Effect of food matrix and fermentation on angiotensin-converting enzyme inhibitory activity and ß-glucan release after in vitro digestion in oat-based products.

The aim of this study was to determine the effects of fermentation and food matrix on the ACE inhibitory activities of the peptides obtained after in vitro gastrointestinal digestion, protein profiles (SDS-PAGE) and ß-glucan amounts of oat products. Furthermore, the physicochemical and microbiological properties of fermented oat drinks and oat yogurt-like product obtained from oat fermentation were evaluated. Oat grains were mixed with a certain ratio of water 1:3 w/v (oat:water, yogurt consistency) and 1:5 w/v (oat:water, drink consistency), and this mixture was fermented with yogurt culture and probiotic Lactobacillus plantarum and fermented drinks and yogurt were produced. The results indicated that the fermented oat drink and the oat yogurt-like product had L. plantarum viability over 107 cfu/g. After the in vitro gastrointestinal digestion of the samples, the hydrolysis levels ranged from 57.70 % to 82.06 %.The hydrolysis level of the samples with fermented-drink consistency was significantly higher than the samples with yogurt consistency (p < 0.05).The SDS-PAGE profiles of the non-digested samples showed that the bands had molecular weights of 12-15 kDa and around 35 kDa. Bands whose molecular weights were around 35 kDA disappeared after gastric digestion. ACE inhibitory activities of the fractions composed of molecular weights of 2 kDa and 2-5 kDa obtained after in vitro gastrointestinal digestion of the oat samples were in the range of 46.93-65.91 %. The effect of fermentation on the ACE inhibitory activities of the peptide mixture with molecular weights between 2 and 5 kDa was not statistically significant, however, fermentation caused an increase in the ACE inhibitory activities of the peptide mixture with a molecular weight<2 kDa (p < 0.05). The ß-glucan amounts of fermented and non-fermented oat products were in the range of 0.57-1.28 %. The ß-glucan amounts detected after gastric digestion decreased considerably and ß-glucan could not be detected in the supernatant after gastrointestinal digestion. This indicated that ß-glucan did not solubilize in the supernatant (bioaccessible) and remained in the pellet. In conclusion, fermentation is a valuable process for releasing peptides with moderately high ACE inhibitory effects from the parent oat proteins.

New applications of advanced instrumental techniques for the characterization of food allergenic proteins.

Current approaches based on electrophoretic, chromatographic or immunochemical principles have allowed characterizing multiple allergens, mapping their epitopes, studying their mechanisms of action, developing detection and diagnostic methods and therapeutic strategies for the food and pharmaceutical industry. However, some of the common structural features related to the allergenic potential of food proteins remain unknown, or the pathological mechanism of food allergy is not yet fully understood. In addition, it is also necessary to evaluate new allergens from novel protein sources that may pose a new risk for consumers. Technological development has allowed the expansion of advanced technologies for which their whole potential has not been entirely exploited and could provide novel contributions to still unexplored molecular traits underlying both the structure of food allergens and the mechanisms through which they sensitize or elicit adverse responses in human subjects, as well as improving analytical techniques for their detection. This review presents cutting-edge instrumental techniques recently applied when studying structural and functional aspects of proteins, mechanism of action and interaction between biomolecules. We also exemplify their role in the food allergy research and discuss their new possible applications in several areas of the food allergy field.

Effects of processing on onion skin powder added extrudates.

It is possible to enhance the functional properties of extruded products with the inclusion of fruit and vegetable by-products. Onion skin, a rich source of quercetin and fiber, is considered as waste in the industry and can be used as an alternative ingredient to improve the nutritional value of the extruded products. Three levels (3, 6, and 9%) of onion skin powder (OSP) were added to wheat flour and compared with control (0% OSP). The effect of the extrusion process on accessible quercetin, total phenolic content, and antioxidant activity of the samples were investigated. In addition, carbohydrate digestibility analyses were conducted for the products. Mass spectrometry (LC-MS/MS) results indicated that increasing the OSP level increased the quercetin content. The process caused the release of the entrapped quercetin from OSP, which was revealed by significantly higher quercetin levels for the extrudates. Some of the quercetin was lost during in vitro digestion process. Increasing the OSP level increased antioxidant activity and total phenolic contents of the samples. Total phenolic contents decreased significantly after the processing, yet antioxidant activities were not affected. The extruded products showed high amounts of rapidly available glucose (69.5 g/100 g). The OSP enhancement did not change the carbohydrate digestibility of products. The results indicated that the extrusion process could increase the level of accessible bioactive ingredients, and the level of functional compound addition can be optimized further.

