Increased O-GlcNAcylation by Upregulation of Mitochondrial O-GlcNAc Transferase (mOGT) Inhibits the Activity of Respiratory Chain Complexes and Controls Cellular Bioenergetics.

O-linked ß-N-acetylglucosamine (O-GlcNAc) is a reversible post-translational modification involved in the regulation of cytosolic, nuclear, and mitochondrial proteins. The interplay between O-GlcNAcylation and phosphorylation is critical to control signaling pathways and maintain cellular homeostasis. The addition of O-GlcNAc moieties to target proteins is catalyzed by O-linked N-acetylglucosamine transferase (OGT). Of the three splice variants of OGT described, one is destined for the mitochondria (mOGT). Although the effects of O-GlcNAcylation on the biology of normal and cancer cells are well documented, the role of mOGT remains poorly understood. In this manuscript, the effects of mOGT on mitochondrial protein phosphorylation, electron transport chain (ETC) complex activity, and the expression of VDAC porins were investigated. We performed studies using normal and breast cancer cells with upregulated mOGT or its catalytically inactive mutant. Proteomic approaches included the isolation of O-GlcNAc-modified proteins of the electron transport chain, followed by their analysis using mass spectrometry. We found that mitochondrial OGT regulates the activity of complexes I-V of the respiratory chain and identified a group of 19 ETC components as mOGT substrates in mammary cells. Furthermore, we observed that the upregulation of mOGT inhibited the interaction of VDAC1 with hexokinase II. Our results suggest that the deregulation of mOGT reprograms cellular energy metabolism via interaction with and O-GlcNAcylation of proteins involved in ATP production in mitochondria and its exchange between mitochondria and the cytosol.

Isolation, structural characterization and biological activity evaluation of melanoidins from thermally processed cocoa beans, carob kibbles and acorns as potential cytotoxic agents.

The aim of this study was to determine the chemical structure and biological activity of melanoidin fractions derived from cocoa beans, carob kibbles, and acorns roasted at different temperature-time conditions. The results showed that plant origin and roasting conditions had significant effects on the chemical composition, structural features, and morphology of melanoidins. All tested melanoidins exhibited significant antioxidant properties in three in vitro assays. In addition, they show significant in vitro anti-inflammatory activity by reducing lipoxygenase. The results from MTT assay showed that the all studied melanoidins had a cytotoxic effect against SW-480 cells in a dose- and time-dependent manner. Furthermore, the most pronounced activity was observed for acorn melanoidins. This is a unique finding, as the specific cytotoxic effect has not been reported for cocoa, carob and acorn melanoidins, and opens up a great opportunity to develop a potential novel cytotoxic agent against deadly colon cancer in the future.

Chemical Composition, Antioxidant Properties and Sensory Aspects of Sponge Cakes Supplemented with Edible Insect Flours.

The chemical composition, antioxidant properties, and sensory aspects of sponge cakes with the addition of flours from edible insects (buffalo worm, cricket, and mealworm) were evaluated. The addition of edible-insect flours increased the protein, fat, and dietary fiber content in all cases. The utilization of edible insects demonstrated a notable augmentation in the phenolic compounds (especially protocatechuic acid and protocatechuic aldehyde, and syringic, ferulic, and sinapic acids). This resulted in an increase in the antioxidant activity measured against the ABTS radical cation, the DPPH radical, and ferric ions. The antioxidant potential, assessed by four different methods, unequivocally confirmed that the aforementioned polyphenolic compounds found in edible insects provide significant radical-scavenging and antioxidant activity in sponge cakes containing them. The polyunsaturated fatty acid contents were significantly lower in cakes with insect flour compared to the standard wheat cakes. Products and raw materials exhibited high values of the n - 6/n - 3 ratio, which may be associated with negative health effects, with a high oleic acid content. The amino acid score (AAS) for the essential amino acids exceeded 100% for all obtained products. The sponge cakes were accepted by consumers and the taste was the most important predictor for overall acceptability, whereas the structure and appearance had less impact.

Effects of In Vitro Digestion of Polyphenols from Coffee on Binding Parameters to Human Topoisomerase II α.

