Tomato waste as feedstock to extract phenolic compounds with anti-oxidant and anti-inflammatory bioactivity

Introduction: Tomato industry is one of the most developed at world level and produces a lot of by-products. Recently tomato peels and seeds (TP) consider as feedstock to extract bioactive fractions like phenolic compounds. In this work two different TP from Italian plant were selected to profile the flavonoids contents and their antioxidant and anti-inflammatory properties. Material & Methods: Tomato peel+seed fraction from conventional (TP-CONV) and biological (TP-BIO) cultivation systems were sampled and dried, and were characterized. Polyphenols were profiled by HPLC, total antioxidant capacity (TAC) was evaluated by TEAC and FRAP methods. Immunomodulating effects were evaluated by using Caco-2 cells transfected with a pNiFty2-Luc vector. Enzyme inhibitory activity was assessed against intestinal \u3b1-glucosidase and pancreatic \u3b1-amylase. Results: Total polyphenol content were the same in both TP-BIO and TP-CONV however identified phenolic compounds showed distinctive distribution. Both sample extracts expressed antioxidant , anti-inflammatory and enzyme inhibitory activities. Conclusion: This study highlights the promising TP extracts antioxidant activity, anti-inflammatory, and inhibition on the activity of enzymes related to release/uptake of glucose. These waste products can be recovered and be regarded as potential nutraceutical sources as low-cost but nutritional supplements either in food production as functional ingredients and nutraceuticals or in cosmetic and pharmaceutical industries

Characterization of pigmented cereal grains and production of functional food: anthocyanins-enriched pasta

This study was aimed at characterizing the anthocyanin profile in different varieties of pigmented corn and wheat and in some of their milling fractions, at assessing the anthocyanins and polyphenol profiles, and at investigating the anti-inflammatory and enzyme-inhibiting activities of these materials. Acid/ethanol extracts were used to assess total anthocyanins, overall antioxidant activity, the overall polyphenol profile, and for evaluating the inhibition of pancreatic a-amylase and of intestinal alpha-glucosidase. Dose-dependent inhibition of both enzymes was evident in all extracts within the same range of bioactives concentration, although with a different efficiency of individual extracts towards each enzyme. Anti-inflammatory response was evaluated by using acid-free extracts and a cellular model based on Caco-2 cells transiently transfected with a luciferase reporter gene responding to cytokine stimulation. The response of interleukin-stimulated cells decreased significantly in a dose-responsive manner in the presence of 20-50 micromol/liter anthocyanins from all grains extracts, again with a different efficiency. By comparing the different inhibitory ability of extracts from the various sources, it appears that the observed effects are in most cases higher than what observed in similar extracts from other sources, and that the activity in each extract may be related to specific synergies between anthocyanins and polyphenols. Therefore, we attempted to incorporate these materials into staple food. Purple pasta was prepared by introducing anthocyanin-rich fractions from purple wheat (PWF1 & PWF2), into soft wheat flour and durum wheat semolina. Bioactive levels were assessed in the different types of pasta after production and after cooking. Acid/ethanol extracts from cooked pasta retained high levels of TAC and TPC, as well as significant anti-oxidant activity, anti-inflammatory properties on cell models, and enzyme inhibitory activities. Thus, in conclusion, it can be stated that foods incorporating a naturally rich source of bioactive compounds. retain beneficial properties for the organism also after cooking

Characterization of Whole Grain Pasta: Integrating Physical, Chemical, Molecular, and Instrumental Sensory Approaches

The consumption of whole-grain foodincluding pastahas been increasing steadily. In the case of whole-grain pasta, given the many different producers, it seems important to have some objective parameters to define its overall quality. In this study, commercial whole-grain pasta samples representative of the Italian market have been characterized from both molecular and electronic-senses (electronic nose and electronic tongue) standpoint in order to provide a survey of the properties of different commercial samples. Only 1 pasta product showed very low levels of heat damage markers (furosine and pyrraline), suggesting that this sample underwent to low temperature dry treatment. In all samples, the furosine content was directly correlated to protein structural indices, since protein structure compactness increased with increasing levels of heat damage markers. Electronic senses were able to discriminate among pasta samples according to the intensity of heat treatment during the drying step. Pasta sample with low furosine content was discriminated by umami taste and by sensors responding to aliphatic and inorganic compounds. Data obtained with this multidisciplinary approach are meant to provide hints for identifying useful indices for pasta quality. Practical ApplicationAs observed for semolina pasta, objective parameters based on heat-damage were best suited to define the overall quality of wholegrain pasta, almost independently of compositional differences among commercial samples. Drying treatments of different intensity also had an impact on instrumental sensory traits that may provide a reliable alternative to analytical determination of chemical markers of heat damage in all cases where there is a need for avoiding time-consuming procedures

