 |
Davide TAGLIAZUCCHI
Professore Associato
Dipartimento di Scienze della Vita sede ex-Agraria
|
Home |
Curriculum(pdf) |
Didattica |
Pubblicazioni
2023
- Effect of Fermentation with Streptococcus thermophilus Strains on In Vitro Gastro-Intestinal Digestion of Whey Protein Concentrates
[Articolo su rivista]
Helal, A.; Pierri, S.; Tagliazucchi, D.; Solieri, L.
abstract
Three Streptococcus thermophilus strains, namely RBC6, RBC20, and RBN16, were proven to release bioactive peptides during whey protein concentrate (WPC) fermentation, resulting in WPC hydrolysates with biological activities. However, these bioactive peptides can break down during gastro-intestinal digestion (GID), hindering the health-promoting effect of fermented WPC hydrolysates in vivo. In this work, the effect of simulated GID on three WPC hydrolysates fermented with S. thermophilus strains, as well as on unfermented WPC was studied in terms of protein hydrolysis, biological activities, and peptidomics profiles, respectively. In general, WPC fermentation enhanced protein hydrolysis compared to unfermented WPC. After in vitro GID, WPC fermented with S. thermophilus RBC20 showed the highest antioxidant activity, whereas WPC fermented with strain RBC06 displayed the highest angiotensin-converting enzyme (ACE)- and dipeptidyl peptidase IV (DPP-IV)-inhibitory activities. Peptidomics analysis revealed that all digested WPC samples were highly similar to each other in peptide profiles, and 85% of the 46 identified bioactive peptides were shared among fermented and unfermented samples. However, semi-quantitative analysis linked the observed differences in biological activities among the samples to differences in the amount of bioactive peptides. The anti-hypertensive peptides VPP and IPP, as well as the DPP-IV-inhibitory peptide APFPE, were quantified. In conclusion, WPC fermentation with S. thermophilus positively impacted protein hydrolysis and bioactive peptide release during GID.
2023
- Effect of Ripening and In Vitro Digestion on Bioactive Peptides Profile in Ras Cheese and Their Biological Activities
[Articolo su rivista]
Helal, A.; Cattivelli, A.; Conte, A.; Tagliazucchi, D.
abstract
The effect of ripening and in vitro digestion on the biological activities, peptide profiles and release of bioactive peptides in Ras cheese has been investigated. Ras cheese ripening largely influenced the extent of protein hydrolysis. The advancement in ripening resulted in an increase in total peptides (from 0.97 to 2.46 mmol leucine/g in samples at 30 and 180 days of ripening, respectively) and bioactive peptides concentration, especially angiotensin-converting enzyme (ACE)-inhibitory, dipeptidyl-peptidase-IV-(DPP-IV)-inhibitory and antioxidant peptides. In vitro gastro-intestinal digestion further promoted protein hydrolysis and the release of bioactive peptides. Digested Ras cheese at 90 and 180 days of ripening displayed the highest bioactive peptides intensity. The variations in bioactive peptides amount during ripening and in vitro digestion were correlated with the changes in ACE-inhibitory, DPP-IV-inhibitory and antioxidant activities. The highest amounts of VPP and IPP were detected in digested Ras cheese at 90 days of ripening (17.44 and 36.50 mg/kg of cheese, respectively), whereas the highest concentrations of APFPE were found in undigested and digested 180-day ripened Ras cheese (82.09 and 52.01 mg/kg of cheese, respectively). The present investigation underlined potential differences in the biological effect after the ingestion of Ras cheese at different ripening times.
2023
- How starter cultures affect the peptidomic profile and bioactive activities of the Asiago-PDO cheese throughout ripening
[Articolo su rivista]
Domingues Galli, Bruno; Nikoloudaki, Olga; Tonini, Stefano; MOHAMED IBRAHIM HELAL, Ahmed; Di Cagno, Raffaella; Gobbetti, Marco; Tagliazucchi, Davide
abstract
Our study investigated the chemical, microbiological, and bioactive peptide profiles of Asiago Protected Designation of Origin (PDO) cheese from two dairies (Dairy I and II) produced over two consecutive days (batches) and analysed during three months of ripening. The effect of different starter cultures was evaluated. The microbiome varied between the dairies and batches, with curds post-salting dominated by the starter culture-associated genera. During ripening, there was an increasing trend in the Lactobacillus genus, especially for Dairy I, which used an industrial starter. Bioactive peptide intensities differed throughout ripening due to the extent of proteolysis, and their intensity or concentration evolved, modifying, and differentiating profiles. The industrial starter used in Dairy I had the highest relative intensity (average value 76.50%) of bioactive peptides after three months of ripening. In contrast, the cheeses made with natural milk starter (Dairy II) had lower total relative intensity (average value 47.75%) but produced ACE-inhibitory peptides through sub-dominant strains and non-starter lactic acid bacteria. The importance of autochthonous strains of each micro-region even within a delimited PDO production area was highlighted.
2023
- Impact of Spontaneous Fermentation and Inoculum with Natural Whey Starter on Peptidomic Profile and Biological Activities of Cheese Whey: A Comparative Study
[Articolo su rivista]
MOHAMED IBRAHIM HELAL, Ahmed; Nasuti, Chiara; Sola, Laura; Sassi, Giada; Tagliazucchi, Davide; Solieri, Lisa
abstract
Fermentation is a promising solution to valorize cheese whey, the main by-product of the dairy industry. In Parmigiano Reggiano cheese production, natural whey starter (NWS), an undefined community of thermophilic lactic acid bacteria, is obtained from the previous day residual whey through incubation at gradually decreasing temperature after curd cooking. The aim of this study was to investigate the effect of fermentation regime (spontaneous (S) and NWS-inoculated (I-NWS)) on biofunctionalities and release of bioactive peptides during whey fermentation. In S and I-NWS trials proteolysis reached a peak after 24 h, which corresponded to the drop out in pH and the maximum increase in lactic acid. Biological activities increased as a function of fermentation time. NWS inoculum positively affected antioxidant activity, whilst S overcame I-NWS in angiotensin-converting enzyme (ACE) and DPP-IV (dipeptidyl peptidase IV) inhibitory activities. Peptidomics revealed more than 400 peptides, mainly derived from β-casein, κ-casein, and α-lactalbumin. Among them, 49 were bioactive and 21 were ACE-inhibitors. Semi-quantitative analysis strongly correlated ACE-inhibitory activity with the sum of the peptide abundance of ACE-inhibitory peptides. In both samples, lactotripeptide isoleucine-proline-proline (IPP) was higher than valine-proline-proline (VPP), with the highest content in S after 24 h of fermentation. In conclusion, we demonstrated the ability of whey endogenous microbiota and NWS to extensively hydrolyze whey proteins, promoting the release of bioactive peptides and improving protein digestibility.
2023
- Impact of cooking methods of red-skinned onion on metabolic transformation of phenolic compounds and gut microbiota changes
[Articolo su rivista]
Cattivelli, A.; Nissen, L.; Casciano, F.; Tagliazucchi, D.; Gianotti, A.
abstract
Herein, we investigated the stability and bioaccessibility of phenolics in differently cooked red-skinned onion (RSO) and consequently their impact on the gut microbiota and metabolism of phenolics. In fact, the different processes used to cook vegetables can modify and re-arrange the molecular profiles of bioactive compounds, such as phenolics in phenolic-rich vegetables, such as RSO. Fried and grilled RSO were compared to raw RSO and a blank control and subjected to oro-gastro-intestinal digestion and subsequent colonic fermentation. For upper gut digestion, the INFOGEST protocol was used, and for lower gut fermentation, a short-term batch model, namely, MICODE (multi-unit in vitro colon gut model), was employed. During the process, phenolic compound profile (through high-resolution mass spectrometry) and colon microbiomics (qPCR of 14 core taxa) analyses were performed. According to the results, the degradation driven by the colon microbiota of RSO flavonols resulted in the accumulation of three main metabolites, i.e., 3-(3 '-hydroxyphenyl)propanoic acid, 3-(3 '-hydroxyphenyl)acetic acid and 3-(3 ',4 '-dihydroxyphenyl)acetic acid. Also, colonic fermentation of raw onions resulted in a substantial increase in beneficial taxa, which was larger compared to the heat-treated onions, particularly Lactobacillales and beneficial clostridia. Also, a higher level of inhibition of opportunistic bacteria was seen for the raw onion samples, namely, Clostridium perfringens group and Escherichia coli. Thus, our results showed that RSO, and especially the raw one, is an excellent dietary source of flavonols that are strongly metabolized by gut bacteria and can positively modulate the gut microbiota. Although additional in vivo studies are necessary, this work is one of the first to explore how RSO processed with different cooking methods can differently impact the phenolic metabolism and microbiota composition in the large intestine of humans, fine-tuning the antioxidant nature of foods.
2023
- OeBAS and CYP716C67 catalyze the biosynthesis of health-beneficial triterpenoids in olive (Olea europaea) fruits
[Articolo su rivista]
Alagna, F.; Reed, J.; Calderini, O.; Thimmappa, R.; Cultrera, N. G. M.; Cattivelli, A.; Tagliazucchi, D.; Mousavi, S.; Mariotti, R.; Osbourn, A.; Baldoni, L.
abstract
center dot The bioactive properties of olive (Olea europaea) fruits and olive oil are largely attributed to terpenoid compounds, including diverse triterpenoids such as oleanolic, maslinic and ursolic acids, erythrodiol, and uvaol. They have applications in the agri-food, cosmetics, and pharmaceutical industries. Some key steps involved in the biosynthesis of these compounds are still unknown.center dot Genome mining, biochemical analysis, and trait association studies have been used to identify major gene candidates controlling triterpenoid content of olive fruits.center dot Here, we identify and functionally characterize an oxidosqualene cyclase (OeBAS) required for the production of the major triterpene scaffold beta-amyrin, the precursor of erythrodiol, oleanolic and maslinic acids, and a cytochrome P450 (CYP716C67) that mediates 2 alpha oxidation of the oleanane- and ursane-type triterpene scaffolds to produce maslinic and corosolic acids, respectively. To confirm the enzymatic functions of the entire pathway, we have reconstituted the olive biosynthetic pathway for oleanane- and ursane-type triterpenoids in the heterologous host, Nicotiana benthamiana. Finally, we have identified genetic markers associated with oleanolic and maslinic acid fruit content on the chromosomes carrying the OeBAS and CYP716C67 genes.center dot Our results shed light on the biosynthesis of olive triterpenoids and provide new gene targets for germplasm screening and breeding for high triterpenoid content.
2023
- Peptidomics Profile, Bioactive Peptides Identification and Biological Activities of Six Different Cheese Varieties
[Articolo su rivista]
MOHAMED IBRAHIM HELAL, Ahmed; Tagliazucchi, Davide
abstract
Several recent published studies reported that cheese consumption may protect against the onset of cardiovascular diseases and type-2 diabetes due to the presence of bioactive peptides. In the present work, six cheese varieties (the Egyptian traditional cheeses Karish, Domiati and Ras as well as Feta-type, Gouda and Edam cheeses) were characterized for their peptidomics profiles with high-resolution mass spectrometry, biological activities and content in bioactive peptides. The highest ACE-inhibitory and DPP-IV-inhibitory activities were found in Gouda cheese, which also displayed the highest antioxidant activity. A total of 809 peptides originating from the major milk proteins were identified, and 82 of them were bioactive. Most of them showed ACE-inhibitory, antioxidant and DPP-IV-inhibitory activities. The highest amount of the in vivo anti-hypertensive tripeptides VPP and IPP was found in Gouda cheese (39.19 ± 1.26 and 17.72 ± 0.89 mg/100 g of cheese, respectively), whereas the highest amount of APFPE was detected in Edam cheese (509.13 ± 20.44 mg/100 g of cheese). These results suggest that the intake of Edam, Domiati and, especially, Gouda cheeses may result in a possible anti-hypertensive effect in hypertensive subjects.