Type II topoisomerase (TOPII) is an enzyme that influences the topology of DNA. DNA breaks generated by TOPII may result in mutagenic or cytotoxic changes in cancer cells. In this study, we characterized interactions of TOPIIα with coffee extracts and individual chlorogenic acids (CHAs) from the extracts by performing isothermal titration calorimetry (ITC) and molecular docking (MD) simulations. The study showed that the highest affinity to TOPIIα was found in green coffee (ΔG = -38.23 kJ/mol) and monochlorogenic acids fraction of coffee extracts (ΔG = -35.80 kJ/mol), resulting from the high content of polyphenols, such as CHAs, which can bind to the enzyme in the active site. Coffee extracts and their fractions maintained a high affinity for TOPIIα after simulated digestion in the presence of probiotic bacteria. It can be concluded that coffee may be a potential TOPIIα inhibitor considered as a functional food for cancer prevention.

Potentiality Assessment of the Acetylcholinesterase-Inhibitory Activity of Olive Oil with an Additive Edible Insect Powder.

Edible insects (Alphitobius diaperinus Panzer, Gryllus campestris, Tenebrio molitor, Chorthippus biguttulus) are rich in nutrients that potentially inhibit acetylcholinesterase (AChE), but also improve cognition. The aim of this study was to evaluate four varied species of freeze-dried edible insects (purchased from a store); their nutrient composition, including fat, total phenolic compounds, vitamins, and antioxidant properties; and the potential inhibitory effect of AChE. An additional goal was to obtain olive oil with the addition of edible insects. Such oil was characterized by high oxidizing properties and showed high affinity to AChE. The results showed that mealworms and grasshoppers had the highest content of fats (PUFA/SFA) and phenolic compounds. These insects also showed a high content of vitamins, which correlated with the highest affinity for AChE. Therefore, they were added as a functional additive to olive oil. Olive oil with the addition of edible insects showed a higher affinity for AChE and enriched the olive oil with vitamin C and B vitamins.

Bioactive Compounds and Antioxidant Composition of Nut Bars with Addition of Various Edible Insect Flours.

Edible insects represent a new functional source of nutrients that can contribute to solving nutritional deficiency problems. The antioxidant potential and bioactive compounds of nut bars with the addition of three edible insects were evaluated. Acheta domesticus L., Alphitobius diaperinus P. and Tenebrio molitor L. flours were used. A 30% share of insect flour in the bars resulted in significantly greater antioxidant activity (TPC increased from 190.19 for standard bars to 309.45 mg catechin/100 g for bars with 30% addition of cricket flour). Insect flour contributed significantly to an increase in 2,5-dihydrobenzoic acid (from 0.12 for bars with a 15% share of buffalo worm flour to 0.44 mg/100 g in the case of bars with a 30% share of cricket flour) and chlorogenic acid in all bars (from 0.58 for bars with a 15% share of cricket flour to 3.28 mg/100 g for bars with a 30% addition of buffalo worm flour), compared to the standard. The highest content of tocopherols was found in bars with cricket flour, compared to standard bars (43.57 and 24.06 mg/100 g of fat, respectively). The dominant sterol in bars enriched with insect powder was cholesterol. The highest amount of it was found in cricket bars, and the lowest in mealworm bars (64.16 and 21.62 mg/100 g of fat, respectively). The enrichment of nut bars with insect flours raises the levels of valuable phytosterols in the final product. The addition of edible insect flours reduced the perception of most sensory attributes of the bars, compared to the standard bar.

Bioactive Compounds, Antioxidant Activity and Sensory Properties of Northern Red Oak (Quercus rubra L., syn. Q. borealis F. Michx) Seeds Affected by Roasting Conditions.