Anti-Inflammatory Effect of Crude Momordica charantia L. Extract on 2,4,6-Trinitrobenzene Sulfonic Acid-Induced Colitis Model in Rat and the Bioaccessibility of its Carotenoid Content.

Momordica charantia L., known as bitter melon (BM), is a plant that belongs to the family Cucurbitaceae. Aims of this study are to investigate the anti-inflammatory effect of crude BM extract on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis model in rat. It was also aimed to determine the content and bioaccessibility of carotenoids of BM. BM was purchased from local markets in Izmir, Turkey. Fruits of BM were lyophilized, powdered, and used in the experiment. Carotenoids were determined by high-performance liquid chromatography. To determine the bioaccessibility of ß-carotene, in vitro digestion was performed. Wistar albino rats were divided into four groups: group A (BM+TNBS), group B (BM), group C (TNBS), and group D (control). BM solution was given 300 mg/(kg·day) for 6 weeks orally. Colitis was induced by 0.25 mL of a solution containing 100 mg/kg 5% (w/v) TNBS in 50% ethanol (w/v) intrarectally after 6 weeks. After sacrification, macroscopic and microscopic evaluations were performed. Myeloperoxidase, cytokines levels (interleukin-17 [IL-17], TNF-alpha, and interleukin-10 [IL-10]) were measured in serum and colonic samples by ELISA test. Institutional Animal Ethics Committee approval was obtained. Total carotenoid content of BM was determined 11.7 mg/g dry weight as ß-carotene equivalents. Bioaccessibility of total carotenoids was determined as 2.1% with in vitro digestion. Pretreatment with crude BM extract significantly reduced weight loss, macroscopic, and microscopic colitis damages in colonic samples (P = .000), (P = .015), and (P = .026), respectively. Serum anti-inflammatory cytokine IL-10 increased significantly in both treatment groups (P = .000). BM is a rich source of carotenoids, but the bioaccessibility of its carotenoids is low. This study displays that BM has protective anti-inflammatory effects on TNBS-induced colitis.

A Review of Factors Affecting Anthocyanin Bioavailability: Possible Implications for the Inter-Individual Variability.

Anthocyanins are dietary bioactive compounds showing a range of beneficial effects against cardiovascular, neurological, and eye conditions. However, there is, as for other bioactive compounds in food, a high inter and intra-individual variation in the response to anthocyanin intake that in many cases leads to contradictory results in human trials. This variability could be caused at two levels, one at the bioavailability level and the other at the effect and mechanisms of action. In this context, we have thoroughly reviewed the scientific literature on anthocyanins variability caused by variation in bioavailability. Based on the literature reviewed, we have concluded that the variability in anthocyanins bioavailability might be produced by the lack of homogeneity introduced at three different levels: food matrix and food processing, enzymes involved in anthocyanin metabolism and transport, and anthocyanin metabolizing gut microbiota. However, it should be noted that the literature on anthocyanins bioavailability considering inter or intra-individual variability is still very scarce, which makes it difficult to reach any firm conclusion on the main metabolizing enzymes or bacteria that would be responsible for the variability in anthocyanin bioavailability.

INFOGEST static in vitro simulation of gastrointestinal food digestion.