2023
- Red-skinned onion phenolic compounds stability and bioaccessibility: A comparative study between deep-frying and air-frying
[Articolo su rivista]
Cattivelli, A.; Di Lorenzo, A.; Conte, A.; Martini, S.; Tagliazucchi, D.
abstract
In this study, the effect of deep-frying and air-frying on the stability and gastro-intestinal release of red-skinned onion phenolic compounds have been assessed by high-resolution mass spectrometry. Forty-four phenolic compounds were identified and quantified in raw onion. Both the frying treatments caused an increase in total phenolic compounds, which was more evident after air-frying (47.4% vs 18.6% increase). This increase was mainly a consequence of the water loss observed during frying. Quercetin was the most important phenolic compound detected both in raw and air-fried samples (99.5 ± 1.7 and 133 ± 2 μmol/100 g, respectively) while deep-fried onion was richer in quercetin-4’-O-glucoside and quercetin-3-O-glucoside-4'-O-glucoside (48.7 ± 0.9 and 41.8 ± 0.3 μmol/100 g, respectively). After digestion, 38.6%, 60.5% and 89.6% of total phenolic compounds were bioaccessible in raw, deep-fried and air-fried onion. Air-fried onion exhibited the highest concentration of bioaccessible phenolic compounds (206 ± 1 μmol/100 g). Oxidative degradation and hydrolysis reactions of quercetin-hexosides occurred during in vitro digestion. Hence, air-frying can be considered a healthy cooking method able to preserve and release after digestion most of the health-promoting compounds found in raw onion and with a lower amount of fats and polar toxic compounds compared to deep-frying.
2022
- Application of a Combined Peptidomics and In Silico Approach for the Identification of Novel Dipeptidyl Peptidase-IV-Inhibitory Peptides in In Vitro Digested Pinto Bean Protein Extract
[Articolo su rivista]
Martini, S.; Cattivelli, A.; Conte, A.; Tagliazucchi, D.
abstract
The conventional approach in bioactive peptides discovery, which includes extensive bioassay-guided fractionation and purification processes, is tedious, time-consuming and not always successful. The recently developed bioinformatics-driven in silico approach is rapid and cost-effective; however, it lacks an actual physiological significance. In this study a new integrated peptidomics and in silico method, which combines the advantages of the conventional and in silico approaches by using the pool of peptides identified in a food hydrolysate as the starting point for subsequent application of selected bioinformatics tools, has been developed. Pinto bean protein extract was in vitro digested and peptides were identified by peptidomics. The pool of obtained peptides was screened by in silico analysis and structure–activity relationship modelling. Three peptides (SIPR, SAPI and FVPH) were selected as potential inhibitors of the dipeptidyl-peptidase-IV (DPP-IV) enzyme by this integrated approach. In vitro bioactivity assay showed that all three peptides were able to inhibit DPP-IV with the tetra-peptide SAPI showing the highest activity (IC50 = 57.7 µmol/L). Indeed, a new possible characteristic of peptides (i.e., the presence of an S residue at the N-terminus) able to inhibit DPP-IV was proposed.
2022
- Attenuation of Pseudomonas aeruginosa Virulence by Pomegranate Peel Extract
[Articolo su rivista]
Peppoloni, S.; Colombari, B.; Tagliazucchi, D.; Odorici, A.; Ventrucci, C.; Meto, A.; Blasi, E.
abstract
Pseudomonas aeruginosa is an opportunistic pathogen often responsible for biofilm-associated infections. The high adhesion of bacterial cells onto biotic/abiotic surfaces is followed by production of an extracellular polysaccharidic matrix and formation of a sessile community (the biofilm) by the release of specific quorum-sensing molecules, named autoinducers (AI). When the concentrations of AI reach a threshold level, they induce the expression of many virulence genes, including those involved in biofilm formation, motility, pyoverdine and pyocyanin release. P. aeruginosa embedded into biofilm becomes resistant to both conventional drugs and the host’s immune response. Accordingly, biofilm-associated infections are a major clinical problem underlining the need for new antimicrobial therapies. In this study, we evaluated the effects of pomegranate peel extract (PomeGr) in vitro on P. aeruginosa growth and biofilm formation; moreover, the release of four AI was assessed. The phenolic profile of PomeGr, exposed or not to bacteria, was determined by high-performance liquid chromatography coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS) analysis. We found that bacterial growth, biofilm production and AI release were impaired upon PomeGr treatment. In addition, the PomeGr phenolic content was also markedly hampered following incubation with bacterial cells. In particular, punicalagin, punicalin, pedunculagin, granatin, di-(HHDP-galloyl-hexoside) pentoside and their isomers were highly consumed. Overall, these results provide novel insights on the ability of PomeGr to attenuate P. aeruginosa virulence; moreover, the AI impairment and the observed consumption of specific phenolic compounds may offer new tools in designing innovative therapeutic approaches against bacterial infections.
2022
- Characterization of Cell-Envelope Proteinases from Two Lacticaseibacillus casei Strains Isolated from Parmigiano Reggiano Cheese
[Articolo su rivista]
Solieri, L; Sola, L; Vaccalluzzo, A; Randazzo, Cl; Martini, S /; Tagliazucchi, D
abstract
2022
- Cooking and In Vitro Digestion Modulate the Anti-Diabetic Properties of Red-Skinned Onion and Dark Purple Eggplant Phenolic Compounds
[Articolo su rivista]
Cattivelli, A.; Conte, A.; Martini, S.; Tagliazucchi, D.
abstract
The intake of phenolic-rich foods is an emerging preventive approach for the management of type 2 diabetes, thanks to the ability of these compounds to inhibit some key metabolic enzymes. In this study, the influence of cooking and in vitro digestion on the α-glucosidase, α-amylase, and dipeptidyl-peptidase IV (DPP-IV) inhibitory activity of red-skinned onion (RSO) and dark purple eggplant (DPE) phenolic fractions was assessed. The applied cooking procedures had different influences on the total and individual phenolic compounds gastrointestinal bioaccessibility. DPE in vitro digested phenolic fractions displayed no inhibitory activity versus α-amylase and DPP-IV, whereas the fried DPE sample exhibited moderate inhibitory activity against α-glucosidase. This sample mainly contained hydroxycinnamic acid amides that can be responsible for the observed effect. Contrariwise, raw and cooked in vitro digested RSO phenolic fractions inhibited all three enzymes but with different effectiveness. Fried and raw RSO samples were the most active against them. Statistical analysis pointed out that quercetin mono-hexosides (mainly quercetin-4′-O-hexoside) were responsible for the inhibition of α-glucosidase, whereas quercetin dihexosides (mainly quercetin-3-O-hexoside-4′-O-hexoside) were responsible for the DPP-IV-inhibitory activity of RSO samples. An accurate design of the cooking methods could be essential to maximize the release of individual phenolic compounds and the related bioactivities.
2022
- Fermentation of whey protein concentrate by Streptococcus thermophilus strains releases peptides with biological activities
[Articolo su rivista]
Solieri, L.; Valentini, M.; Cattivelli, A.; Sola, L.; Helal, A.; Martini, S.; Tagliazucchi, D.
abstract
Cheese whey is the principal by-product of cheese making and is perceived as an environmental pollutant. Different technological procedures have been applied to convert whey into value-added products including whey protein concentrate (WPC). In the present study, 2 Lactobacillus helveticus strains and 5 Streptococcus thermophilus strains, isolated from natural whey starter used in Parmigiano Reggiano production, have been evaluated for their capacity to ferment WPC and to release bioactive peptides. WPC fermented with S. thermophilus RBC06 showed the highest antioxidant activity as well as angiotensin-converting enzyme- and dipeptidyl-peptidase-IV-inhibitory activities followed by WPC fermented by S. thermophilus RBC20 and RBN16. An untargeted peptidomics approach was applied to profile the peptides released after WPC fermentation. Several bioactive peptides, mainly angiotensin-converting enzyme inhibitors, were identified and the anti-hypertensive lactotripeptides valine-proline-proline (VPP) and isoleucine-proline-proline (IPP) were quantified. S. thermophilus RBC06 produced the highest amount of VPP (12.80 ± 0.45 mg/L) whereas the highest quantity of IPP (0.56 ± 0.02 mg/L) was detected in WPC fermented with S. thermophilus RBN16. The present study paved the way for the further exploitation of S. thermophilus RBC06 as new bioactive-peptide producer and for the application of WPC hydrolysates as additives for designing whey-based beverages with improved healthy properties.
2022
- In Vitro Bioaccessibility and Antioxidant Activity of Phenolic Compounds in Coffee-Fortified Yogurt
[Articolo su rivista]
Helal, A.; Cattivelli, A.; Conte, A.; Tagliazucchi, D.
abstract
Yogurt is considered one of the most popular and healthy dairy products, and has been exploited as a delivery matrix for phenolic compounds. In this study, coffee powder was added to yogurt as a functional ingredient to produce coffee-fortified yogurt. Total phenolic compounds, antioxidant activity and individual hydroxycinnamic acids have been identified and quantified through mass spectrometry. The results from coffee-fortified yogurt were compared with fermented coffee and plain yogurt. Coffee-fortified yogurt had higher total phenolic content and antioxidant activity compared to plain yogurt. However, the total phenolic compounds found in coffee-fortified yogurt represented only 38.9% of the original content in coffee. Caffeoylquinic acids were the most abundant phenolic compounds in coffee. Fermented coffee and coffee-fortified yogurt displayed lower amounts of individual phenolic compounds with respect to coffee (69.8% and 52.4% of recovery, respectively). A protective effect of the yogurt matrix on total and individual coffee phenolic compounds has been observed after in vitro digestion, resulting in a higher bioaccessibility in comparison with digested fermented coffee. Moreover, coffee-fortified yogurt showed the highest antioxidant values after digestion. These findings clearly demonstrate that coffee-fortified yogurt can be considered a significant source of bioaccessible hydroxycinnamic acids, besides its health benefits as a fermented dairy product.
2022
- Metabolomic and Transcriptional Profiling of Oleuropein Bioconversion into Hydroxytyrosol during Table Olive Fermentation by Lactiplantibacillus plantarum
[Articolo su rivista]
Vaccalluzzo, A; Solieri, L; Tagliazucchi, D; Cattivelli, A; Martini, S; Pino, A; Caggia, C; Randazzo, Cl
abstract
2022
- Pomegranate extract affects fungal biofilm production: consumption of phenolic compounds and alteration of fungal autoinducers release
[Articolo su rivista]
Colombari, B; Tagliazucchi, D; Odorici, A; Pericolini, E; Foltran, I; Pinetti, D; Meto, A; Peppoloni, S; Blasi, E.
abstract
Candida albicans expresses numerous virulence factors that contribute to pathogenesis, including its dimorphic transition and even biofilm formation, through the release of specific quorum sensing molecules, such as the autoinducers (AI) tyrosol and farnesol. In particular, once organized as biofilm, Candida cells can elude conventional antifungal therapies and the host’s immune defenses as well. Accordingly, biofilm-associated infections become a major clinical challenge underlining the need of innovative antimicrobial approaches. The aim of this in vitro study was to assess the effects of pomegranate peel extract (PomeGr) on C. albicans growth and biofilm formation; in addition, the release of tyrosol and farnesol was investigated. The phenolic profile of PomeGr was assessed by high-performance liquid chromatography coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS) analysis before and after exposure to C. albicans. Here, we showed that fungal growth, biofilm formation and AI release were altered by PomeGr treatment. Moreover, the phenolic content of PomeGr was substantially hampered upon exposure to fungal cells; particularly pedunculagin, punicalin, punicalagin, granatin, di-(HHDP-galloyl-hexoside)-pentoside and their isomers as well as ellagic acid–hexoside appeared highly consumed, suggesting their role as bioactive molecules against Candida. Overall, these new insights on the anti-Candida properties of PomeGr and its potential mechanisms of action may represent a relevant step in the design of novel therapeutic approaches against fungal infections.