The nutritional value and health-promoting properties cause the fruits (acorns) of Quercus spp. to have great potential for use in the food industry as functional ingredients and antioxidants source. The aim of this study was to examine the bioactive compound's composition, antioxidant potential, physicochemical properties and taste characteristics of northern red oak (Quercus rubra L.) seeds subjected to roasting at different temperatures and times. The results indicate that the roasting markedly affects the composition of bioactive components of acorns. In general, the use of roasting temperatures greater than 135 °C causes a decrease in the total phenolic compound content of Q. rubra seeds. Furthermore, along with an increase in temperature and thermal processing time, a remarkable increase in melanoidins, which are the final products of the Maillard reaction, was also observed in processed Q. rubra seeds. Both unroasted and roasted acorn seeds had high DPPH radical scavenging capacity, ferric reducing antioxidant power (FRAP) and ferrous ion chelating activity. Roasting at 135 °C caused negligible changes in total phenolics content and antioxidant activity of Q. rubra seeds. Almost all samples had lower antioxidant capacity along with an increase in the roasting temperatures. Additionally, thermal processing of acorn seeds contributes to the development of the brown color and the reduction of bitterness, and the creation of a more pleasant taste of the final products. Overall, the results of this study show that both unroasted and roasted Q. rubra seeds may be an interesting source of bioactive compounds with high antioxidant activity. Therefore, they can be used as a functional ingredient of beverages or food.

Quality Assessment of Waste from Olive Oil Production and Design of Biodegradable Packaging.

The use of olive pomace from olive oil production is still insufficient. The lingering olive pomace is harmful to the environment. On the other hand, the world is increasingly polluted with plastic or by-products from the production of biodegradable products. Considering these two problems, the aim of this work was to develop a mixture and create biodegradable disposable tableware characterized by high antioxidant activity. The disposable tableware was made by mixing olive pomace with teff flour or/and sorghum groats and lecithin. Baking was carried out at the temperature of 180 °C. The best variant of the mixture for the preparation of disposable tableware was olive pomace, teff flour, sorghum groats and lecithin. These vessels were the toughest, with low water absorption and had a high antioxidant potential due to the high content of polyphenols and omega acids. Protecting the cups and bowls with beeswax had a positive effect on reducing water absorption.

Bioactive Properties of Extracts from Plectranthus barbatus (Coleus forskohlii) Roots Received Using Various Extraction Methods.

The aim of this study was to verify various extraction methods: shaking water bath extraction (SWA), ultrasound-assisted extraction (UAE) and microwave assisted extraction (MAE), and their parameters to optimize the extraction yield as well as maximize the concentration of polyphenols in Plectranthus barbatus extracts. Extracts were obtained from dried roots of P. barbatus in various degrees of fragmentation and analyzed for content of polyphenols, antioxidant capacity and flavonoids. Additionally, phenolic compounds in extracts were analyzed using the UHPLC-DAD-ESI-MS/MS method. The conducted research showed that roots of P. barbatus are rich in polyphenolic compounds. A total of 15 phenolic compounds, belonging to the group of phenolic acids and their derivatives, were identified. The extraction yield was similar for all extraction methods and averaged 31%. Irrespective of the extraction method, the yield was the lowest in the case of using 80% ethanol as the solvent. The extracts obtained from the finer fraction were characterized by a higher antioxidant capacity as well as a higher concentration of polyphenolic compounds including flavonoids. UAE seems to be the most effective method for extraction of polyphenols from P. barbatus roots. Regardless of the extraction method, ethanol was a better extractant than distilled water. All ethanolic extracts were characterized by a high antioxidant capacity. The 80% ethanol solution was considered the best solvent for the extraction of flavonoids, while the 40% and 60% ethanol solutions were sufficient for the effective extraction of polyphenolic compounds in general.

Influence of Freeze-Dried Phenolic-Rich Plant Powders on the Bioactive Compounds Profile, Antioxidant Activity and Aroma of Different Types of Chocolates.

In this study, the blueberries (BLUB), raspberries (RASB), blackberries (BLCB), pomegranates pomace (POME) and beetroots (BEET) freeze-dried powders were used as the sources of phenolic compounds to enrich different types of chocolates, substituting a part of the sweetener. It was found that 1% addition of fruit or vegetable powders to chocolates increased the content of total phenolic compounds (flavan-3-ols, phenolic acids and anthocyanins) of enriched dark and milk chocolates compared to the control ones dependent on the powder used. Among the enriched chocolates, the chocolates with the addition of BLUB powder were characterized by the highest total polyphenol content. The highest percentage increase (approximately 80%) in the total polyphenol content was observed in MCH chocolate enriched with BLUB powder. Chocolates incorporated with BLUB, RASB, BLCB and POME powders presented a richer phenolic compound profile than control counterparts. The highest DPPH radical-scavenging capacity was exhibited by the DCH98S chocolate enriched with BEET powder. However, the DCH98ESt chocolates enriched with POME and BEET powders demonstrated the highest FRAP values. An electronic nose analysis confirmed the existence of differences between the profiles of volatile compounds of various types of chocolates enriched with fruit or vegetable powders. Thus, the enrichment of dark and milk chocolates with BLUB, RASB, BLCB, POME and BEET powders seemed to be an interesting approach to enhance bioactivity and to enrich the sensory features of various chocolate types.