Developing a mechanistic understanding of the impact of food structure and composition on human health has increasingly involved simulating digestion in the upper gastrointestinal tract. These simulations have used a wide range of different conditions that often have very little physiological relevance, and this impedes the meaningful comparison of results. The standardized protocol presented here is based on an international consensus developed by the COST INFOGEST network. The method is designed to be used with standard laboratory equipment and requires limited experience to encourage a wide range of researchers to adopt it. It is a static digestion method that uses constant ratios of meal to digestive fluids and a constant pH for each step of digestion. This makes the method simple to use but not suitable for simulating digestion kinetics. Using this method, food samples are subjected to sequential oral, gastric and intestinal digestion while parameters such as electrolytes, enzymes, bile, dilution, pH and time of digestion are based on available physiological data. This amended and improved digestion method (INFOGEST 2.0) avoids challenges associated with the original method, such as the inclusion of the oral phase and the use of gastric lipase. The method can be used to assess the endpoints resulting from digestion of foods by analyzing the digestion products (e.g., peptides/amino acids, fatty acids, simple sugars) and evaluating the release of micronutrients from the food matrix. The whole protocol can be completed in ~7 d, including ~5 d required for the determination of enzyme activities.

Changes in functional properties and in vitro bioaccessibilities of ß-carotene and lutein after extrusion processing.

In this research, carrot pulp was added to traditional snack made from corn-grit. As the biological activity of carotenoids in the body depends on their bioaccessibility, change in carotenoid bioaccessibility during extrusion processing was investigated. In addition, phenolic content, antioxidant activity, ß-carotene and lutein contents were investigated before and after the extrusion process. Two different temperature profiles were used for extrusion process. In-vitro bioaccessibilities of ß-carotene and lutein increased by extrusion, ß-carotene at both temperature profiles while lutein only at higher temperature profile. Extrusion decreased the antioxidant activity, total phenolic, ß-carotene and lutein contents. Results suggest that even though amount of functional components decrease, in vitro bioaccessibility could be enhanced by extrusion. Therefore, carrot pulp can successfully be added as a functional ingredient to extrudated.

Protease inhibitors in various flours and breads: Effect of fermentation, baking and in vitro digestion on trypsin and chymotrypsin inhibitory activities.

In this study trypsin (TIA) and chymotrypsin inhibitory (CIA) activities were determined in the extracts of wheat, rye mix, mixed cereals and, whole wheat flours and, breads made with these flours. In addition, effects of fermentation, baking and in vitro digestion on TIA and CIA were studied. Whole wheat flour, dough, and bread did not show any TIA. Other flours displayed TIA. Contrary to, all flours, doughs, and breads exhibited CIA. Although TIA was not detected in the protein extracts obtained from wheat and rye mix breads, protein extract of rye mix flour exhibited TIA. Following in vitro digestion process, TIA of wheat bread was found as 5.91units/mL gastric dialysate and 9.17units/mL intestine dialysate. CIA was determined in dialysates obtained from wheat (2.12CI/mL and 3.78CI/mL for gastric and intestinal dialysate respectively) and rye breads (4.16CI/mL and 2.46CI/mL for gastric and intestinal dialysate respectively).

The food matrix affects the anthocyanin profile of fortified egg and dairy matrices during processing and in vitro digestion.

The aim of the present study was to understand to what extent the inclusion of anthocyanins into dairy and egg matrices could affect their stability after processing and their release and solubility during digestion. For this purpose, individual and total anthocyanin content of four different enriched matrices, namely custard dessert, milkshake, pancake and omelettete, was determined after their manufacturing and during in vitro digestion. Results showed that anthocyanin recovery after processing largely varied among matrices, mainly due to the treatments applied and the interactions developed with other food components. In terms of digestion, the present study showed that the inclusion of anthocyanins into food matrices could be an effective way to protect them against intestinal degradation, and also the incorporation of anthocyanins into matrices with different compositions and structures could represent an interesting and effective method to control the delivery of anthocyanins within the different compartments of the digestive tract.

Stability and bioaccessibility of anthocyanins in bakery products enriched with anthocyanins.

