Inhibitory effect of Ginkgo biloba seeds peptides on methylglyoxal-induced glycations.

The aim of this study was to investigate the antiglycation activity and mechanism of two identified peptides, Valine-Valine-Phenylalanine-Proline-Glycine-Cysteine-Proline-Glutamic acid (VVFPGCPE) and Serine-Valine-Aspartic acid-Aspartic acid-Proline-Arginine-Threonine-Lysine (SVDDPRTL), from Ginkgo biloba seeds protein hydrolysates. Both VVFPGCPE and SVDDPRTL were efficient in bovine serum albumin (BSA)-methylglyoxal (MGO) model to inhibit BSA glycation, while VVFPGCPE showed higher antiglycation activity than SVDDPRTL. In antioxidant assays, VVFPGCPE scavenged more hydroxyl and super anion radicals, and chelated more Fe2+. Moreover, VVFPGCPE was more efficient in alleviating glycoxidation since it retained higher content of tryptophan and reduced dityrosine and kynurenine generation. Compared with SVDDPRTL, VVFPGCPE showed better performance in inhibiting protein aggregation and amyloid-like fibrillation formation. Therefore, VVFPGCPE was selected for further mechanism study. The circular dichroism analysis suggested VVFPGCPE could preserve α-helix structure and stabilize protein structure. The MGO trapping assay indicated VVFPGCPE (5 mg/mL) could capture 66.25% MGO within 24 h, and the mass spectrometry revealed VVFPGCPE could trap MGO by forming VVFPGCPE-mono-MGO adducts. Besides, molecular simulations suggested VVFPGCPE could interact with key glycation residues, arginine and lysine residues, of BSA mainly through van der Waals and hydrogen bonds. This study might supply a theoretical basis for the development of VVFPGCPE as an effective antiglycation agent.

Prevention of Enzymatic Browning by Natural Extracts and Genome-Editing: A Review on Recent Progress.

Fresh fruits and vegetable products are easily perishable during postharvest handling due to enzymatic browning reactions. This phenomenon has contributed to a significant loss of food quality and appearance. Thus, a safe and effective alternative method from natural sources is needed to tackle enzymatic browning prevention. The capabilities of natural anti-browning agents derived from plant- and animal-based resources in inhibiting enzymatic activity have been demonstrated in the literature. Some also possess strong antioxidants properties. This review aims to summarize a recent investigation regarding the use of natural anti-browning extracts from different sources for controlling the browning. The potential applications of genome-editing in preventing browning activity and improving postharvest quality is also discussed. Moreover, the patents on the anti-browning extract from natural sources is also presented in this review. The information reviewed here could provide new insights, contributing to the development of natural anti-browning extracts and genome-editing techniques for the prevention of food browning.

The Effect of Ethanol Treatment on the Quality of a New Table Grape Cultivar It 681-30 Stored at Low Temperature and after a 7-Day Shelf-Life Period at 20 °C: A Molecular Approach.

Despite the fact that many studies have examined the effectiveness of different gaseous postharvest treatments applied at low temperature to maintain table grape quality, the use of ethanol vapor has hardly been investigated. Thus, this work has studied the effectiveness of ethanol vapor-generating sachets in the maintenance of It 681-30 table grape quality, a new cultivar, during storage at low temperature and after the shelf-life period at 20 °C. To this end, various quality assessments have been carried out and the effect of the ethanol treatment on the expression of different genes (phenylpropanoids, transcription factors, PRs, and aquaporins) was determined. The results indicated that the application of ethanol vapor reduced the total decay incidence, weight loss, and the rachis browning index in It 681-30 grapes stored at 0 °C and after the shelf-life period at 20 °C, as compared to non-treated samples. Moreover, the modulation of STS7 and the different PR genes analyzed seems to play a part in the molecular mechanisms activated to cope with fungal attacks during the postharvest of It 681-30 grapes, and particularly during the shelf-life period at 20 °C. Furthermore, the expression of aquaporin transcripts was activated in samples showing higher weight loss. Although further work is needed to elucidate the role of ethanol in table grape quality, the results obtained in this work provide new insight into the transcriptional regulation triggered by ethanol treatment.

Use of Fatty Acid Metal Salts for Preventing Maillard Reaction-Driven Browning of Lecithins.