2022
- Roasting and frying modulate the phenolic profile of dark purple eggplant and differently change the colon microbiota and phenolic metabolites after in vitro digestion and fermentation in a gut model
[Articolo su rivista]
Nissen, L.; Cattivelli, A.; Casciano, F.; Gianotti, A.; Tagliazucchi, D.
abstract
The way of cooking vegetables could differently affect the phenolic profiles of foods and their impact on human colon microbiota. In this work, we investigated the stability and bioaccessibility as well as the impact and fate of dark purple eggplant (DPE) phenolic compounds in the gut microbiota after grilling or frying in comparison to the raw one. After cooking, DPE underwent a gastro-intestinal digestion along with a proximal colon fermentation using the short-term batch model MICODE (multi-unit in vitro colon gut model). During the process, the phenolic compounds profiles (through high-resolution mass spectrometry) and microbiomics (qPCR of 14 core taxa) analyses were performed. Results showed that thermal treatments increased the amount of extractable phenolic compounds as well as their bioaccessibility. The highest gastro-intestinal release was observed in fried DPE (2468.46 ± 13.64 μmol/100 g), followed by grilled DPE (1007. 96 ± 12.84 μmol/100 g) and raw DPE (900.93 ± 10.56 μmol/100 g). Mass spectrometry analysis confirmed that colonic bacteria were able to metabolize DPE phenolic compounds mainly to 3-(3′-hydroxyphenyl)propanoic acid. Furthermore, results indicated that frying was better than grilling in terms of fostering more the growth of beneficial bacterial taxa and limiting that of opportunistic taxa. For example, fried DPE determined an increase in abundance of Bifidobacteriaceae Lactobacillales of 2.66 and 3.80 times. This work is one of the first exploring how cooking methods can affect the phenolic composition of DPE and differently impact on the colon microbiota tuning and modifying the food functionalities.
2021
- An integrated peptidomics and in silico approach to identify novel anti-diabetic peptides in parmigiano-reggiano cheese
[Articolo su rivista]
Martini, S.; Solieri, L.; Cattivelli, A.; Pizzamiglio, V.; Tagliazucchi, D.
abstract
Inhibition of key metabolic enzymes linked to type-2-diabetes (T2D) by food-derived compounds is a preventive emerging strategy in the management of T2D. Here, the impact of Parmigiano- Reggiano (PR) cheese peptide fractions, at four different ripening times (12, 18, 24, and 30 months), on the enzymatic activity of α-glucosidase, α-amylase, and dipeptidyl peptidase-IV (DPPIV) as well as on the formation of fluorescent advanced glycation end-products (fAGEs) was assessed. The PR peptide fractions were able to inhibit the selected enzymes and fAGEs formation. The 12-month-ripening PR sample was the most active against the three enzymes and fAGEs. Mass spectrometry analysis enabled the identification of 415 unique peptides, 54.9% of them common to the four PR samples. Forty-nine previously identified bioactive peptides were found, mostly characterized as angiotensin-converting enzyme-inhibitors. The application of an integrated approach that combined peptidomics, in silico analysis, and a structure–activity relationship led to an efficient selection of 6 peptides with potential DPP-IV and α-glucosidase inhibitory activities. Peptide APFPE was identified as a potent novel DPP-IV inhibitor (IC50 = 49.5 ± 0.5 μmol/L). In addition, the well-known anti-hypertensive tripeptide, IPP, was the only one able to inhibit the three digestive enzymes, highlighting its possible new and pivotal role in diabetes management.
2021
- Antioxidant and Angiotensin I-Converting Enzyme (ACE) Inhibitory Peptides Obtained from Alcalase Protein Hydrolysate Fractions of Hemp (Cannabis sativa L.) Bran
[Articolo su rivista]
Samaei, S. P.; Martini, S.; Tagliazucchi, D.; Gianotti, A.; Babini, E.
abstract
Proteins from hemp bran (HPB), a byproduct of the hemp seed food-processing chain, were chemically extracted, hydrolyzed by Alcalase, and separated by membrane ultrafiltration into four fractions (MW <1, 1-3, 3-5, and >5 kDa). The antioxidant and antihypertensive properties of the initial extract and the fractions were evaluated by in vitro assays for their ability to scavenge radical species, bind with metal ions, reduce ferric ions, and inhibit angiotensin-converting enzyme (ACE) activity. Bioactive peptides were identified by high-resolution mass spectrometry and sequence comparison with BIOPEP and BioPep DB databases. The hydrolysate was strongly antioxidant and ACE-inhibiting; the most bioactive peptides were further concentrated by ultrafiltration. Of the 239 peptides identified, 47 (12 antioxidant and 35 ACE-inhibitory) exhibited structural features correlated with the specific bioactivity. These results highlight the promise of hydrolysate and size-based HPB fractions as natural functional ingredients for the food or pharmaceutical industry.
2021
- Black, green, and pink pepper affect differently lipid oxidation during cooking and in vitro digestion of meat
[Articolo su rivista]
Martini, S.; Cattivelli, A.; Conte, A.; Tagliazucchi, D.
abstract
Lipid oxidation products generated during meat digestion may contribute to the apparent epidemiological link between red meat intake and the risk of cardiovascular diseases and colorectal cancer. The aim of this work was to assess the lipid oxidation inhibitory activity of black, green, and pink pepper during cooking and in vitro digestion of meat. Peppers were characterized for their phenolic profiles by LC-ESI-MS and the antioxidant properties. Pink pepper showed the highest phenolic content and antioxidant activities. Then, the peppers were added to meat either before or after cooking, and the meat was subjected to in vitro digestion. Pink pepper added before cooking was the most effective, with an inhibition of 80% and 72% in lipid hydroperoxides and TBA-RS formation after digestion, respectively. These findings suggest that peppers, particularly pink pepper, can be used to minimize lipid oxidation in the gastro-intestinal tract and for the design of healthy dietary patterns.
2021
- Characterization of yeasts isolated from parmigiano reggiano cheese natural whey starter: From spoilage agents to potential cell factories for whey valorization
[Articolo su rivista]
Martini, S.; Bonazzi, M.; Malorgio, I.; Pizzamiglio, V.; Tagliazucchi, D.; Solieri, L.
abstract
Whey is the main byproduct of the dairy industry and contains sugars (lactose) and proteins (especially serum proteins and, at lesser extent, residual caseins), which can be valorized by the fermentative action of yeasts. In the present study, we characterized the spoilage yeast population inhabiting natural whey starter (NWS), the undefined starter culture of thermophilic lactic acid bacteria used in Parmigiano Reggiano (PR) cheesemaking, and evaluated thermotolerance, mating type, and the aptitude to produce ethanol and bioactive peptides from whey lactose and proteins, respectively, in a selected pool of strains. PCR-RFLP assay of ribosomal ITS regions and phylogenetic analysis of 26S rDNA D1/D2 domains showed that PR NWS yeast population consists of the well-documented Kluyveromyces marxianus, as well as of other species (Saccharomyces cerevisiae, Wickerhamiella pararugosa, and Torulaspora delbrueckii), with multiple biotypes scored within each species as demonstrated by (GTG)5-based MSP-PCR. Haploid and diploid K. marxianus strains were identified through MAT genotyping, while thermotolerance assay allowed the selection of strains suitable to grow up to 48◦C. In whey fermentation trials, one thermotolerant strain was suitable to release ethanol with a fermentation efficiency of 86.5%, while another candidate was able to produce the highest amounts of both ethanol and bioactive peptides with potentially anti-hypertensive function. The present work demonstrated that PR NWS is a reservoir of ethanol and bioactive peptides producer yeasts, which can be exploited to valorize whey, in agreement with the principles of circularity and sustainability.
2021
- Domestic cooking methods affect the stability and bioaccessibility of dark purple eggplant (Solanum melongena) phenolic compounds
[Articolo su rivista]
Martini, S.; Conte, A.; Cattivelli, A.; Tagliazucchi, D.
abstract
Eggplant is an important component of the Mediterranean Diet, which becomes edible after cooking. This study determined the fate of dark purple eggplant phenolic compounds after baking, boiling, frying, grilling and digestion. Thirty-seven phenolic compounds were identified and quantified in raw eggplant. Frying determined a 74% increase in total hydroxycinnamic acids whereas a decrease was observed after boiling (27%), grilling (51%), and baking (60%). After digestion, 45%, 33% and 22% of total phenolic compounds resulted bioaccessible in baked, grilled and fried dark purple eggplant. Fried eggplant displayed the highest amount of phenolic compounds (751.46 mg/100 g) after digestion. The cooking methods differently affected the release of individual phenolic compounds. Baking and grilling resulted in higher amount of bioaccessible caffeoylquinic acids whereas frying in di-caffeoylquinic acids and hydroxycinnamic acid-amides. A careful design of the cooking method may be pivotal to modulate the release of specific phenolic compounds.
2021
- Influence of cooking methods on onion phenolic compounds bioaccessibility
[Articolo su rivista]
Cattivelli, A.; Conte, A.; Martini, S.; Tagliazucchi, D.
abstract
The impact of domestic cooking (baking, boiling, frying and grilling) and in vitro digestion on the stability and release of phenolic compounds from yellow-skinned (YSO) and red-skinned onions (RSO) have been evaluated. The mass spectrometry identification pointed out flavonols as the most representative phenolic class, led by quercetin-derivatives. RSO contained almost the double amount of phenolic compounds respect to YSO (50.12 and 27.42 mg/100 g, respectively). Baking, grilling and primarily frying resulted in an increased amount of total phenolic compounds, especially quercetin-derivatives, in both the onion varieties. Some treatments promoted the degradation of quercetin-3-O-hexoside-4′-O-hexoside, the main compound present in both the onion varieties, leading to the occurrence of quercetin-4′-O-hexoside and protocatechuic acid-4-O-hexoside. After in vitro digestion, the bioaccessibility index for total phenolic compounds ranged between 42.6% and 65.5% in grilled and baked YSO, respectively, and between 39.8% and 80.2% in boiled and baked RSO, respectively. Baking contributed to the highest amount of bioaccessible phenolic compounds for both the onion varieties after in vitro digestion. An in-depth design of the cooking process may be of paramount importance in modulating the gastro-intestinal release of onion phenolic compounds.
2021
- Peptidomics: new trends in food science
[Articolo su rivista]
Martini, S.; Solieri, L.; Tagliazucchi, D.
abstract
In recent years, peptidomics is gaining ever-growing relevance in food science. The emerging field of food peptidomics is defined as the whole pool of peptides existing in food products or generated during food processing, storage or digestion. The recent advancement in high-resolution mass spectrometry techniques provides a more detailed picture, although not yet exhaustive, of the peptides present or derived from food. Food peptidomics techniques have been successfully applied in understanding food protein digestion, in the study of the microbial contribution to food protein hydrolysis, in the identification of food-derived bioactive peptides and peptide biomarkers. Ad-hoc efforts are still needed to improve the identification of short peptides and the analysis of the large data set generated.