Anthocyanins, water soluble polyphenols, have been associated with several beneficial health effects. The aim of this study was to determine how the baking process and food matrix affect anthocyanin stability and bioaccessibility in bakery products in order to develop functional foods. Three well known regularly consumed bakery products (buns, breadsticks and biscuits) were enriched with anthocyanin (AC) isolated from grape skin alone or in combination with docosahexaenoic acid (AC + DHA) to reveal knowledge on AC as active ingredients in real food systems rather than pure compounds. Anthocyanin amounts added to the formulations of buns, breadsticks and biscuits were 34 mg per 100 g, 40 mg per 100 g and 37 mg per 100 g, respectively. The effect of processing, storage and the food matrix on AC stability and bioaccessibility was investigated. In addition, the sensory properties of bakery products were evaluated. Breadsticks enriched with AC and AC + DHA received the lowest scores in the pre-screening sensory test. Therefore breadsticks were excluded from further analysis. AC retentions, which were monitored by determination of malvidin 3-O-glucoside, in the bun and biscuit after baking were 95.9% (13.6 mg per 100 g) and 98.6% (15.2 mg per 100 g), respectively. Biscuits and buns enriched only with AC showed significantly higher anthocyanin bioaccessibilities (57.26% and 57.30%, respectively) than the same ones enriched with AC + DHA. AC stability in enriched products stored for 21 days was significantly lower than in products stored for 7 days (p < 0.05). However, this loss can be accepted as negligible since more than 70% of AC was retained in all the products.

Vegetable product containing caseinomacropeptide and germinated seed and sprouts.

In this study vegetable product containing germinated seed and sprouts of lentils and cowpeas, and caseinomacropeptide isolated from whey is produced. Three different forms of vegetable product namely puree (VP), freeze-dried (FD) and drum-dried (DD) are produced. Freeze-dried and DD forms are produced to diversify forms of utilization and to improve functionality such as increased shelf life and decreased storage space. Their beneficial effects on health are determined using in vitro methods. All forms displayed antioxidant activities against DPPH radical and oxygen radical, α-amylase inhibitory activities, bile acid binding capacities, and angiotension converting enzyme (ACE) inhibitory activities. Freeze-dried product exhibited the strongest inhibition on α-amylase and ACE with the IC50 value 0.09 µM total phenolic and 0.82 mg protein/g sample, respectively when evaluated on the basis of serving size.

The effect of extrusion on the functional components and in vitro lycopene bioaccessibility of tomato pulp added corn extrudates.

The effect of processing on functional ingredients and their in vitro bioaccessibility should be investigated to develop better food products. Tomato pulp was added as a functional ingredient to extrudates. The effects of extrusion on the functional properties of the extrudates and the in vitro bioaccessibility of lycopene were investigated. Two different temperature sets were applied during extrusion: 80 °C, 90 °C, 100 °C and 130 °C and 80 °C, 100 °C, 130 °C and 160 °C. Screw speed and feed rate were kept constant at 225 rpm and 36 ± 1 g min(-1), respectively. The feed moisture content was adjusted to 30 ± 1% by mixing the tomato pulp to the corn grit. Antioxidant activity and the total phenolic content decreased after the extrusion process. High performance liquid chromatography (HPLC) analysis indicated that the lycopene content decreased after the extrusion process when feed and extrudates were compared. In vitro bioaccessibility of lycopene for the extruded samples with 160 °C last zone treatment temperature was higher than the feed and extruded samples with 130 °C last zone treatment temperature. The results indicate that extrusion affects the food matrix and the release of functional components.

In vitro digestibility of goat milk and kefir with a new standardised static digestion method (INFOGEST cost action) and bioactivities of the resultant peptides.

The hydrolysis degrees of goat milk and kefir during simulated gastrointestinal digestion and some bioactivities of the resulting peptides after fermentation and digestion were studied. A static in vitro digestion method by the COST FA1005 Action INFOGEST was used and goat milk and kefir were partially hydrolyzed during the gastric phase and had above 80% hydrolysis after duodenal digestion. There were no differences between the digestibility of goat milk and kefir (p > 0.05). Goat milk and kefir displayed about 7-fold antioxidant activity after digestion (p < 0.05). Fermentation showed no effect on the calcium-binding capacity of the samples (p > 0.05), however, after in vitro digestion calcium-binding capacity of the goat milk and kefir increased 2 and 5 fold, respectively (p < 0.05). Digested goat milk and kefir showed a higher dose-dependent inhibitory effect on α-amylase compared to undigested samples (p < 0.05). α-Glucosidase inhibitory activities and in vitro bile acid-binding capacities of the samples were not determined at the studied concentrations.