We previously reported that fluid soybean lecithin (SL) undergoes heat deterioration by the newly reported pseudo-Maillard rearrangement reaction. To inhibit heat deterioration, SLs were treated with metal silicates, such as magnesium silicate and calcium silicate. When soybean fatty acid was added to SL before treatment with calcium silicate, the color index after heating improved significantly as the acid value increased from 10 to 35 mg KOH/g. To elucidate the role of soybean fatty acid, calcium silicate treatment was carried out by adding several fatty acids to SL. Although saturated fatty acids had no effect on the heat deterioration of SL, unsaturated fatty acids were significantly more effective at inhibiting heat deterioration. Furthermore, for unsaturated fatty acids, it was confirmed that the calcium concentration increased in SL. Based on these results, several fatty acid metal salts were added to confirm whether heat deterioration while heating SL could be inhibited. It was observed that the heat deterioration of SL could be inhibited with fatty acid metal salts, regardless of whether the fatty acids were saturated or unsaturated and whether the metal was monovalent, divalent, or trivalent. Therefore, in this study, we clarified that the heat deterioration of SL could be inhibited by adding fatty acid metal salts to SL. Among sodium stearate, calcium stearate, magnesium stearate, barium stearate, and aluminum tristearate, the divalent fatty acid metal salts had a stronger inhibitory effect on heat deterioration than the monovalent and trivalent salts.

l-Theanine Activates the Browning of White Adipose Tissue Through the AMPK/α-Ketoglutarate/Prdm16 Axis and Ameliorates Diet-Induced Obesity in Mice.

l-Theanine is a nonprotein amino acid with much beneficial efficacy. We found that intraperitoneal treatment of the mice with l-theanine (100 mg/kg/day) enhanced adaptive thermogenesis and induced the browning of inguinal white adipose tissue (iWAT) with elevated expression of Prdm16, Ucp1, and other thermogenic genes. Meanwhile, administration of the mice with l-theanine increased energy expenditure. In vitro studies indicated that l-theanine induced the development of brown-like features in adipocytes. The shRNA-mediated depletion of Prdm16 blunted the role of l-theanine in promoting the brown-like phenotypes in adipocytes and in the iWAT of mice. l-theanine treatment enhanced AMPKα phosphorylation both in adipocytes and iWAT. Knockdown of AMPKα abolished l-theanine-induced upregulation of Prdm16 and adipocyte browning. l-Theanine increased the α-ketoglutarate (α-KG) level in adipocytes, which may increase the transcription of Prdm16 by inducing active DNA demethylation on its promoter. AMPK activation was required for l-theanine-induced increase of α-KG and DNA demethylation on the Prdm16 promoter. Moreover, intraperitoneal administration with l-theanine ameliorated obesity, improved glucose tolerance and insulin sensitivity, and reduced plasma triglyceride, total cholesterol, and free fatty acids in the high-fat diet-fed mice. Our results suggest a potential role of l-theanine in combating diet-induced obesity in mice, which may involve l-theanine-induced browning of WAT.

Effect directed synthesis of a new tyrosinase inhibitor with anti-browning activity.

The inhibition of enzymatic browning is an attractive target to elevate the quality of foods. The objective of this work is to describe a novel platform for the discovery of tyrosinase inhibitors, based on (a) one-pot preparation of a library of thiosemicarbazide compounds, (b) biological evaluation using tyrosinase TLC bioautography, (c) inhibitor identification via mass spectrometry coupled to bioautography. During these proof-of-concept experiments, the approach led to the straightforward identification of a new thiosemicarbazone with improved tyrosinase inhibition properties and fresh-cut apple slices antibrowning effect when compared to kojic acid. In conclusion, the platform represents an interesting strategy for the discovery of this type of inhibitors.

Dietary Advanced Glycation Endproducts and the Gastrointestinal Tract.

The prevalence of inflammatory bowel diseases (IBD) is increasing in the world. The introduction of the Western diet has been suggested as a potential explanation of increased prevalence. The Western diet includes highly processed food products, and often include thermal treatment. During thermal treatment, the Maillard reaction can occur, leading to the formation of dietary advanced glycation endproducts (dAGEs). In this review, different biological effects of dAGEs are discussed, including their digestion, absorption, formation, and degradation in the gastrointestinal tract, with an emphasis on their pro-inflammatory effects. In addition, potential mechanisms in the inflammatory effects of dAGEs are discussed. This review also specifically elaborates on the involvement of the effects of dAGEs in IBD and focuses on evidence regarding the involvement of dAGEs in the symptoms of IBD. Finally, knowledge gaps that still need to be filled are identified.