Mitochondrial O-GlcNAc Transferase Interacts with and Modifies Many Proteins and Its Up-Regulation Affects Mitochondrial Function and Cellular Energy Homeostasis.

O-GlcNAcylation is a cell glucose sensor. The addition of O-GlcNAc moieties to target protein is catalyzed by the O-Linked N-acetylglucosamine transferase (OGT). OGT is encoded by a single gene that yields differentially spliced OGT isoforms. One of them is targeted to mitochondria (mOGT). Although the impact of O-GlcNAcylation on cancer cells biology is well documented, mOGT's role remains poorly investigated. We performed studies using breast cancer cells with up-regulated mOGT or its catalytic inactive mutant to identify proteins specifically modified by mOGT. Proteomic approaches included isolation of mOGT protein partners and O-GlcNAcylated proteins from mitochondria-enriched fraction followed by their analysis by mass spectrometry. Moreover, we analyzed the impact of mOGT dysregulation on mitochondrial activity and cellular metabolism using a variety of biochemical assays. We found that mitochondrial OGT expression is glucose-dependent. Elevated mOGT expression affected the mitochondrial transmembrane potential and increased intramitochondrial ROS generation. Moreover, mOGT up-regulation caused a decrease in cellular ATP level. We identified many mitochondrial proteins as mOGT substrates. Most of these proteins are localized in the mitochondrial matrix and the inner mitochondrial membrane and participate in mitochondrial respiration, fatty acid metabolism, transport, translation, apoptosis, and mtDNA processes. Our findings suggest that mOGT interacts with and modifies many mitochondrial proteins, and its dysregulation affects cellular bioenergetics and mitochondria function.

Polyphenols and Other Bioactive Compounds of Sideritis Plants and Their Potential Biological Activity.

Due to the growing problem of obesity associated with type 2 diabetes and cardiovascular diseases, causes of obesity are extensively investigated. In addition to a high caloric diet and low physical activity, gut microbiota disturbance may have a potential impact on excessive weight gain. Some reports indicate differences in the composition of the intestinal microflora of obese people in comparison to lean. Bioactive compounds of natural origin with beneficial and multifaceted effects on the body are more frequently used in prevention and treatment of many metabolic diseases including obesity. Sideritis scardica is traditionally consumed as mountain tea in the Balkans to strengthen the body and improve mood. Many reports indicate a positive effect on digestive system, weight loss, and prevention of insulin resistance. Additionally, it exhibits antioxidant activity and anti-inflammatory effects. The positive effect of Sideritis scardica extracts on memory and general cognitive abilities is indicated as well. The multilevel positive effect on the body appears to originate from the abundant occurrence of phenolic compounds, especially phenolic acids in Sideritis scardica extracts. However, mechanisms underlying their action require careful discussion and further research. Therefore, the objective of this review is to summarize the available knowledge on the role and mechanism of action of biologically active compounds of Sideritis scardica and other related species from the genus Sideritis.

Effects of Raw and Roasted Cocoa Bean Extracts Supplementation on Intestinal Enzyme Activity, Biochemical Parameters, and Antioxidant Status in Rats Fed a High-Fat Diet.