Molecular characterization of pasta enriched in fiber and antioxidants: investigation of their functional role by in vitro and in vivo approaches=caratterizzazione molecolare di pasta arricchita in fibra e antiossidanti: approcci in vitro ed in vivo al loro ruolo funzionale

Questo progetto di ricerca intende valutare alcuni aspetti connessi alla produzione ed al consumo di pasta arricchita in fibra ed antiossidanti. Il progetto nasce dal largo utilizzo della pasta, dalla possibilit\ue0 di utilizzarla su larga scala come veicolante di molecole nutrizionalmente interessanti, ma anche dalla necessit\ue0 di valutare l'impatto del processo di produzione e il destino metabolico dei componenti naturali usati per l'arricchimento che, in questa fase del progetto, sono frazioni arricchite derivate da grano e grano saraceno mediante l'esclusivo utilizzo di metodi fisici.Cereal products are consumed daily by a vast majority of the world population. Functional foods offer an outstanding opportunity to improve the quality of these products. Common buckwheat (Fagopyrum esculentum Moench) is recognized as an important functional food in many countries. The nutrient profile of buckwheat is of high interest due to its high protein quality and the high levels of fiber, vitamins and minerals. Buckwheat is also a good source of bioactive compounds like flavonoids and phyosterols. Phenolics in buckwheat have been shown to possess antioxidant activity (\u15eensoy, Rosen et al. 2006). Flavonol glycosides include rutin, quercetin, kaemferol-3-rutinoside and minor amounts of a \ufb02avanol triglycoside (Tian, Li et al. 2002). Rutin (quercetin-3-rutinoside) is highly represented, and has reported antioxidant, anti-in\ufb02ammatory and anti-carcinogenic effects (Baumgertel, Grimm et al. 2003). Whole buckwheat grains also can be considered as a prebiotic food because of their effect on decreasing Enterobacteria, while increasing lactic acid bacteria (Krko\u161kov\ue1 and Mr\ue1zov\ue1 2005)

Molecular characterization of pasta enriched in antioxidants: investigation of their functional role by in vitro and in vivo approaches

1-Introduction: In agreement with the PhD thesis project described last year, this poster reports the main results of the first two activities concerning: (A1) Characterization and determination of the amount of starch, proteins and phenolics in different fractions of pigmented grains yielded by mechanical treatments. Monitoring the properties of the polysaccharides and proteins in the various pasta samples through novel methodologies (Bonomi et al., 2012). Studying the effect of extracts on glycaemia and on biomarkers of oxidative stress and inflammation by using inflamed Caco-2 line as a model. (A2) Producing pasta by incorporating pigmented grain flour in the nutritionally relevant amounts of fiber and anthocyanins, determining the nutritional/functional properties of the developed food products. 2-Materials and Methods: Pigmented grains were de-branned and milled in a lab-scale milling system. Extraction was conducted with ethanol and water (65:35 v/v) containing Hcl (%0.01). Analytical profiling of the phenolics in bran components/fractions separated by physical techniques was carried out by HPLC method. Free radical scavenging activity and the antioxidant capacity was measured by DPPH and FRAP method respectively. To study the anti-inflammatory effects we used a convenient way which relies on a reporter gene system in which Caco-2 cells are transfected with a vector (pNiFty2-Luc) which that enables facile monitoring of inflammatory state. The reporter gene encodes for a luciferase-labeled product, and is under control of a native promoter of the transcription regulator NF- B, which has a key role in the inflammatory response. In this system, the light emission (bioluminescence) is directly connected to the activation or repression of the promoter (Huang, Kim et al. 2010). Cells were supplemented with in vitro at different concentration of anthocyanin rich extracts in the presence/absence of interleukin (IL)-1 f, an important pro-inflammatory cytokine working as an activator of NF-B expression. Monitoring bioluminescence allows us for relatively easy estimation of the role of anthocyanins and/or metabolites with respect to the cellular response to stimulation. The \u3b1-glucosidase inhibitory activity was determined to measure antidiabetic effect of the pigmented grains extracts. 3-Results and discussion 3-1: Analytical profiling of the phenolics in the various fractions separated by physical techniques indicated that the outermost fraction obtained through debranning of purple wheat represents a very rich (and convenient) source of phenolics (and fiber, as assessed by independent measurements). The major anthocyanins found are cyanidin-3-o-glucoside and Petunidin-3-O- glucoside. DPPH and FRAP showed a relevant antioxidant activity with the anthocyanins amount in different fractions. The ACN-rich fraction from purple wheat decreased the immune response of Caco-2 cells in a dose-responsive manner, and wheat phenolics were apparently more effective than Cyanidines, that reportedly possess a strong anti-inflammatory activity (Yu and Ahmedna 2013).3-2: Enriched pasta were produced from different fractions of wheat flour and purple wheat bran, with fiber content in excess of 10%, and a content in anthocyanins in the 100 mg/kg range.Our results indicate that the phenolics-rich fraction from purple wheat at the same concentration to Cyanidine appears to be more effective in impairing the expression of NF-\u3baB in transformed cells. Finding cyanidin-3-o-glucoside and Petunidin-3-O- glucoside as the main anthocyanin in purple wheat is in parallel with studding by Chen (Chen, Mu\u308ller et al. 2013). These bioactive-rich fractions were incorporated in making pasta as staple food for human consumption to study its effectiveness on reducing the inflammation and hyperglycemia (Tsuda 2012, Cassidy, Mukamal et al. 2013). Although the bioavailability of anthocyanins and the amount of metabolites formed in the gastrointestinal tract, in order to introduce the desired anthocyanin efficacy should be considered during the following year of this PhD project, and also further studies will address the bioavailability of anthocyanins in pasta and the amount of metabolites they form or release in the gastrointestinal tract through studding their effects on the local microflora, and of the gut microflora interactions