Mind the gap-deficits in our knowledge of aspects impacting the bioavailability of phytochemicals and their metabolites--a position paper focusing on carotenoids and polyphenols.

Various secondary plant metabolites or phytochemicals, including polyphenols and carotenoids, have been associated with a variety of health benefits, such as reduced incidence of type 2 diabetes, cardiovascular diseases, and several types of cancer, most likely due to their involvement in ameliorating inflammation and oxidative stress. However, discrepancies exist between their putative effects when comparing observational and intervention studies, especially when using pure compounds. These discrepancies may in part be explained by differences in intake levels and their bioavailability. Prior to exerting their bioactivity, these compounds must be made bioavailable, and considerable differences may arise due to their matrix release, changes during digestion, uptake, metabolism, and biodistribution, even before considering dose- and host-related factors. Though many insights have been gained on factors affecting secondary plant metabolite bioavailability, many gaps still exist in our knowledge. In this position paper, we highlight several major gaps in our understanding of phytochemical bioavailability, including effects of food processing, changes during digestion, involvement of cellular transporters in influx/efflux through the gastrointestinal epithelium, changes during colonic fermentation, and their phase I and phase II metabolism following absorption.

Microwave-assisted hydrodistillation of essential oil from rosemary.

Effects of microwave assisted hydrodistillation (MAHD) and conventional hydrodistillation (HD) methods on yield, composition, specific gravity, refractive index, and antioxidant and antimicrobial activities of essential oil of Rosmarinus officinalis L were studied. The main aroma compounds of rosemary essential oil were found as 1,8-cineole and camphor. Trolox equivalent antioxidant capacity (TEAC) values for essential oils extracted by MAHD and HD were 1.52 mM/ml oil and 1.95 mM/ml oil, respectively. DPPH radical scavenging activity of the oils obtained by MAHD and HD were found as 60.55% and 51.04% respectively. Inhibitory effects of essential oils obtained by two methods on linoleic acid peroxidation were almost the same. Essential oils obtained by two methods inhibited growth of Esherichia coli O157:H7, Salmonella typhimurium NRRLE 4463 and Listeria monocytogenes Scott A with the same degree. However, inhibitory activity of essential oil obtained by MAHD on Staphylococcus aureus 6538P was stronger than that of obtained by HD (p < 0.05).

Vegetable and fermented vegetable juices containing germinated seeds and sprouts of lentil and cowpea.

Health-promoting effects of vegetable juice (VJ) and fermented vegetable juice (FVJ) were determined. Both juices displayed antioxidant activity against DPPH radical, and ORAC values obtained for both juices were not statistically different. The α-glucosidase inhibitory activities of the VJ and FVJ were not different. However, α-amylase inhibitory effect of the VJ (IC50: 41µM) was higher than that of FVJ (IC50: 149µM) (p<0.05). In vitro bile acid-binding capacity of FVJ was about 4.30times higher than that of VJ (p<0.05). Although in vitro ACE inhibitory activity of VJ was below 50%, FVJ displayed ACE inhibition (80.2%) with an IC50 value of 50µgprotein/ml. Even though ACE inhibitory activities of digested and undigested FVJ were similar, there was a 42-fold decrease in the IC50 value of FVJ after small intestinal digestion (p<0.05). FVJ, diluted by one half, displayed hemagglutinating activity whilst VJ did not display any hemagglutinating activity.

Angiotensin-converting enzyme (ACE)-inhibitory activity of boza, a traditional fermented beverage.