Inhalation of odors containing DMHF generated by the Maillard reaction affects physiological parameters in rats.

The effects of odors generated by the Maillard reaction from amino acids and reducing sugars on physiological parameters (blood pressure, heart rate, and oxidative stress levels) in Wistar rats were investigated in the present study. The Maillard reaction samples were obtained from glycine, arginine, or lysine of 1.0 mol/L and glucose of 1.0 mol/L with heat treatment. The odor-active compounds in the Maillard reaction samples were identified using the aroma extract dilution analysis. Among the odor-active compounds identified, 2,5-dimethyl-4-hydroxy-3(2H)-furanone (DMHF, FURANEOL and strawberry furanone) had the highest odor activity and its concentration was affected by amino acid types. The Maillard reaction odors generated from glycine or arginine significantly decreased systolic blood pressure and heart rate in rats when inhaled. These physiological effects were associated with DMHF. Furthermore, oxidative stress marker levels in rat plasma were decreased by the inhalation of DMHF. The inhalation of DMHF appears to at least partly affect physiological parameters by decreasing oxidative stress.

Recent Trends in Controlling the Enzymatic Browning of Fruit and Vegetable Products.

Enzymatic browning because of polyphenol oxidases (PPOs) contributes to the color quality of fruit and vegetable (FV) products. Physical and chemical methods have been developed to inhibit the activity of PPOs, and several synthetic chemical compounds are commonly being used as PPO inhibitors in FV products. Recently, there has been an emphasis on consumer-oriented innovations in the food industry. Consumers tend to urge the use of natural and environment-friendly PPO inhibitors. The purpose of this review is to summarize the mechanisms underlying the anti-browning action of chemical PPO inhibitors and current trends in the research on these inhibitors. Based on their mechanisms of action, chemical inhibitors can be categorized as antioxidants, reducing agents, chelating agents, acidulants, and/or mixed-type PPO inhibitors. Here, we focused on the food ingredients, dietary components, food by-products, and waste associated with anti-browning activity.

Inhibition of Maillard reaction in production of low-molecular-weight chitosan by enzymatic hydrolysis.

A low-molecular-weight chitosan (LMWC) sample was prepared by enzymatic hydrolysis, and used for investigation of special Maillard reaction products (MRPs) and factors affecting LMWC bioactivities. After undergoing MR, LMWC turned to brown color (termed BLMWC), showed reduction of several indices of rice growth promotion. This alteration of bioactivities was attributable to MRPs in BLMWC. A special MRP, 5-hydroxy-2-pyridine methanol isomer (5-H-2PMIS), was identified by HPLC and LC-MS. Analysis of key factors affecting MR, using this MRP as monitoring target compound and OD420 value, suggested that MR process can be minimized by storing LMWC under vacuum in a dry, low-temperature, neutral-pH environment. Na2SO3 was effective for inhibition of MR, at optimal concentration 0.5 %. Chemical and FTIR analyses showed that Na2SO3-treated sample conformed to the Chinese National Standard of chitosan (GB 29941-2013). Control of MR is essential for application of LMWC in food, pharmaceutical, and other industries.

Hydroxyphenyl thiosemicarbazones as inhibitors of mushroom tyrosinase and antibrowning agents.

Tyrosinase is a metalloenzyme involved in o-hydroxylation of monophenols and oxidation of o-diphenols to o-quinones, with formation of brown or black pigments (melanines). Tyrosinase inhibitors are of great interest in medicine and cosmetics (skin whitening compounds), but also in food and beverage industry (antibrowning agents). Here we report on the activity as mushroom tyrosinase inhibitors of a series of hydroxyphenyl thiosemicarbazones (1-5): one of them revealed an inhibitory activity stronger than kojic acid, used as reference. Enzymatic inhibition activity was confirmed by colorimetric measurements on small wheels of Fuji apples treated with the hydroxyphenyl thiosemicarbazones. The mechanism of action of compounds 1-5 was investigated by molecular modelling and by studying in solution their speciation with Cu(II) ions, the ions in the active site of the enzyme. Finally, compounds 1-5 were tested on human fibroblasts: they are not cytotoxic and they do not activate cells in a pro-inflammatory way.

Advanced Glycation End Products of Bovine Serum Albumin Suppressed Th1/Th2 Cytokine but Enhanced Monocyte IL-6 Gene Expression via MAPK-ERK and MyD88 Transduced NF-κB p50 Signaling Pathways.

