-acetyl aspartate (NAA), an osmolyte derived from L-aspartic acid, is one of the most abundant metabolites in the mammalian brain. Although NAA is supposed to be a substitute for a neuronal marker, its function is not fully elucidated. Herein, we have investigated the effect of NAA on glycation, AGEs formation and aggregation of irisin. AGE-specific fluorescence showed strong inhibition of AGEs formation in the presence of NAA, demonstrating its anti-glycating property. The aggregates present in MG-modified irisin were also reduced by NAA, which was confirmed by Thioflavin T fluorescence and fluorescence microscopy. Further, for the explanation of the strong anti-glycating potential of NAA, the interaction between irisin and NAA was also examined. Interaction studies involving steady-state fluorescence and molecular docking demonstrated that hydrogen bonding and salt bridges by NAA stabilize the irisin. It was found that glycation-prone residues i.e., lysine and arginine are specifically involved in the interaction which might prevent them from getting modified during the process of glycation. This study for the first time reported the antiglycating potential of NAA which can be implicated in the therapeutic management of various glycation-related complications., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Dicarbonyls are reactive precursors of advanced glycation end-products (AGEs). Dicarbonyls are formed endogenously, but also during food processing. Circulating dicarbonyls are positively associated with insulin resistance and type 2 diabetes, but the consequences of dietary dicarbonyls are unknown., Objectives: We aimed to examine the associations of dietary intake of dicarbonyls with insulin sensitivity, β-cell function, and the prevalence of prediabetes or type 2 diabetes., Methods: In 6282 participants (aged 60 ± 9 y; 50% men, 23% type 2 diabetes [oversampled]) of the population-based cohort the Maastricht Study, we estimated the habitual intake of the dicarbonyls methylglyoxal (MGO), glyoxal (GO), and 3-deoxyglucosone (3-DG) using food frequency questionnaires. Insulin sensitivity (n = 2390), β-cell function (n = 2336), and glucose metabolism status (n = 6282) were measured by a 7-point oral glucose tolerance test. Insulin sensitivity was assessed as the Matsuda index. Additionally, insulin sensitivity was measured as HOMA2-IR (n = 2611). β-cell function was assessed as the C-peptidogenic index, overall insulin secretion, glucose sensitivity, potentiation factor, and rate sensitivity. Cross-sectional associations of dietary dicarbonyls with these outcomes were investigated using linear or logistic regression adjusting for age, sex, cardiometabolic risk factors, lifestyle, and dietary factors., Results: Higher dietary MGO and 3-DG intakes were associated with greater insulin sensitivity after full adjustment, indicated by both a higher Matsuda index (MGO: Std. β [95% CI] = 0.08 [0.04, 0.12]; 3-DG: 0.09 [0.05, 0.13]) and a lower HOMA2-IR (MGO: Std. β = -0.05 [-0.09, -0.01]; 3-DG: -0.04 [-0.08, -0.01]). Moreover, higher MGO and 3-DG intakes were associated with a lower prevalence of newly diagnosed type 2 diabetes (OR [95% CI] = 0.78 [0.65, 0.93] and 0.81 [0.66, 0.99]). There were no consistent associations of MGO, GO, and 3-DG intakes with β-cell function., Conclusion: Higher habitual consumption of the dicarbonyls MGO and 3-DG was associated with better insulin sensitivity and lower prevalence of type 2 diabetes, after excluding individuals with known diabetes. These novel observations warrant further exploration in prospective cohorts and intervention studies., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Human serum albumin (HSA) is a versatile and important protein for the pharmaceutical industry (Fanali et al., Mol. Aspects Med. 33(3) (2012) 209–290). Due to the potential transmission of pathogens from plasma sourced albumin, numerous expression systems have been developed to produce recombinant HSA (rHSA) (Chen et al., Biochim. Biophys. Acta (BBA)—Gen. Subj. 1830(12) (2013) 5515–5525; Kobayashi, Biologicals 34(1) (2006) 55–59). Based on our previous study showing increased glycation of rHSA expressed in Asian rice (Frahm et al., J. Phys. Chem. B 116(15) (2012) 4661–4670), both supplier-to-supplier and lot-to-lot variability of rHSAs from a number of expression systems were evaluated using reversed phase liquid chromatography linked with MS and MS/MS analyses. The data are associated with the research article ‘Determination of Supplier-to-Supplier and Lot-to-Lot Variability in Glycation of Recombinant Human Serum Albumin Expressed in Oryza sativa’ where further analysis of rHSA samples with additional biophysical methods can be found (Frahm et al., PLoS ONE 10(9) (2014) e109893). We determined that all rHSA samples expressed in rice showed elevated levels of arginine and lysine hexose glycation compared to rHSA expressed in yeast, suggesting that the extensive glycation of the recombinant proteins is a by-product of either the expression system or purification process and not a random occurrence.

This study was designed to evaluate the protective activity of gallic acid (GA) against glyoxal (GO) an advanced glycation intermediate-induced renal fibrosis in experimental rats. Glyoxal (i.p) at a dose of 15 mg/Kg body weight/day for 4 weeks induces renal fibrosis. GA was administered orally (100 mg/Kg body weight/day) along with GO for 4 weeks. The anti-fibrotic activity of GA was analyzed by measuring the collagen synthesis and deposition in renal tissues using mRNA expression analysis and Masson trichrome staining (MTS), respectively. The nephroprotective potential of GA was assessed by quantifying the markers of kidney damage such as serum blood-urea-nitrogen (BUN), creatinine (CR) and alkaline phosphatase (AP). Moreover, basement membrane damage in renal tissues was analysed by periodic acid Schiff’s (PAS) staining. GA co-treatment markedly suppressed the GO-induced elevation in mRNA expression of collagen I and III, MMP-2, MMP-9 and NOX (p

Glycation of human low-density protein (LDL) has an essential contribution to cardiovascular diseases. Natural compounds like rutin have been extensively studied in preventing glycation-induced oxidative stress. This study examined rutin's anti-glycation potential with glycated LDL utilizing spectroscopic and in silico methods. Glycated LDL treated with rutin, showed around 80 % inhibition in advanced glycation end-product production. Carbonyl content and lipid peroxidation like assays were used to establish the development of oxidative stress. Rutin was seen to lower the generation of oxidative stress in a dose-dependent manner. Using thioflavin-T assay and electron microscopy, rutin was suggested to restore the structural disturbances in glycated LDL. Moreover, CD spectroscopy suggested reinstation of secondary structure of glycated LDL treated with rutin. Mechanistic insights between rutin and LDL were observed through spectroscopic measures. Molecular docking study confirmed the LDL-rutin binding with a binding energy of -10.0 kcal/mol. The rutin-LDL complex was revealed to be highly stable by molecular dynamics simulation, with RMSD, RMSF, Rg, SASA, and the secondary structure of LDL remaining essentially unchanged during the simulation period. Our study suggests that rutin possesses strong anti-glycating properties, which can be useful in therapeutics, as glycated LDL has an important role in atherosclerotic cardiovascular diseases., Competing Interests: Declaration of competing interest The authors of this paper declare no social or financial conflict of interest in this research., (Copyright © 2023 Elsevier B.V. All rights reserved.)