2020
- Cultivable non-starter lactobacilli from ripened Parmigiano Reggiano cheeses with different salt content and their potential to release anti-hypertensive peptides
[Articolo su rivista]
Tagliazucchi, Davide; Baldaccini, Andrea; Martini, Serena; Bianchi, Aldo; Pizzamiglio, Valentina; Solieri, Lisa
abstract
The impact of salt and fat intake on human health drives the consumer's attention towards dairy food with reduced salt and fat contents. How changes in salt and fat content modulate dairy LAB population and the associated proteolytic activities have been poorly studied. Here, non-starter LAB populations from 12 Parmigiano Reggiano (PR) cheeses (12-month ripened), clustered in low salt and fat content (LL-PR) and high salt and fat content (HH-PR) groups, were investigated and identified at specie-level with molecular assays. Lactobacillus rhamnosus was dominant in HH-PR samples, whereas Lactobacillus paracasei in LL-PR samples. (GTG)5 rep-PCR analysis discriminated 11 and 12 biotypes for L. rhamnosus and L. paracasei isolates, respectively. Screening for proteolytic activity identified L. rhamnosus strains more proteolytic than L. paracasei, and, within L. rhamnosus species, HH-PR strains were generally more proteolytic than LL-PR strains. Two L. rhamnosus representatives, namely strain 0503 from LL-PR and strain 2006 from HH-PR, were functionally characterized in cow milk fermentation assay. HH-PR strain 2006 overcame LL-PR strain 0503 in acidification performance, leading to a fermented milk with higher angiotensin I-converting enzyme inhibitory and antioxidant activities. L. rhamnosus 2006 was more prone to release VPP, while L. rhamnosus 0503 released higher amount of IPP. This study provides evidences that salt/fat content affects NSLAB cultivable fraction and the associated proteolytic ability resulting in a complex occurrence of bioactive peptides featuring health-promoting properties.
2020
- Effect of ripening and in vitro digestion on the evolution and fate of bioactive peptides in Parmigiano-Reggiano cheese
[Articolo su rivista]
Martini, Serena; Conte, Angela; Tagliazucchi, Davide
abstract
The influence of ripening and in vitro digestion on the peptidomic profile of Parmigiano-Reggiano (PR) cheeses was investigated. Ripening and in vitro digestion thoroughly modified the peptidomic profile of the three cheeses. Twenty-six bioactive peptides were identified in undigested PR. Some peptides were degraded and others released during ripening. After digestion, 52 bioactive peptides were identified. Semi-quantitative data suggested that bioactive peptides released after digestion can be clustered in 5 groups according to the ripening time. VPP and IPP peptide levels in undigested samples were in the range of 4.52–11.34 and 0.66–4.24 mg kg−1, with the highest amounts found in 18-month ripened PR. YPFPGPI peptide was absent in undigested PRs but was released after digestion, especially in the 12-month-old sample (20.18 mg kg−1). The present study suggests possible differences in bioactive peptide levels after digestion as a function of the duration of ripening of PR cheese.
2020
- Functional, nutritional, antioxidant, sensory properties and comparative peptidomic profile of faba bean (Vicia faba, L.) seed protein hydrolysates and fortified apple juice
[Articolo su rivista]
Samaei, S. P.; Ghorbani, M.; Tagliazucchi, D.; Martini, S.; Gotti, R.; Themelis, T.; Tesini, F.; Gianotti, A.; Gallina Toschi, T.; Babini, E.
abstract
Enzymatic hydrolysis of plant-derived proteins can improve their quality by offering opportunities for food applications. In this study, three proteolytic enzymes (pepsin, trypsin, Alcalase®) were used, alone or combined, to produce faba bean protein hydrolysates (PHs). Their functional, nutritional and antioxidant properties were evaluated, and the peptidomic profile was assessed by LC–MS/MS. Hydrolysis improved solubility of faba proteins at acidic and neutral pH, and their antioxidant properties. Peptidomic analysis identified 2031 peptides in the different PHs. Among them, 9 showed 100% homology with previously known antioxidant peptides and several others had antioxidant motifs in their sequences. Sensory data analysis showed that after addition of PHs to apple juice, no significant differences were perceived between control and some of the PHs. This study demonstrates that enzymatic hydrolysis enhances the functional and antioxidant properties of faba bean proteins. Specifically, hydrolysates can be used as functional food ingredients to produce fortified beverages.
2020
- Influence of linseed and antioxidant-rich diets in pig nutrition on lipidoxidation during cooking and in vitro digestion of pork
[Articolo su rivista]
Martini, S.; Tagliazucchi, D.; Minelli, G.; Lo Fiego, D. P.
abstract
Enrichment of pig diets with polyunsaturated fatty acids (PUFA) is considered an emerging strategy to increasetheir intake in the human diet. However, PUFA are particularly vulnerable to oxidative reactions leading to thegeneration of toxic compounds. The aim of this study was to evaluate the effect of supplementation of pig dietswith extruded linseed (L), either or not in combination with synthetic antioxidants (E, tocopheryl-acetate andselenium) or natural extracts (P, grape-skin and oregano), and basal diet (C, without linseed) on the oxidativestability in raw, grilled andin vitrodigested pork. The diet supplementation with antioxidant-rich ingredientsresulted in the accumulation of specific metabolites in meat. Actually, 11 different phenolic- and 6 tocopherol-derived metabolites were identified by UHPLC/HR-MS. These metabolites were potentially correlated with thereduction in the oxidative phenomena occurring during meat cooking and digestion. Specifically, 16% and 35%reduction in the amounts of lipid hydroperoxides and TBA-RS were assessed after cooking of meat from P diet,respect to the L diet. Diet supplementations withα-tocopheryl acetate and selenium reduced the oxidativereactions only during meat cooking. A significant reduction was attended at the end ofin vitrodigestion, showingabout 24% and 34% hydroperoxides and TBA-RS concentration reductions, respectively, in P diet samples re-spect to the L ones. Thus, our study suggests that the appearance of phenolic metabolites in meat could beassociated to a reduction in the oxidative phenomena during meat cooking and digestion.
2020
- Mediterranean diet vegetable foods protect meat lipids from oxidation during in vitro gastro-intestinal digestion
[Articolo su rivista]
Martini, S.; Conte, A.; Bottazzi, S.; Tagliazucchi, D.
abstract
Meat lipids oxidation during digestion gives rise to a post-prandial oxidative stress condition, which negatively affects human health. Mediterranean Diet vegetables contain high amount of phenolic compounds, which potentially may reduce the oxidative phenomena during digestion. In vitro co-digestion of turkey meat with a typical Mediterranean Diet salad containing tomato, onion, black olives, extra-virgin olive oil (EVOO) and basil, dose-dependently reduced lipid peroxidation. Onion and EVOO were more effective in limiting oxidation than the other foods, resulting in negligible concentrations of lipid hydroperoxides after digestion. Specific phenolic classes dominated the phenolic profile of the different foods, such as flavonols and anthocyanins in onion, phenolic acids in tomato and basil, and tyrosol-derivatives in black olives and EVOO. The correlation between lipid peroxidation inhibition, phenolic constituents and antioxidant properties was evaluated by principal component analysis (PCA). Flavonols and anthocyanin were the major contributors to the bioactive response of vegetable foods.
2020
- Microbial fermentation of industrial rice-starch byproduct as valuable source of peptide fractions with health-related activity
[Articolo su rivista]
Babini, E.; Taneyo-Saa, D. L.; Tassoni, A.; Ferri, M.; Kraft, A.; Gran-Heedfeld, J.; Bretz, K.; Roda, A.; Michelini, E.; Calabretta, M. M.; Guillon, F.; Tagliazucchi, D.; Martini, S.; Nissen, L.; Gianotti, A.
abstract
The rice-starch processing industry produces large amounts of a protein-rich byproducts during the conversion of broken rice to powder and crystal starch. Given the poor protein solubility, this material is currently discarded or used as animal feed. To fully exploit rice’s nutritional properties and reduce this waste, a biotechnological approach was adopted, inducing fermentation with selected microorganisms capable of converting the substrate into peptide fractions with health-related bioactivity. Lactic acid bacteria were preferred to other microorganisms for their safety, efficient proteolytic system, and adaptability to different environments. Peptide fractions with different molecular weight ranges were recovered from the fermented substrate by means of cross-flow membrane filtration. The fractions displayed in vitro antioxidant, antihypertensive, and anti-tyrosinase activities as well as cell-based anti-inflammatory and anti-aging effects. In the future, the peptide fractions isolated from this rice byproduct could be directly exploited as health-promoting functional foods, dietary supplements, and pharmaceutical preparations. The suggested biotechnological process harnessing microbial bioconversion may represent a potential solution for many different protein-containing substrates currently treated as byproducts (or worse, waste) by the food industry.
2020
- Peptide profiling and biological activities of 12- month ripened parmigiano reggiano cheese
[Articolo su rivista]
Tagliazucchi, D.; Solieri, L.; Baldaccini, A.; Martini, S.; Bianchi, A.; Pizzamiglio, V.
abstract
Proteolysis degree, biological activities, and water-soluble peptide patterns were evaluated in 12 month-ripened Parmigiano Reggiano (PR) cheeses collected in different dairy farms and showing different salt and fat content. Samples classified in high-salt and high-fat group (HH) generally showed lower proteolysis degree than samples having low-salt and low-fat content (LL). This positive correlation between salt/fat reduction and proteolysis was also confirmed by the analysis of biological activities, as the LL group showed higher average values of angiotensin-converting enzyme (ACE)-inhibitory and antioxidant activities. UHPLC/HR-MS allowed the identification of 805 unique peptides: LL and HH groups shared 59.3% of these peptides, while 20.9% and 19.9% were LL and HH specific, respectively. Frequency analysis of peptides identified a core of 183 peptides typical of 12-month ripened PR cheeses (corresponding to the 22.7% of total peptides), but no significant differences were detected in peptide patterns between LL and HH groups. Forty bioactive peptides, including 18 ACE-inhibitors and 12 anti-microbial peptides, were identified, of which 25 firstly found in PR cheese. Globally, this work contributed to unraveling the potentially healthy benefits of peptides fraction in PR cheese and provided prior evidence that PR with reduced at/salt content showed the highest antihypertensive and antioxidant activities.
2019
- Activity of extracts from three tropical plants towards fungi pathogenic to tomato (Solanum lycopersicum)
[Articolo su rivista]
Mekam, Pascal Noel; Martini, Serena; Nguefack, Julienne; Tagliazucchi, Davide; Mangoumou, Ghislaine Ndonkeu; Stefani, Emilio
abstract
Antifungal properties were assessed of water and ethanol extracts from the pan-tropical plants Oxalis barrelieri L., Stachytarpheta cayennensis L., and Euphorbia hirta L. against Fusarium oxysporum f. sp. vasinfectum, Alternaria solani Sorauer, and Rhizoctonia solani Kuhn. The plant extracts inhibited fungal growth in vitro at 1.25-20 mg mL-1, and the degrees of inhibition increased in a dose-dependent manner. Ethanol extracts from the plants inhibited fungal growth by 80-100%, while water extracts showed less antifungal activity, with maximum growth inhibition of 62%. Growth inhibition from ethanol extracts was two- to three-fold greater than for water extracts at equivalent concentrations. Antifungal activity of the extracts varied with their content and composition of phenolics, flavonoids, tannins, and alkaloids. In greenhouse experiments, spraying tomato plants (Solanum lycopersicum L.) with ethanol extract from E. hirta at 2.5 mg mL-1 did not cause phytotoxicity, and increased plant size, when compared to untreated plants. Spraying E. hirta ethanol extract on tomato plants infected by R. solani reduced disease severity up to 80%, when compared to non-sprayed plants. These results demonstrate potential of leaf extracts from E. hirta, O. barrelieri, and S. cayennensis as biofungicides for the control of R. solani, A. solani, and F. oxysporum, which are among the most important causal agents of tomato diseases.