Advanced glycation end products (AGE), the most known aging biomarker, may cause "inflamm-aging" (i.e., chronic low-grade inflammation that develops with aging) in both aged and diabetes groups. However, the molecular bases of inflamm-aging remain obscure. We prepared AGE by incubating BSA (0.0746 mmol/L) + glucose (0.5 mol/L) at 37 °C in 5% CO2-95% air for 1-180 days. The lysine glycation in BSA-AGE reached 77% on day 30 and 100% after day 130, whereas the glycation of arginine and cysteine was minimal. The Nε-(carboxymethyl)-lysine content in BSA-AGE was also increased with increasing number of incubation days. The lectin-binding assay revealed that the glycation of BSA not only altered the conformational structure, but lost binding capacity with various lectins. An immunological functional assay showed that BSA-AGE > 8 µg/mL significantly suppressed normal human Th1 (IL-2 and IFN-γ) and Th2 (IL-10) mRNA expression, whereas AGE > 0.5 µg/mL enhanced monocyte IL-6 production irrelevant to cell apoptosis. The AGE-enhanced monocyte IL-6 production was via MAPK-ERK and MyD88-transduced NF-κBp50 signaling pathways. To elucidate the structure-function relationship of BSA-AGE-enhanced IL-6 production, we pre-preincubated BSA-AGE with different carbohydrate-degrading, protein-degrading, and glycoprotein-degrading enzymes. We found that trypsin and carboxypeptidase Y suppressed whereas ß-galactosidase enhanced monocyte IL-6 production. In conclusion, BSA-AGE exerted both immunosuppressive and pro-inflammatory effects that are the molecular basis of inflamm-aging in aged and diabetes groups.

Nullifying phosphatidic acid effect and controlling phospholipase D associated browning in litchi pericarp through combinatorial application of hexanal and inositol.

The role of phospholipid modification initiated by phospholipase D (PLD) in enzymatic browning has been revoked through this study. Various alcohols and aldehydes were tried to read out their PLD controlling behaviour. Based on in-vitro results, reagents like hexanal and inositol were used to regulate PLD activity of litchi fruit stored at ambient temperature and their effects on fruit quality and physiological characteristics were also investigated. The results showed that combinatorial chemical treatment was successful in maintaining freshness of fruit through delayed physiological loss in weight and hence maintaining firmness. Combinatorial treated fruit had lower browning index than control by day 7. This novel treatment also maintained comparable levels of total phenolics and lowered the level of malondialdehyde. Evaluation of antioxidative enzymatic profile also confirmed the alleviation of oxidative stress of litchi fruit at ambient temperature. Thus, this strategy of enzyme regulation could play a vital role in overall quality maintenance of litchi fruit.

Full inhibition of Whangkeumbae pear polyphenol oxidase enzymatic browning reaction by l-cysteine.

The enzymatic browning reaction caused by polyphenol oxidase (PPO) has a negative effect on the processing of fruits and vegetables. However, some chemical inhibitors have been used to prevent enzymatic browning reaction, although they may be toxic and potentially hazardous to use in food. In this study, PPO was isolated and purified from the Whangkeumbae pear, and four food grade inhibitors were used to prevent the enzymatic browning reaction. The results showed that the PPO activity increased by 32.93-fold; its yield was 0.2 U/100 U, and the specific activity was 519,895.73 U/mg protein. The molecular weight of the PPO was approximately 44 kDa. The most potent inhibitor was l-cysteine, which fully inhibited the PPO activity at a concentration of 0.8 mg/mL. The type of inhibition of l-cysteine was noncompetitive. It suggests that l-cysteine can be utilized to prevent enzymatic browning reaction during the processing of pear juice.

Synthesis of Some Novel Fused Pyrimido[4″,5″:5',6']-[1,2,4]triazino[3',4':3,4] [1,2,4]triazino[5,6-b]indoles with Expected Anticancer Activity.