Cereali pigmentati: proprietà funzionali della componente polifenolica

Numerosi studi hanno evidenziato come la componente polifenolica presente nel pericarpo di cereali pigmentati abbia attivit\ue0 antiinfiammatoria (1,2) e antidiabetica (3,4). Scopo di questo studio \ue8 stato quello di indagare alcune propriet\ue0 biofunzionali della componente fenolica in diversi cereali pigmentati. Nello specifico \ue8 stato indagato il ruolo di questi composti nell\u2019espressione di specifici marcatori di infiammazione in sistemi cellulari e sull\u2019attivit\ue0 di enzimi pancreatici e dell\u2019orletto a spazzola intestinali. In questo studio sono stati pressi in esame cinque cereali pigmentati quali tre variet\ue0 di mais (due variet\ue0 blu (T, MF) ed una non-blu (Rostrato)) e due variet\ue0 di frumento (Purple Wheat, Skorpion). La quantit\ue0 di antocianine \ue8 risultata massima in Purple Wheat (0. 69 mg/g), seguita dalle due variet\ue0 di blue corn (T, MF, con 0.66 e 0.53 mg/g rispettivamente). La capacit\ue0 antiossidante \ue8 risultata invece maggiore nel campione di mais Rostrato e in quella di Purple Wheat. Il profilo polifenolico, indagato mediante RP-HPLC, ha evidenziato la presenza in proporzione diversa di almeno cinque componenti principali. Tutti i cereali considerati hanno evidenziato attivit\ue0 antinfiammatoria, dose dipendente, in modelli cellulari basati su cellule Caco2. Le due variet\ue0 di frumento pigmentato hanno evidenziata un\u2019attivit\ue0 antiinfiammatoria maggiore rispetto a quella delle variet\ue0 di mais, e paragonabile a quella osservata con cianidina 3-glucoside purificata. L\u2019inattivazione dell\u2019attivit\ue0 di alfa amilasi e alfa-glucosidasi \ue8 stata invece osservata invece nel caso di Purple Wheat e di blue corn, variet\ue0 T. Nel loro complesso questo studio ha evidenziato la presenza di attivit\ue0 antiinfiammatoria ed una inibizione di enzimi pancreatici ed intestinali molto interessanti. In questo contesto si intende completare la caratterizzazione biofunzionale di questi cereali indagando il loro possibile effetto nei confronti sia di fattori di rischio cardiovascolare e del microbiota intestinale

Interaction with Semolina Proteins Allows Retention of Bioactives in Pasta From Purple Wheat