2019
- Antiproliferative Activity and Cell Metabolism of Hydroxycinnamic Acids in Human Colon Adenocarcinoma Cell Lines
[Articolo su rivista]
Martini, Serena; Conte, Angela; Tagliazucchi, Davide
abstract
In this study, we investigated the antiproliferative activity and the stability and metabolic fate of the main dietary hydroxycinnamates, using two colonic adenocarcinoma cell models (Caco-2 and SW480). Dihydrocaffeic and dihydroferulic acids were the most effective against cell proliferation in both cell lines with IC50 values of 71.7 ± 1.1 and 83.1 ± 1.1 μmol/L, respectively (P < 0.05) in Caco-2. At 200 μmol/L, caffeic and ferulic acids inhibited SW480 proliferation by 40.8 ± 1.6 and 59.9 ± 1.3%, respectively. Hydroxycinnamic acids with a catechol-type structure were degraded in Caco-2 cell medium, resulting in the production of H2O2. Intracellular Caco-2 UDP-glucuronosyltransferases and catechol-O-methyltransferases were able to form glucuronide and methyl conjugates. However, only the sulfate conjugates were detected after incubation with SW480. In addition, simple hydroxycinnamates were released from quinic and aspartic conjugates. The remarkable effect of dihydrocaffeic and dihydroferulic acids against cell proliferation is of paramount importance, since these compounds are the main metabolites detectable at the colonic level.
2019
- Bioactivity and cell metabolism of in vitro digested sweet cherry (Prunus avium) phenolic compounds
[Articolo su rivista]
Martini, Serena; Conte, Angela; Tagliazucchi, Davide
abstract
In this study, the bioaccessibility of phenolic compounds after in vitro gastrointestinal digestion of two cherry cultivars was assessed. The phenolic profile was modified during in vitro digestion, with a considerable decrease of total and individual phenolic compounds. Hydroxycinnamic acids and especially coumaroylquinic acids showed the highest bioaccessibility. Isomerisation of caffeoylquinic and coumaroylquinic acids was observed after in vitro digestion. Modification of the phenolic profile after digestion resulted in an increased or decreased scavenging activity depending on the assay. In vitro digested phenolic-rich fractions also showed antiproliferative activity against SW480 but no effect against Caco-2 cell lines. Both Caco-2 and SW480 cell lines were able to metabolise cherry phenolic compounds with remarkable differences. An accumulation of glycosylated flavonols was observed in SW480 medium. In conclusion, phenolic compounds from cherries and especially hydroxycinnamic acids were efficiently released and remained bioaccessible after in vitro digestion, resulting in antioxidant and antiproliferative activities.
2019
- Bioprospecting for Bioactive Peptide Production by Lactic Acid Bacteria Isolated from Fermented Dairy Food
[Articolo su rivista]
Tagliazucchi, Davide; Martini, Serena; Solieri, Lisa
abstract
With rapidly ageing populations, the world is experiencing unsustainable healthcare from chronic diseases such as metabolic, cardiovascular, neurodegenerative, and cancer disorders. Healthy diet and lifestyle might contribute to prevent these diseases and potentially enhance health outcomes in patients during and after therapy. Fermented dairy foods (FDFs) found their origin concurrently with human civilization for increasing milk shelf-life and enhancing sensorial attributes. Although the probiotic concept has been developed more recently, FDFs, such as milks and yoghurt, have been unconsciously associated with health-promoting effects since ancient times. These health benefits rely not only on the occurrence of fermentation-associated live microbes (mainly lactic acid bacteria; LAB), but also on the pro-health molecules (PHMs) mostly derived from microbial conversion of food compounds. Therefore, there is a renaissance of interest toward traditional fermented food as a reservoir of novel microbes producing PHMs, and “hyperfoods” can be tailored to deliver these healthy molecules to humans. In FDFs, the main PHMs are bioactive peptides (BPs) released from milk proteins by microbial proteolysis. BPs display a pattern of biofunctions such as anti-hypertensive, antioxidant, immuno-modulatory, and anti-microbial activities. Here, we summarized the BPs most frequently encountered in dairy food and their biological activities; we reviewed the main studies exploring the potential of dairy microbiota to release BPs; and delineated the main effectors of the proteolytic LAB systems responsible for BPs release
2019
- Comparative peptidomic profile and bioactivities of cooked beef, pork, chicken and turkey meat after in vitro gastro-intestinal digestion
[Articolo su rivista]
Martini, Serena; Conte, Angela; Tagliazucchi, Davide
abstract
This study was designed to investigate the potential contribution of bioactive peptides to the biological activities related to the consumption of pork, beef, chicken and turkey meat following in vitro gastro-intestinal digestion. After extraction of the peptidic fractions from digested samples, the bioactivities were evaluated by in vitro antioxidant activity as well as angiotensin-converting enzyme (ACE) and dipeptidyl peptidase-IV (DPP-IV) inhibition assays. Pork and turkey meat appeared to be the best sources of antioxidant peptides. Pork was found to be the best source of DPP-IV-inhibitory peptides whereas chicken meat supplied peptides with the highest ACE-inhibitory activity. The comprehensive analysis of the peptidomic profile of digested samples was performed by nano-LC-ESI-QTOF MS/MS analysis. A total of 217, 214, 257 and 248 peptides were identified in digested pork, beef, chicken and turkey meat, respectively. Chicken and turkey meat showed the highest similarity in peptide sequences with 202 common peptides. Sixty-two peptides matched with sequences with previously demonstrated biological activity. In particular, 35 peptides showed ACE-inhibitory activity and 23 DPP-IV inhibitory activity. Twenty-two bioactive peptides were commonly released from the different types of meat. The relative amount of identified bioactive peptides were positively correlated to the biological activities of the different digested meats.
Biological significance: The present study describes for the first time a comprehensive peptide profile of four types of meat after in vitro gastro-intestinal digestion. The peptide inventory was used to identify 62 bioactive peptides with ACE- and DPPIV-inhibitory and antioxidant activities. The bioactivity analysis revealed interesting and significant differences between the studied meats. The originality of this work lay in the description of intrinsic differences in physiological functions after the ingestion of meat proteins from different species. In a context in which the current research scene relates meat consumption to the onset of chronic pathologies, this peptide profiling and bioactivity analysis shed light on the possible health benefits of peptides released from meat proteins. In fact, this paper represents a sort of detailed peptide list that may help to predict which peptides could be generated after meat intake and detectable at gastro-intestinal level. It also provides a thorough investigation of novel biological activities associated to meat protein hydrolysates, giving a new positive aspect to meat consumption.
2019
- Phenolic compounds profile of water and ethanol extracts of Euphorbia hirta L. leaves showing antioxidant and antifungal properties
[Articolo su rivista]
Mekam, Pascal Noel; Martini, Serena; Nguefack, Julienne; Tagliazucchi, Davide; Stefani, Emilio
abstract
The bioactive chemical constituents of water and ethanol extracts of Euphorbia hirta L. leaves have been identified and quantified using an un-targeted mass spectrometric approach. The study allowed the tentative identification of 123 individual phenolic compounds and 18 non-phenolic phytochemicals, most of them described in Euphorbia hirta L. leaves for the first time. Gallotannins, hydroxybenzoic and hydroxycinnamic acids were the most abundant phenolic classes in Euphorbia hirta L. leaves, representing together the 71.5% (26.3%, 25.2% and 20%, respectively) of the total amount of identified phenolics. The main phenolic compounds detected were tri-O-galloyl-glucose isomers, feruloyl-coniferin, tetra-O-galloyl-glucose isomers, di-O-galloyl-glucose isomers, ethyl-gallic acid, protocatechuic acid-O-pentoside-O-hexoside, 5-O-caffeoyl-quinic acid trans isomer and digalloyl-quinic acid. Feruloyl-coniferin was found to be approximately six times more concentrated in the ethanol extract with respect to the water extract. The ethanol extract exhibited higher ABTS (1338.3 ± 85.3 and 802.3 ± 91.0 μmol ascorbic acid equivalent/gram of dry extract, respectively) and superoxide anion (2014.6 ± 78.6 and 1528.0 ± 111.7 μmol ascorbic acid equivalent/gram of dry extract, respectively) scavenging abilities than the water extract. Additionally, the ethanol extract also showed a remarkable anti-fungal effect against Fusarium oxysporum f. sp. vasinfectum, Alternaria solani and Rhizoctonia solani. This study provides new information about Euphorbia hirta L., offering reasons to promote this plant species as rich source of phenolics and an excellent source of antifungal molecules that might have a prospective use in controlling fungal diseases of vegetable crops.
2019
- Protocatechuic and 3,4-Dihydroxyphenylacetic Acids Inhibit Protein Glycation by Binding Lysine through a Metal-Catalyzed Oxidative Mechanism
[Articolo su rivista]
Tagliazucchi, Davide; Martini, Serena; Conte, Angela
abstract
The mechanism of inhibition of advanced glycation end product (AGE) formation by protocatechuic acid and 3,4-dihydroxyphenylacetic acid (DHPA) has been studied using a widespread applied in vitro model system composed of bovine serum albumin (BSA) and supraphysiological glucose concentrations. Protocatechuic acid and DHPA inhibited the formation of Amadori compounds, fluorescent AGEs (IC50 = 62.1 ± 1.4 and 155.4 ± 1.1 μmol/L, respectively), and Nε-(carboxymethyl)lysine (IC50 = 535.3 ± 1.1 and 751.2 ± 1.0 μmol/L, respectively). BSA was pretreated with the two phenolic acids, and the formation of BSA–phenolic acid adducts was estimated by nanoflow liquid chromatography–electrospray ionization–quadrupole time-of-flight mass spectrometry. Results showed that the tested phenolic acids bound key sites of glycation in BSA through a metal-catalyzed oxidative mechanism. The antiglycative activity mechanism involved the formation of BSA–phenolic acid adducts, and it is unlikely that this occurs in vivo. These results raise the problem to design in vitro models closer to physiological conditions to reach biologically sound conclusions.
2018
- Bioaccessibility, bioactivity and cell metabolism of dark chocolate phenolic compounds after in vitro gastro-intestinal digestion
[Articolo su rivista]
Martini, Serena; Conte, Angela; Tagliazucchi, Davide
abstract
The bioaccessibility of phenolic compounds after in vitro gastro-intestinal digestion of dark chocolate, dark chocolate enriched with Sakura green tea and dark chocolate enriched with turmeric powder was studied. The phenolic profile, assessed by accurate mass spectrometry analysis, was modified during in vitro gastro-intestinal digestion, with a considerable decrease of total and individual phenolic compounds. Phenolic acids showed the highest bioaccessibility with hydroxycinnamic acids displaying higher bioaccessibility (from 41.2% to 45.1%) respect to hydroxybenzoic acids (from 28.1% to 43.5%). Isomerisation of caffeoyl-quinic acids and galloyl-quinic acids as well as dimerization of (epi)gallocatechin were also observed after in vitro gastro-intestinal digestion. Antioxidant activity increased after the gastric step and rose further at the end of the digestion. Furthermore, in vitro digested phenolic-rich fractions showed anti-proliferative activity against two models of human colon adenocarcinoma cell lines. Cell metabolism of digested phenolic compounds resulted in the accumulation of coumaric and ferulic acids in the cell media.
2018
- Biological activities and peptidomic profile of in vitro-digested cow, camel, goat and sheep milk
[Articolo su rivista]
Tagliazucchi, Davide; Martini, Serena; Shamsia, Sherif; MOHAMED IBRAHIM HELAL, Ahmed; Conte, Angela
abstract
In vitro digestibility, selected biological activities and digested products of proteins from skimmed cow, camel, goat and sheep milk were compared. The harmonised in vitro INFOGEST digestion model and mass spectrometry were combined to identify peptides. Goat milk had the highest digestibility, sheep milk the highest angiotensin-converting enzyme (ACE)-inhibitory activity after digestion and cow milk the highest dipeptidyl peptidase-IV (DPP-IV)-inhibitory activity. After in vitro digestion of the milk samples, 522 peptides were identified. Goat and sheep milk showed the highest peptide sequence similarity with 151 common peptides. Thirteen, forty-three and twenty peptides with previously demonstrated antioxidant, ACE-inhibitory and DPP-IV-inhibitory activities, respectively, were found. Nineteen bioactive peptides in common were released from the different milk types. Despite limitations related to analysis of one milk sample for each species, possible differences in physiological functions after ingestion of milk from different species are suggested; this requires confirmation by in vivo testing.