The aim of the study was to analyze the influence of diet containing the polyphenol-rich material on intestinal enzyme activity, oxidative stress markers, lipid metabolism and antioxidant status of laboratory rats. The animals were fed high-fat diet supplemented with freeze-dried water extracts of raw and roasted cocoa beans of Forastero variety. The observed changes indicated the biological activity of polyphenols and other components of the prepared cocoa beans extracts (CBEs). The presence of raw and roasted CBEs in the diets diversified the activity of the enzymes of the cecal microflora of rats. Both CBEs beneficially affect the antioxidant status of the serum, even in relation to the control standard group. The experimental cocoa bean preparations showed no significant effect on the mass of rats' liver, heart, and kidneys, but varied some parameters of the antioxidant status of their organisms. The raw CBE in rats fed with the high-fat diet shows a high ability to inhibit lipid peroxidation in heart and more effectively increases hepatic reduced glutathione (GSH) concentrations compared to the roasted CBE, which did not show any significant effect. Moreover, supplementation with both CBEs significantly affects the volatile fatty acids concentration in the rats' cecum. Results of this study contribute to the evidence that dietary supplementation with raw and roasted CBEs can exert health-promoting effects, however further studies are necessary.

Bioavailability and metabolism of selected cocoa bioactive compounds: A comprehensive review.

Cocoa beans and their co-products are a rich source of beneficial compounds for health promotion, including polyphenols and methylxanthines. Knowledge of bioavailability and in vivo bioactivity of these phytochemicals is crucial to understand their role and function in human health. Therefore, many studies concerning bioavailability and bioactivity of cocoa bioactive compound have been done in both in vivo animal models and in humans. This critical review comprehensively summarizes the existing knowledge about the bioavailability and the major metabolic pathways of selected cocoa bioactive compounds (i.e. monomeric flavan-3-ols, procyanidins, anthocyanins, flavonols, phenolic acids, N-phenylpropenoyl-L-amino acids, stilbenes, and methylxanthines). The compiled results indicated that many of these compounds undergo extensive metabolism prior to absorption. Different factors have been suggested to influence the bioavailability of polyphenols and methylxanthines among them the role of gut microbiota, structure of these compounds, food matrix and occurrence of other substances were the most often considered. Aforementioned factors decided about the site where these bioactive compounds are digested and absorbed from the alimentary tract, as well as the pathway by which they are metabolized. These factors also determine of the type of transport through the intestine barrier (passive, involving specific enzymes or mediated by specific transporters) and their metabolic path and profile.

Influence of Diet Enriched with Cocoa Bean Extracts on Physiological Indices of Laboratory Rats.

Cocoa bean is a rich source of polyphenols, mainly flavonoids which have a wide range of biological properties. The aim of the study was to determine the physiological indices of laboratory rats as a response to diets containing water extracts of raw or roasted cocoa beans of Forastero variety, as well as purified monomeric flavan-3-ols fraction isolated from them. The influence of these extracts on selected parameters was studied during 4 weeks feeding. The samples of rats feces were collected throughout the experiment and after its completion, biological samples (intestines content, blood, and organs) were retrieved individually from each rat and subjected to analyses. The observed changes in the gastrointestinal tract functioning indices and metabolism indicators, determined throughout the study and after its completion, confirm to some extent the biological activity of polyphenol extracts of cocoa beans. The differences in the results obtained for the analyzed parameters of the gastrointestinal tract revealed that the cocoa bean extracts differently affected the physicochemical properties of rats' intestines. The results indicate the beneficial effects of the applied nutrition treatment on the activity of cecal enzymes and the content of volatile fatty acids in the gut.

The effect on bioactive components and characteristics of chocolate by functionalization with raw cocoa beans.

Chocolate manufacturers often develop their own original blends of cocoa beans, which, after processing, will give an unique sensory properties to their chocolates. In recent years, there has been a trend in the production of an increasing amount of low-processed or unprocessed food. Therefore, we carried out the research in which we obtained and analyzed chocolates with different blends of cocoa liquors prepared from roasted and unroasted cocoa beans of Criollo and Forastero varieties.Studies have shown that viscosity and yield value increased and hardness of chocolates lowered with the increase content of cocoa liquor from unroasted beans. Chocolates with cocoa liquor from roasted beans were characterized by the largest ability to scavenge DPPH radicals. Antioxidant activity determined by the ORAC-FL method increased with increasing polyphenol content in the chocolates. Type of the cocoa liquor used induced minor changes in the polyphenol and fatty acid composition. Asparagine was determined at the highest concentration, followed by alanine, glutamine, and serine. The concentration of acetic acid was the higher the higher the content of unroasted beans. The increased content of cocoa liquor from unroasted beans caused the color of the chocolate to brighten, more red and yellow. All tested chocolates were characterized by good organoleptic properties and microbial stability.