Abstract: Postprandial hyperglycemia, is the earliest metabolic abnormality occur in type-2 Diabetes Mellitus. The major source of glucose in the body comes from the hydrolysis of dietary carbohydrates, like starch by the action of pancreatic \u251-amylase and intestinal \u251 -glucosidase enzymes which hydrolyze the glycosidic bonds in starch components from the diet. Therefore, the inhibition of these enzymes can be an important concept for management of type 2 diabetes (Rios, Francini et al. 2015) Grains and cereals containing high amounts of dietary fibers are generally recommended for diabetic patients to control their blood glucose level. Recently food-based active ingredients, such as polyphenols, have shown the ability to modulate blood glucose level (Thondre 2013). Colored cereal grains contain higher amount of phenolics and flavonoids, including anthocyanins than common grains (Liu, Qiu et al. 2010). Total polyphenols and anthocyanins in different cereal grains showed positive correlation with \u251 -glucosidase inhibition activities (Yao, Sang et al. 2010). Previous studies indicate that anthocyanins such as cyanidin-3-rutinoside exhibits an inhibitory effect on \u251-glucosidase in a non-competitive manner, although other anthocyanins may inhibit the \u251-amylase and \u251 -glucosidase enzymes activity in different ways (Akkarachiyasit, Charoenlertkul et al. 2010). The structure of different anthocyanins that affect the inhibitory activity against these enzymes has been recently studied (Xiao, Ni et al. 2013). Anthocyanins also significantly protect beta-cells against glucose-induced oxidative stress (Yan, Zheng et al. 2017). In this study we aimed at characterizing the anthocyanin profile in different varieties of pigmented corn and wheat and in some of their milling fraction. In the second phase of study four different types of purple pasta was prepared incorporating debranned fractions, wholegrain flour of purple wheat and semolina flour. Acid/ethanol extracts were used to assess total anthocyanins, overall antioxidant activity, the overall polyphenol profile, and for evaluating the inhibition of pancreatic \u251-amylase and of intestinal \u251-glucosidase both in corn/wheat flours, and in the extracts from produced pasta (before and after cooking procedure)

Molecular Aspects of the Functional Activities of Pigmented Grains

I cereali colorati sono ricche fonti di antociani con attivit\ue0 antiossidante, ed hanno un grande potenziale non solo come coloranti alimentari ma anche come ingredienti alimentari in grado di esercitare effetti benefici per la salute. La quantificazione del tenore di antocianine e polifenoli nei cereali \ue8 cruciale per valutare in modo appropriato i loro benefici nutrizionali e per consentire la loro applicazione come materie prime alimentari con propriet\ue0 funzionali. Il nostro obiettivo era studiare le attivit\ue0 anti-infiammatorie di estratti ricchi di antociani da diversi grani pigmentati utilizzando cellule Caco-2 transfettate con un vettore pNiFty2-Luc, e di analizzare la capacit\ue0 di questi estratti nell\u2019inibire l\u2019attivit\ue0 dell'alfa-amilasi pancreatica e delle alfa-glucosidasi intestinali

Pigmented cereals have remarkably anti-inflammatory and anti-diabetic effects

Anthocyanins in grains are expressed in either the pericarp or aleuronic layer of different cereal varieties. Many studies have shown the health-promoting effects of anthocyanins, that have anti-inflammatory and antidiabetic activities. In this study we have investigated the profile of phenolics in pigmented grains and tested their bioactive molecules as for efficacy on inflammation markers (by using Caco-2cells transfected with a pNiFty2-Luc vector), and as for their inhibitory activities against alpha-glucosidase and alpha-amylase. Analytical profiling of the anthocyanins in five different colored grains (three varieties of corn and two of wheat) was carried out by advanced LC methods. Total anthocyanins amount was determined using a pH differential method. Antioxidant capacity was measured as FRAP. Immunomodulating effects were assessed by testing cytokine-stimulated synthesis of a luciferase-labeled reporter of gene activation in transformed Caco-2 cell lines. Inhibition of alpha-amylase and alpha -lucosidase was assessed according to standard methods. Four major anthocyanins were characterized from HPLC profile. Cyanidine-3-glucoside was predominant both in corn and wheat samples. Purple wheat and Blue corn (T) had the highest anthocyanin contents (0.74 and 0.66 mg/g, respectively), followed by blue corn (MF) (0.53 mg/g) and a fraction from de-hulling of Skorpion wheat (0.18mg/g), that had the highest rutin content. The Rostrato variety of corn showed the highest antioxidant capacity (47.2 \u3bcmolFe(II)/g) between all samples. The highest amount of ferulic acid was observed in Blue corn (T) and Rostrato. The immune response of Caco-2 cells decreased significantly in a dose-responsive manner for all grains extracts. Extracts from blue wheat (cultivar SK) and purple wheat where effective in decreasing inflammatory response at anthocyanin concentrations comparable to those of pure cyanindin-3-glucoside. Purple wheat and blue corn (cultivar T) extracts also displayed significant inhibition of \u3b1-amylase activity. Values ranged between 78% (wheat) and 95.4% (corn) at final concentrations of extract solids in the 100-130 \u3bcg/ml range. Under comparable conditions, brush border \u3b1-glucosidase inhibition from the same extracts was 57.8 % (wheat) and 65.4% (corn). The present work highlights the potential health promoting effects of phenolic-rich extracts of pigmented grains. The enriched cereal fractions studied here can be stably and directly incorporated into staple food material like pasta, bread, noodles or snacks. Work is in progress to assess their possible role in regulating the intestinal microbiota