2018
- Comprehensive evaluation of phenolic profile in dark chocolate and dark chocolate enriched with Sakura green tea leaves or turmeric powder
[Articolo su rivista]
Martini, Serena; Conte, Angela; Tagliazucchi, Davide
abstract
Recently, a huge number of studies have confirmed the important role of chocolate polyphenols in human health, underlining its beneficial effects especially in the treatment of cardiovascular diseases. However, a thorough evaluation of chocolate phenolic profile is still lacking. This study aimed at a comprehensive characterization of dark chocolate phenolic profile, using non-targeted mass spectrometry identification. This approach allowed a tentative identification of 158 individual phenolic compounds: 67 were newly detected in dark chocolate, among these 38 were observed for the first time in chocolate as well as in cocoa beans or products. Ellagitannins, which have never been reported in cocoa or chocolate, represented about the 10% of the phenolic profile of dark chocolate. The enrichment of dark chocolate with Sakura green tea leaves or turmeric powder influenced and modified the phenolic profile, resulting in a phenolic concentration increase. In this way, this functional chocolate might maximize the beneficial effect of chocolate consumption, combining the positive health effects of chocolate, turmeric and green tea and, at the same time, reducing the amount of sugars and calories introduced with chocolate.
2018
- Impact of in-vitro gastro-pancreatic digestion on polyphenols and cinnamaldehyde bioaccessibility and antioxidant activity in stirred cinnamon-fortified yogurt
[Articolo su rivista]
MOHAMED IBRAHIM HELAL, Ahmed; Tagliazucchi, Davide
abstract
In this study, cinnamon powder was supplemented into yogurt as a functional ingredient. The total phenolic
compounds, individual phytochemicals and radical scavenging activity of the yogurts were measured and
compared with a cinnamon water extract treated in the same way as the fortified yogurt. Cinnamon-fortified
yogurt displayed higher total phenolic content (P < 0.05) and higher radical scavenging activity (P < 0.05)
compared to plain yogurt. Phenolic acids, flavonols and cinnamaldehyde were identified in the cinnamon-fortified yogurt. Results showed that only the 34.7% of the total phenolic compounds present in the cinnamon
water extract were found in the cinnamon-fortified yogurt, the remaining being bound to milk proteins. A low
recovery was also found for the individual phytochemicals. However, in-vitro digestion of the cinnamon-fortified
yogurt resulted in the release of phenolic compounds from milk proteins so that at the end of the digestion the
amount of phenolic compounds recovered in the cinnamon-fortified yogurt was higher than that found in the
digested cinnamon water extract (P < 0.05). These results clearly showed that yogurt matrix enhance the
gastro-intestinal stability and the bioaccessibility of cinnamon polyphenols. Cinnamon-fortified yogurt can be
considered an important source of dietary bioaccessible polyphenols.
2018
- Peptidomic study of casein proteolysis in bovine milk by Lactobacillus casei PRA205 and Lactobacillus rhamnosus PRA331
[Articolo su rivista]
Solieri, Lisa; DE VERO, Luciana; Tagliazucchi, Davide
abstract
Lactobacilli contain different cell envelope proteinases (CEPs) responsible for the hydrolysis of caseins and the release of various bioactive peptides. In this work, we explored the CEP activity of Lactobacillus casei PRA205 and Lactobacillus rhamnosus PRA331 whole cells towards β-, αS1-, κ- and αS2-caseins in bovine milk. Mass spectrometry analysis of fermented milk hydrolysates identified a total of 331 peptides, which were mainly derived from β-caseins (59.0 and 60.1% for PRA205 and PRA331, respectively). Analysis of αS1-casein (f1–23) cleavage site specificity congruently supports that Lb. casei PRA205 and Lb. rhamnosus PRA331 exhibited a mixed-type CEPI/III activity. PRA205 and PRA331 CEPs also showed cleavage site specificity toward β-casein. These CEPs cleaved the peptide bond preferentially when hydrophobic or negatively charged amino acids were present. 13.5% and 13.7% of peptides released by Lb. casei PRA205 and Lb. rhamnosus PRA331 CEPs were found to have 100% homology with previously identified bioactive peptides.
2018
- The paradoxical effect of extra-virgin olive oil on oxidative phenomena during in vitro co-digestion with meat
[Articolo su rivista]
Martini, Serena; Cavalchi, Martina; Conte, Angela; Tagliazucchi, Davide
abstract
Extra-virgin olive oil is an integral part of the Mediterranean diet and its consumption has been associated with a reduction risk of chronic diseases. Here we tested the potential of extra-virgin olive oil to limit the oxidative phenomena during in vitro gastro-intestinal co-digestion with turkey breast meat. The extra-virgin olive oil was particularly rich in oleuropein aglycone isomers, which represented the 66.8% of total phenolic determined with MS/MS experiments. Meals supplemented with extra-virgin olive oil equivocally affected lipid peroxidation. At low concentration (2.5% respect to meat), a significant inhibition of lipid oxidation was observed, whereas lipid peroxidation was greatly enhanced when the amount of extra-virgin olive oil was increased in the gastro-intestinal system. The inhibitory effect observed at 2.5% extra-virgin olive oil was due to the antioxidant properties of extra-virgin olive oil phenolic compounds. At high concentration, extra-virgin olive oil phenolic compounds (especially hydroxytyrosol-derivative) behaved as pro-oxidants increasing the generation of lipid hydroperoxides from meat. At the same time, the presence in the digestive system of catalysers from meat induced the peroxidation of extra-virgin olive oil fatty acids, which was further intensified by the pro-oxidant activity of extra-virgin olive oil phenolic compounds. Our study underlined the importance of the timing and amount of consumption of extra-virgin olive oil as well as its phenolic composition in limiting the peroxidative phenomena on meat lipids during digestion.
2017
- ANTIFUNGAL ACTIVITY OF AQUEOUS AND ETHANOL EXTRACTS OF TROPICAL PLANTS AGAINST TOMATO FUNGAL PATHOGENES.
[Abstract in Atti di Convegno]
Mekam, Pascal Noel; Martini, Serena; Tagliazucchi, Davide; Nguefack, Julienne; Stefani, Emilio
abstract
Several plant extracts may have a strong antifungal activity
that can be exploited in the management of fungal diseases as an
alternative to synthetic fungicides, the abuse of which may have
consequences on the environment and health. Ethanol extracts
(EE) and water extracts (WE) of three tropical plants, Stachytarpheta
cayennensis (Verbenaceae), Oxalis barrelieri (Oxalidaceae) and
Euphorbia hirta (Euphorbiaceae) were obtained and screened for
their antifungal activity against three major phytopathogenic fungi
of tomato: Fusarium sp., Alternaria sp. and Colletotrichum sp. The
phytopathogenic fungi used across the experiments were isolated in
Cameroon on severely affected tomatoes and their molecular identification
and characterization is ongoing. The antifungal activity was
checked in vitro on different media supplemented with different
concentrations of the extracts. O. barrelieri EE, at the concentration
of 25 mg/ml, inhibited the mycelium growth of Fusarium sp. by
26.8% and remarkably modified its morphology. WE and EE of the
same O. barrelieri inhibited the mycelium growth of Colletotrichum sp. by 43.7% and 50.3% respectively, but showed no inhibition on
Alternaria sp. The phytochemical analysis of these plant extracts
revealed that EE of S. cayennensis was the richest in polyphenols
and flavonoids. EE from O. barrelieri was particularly rich in alkaloids.
The inhibitory effects on the phytopathogenic fungi were possibly
related to the amount of polyphenols and alkaloids obtained
through the extraction. Field experiments are being conducted on
tomato to confirm the action of such extracts in vivo. These findings
may contribute to develop new biofungicides to protect tomato
from some fungal pathogens.
2017
- Angiotensin-converting enzyme inhibitory peptides from goats' milk released by in vitro gastro-intestinal digestion
[Articolo su rivista]
Tagliazucchi, Davide; Shamsia, Sherif; Helal, Ahmed; Conte, Angela
abstract
The aim of this study was to identify the angiotensin-converting enzyme (ACE) inhibitory peptides released after in vitro gastro-intestinal digestion of skimmed goats' milk. The experimental approach involved a recently developed harmonised static in vitro digestion model, with mass spectrometry (MS) to identify bioactive peptides. Peptides in the post-pancreatic digest were extracted by ultrafiltration and isolated by reversed-phase high-performance liquid chromatography followed by MS. Among the
identified sequences, eighteen were identical to known bioactive peptides with ACE-inhibitory activity. Peptides with dipeptidyl peptidase IV-inhibitory and antioxidant activities were also identified. The antihypertensive tripeptides valine-proline-proline and isoleucine-proline-proline were released from goats’ milk protein during in vitro gastro-intestinal digestion at concentrations of 1829.8 ± 216.4 and 141.4 ± 15.1 ug L-1, respectively. This research underlines the suitability of the harmonised digestive model system to study the release of short bioactive peptides during gastro-intestinal transit.
2017
- Chocolate melanoidins as a carrier for the delivery of phenolic compounds in the gut
[Poster]
Martini, S.; Conte, A.; Tagliazucchi, D.
abstract
Emerging evidence from in vitro and in vivo studies suggested that the gastro-intestinal tract may be the key site for the biological action of food melanoidins [1]. In this work, we tested high molecular weight melanoidins (HMWM) extracted from dark chocolate for their ability to release low molecular weight polyphenols following in vitro digestion.
Water soluble HMWM were extracted by ultrafiltration (10 kDa) from three different types of dark chocolate (dark 70% cocoa (DC), dark 70% cocoa and 8% turmeric (TDC), dark 62% cocoa and 2% Sakura green tea (GTDC)). HMWM have been chemically characterized for their content in total polyphenols, protein, and polysaccharides [2]. HMWM were in vitro digested and separated from low molecular weight fractions (LMWF) by ultrafiltration. Digested HMWM were chemically characterized and LMWF were characterized for their total phenolic content. Individual phenolic compounds in the LMWF were identified and quantified by HPLC-ESI-QTOF-MS/MS.
Results showed that melanoidins are carbohydrate-based structures with the phenolic content ranging from 8.62 ± 1.0 in TDC to 12.79 ± 0.4 g/100 g HMWM in GTDC. The in vitro digestion caused a general decrease in the polysaccharide, protein and phenolic content. Digestion resulted in the release of phenolic compounds from HMWM to LMWF. The highest amount was recovered in the LMWF of GTDC (436.2 ± 87.6 mg/L), whereas the lowest amount in the LMWF of TDC (242.1 ± 17.1 mg/L). The most representative classes of polyphenols released from HMWM were stilbenes (resveratrol and derivatives) and phenolic acids (hydroxybenzoic, benzoic, quinic, cinnamic, coumaric and ferulic acids). As expected, food matrices influenced the chemical composition of HMWM. Specific phenolic compounds were detected in the LMWF from GTDC (quercetin-3-rutinoside) and TDC (curcumin, cathecol and protocatechuic acid).
2017
- Identification and in vitro bioaccessibility of dark chocolate phenolic compounds
[Poster]
Martini, S.; Conte, A.; Tagliazucchi, D.
abstract
Polyphenols have become an intense focus of research interest because of their perceived health-beneficial effects such as anti-atherogenic, anti-inflammatory and anti-tumoral effects. Cocoa (Theobroma cacao) is a major economically important international crop and has been associated with several nutritional benefits [1]. The aim of the present study was to identify and quantify phenolic compounds from three different types of dark chocolate using HPLC-ESI-QTOF-MS/MS. Phenolic compounds were also identified and quantified after in vitro gastro-intestinal digestion.