Influence of diet based on bread supplemented with raw and roasted cocoa bean extracts on physiological indices of laboratory rats.

The aim of the study was to determine the physiological indices of laboratory rats as a response to a diet consisting of a bread supplemented with freeze-dried water extracts of raw and roasted cocoa beans of Forastero variety. The influence of different cocoa extracts on oxidative stress, hyperglycemia, and lipid metabolism was studied during long-term feeding of laboratory rats with bread supplemented with cocoa extracts. During the 4-week experimental feeding samples of rats feces were subjected to analysis and after its completion, biological samples were collected i.e. intestines content, blood, and organs retrieved individually from each rat. The observed changes indicate the biological activity of polyphenol extracts and other components of cocoa beans present in the prepared extracts. The differences in the results obtained for the analyzed parameters of the gastrointestinal tract revealed that the cocoa bean extracts being the subject of this investigation differently affected the physicochemical properties of rats' intestines, including intestinal microflora. The results show the influence of the diet on the activity of both fecal and cecal enzymes. Applied nutrition treatment varied the concentration and total production of volatile fatty acids in the cecum. Furthermore, the diverse composition of the applied diet affected the level of some biochemical indicators of blood serum of rats. Additionally, to a small extent influenced the analyzed indicators of the antioxidative status of rats as characterized by the concentration of substances reacting with thiobarbituric acid in internal organs.

Phenolic Profile and Antioxidant Potential of Leaves from Selected Cotoneaster Medik. Species.

The antioxidant efficiency of 70% aqueous methanolic extracts from the leaves of twelve selected Cotoneaster Medik. species was evaluated using four complementary in vitro tests based on SET- (single electron transfer) and HAT-type (hydrogen atom transfer) mechanisms (DPPH, FRAP, O2(•-) and H2O2 scavenging assays). The samples exhibited the dose-dependent responses in all assays with activity parameters of EC50 = 18.5-34.5 µg/mL for DPPH; 0.9-3.8 mmol Fe(2+)/g for FRAP; SC50 = 27.7-74.8 µg/mL for O2(•-); and SC50 = 29.0-91.3 µg/mL for H2O2. Significant linear correlations (|r| = 0.76-0.97, p < 0.01) between activity parameters and total contents of phenolics (5.2%-15.4% GAE) and proanthocyanidins (2.1%-15.0% CYE), with weak or no effects for chlorogenic acid isomers (0.69%-2.93%) and total flavonoids (0.28%-1.40%) suggested that among the listed polyphenols, proanthocyanidins are the most important determinants of the tested activity. UHPLC-PDA-ESI-QTOF-MS analyses led to detection of 34 polyphenols, of which 10 B-type procyanidins, 5 caffeoylquinic acids and 14 flavonoids were identified. After cluster analysis of the data matrix, the leaves of Cotoneaster zabelii, C. splendens, C. bullatus, C. divaricatus, C. hjelmqvistii and C. lucidus were selected as the most promising sources of natural antioxidants, exhibiting the highest phenolic levels and antioxidant capacities, and therefore the greatest potential for pharmaceutical applications.

Effect of inclusion of hydroxycinnamic and chlorogenic acids from green coffee bean in ß-cyclodextrin on their interactions with whey, egg white and soy protein isolates.

The aim of the study was to characterise the interactions of hydroxycinnamic and chlorogenic acids (CHAs) from green coffee, with isolates of proteins from egg white (EWP), whey (WPC) and soy (SPI), depending on pH and temperature. The binding degree was determined by liquid chromatography coupled to a diode array detector and an ultrahigh resolution hybrid quadruple-time-of-flight mass spectrometer with ESI source (LC-QTOF-MS/MS). As a result of binding, the concentration of CHAs in proteins ranged from 9.44-12.2, 11.8-13.1 and 12.1-14.4g/100g for SPI, WPC and EWP, respectively. Thermodynamic parameters of protein-ligand interactions were determined by isothermal titration calorimetry (ITC) and energetics of interactions at the atomic level by molecular modelling. The amount of CHAs released during proteolytic digestion was in the range 0.33-2.67g/100g. Inclusion of CHAs with ß-cyclodextrin strongly limited these interactions to a level of 0.03-0.06g/100g.