Bioactives in pigmented grains: properties, activities, and incorporation into functional foods

Background: Grains as staples are good source of nutrients like carbohydrates, protein, lipids, vitamins, minerals and dietary phytochemicals such as phenolics, carotenoids, etc. Phenolic compounds have shown a broad range of biological activities. In this study, we have aimed to investigate the anthocyanin profile in diverse pigmented grains and to examine their bioactive molecules. to test on inflammation markers by using transfected Caco-2cells (with a vector pNiFty2-Luc) as a cell model, detecting their inhibitory activities against a-glucosidase and a-amylase enzymes, in order to select the appropriate grain to formulate functional food. Methods: Samples: Three varieties of corn and two varieties of wheat were used in this study. Methods: Ethanolic extracts acidified with 1.0 N HCl (65:35, v/v) was prepared. TAC and TPC was determined (Abdel Aal et al. (2006)). The FRAP assay was performed (Benzie and Strain (1996)). The major anthocyanins components were separated by HPLC (Hosseinian et al., 2008). The immunomodulatory activity of ACN-rich fractions was tested by using Caco-2 cell transfected model (Taverniti et al.,2014). Pancreatic \u3b1-amylase and intestinal \u3b1-glucosidase inhibitory activities were detected (Lavelli et al., 2016). Pasta making: Bioactive compounds-enriched pasta was prepared by adding 14.2 g/100g of anthocyanin-rich fraction of purple wheat to commercial flour, given us a pasta with greater nutritional value than the commercial durum wheat semolina pasta. Results: Purple wheat showed the highest TAC (690\ub10.02 \ub5g/g), TPC (736\ub10.44 \ub5g/g) and antioxidant activity (47.5 \ub110.26 \u3bcmol Fe(II)/g) among all samples. The immune response of Caco-2 cells decreased for all grains extracts. Purple wheat and blue corn (cultivar T) extracts were much active to inhibit \u3b1-amylase enzyme. Enriched purple wheat pasta showed a relatively high antioxidant capacity. The anthocyanins content value ranged from 67.9\ub10.9 \ub5g/g which is higher than values found by Ficco et al. (2016) in cooked pasta prepared from purple wholemeal semolina (16.89 \ub5g/g). Extracts from cooked pasta showed noticeable Immunomodulatory effects. Conclusion: Naturally functional pasta prepared from purple wheat showed antioxidant and anti-inflammatory effects. Their use as daily food deserves further studies on their reported effects on cardiovascular risk factors, as well as their possible effects on the gut microflora. References: Nankar, Amol N., et al. "Quantitative and qualitative evaluation of kernel anthocyanins from southwestern United States blue corn." Journal of the Science of Food and Agriculture 96.13 (2016): 4542-4552. Huang, Bo, et al. "Anti-diabetic effect of purple corn extract on C57BL/KsJ db/db mice." Nutrition research and practice 9.1 (2015): 22-29. Tsuda, Takanori, et al. "Dietary cyanidin 3-O-\u3b2-D-glucoside-rich purple corn color prevents obesity and ameliorates hyperglycemia in mice." The Journal of nutrition 133.7 (2003): 2125-2130. Kim, Eun Ok, et al. "Anti-inflammatory activity of hydroxycinnamic acid derivatives isolated from corn bran in lipopolysaccharide-stimulated Raw 264.7 macrophages." Food and Chemical Toxicology 50.5 (2012): 1309-1316. Miguel, Maria Gra\ue7a. "Anthocyanins: Antioxidant and/or anti-inflammatory activities." (2011). Ficco, D. B. M., De Simone, V., De Leonardis, A. M., Giovanniello, V., Del Nobile, M. A., Padalino, L., ... & De Vita, P. (2016). Use of purple durum wheat to produce naturally functional fresh and dry pasta. Food chemistry, 205, 187-195