Three different types of dark chocolate (dark 70% cocoa (DC), dark 70% cocoa and 8% turmeric (TDC), dark 62% cocoa and 2% Sakura green tea (GTDC)) were in vitro digested with a simulated gastro-intestinal procedure [2]. Polyphenols were extracted from un-digested samples with methanol-water-formic acid solution (70:28:2 v/v) followed by a second extraction step with acetone. Total polyphenols were determined by Folin-Ciocalteau assay whereas antioxidant activity by FRAP methods. Individual phenolic compounds were identified and quantified using mass spectrometry.
Total polyphenol concentration in the un-digested samples varied from 2.6 ± 0.1 in DC to 3.4 ± 0.2 g/100g of chocolate in GTDC. The antioxidant activity ranged between 6.3 ± 0.1 to 15.4 ± 0.7 mmol trolox/100g in DC and GTDC, respectively. As expected, monomeric and oligomeric flavan-3-ols were the most representative compounds in all the three samples, followed by resveratrol and derivative. In addition, some phenolic acids (caffeoylquinic, protocatechuic, vanillic acids) and ellagic acid derivative (ellagic acid, ellagic acid hexoside) were also identified. The addition of green tea and turmeric resulted in the presence of gallate flavan-3-ols (such as epigallocatechin-3-gallate, epigallocatechin and epicatechin gallate) and curcuminoids, respectively. In vitro digestion resulted in a decrease in total polyphenols and in the individual phenolic profile modification.
2017
- LC-ESI-QTOF-MS identification of novel antioxidant peptides obtained by enzymatic and microbial hydrolysis of vegetable proteins
[Articolo su rivista]
Babini, Elena; Tagliazucchi, Davide; Martini, Serena; Più, Lucilla Dei; Gianotti, Andrea
abstract
Bioactive antioxidant peptides are more and more attracting the attention of food manufacturers for their potential to transform food in functional food, able to prevent a variety of chronic diseases associated with oxidative stress. In the present study proteins extracted from different vegetable sources (KAMUT khorasan wheat, emmer, lupine and pea) were hydrolyzed with commercial enzymes and Lactobacillus spp. strains. Hydrolysates were separated by size exclusion chromatography and purified fractions were analyzed for their antioxidant activity. Peptides from the fractions with the highest activity were identified by nanoLC-ESI-QTOF-MS and thirteen peptides were selected for synthesis on the basis of their sequence. Four peptides (VLPPQQQY, TVTSLDLPVLRW, VTSLDLPVLRW, FVPY) were found able to scavenge superoxide anion and hydroxyl radicals, organic nitro-radicals (ABTS, DPPH) and to inhibit lipid peroxidation. The impact of this work is targeted to add hydrolysed vegetable proteins
to reformulated functional food or to produce health-promoting ingredients and nutraceuticals.
2017
- PEPTIDOMIC PROFILE OF IN VITRO DIGESTED BOVINE, CAMEL, GOAT AND SHEEP MILK
[Abstract in Atti di Convegno]
Tagliazucchi, Davide; Martini, Serena; Sherif, Shamsia; Ahmed, Helal; Conte, Angela
abstract
Milk and dairy products have long traditions in human nutrition since they are a rich source of
nutrients such lipids, proteins, amino acids, vitamins and minerals. Nowadays, milk may also be considered
as a source of health-promoting compounds. Bioactive peptides deriving from milk proteins may play an
important role in the prevention and treatment of metabolic syndrome and its complications [1].
This study aimed to characterize the peptidomic profile of in vitro digested skimmed bovine, camel,
goat and sheep milk. Milk samples were in vitro digested following the harmonized INFOGEST protocol [2].
The peptide fractions were extracted from fully digested milk by ultrafiltration (cut-off 3 kDa) and the
permeate were characterized by nanoflow-LC-ESI-QTOF MS/MS analysis. Digestibility and biological activities
of the peptide fractions were also determined at the end of the digestion.
The results showed a faster and more efficient gastric and duodenal degradation of goat milk proteins
than camel, bovine and sheep milk. More than 100 peptides were identified in the permeate of digested milk,
most of them arising from beta-casein. Goat, sheep and bovine milk showed the highest similarity in peptide
sequences than camel milk. Peptides with previously demonstrated biological activities were found in the
permeate of all the milk samples after in vitro digestion. Biological activities analysis showed that bovine milk
peptides were the most effective in scavenging hydroxyl radical and in the inhibition lipid peroxidation.
Bovine milk was also the best source of DPPIV-inhibitory peptides (IC50=6.87 mg of peptides/mL) whereas
sheep milk was the best source of ACE-inhibitory peptides (IC50=625.4 μg of peptides/mL). The quantitative
diversity in the identified bioactive peptides explained the observed differences in the biological activities.
2017
- Phenolic compounds profile and antioxidant properties of six sweet cherry (Prunus avium) cultivars
[Articolo su rivista]
Martini, Serena; Conte, Angela; Tagliazucchi, Davide
abstract
Sweet cherry (Prunus avium) fruits are a nutritionally important food rich in dietary phenolic compounds. The aim of this study was to investigate the phenolic pro file and chemometric discrimination of fruits from six cherry cultivars using a quantitative metabolomics approach, which combine non-targeted mass spectrometry and chemometric analysis. The assessment of the phenolic fingerprint of cherries allowed the tentative identi fication of 86 compounds. A total of 40 chlorogenic acids were identi fied in cherry fruit, which pointed out hydroxycinnamic acid derivatives as the main class of phenolics by number of compounds. Among the compounds detected, 40 have been reported for the first time in sweet cherry fruit. Hydroxycinnamic acids are also the quantitatively most represented class of phenolic compounds in the cherry cultivars with the exception of Lapins and Durone della Marca where the most representative class of phenolic compounds were anthocyanins and flavan-3-ols, respectively. This non-targeted approach allowed the tentative identi fication of the cultivar-compound relationships of these six cherry cultivars. Both anthocyanins and colorless phenolic compounds profile appeared to be cultivar-dependent. In detail, anthocyanins and flavonols patterns have the potential to be used for the determination of a varietal assignment of cherries
2016
- Bioaccesssibility and bioactivity of polyphenols extracted from six cherry cultivars
[Poster]
Martini, S.; Tagliazucchi, D; Conte, A
abstract
Introduction
There is a possible association between consumption of fruit and vegetables and reduced risk of cancer, particularly cancer of the digestive tract. This anti-cancer activity has been attributed in part to polyphenols present in foods [1]. Cherries in particular are a rich source of polyphenols, especially anthocyanins and hydroxycinnamic acids [2].
Method
Cherries from six different cultivars (Durone della Marca, Celeste, Durone Nero I, Bigarreau, Lapins and Moretta) were in vitro digested with a procedure that mimicked the physiochemical conditions of the gastro-intestinal tract [3]. After digestion, polyphenol-rich extracts were obtained by preparative chromatography on C18 column. Polyphenols were also extracted from un-digested cherries with methanol-water-formic acid solution (70:28:2 v/v) and chromatographed on C18 column. Total polyphenols and anthocyanins concentration were determined by Folin-Ciocalteau assay and pH differential method, respectively [3]. Polyphenols were identified using liquid chromatography mass spectrometry. The digested and un-digested preparations were assessed for their cytotoxic and anti-proliferative activities using two human colon adenocarcinoma cell lines (Caco-2 and SW480) and IC50 values of anti-proliferative activity were determined.
Results / Discussion / Conclusion
The total polyphenol concentration in the un-digested samples varied from 70.2mg/100g of cherry in the cultivar Durone della Marca to 280.6mg/100g of cherries in the cultivar Durone Nero I. The total anthocyanin content varied from 0.12mg/100g of cherry in the cultivar Durone della Marca to about 17mg/100g of cherry in the two darker cultivars (Lapins and Moretta). In vitro digestion resulted in a decrease in both total polyphenols and anthocyanins. Total polyphenols decrease ranged from 56 to 80%, whereas total anthocyanin decrease was more than 90% in all the tested cultivars. Mass spectrometry showed that the gastro-intestinal digestion decreased the content of hydroxycinnamic acids and anthocyanins. Other classes of polyphenols are more stable to digestion. Breakdown products of polyphenols are also present. Digested and un-digested polyphenol-rich extracts showed no cytotoxic activity. Un-digested polyphenol-rich extract from the cultivar Bigarreau showed the highest anti-proliferative effect (IC50 of 7.1 ± 1.1 µg polyphenols/mL) on the SW480 cells. The in vitro digestion increased the anti-proliferative activity of cherry extracts. Polyphenol-rich extracts from digested Durone della Marca and Celeste were the most active with IC50 values of 1.7 ± 0.8 and 3.5 ± 1.2 µg polyphenols/mL, respectively. All of the tested extracts showed no anti-proliferative activity on Caco-2 cells.
Our results showed that cherry polyphenols were effective anti-proliferative agents on the more undifferentiated colon adenocarcinoma cells SW480 and that their digestion increased the effect. The increase may be due to the formation of polyphenol metabolites. A great difference was observed between the cultivars used. Further research is required to identify the compounds responsible for the observed anti-proliferative effect.
2016
- Bioactive peptides from vegetable food matrices: Research trends and novel biotechnologies for synthesis and recovery
[Articolo su rivista]
Rizzello, Carlo G.; Tagliazucchi, Davide; Babini, Elena; Rutella, Giuseppina Sefora; Saa, Danielle L. Taneyo; Gianotti, Andrea
abstract
Currently, the interest for health-promoting functional foods, dietary supplements and pharmaceutical preparations containing bioactive peptides deriving from food proteins, is increasing. Despite the large literature concerning peptides derived from animal proteins, only recently the scientific community investigated the possibility to obtain bioactive peptides from vegetable sources, also discovering novel functional features. In this article, the functional effects of vegetable-derived peptides, including antihypertensive, antioxidant, antitumoral, antiproliferative, hypocholesterolemic, antinflammatory activities, are described.
The novel biotechnologies for the release of bioactive peptides from vegetable matrices, including microbial fermentation and the use of microbial enzymes are investigated. Moreover, the modern technologies for their recovery, purification, and analysis, are reviewed and discussed.
2016
- Bovine milk antioxidant properties: effect of in vitro digestion and identification of antioxidant compounds
[Articolo su rivista]
Tagliazucchi, Davide; Helal, Ahmed; Verzelloni, Elena; Conte, Angela
abstract
Milk proteins contained encrypted in their sequence biologically active components that can be released by enzymatic hydrolysis. Among the biological activities recognized in milk components, the antioxidant activity is of great interest.
The objective of the present study was to analyse the antioxidant properties of whole, semi-skimmed and skimmed milk during simulated gastrointestinal digestion and to identify the compounds responsible for the antioxidant activity. Simulated digestion
increased the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS)+ radical scavenging activity of milk. In digested whole milk, the main contribution to ABTS+ radical scavenging activity was due to high molecular weight fraction (>3 kg.mol−1).
For semi-skimmed and skimmed milk, the main contribution was due to low molecular weight fraction (<3 kg.mol−1). Twelve major peaks were collected from low molecular weight fraction of digested skimmed milk by reversed-phase high-performance liquid chromatography and evaluated for their ABTS+ radical scavenging activity. Among the different fractions, three (F2, F3 and F5) showed high ABTS+ and hydroxyl radical scavenging activity and lipid peroxidation inhibitory capacity. The compounds (free
amino acids and peptides) present in these fractions were identified with nano-LCQTOF MS/MS analysis. The amino acids tryptophan and tyrosine seemed fundamental in the ABTS+ and hydroxyl radical scavenging capacities whereas the amino acids
phenylalanine and histidine played an important role in the lipid peroxidation inhibitory activity of the peptides. The results reported in this study suggested that milk proteins could act as a carrier for the delivery of antioxidant compounds in the gastrointestinal tract possibly protecting the gastrointestinal tract itself from the oxidative damage.