BACKGROUND: In this study the angiotensin-converting enzyme (ACE)-inhibitory activity of boza and protein fractions of boza separated according to molecular weight was determined. In addition, the effect of in vitro digestion on ACE-inhibitory activity was investigated. RESULTS: The protein content, ACE-inhibitory activity and IC(50) value of boza were 1.0896 ± 0.08%, 76.76 ± 14.93% and 7.2 ± 0.28 µg protein mL(-1) respectively. The protein hydrolysate was separated into three fractions according to molecular weight (MW), i.e. MW < 5000 Da, 5000 < MW < 10 000 Da and 10 000 < MW < 20 000 Da. The lowest IC(50) value (0.268 ± 0.07 µg protein mL(-1)) was found for the fraction with 5000 < MW < 10 000 Da (P < 0.05). After in vitro digestion the ACE-inhibitory activities of stomach and intestine dialysates were almost the same (P > 0.05). The IC(50) value of stomach digest was determined as 2.06 ± 0.32 (µg protein mL(-1)). However, the IC(50) value of intestine digest could not be determined, because all dialysates with different protein concentrations displayed ACE-inhibitory activity greater than 50%. CONCLUSION: The findings indicate that boza, protein hydrolysate, fractionated hydrolysates and dialysates obtained after in vitro digestion contain bioactive compounds with different ACE-inhibitory activities. Based on these results, boza can be considered as a good source of ACE-inhibitory peptides.

Antioxidant and antimicrobial activities of essential oils obtained from oregano (Origanum vulgare ssp. hirtum) by using different extraction methods.

In this study, antioxidant and antimicrobial activities of essential oils obtained from oregano (Origanum vulgare ssp. hirtum) were determined by using solvent-free microwave extraction (SFME), supercritical fluid extraction, and conventional hydrodistillation (CH) methods. The inhibitory effects on the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical of essential oils obtained from oregano by using SFME and CH were similar. However, essential oil extracted by CH showed greater (2.69 µmol/µL of oil) Trolox equivalent antioxidant capacity (TEAC) than oregano oils obtained by SFME (P < .05). The difference between percentage inhibition and TEAC values most probably is due to the fact that undiluted and diluted samples are used in the percentage inhibition assay and the TEAC assay, respectively. TEAC values of oregano essential oils obtained by SFME at different microwave power levels were found to be similar and ranged from 0.72 to 0.84 µmol/µL of oil. Essential oils obtained by CH and SFME at different microwave powers inhibited the survival of Listeria monocytogenes, Salmonella typhimurium, and Escherichia coli O157:H7, whereas survival of Staphylococcus aureus was not influenced. In addition, oregano oil obtained by SFME at 40% power level did not show any inhibitory effect on E. coli O157:H7.

Olive tree (Olea europaea) leaves: potential beneficial effects on human health.

Olive tree (Olea europaea L.) leaves have been widely used in traditional remedies in European and Mediterranean countries such as Greece, Spain, Italy, France, Turkey, Israel, Morocco, and Tunisia. They have been used in the human diet as an extract, an herbal tea, and a powder, and they contain many potentially bioactive compounds that may have antioxidant, antihypertensive, antiatherogenic, anti-inflammatory, hypoglycemic, and hypocholesterolemic properties. One of these potentially bioactive compounds is the secoiridoid oleuropein, which can constitute up to 6-9% of dry matter in the leaves. Other bioactive components found in olive leaves include related secoiridoids, flavonoids, and triterpenes. The evidence supporting the potentially beneficial effects of olive leaves on human health are presented in this brief review.

Primary oral cancer in a Turkish population sample: association with sociodemographic features, smoking, alcohol, diet and dentition.

The aim of this multicentre case-control study was to investigate the association of a variety of factors with oral cancer in a group of Turkish patients. Questionnaires were used to investigate the sociodemographic features, smoking and alcohol consumptions, dietary habits and dental status of 79 primary oral cancer patients and 61 controls. Data were statistically analysed with Mann-Whitney U-test, Pearson Chi-square and binary logistic regression analyses to determine the odds ratios. Low level of education, gender, dietary habits, having poor oral hygiene and denture sores were associated with primary oral cancer in this patient sample; but eating salads and raw vegetables, fish, and drinking red wine were related with healthy status. Determination of the factors associated with oral cancer and of the high-risk groups would be beneficial to provide efficient screening protocols and prevention programmes for oral cavity cancers.