Our current goal is the synthesis of polyheterocyclic compounds starting from 3-amino-[1,2,4]triazino[5,6-b]indole 1 and studying their anticancer activity to determine whether increasing of the size of the molecules increases the anticancer activity or not. 1-Amino[1,2,4]triazino[3',4':3,4]-[1,2,4]triazino[5,6-b]indole-2-carbonitrile (4) was prepared by the diazotization of 3-amino[1,2,4]-triazino[5,6-b]indole 1 followed by coupling with malononitrile in basic medium then cyclization under reflux to get 4. Also, new fused pyrimido[4″,5″:5',6'][1,2,4]triazino-[3',4':3,4][1,2,4]triazino[5,6-b]indole derivative 6 was prepared and used to obtain polycyclic heterocyclic systems. Confirmation of the synthesized compounds' structures was carried out using elemental analyses and spectral data (IR, ¹H-NMR and 13C-NMR and mass spectra). The anticancer activity of some of the synthesized compounds was tested against HepG2, HCT-116 and MCF-7 cell lines. The anticancer screening results showed that some derivatives display good activity which was more potent than that of the reference drug used. Molecular docking was used to predict the binding between some of the synthesized compounds and the prostate cancer 2q7k hormone and breast cancer 3hb5 receptors.

Effect of whey protein isolate films incorporated with montmorillonite and citric acid on the preservation of fresh-cut apples.

The objective of this paper was to evaluate the effect of bioactive whey protein isolate/montmorillonite films containing citric acid on the inhibition of enzymatic browning and physicochemical properties in minimally processed apples. Whey protein isolate films incorporated with montmorillonite (3 g/100 g) and citric acid (5 and 10 g/100 g) were applied to the apples slices. All samples were packaged in polypropylene trays (14.6 cm × 11.4 cm × 6.5 cm) and stored at 5 ±â€¯2 °C and 85 ±â€¯3% RH for eight days. Every two days, the apples samples were evaluated for color, acidity, pH, soluble solids, water activity and polyphenol oxidase and peroxidase enzyme activity. The enzymatic browning of the apples slices was reduced for all films during storage. However, the films containing citric acid maintained the color characteristics, reducing the loss of quality associated the maintenance of acidity, soluble solids, water activity, reduction of polyphenol oxidase and peroxidase activity, thus prolonging the shelf life of the apples.

Alpha-Synuclein Glycation and the Action of Anti-Diabetic Agents in Parkinson's Disease.

Parkinson's disease (PD) is a neurodegenerative disorder with complex etiology and variable pathology. While a subset of cases is associated with single-gene mutations, the majority originates from a combination of factors we do not fully understand. Thus, understanding the underlying causes of PD is indispensable for the development of novel therapeutics. Glycation, the non-enzymatic reaction between reactive dicarbonyls and amino groups, gives rise to a variety of different reaction products known as advanced glycation end products (AGEs). AGEs accumulate over a proteins life-time, and increased levels of glycation reaction products play a role in diabetic complications. It is now also becoming evident that PD patients also display perturbed sugar metabolism and protein glycation, including that of alpha-synuclein, a key player in PD. Here, we hypothesize that anti-diabetic drugs targeting the levels of glycation precursors, or promoting the clearance of glycated proteins may also prove beneficial for PD patients.

The influence of proteases on the browning of dried squid products processed by air-drying.

To clarify why mantle meat from Japanese common squid (Todarodes pacificus) dried products discolor dramatically when processed by air-drying, the role of endogenous protease(s) on browning during the processing of dried squid products was studied. The involvement of endogenous protease(s) in the generation of Free Amino Acids (FAAs) participating in the Maillard reaction was characterized. The browning and myosin heavy chain degradation during air-drying were obviously mitigated by the addition of metalloprotease inhibitors, especially EGTA and 1,10-phenanthroline, followed by serine proteinase inhibitor, especially PMSF. The amount of total FAAs in dried products increased by only 0.17% with the addition of 1,10-phenanthroline and 5.0% with added EGTA. In autolysis models, protein autolysis was almost completely inhibited by 1,10-phenanthroline. The addition of, 1,10-phenanthroline inhibited the increase in total FAAs including arginine (Arg), followed by EGTA and PMSF. The results indicated that endogenous metalloproteases and serine proteases might influence the generation of FAAs including Arg, and contribute to product discoloration during air-drying.

Dicarbonyls and Advanced Glycation End-Products in the Development of Diabetic Complications and Targets for Intervention.

Advanced glycation end-products (AGEs) are non-enzymatic protein and amino acid adducts as well as DNA adducts which form from dicarbonyls and glucose. AGE formation is enhanced in diabetes and is associated with the development of diabetic complications. In the current review, we discuss mechanisms that lead to enhanced AGE levels in the context of diabetes and diabetic complications. The methylglyoxal-detoxifying glyoxalase system as well as alternative pathways of AGE detoxification are summarized. Therapeutic approaches to interfere with different pathways of AGE formation are presented.