2016
- Colon-available dietary melanoidins exhibit cytotoxic activity on human colon adenocarcinoma cell lines
[Poster]
Tagliazucchi, D.; Martini, S.; Conte, A
abstract
Introduction
Emerging evidence from in vitro and in vivo studies suggested that the gastro-intestinal tract might be the key site for the biological action of food melanoidins [1]. Recently, Vitaglione et al. [2] reviewed the possible mechanisms by which coffee melanoidins may influence the risk of colorectal cancer development. In this work, we tested the water-soluble high molecular weight melanoidins (HMWM) obtained from different food sources for their cytotoxic activity against colon cancer cell lines.
Method
HMWM were obtained by ultrafiltration (cut-off 10 kDa) from instant coffee, cocoa brew, dark beer and instant barley coffee beverages [2]. HMWM have been chemically characterized for their content in total polyphenols, protein, and polysaccharides as well as their spectroscopic properties. The value of Kmix 420nm (defined as the absorption at 420nm of a solution of HMWM at a concentration of 1 g/L) provides information on the relative amount of melanoidins (Kmix 420nm) respect to other compounds found in the HMWM of beverages. Cytotoxic activity against two colon cancer cell lines (Caco-2 and SW480) was assayed by MTT assay. HMWM obtained from the above beverages submitted to in vitro gastro-intestinal digestion [3], were subjected to the same chemical analysis and assayed for their cytotoxic activity.
Results / Discussion / Conclusion
Barley coffee had the highest HMWM content (83.3g/100g of dry matter) followed by coffee (34.4g/100g of dry matter), cocoa (28.1g/100g of dry matter) and dark beer (20.0g/100g of dry matter). The HMWM of coffee had the highest Kmix 420nm value, suggesting that this fraction is relatively rich in melanoidins. On the contrary, despite the highest content in HMWM, barley coffee was the sample with the lowest melanoidins content. Protein content of HMWM ranged from 6.1% (dark beer) to 10.6% (coffee) whereas polysaccharides content ranged from 42% (dark beer) to 61% (cocoa). Cocoa and coffee HMWM had the highest phenolic content (6 and 12%, respectively). The in vitro digestion caused a general decrease in the polysaccharide and protein content of all the analysed HMWM because of the action of amylolytic and proteolytic enzymes. The analysis of the content of polyphenols showed a decrease in these components as a result of the digestion. On the contrary, the in vitro digestion caused an increase in the Kmix 420nm of all the HMWM investigated suggesting that this process resulted in an enrichment in melanoidins. The effect of HMWM on the human colon adenocarcinoma Caco-2 and SW480 cell cultures was investigated. Un-digested HMWM had no significant cytotoxic effect against both the cell lines. However, the in vitro digestion greatly increased the cytotoxic activity of HMWM against both the cell lines. Coffee HMWM were the most active against Caco-2 cells whereas cocoa HMWM were found to be most active against SW480 cells. Considering that the colon accumulates its content over at least 24 h in a maximum volume of 2 l, it is possible to estimate the concentrations of HMWM reaching daily the colon with diet according to the different food matrices [1]. Data showed that, the consumption of coffee, cocoa and barley coffee beverages may result in intestinal concentrations of HMWM, which are cytotoxic against Caco-2 and SW480 cell lines.
2016
- Composition and properties of peptides that survive standardised in vitro gastro-pancreatic digestion of bovine milk
[Articolo su rivista]
Tagliazucchi, Davide; Helal, Ahmed; Verzelloni, Elena; Bellesia, Andrea; Conte, Angela
abstract
There are conflicting results on the release and conservation of bioactive protein sequences during in vitro digestion. Only the peptides that are released and survive may exert their action. Peptides <3 kDa were determined by mass spectrometry after standardised in vitro gastro-pancreatic digestion of bovine milk. At a degree of digestion of 30.7%, 384 peptides were obtained from 2 to 25 residues in length. Five other peptides were recovered after reduction of disulphide bonds. Due to the presence of proline residues at the last or second last position at the C-terminus, a high number of peptides with ACE-inhibitory activity were obtained. Peptides with opioid, antimicrobial, immunomodulatory and antithrombotic activities were recovered, with many precursors and degradative products. Peptides with opioid and antimicrobial activities may be the result of evolutionary processes. The peptides obtained were similar to those released in the human jejunum.
2016
- Release of angiotensin converting enzyme-inhibitory peptides during in vitro gastro-intestinal digestion of camel milk
[Articolo su rivista]
Tagliazucchi, Davide; Shamsia, Sherif; Conte, Angela
abstract
Angiotensin-converting enzyme (ACE)-inhibitory peptides released from camel milk after simulated gastro-intestinal digestion were identified. The hydrolysis degree increased during digestion. The highest ACE-inhibitory activity was found in the post-pancreatic <3 kDa fraction. Peptides responsible for the biological activity were isolated by reversed-phase high-performance liquid chromatography and identified by mass spectrometry. Among the identified sequences, 17 were identical to known bioactive peptides with ACE-inhibitory activity. Based on previous structureeactivity relationship studies, the sequence of some peptides allowed us to anticipate the presence of biological activities. The antihypertensive tripeptide isoleucine-proline-proline (IPP) was identified and quantified in digested camel milk. The amount of released IPP was 2.56 ± 0.15 mg L1 of milk. For the first time, we showed that IPP is released during the gastro-intestinal digestion of camel milk k-casein. This research provides the
basis to increase the exploitation of the health benefits of camel milk.
2016
- Release of the Antihypertensive Tripeptides Valine-Proline-Proline and Isoleucine-Proline-Proline from Bovine Milk Caseins during in Vitro Gastrointestinal Digestion
[Articolo su rivista]
Rutella, GIUSEPPINA SEFORA; Solieri, Lisa; Martini, Serena; Tagliazucchi, Davide
abstract
The aim of this study was to identify and quantify the release of antihypertensive tripeptides valine-proline-proline (VPP) and isoleucine-proline-proline (IPP) during in vitro oro-gastro-intestinal (OGI) digestion of bovine skimmed milk. The experimental approach combined the recently developed harmonized static in vitro digestion (IVD) model and targeted mass spectrometry to monitor peptide generation. We first demonstrated that VPP and IPP are released from bovine milk proteins during in vitro OGI digestion at final concentrations of 354.3 ± 29.8 and 973.8 ± 155.7 μg/L, respectively. In silico analysis of cleavage sites and mass spectrometry revealed that tetrapeptides VPPF, IPPL, and IPPK are precursors of VPP and IPP. The release of other ACE-inhibitory peptides, such as FVAP, VAP, AW, and VY, was demonstrated, and their fate and the time course were investigated. This research underlines the suitability of an IVD system to study the release of short bioactive peptides during OGI transit.
2016
- Survival and bioactivities of selected probiotic lactobacilli in yogurt fermentation and cold storage: New insights for developing a bi-functional dairy food
[Articolo su rivista]
Rutella, GIUSEPPINA SEFORA; Tagliazucchi, Davide; Solieri, Lisa
abstract
In previous work , we demon strated that two probiotic strains, name ly Lactobacillus casei PRA205 and Lactobacillus rhamnosus PRA331, produce fermented milks with potent angiotensin-converting enzyme (ACE)-inhibitory and antioxidant activities. Here, we tested these strains for the survivability and the release of antihypertensive and antioxidant peptides in yogurt fermentation and cold storage. For these purposes three yogurt batches were compared: one prepared using yogurt starters alone (Lactobacillus delbrueckii subspecies bulgaricus 1932 and Streptococcus thermophilus 99), and the remaining two containing either PRA205 or PRA331 in addition to yogurt starters. Despite the lower viable counts at the fermentation end compared to PRA331, PRA205 overcame PRA331 in survivability during refrigerated storage for 28 days, leading to viable counts (>108 CFU/g) higher than the minimum therapeutic threshold (106 CFU/g). Analyses of in vitro ACE-inhibitory and antioxidant activities of peptide fractions revealed that yogurt supplemented with PRA205 displays higher amounts of antihypertensive and antioxidant peptides than that produced with PRA331 at the end of fermentation and over storage. Two ACE-inhibitory peptides, Valine-Proline-Proline (VPP) and Isoleucine-Proline-Proline (IPP), were identified and quantified. This study demonstrated that L. casei PRA205 could be used as adjunct culture for producing bi-functional yogurt enriched in bioactive peptides and in viable cells, which bring health benefits to the host as probiotics.
2015
- Characterization of a Combined Treatment with Alpha-Lipoic Acid for the Control of Enzymatic Browning in Fresh-Cut Golden Delicious Apples
[Articolo su rivista]
Musetti, Alessandro; Tagliazucchi, Davide; Montevecchi, Giuseppe; Verzelloni, Elena; Antonelli, Andrea; Fava, Patrizia
abstract
Enzymatic browning is the main cause of quality loss in minimally processed fresh-cut apples and this study was aimed to control it by evaluating the use of a treatment with α-lipoic acid and ascorbic acid on Golden Delicious apples. Also hexanal, an effective natural enhancer of apple odor, was added to the treatment.At the end of the preservation period, colorimetric analysis demonstrated that α-lipoic acid exerts an anti-browning effect comparable with that of ascorbic acid about 60 times more concentrated and, moreover, chemical assays demonstrated its synergistic interaction with this substance. The application of the “anti-browning mix” does not substantially change the volatile profile of the apple slices while the addition of hexanal (0.076 mmol per liter of air) improves it primarily with an increased production of hexyl acetate (about 86% of the average aromatic component), without modifying the fruit respiration.Practical ApplicationsThe combined treatment has proved to be useful to preserve and improve Golden Delicious apples quality over the span of 8 days (1 day more than the average shelf life recommended by many producers for this kind of product).The presence of hexanal makes the treatment very interesting in the context of minimally processed fruit, where apples are used in an early stage of ripening that best tolerates the processing chain but that is characterized by a low volatile biosynthesis.Moreover, the presence of α-lipoic acid gives to the fruit the connotation of “nutraceutical food.”
2015
- Characterization of dietary melanoidins and evaluation of the cytotoxic activity against colon cancer cells
[Poster]
Tagliazucchi, Davide; Bellesia, Andrea; Martini, Serena; Conte, Angela
abstract
Emerging evidence from in vitro and in vivo studies suggested that the gastro-intestinal tract may be the key site for the biological action of food melanoidins. Recently, Vitaglione et al. reviewed the possible mechanisms by which coffee melanoidins may influence the risk of colorectal cancer development. In this work, we tested high molecular weight melanoidins (HMWM) extracted from different food sources for their cytotoxic activity against colon cancer cell lines. Water soluble HMWM were extracted by ultrafiltration (cut-off 10 kDa) from instant coffee, cocoa brew, dark beer and instant barley coffee. HMWM have been chemically characterized for their content in total polyphenols, protein, and polysaccharides, antioxidant activity as well as their spectroscopic properties. These extracts were assessed using two colon cancer cell lines: Caco-2 and SW480. Cytotoxic activity was assayed by MTT and data are presented as the dose that inhibited 50% control growth (IC50). All of the assessed HMWM were cytotoxic against Caco-2 cell line. Coffee HMWM were the most effective (IC50 of 1042 and 857 µg/mL after 24 and 48h of treatment, respectively) followed by dark beer, barley coffee and cocoa HMWM. Meanwhile, they are less effective against SW480 with coffee HMWM that were the most effective (IC50 of 2038 µg/mL after 48h) followed by cocoa, dark beer and barley coffee. Considering that the colon accumulates its content over at least 24h in a maximum volume of 2 litres, and that the daily intake of coffee melanoidins range between 0.5 and 2.0 g , it is possible to calculate a hypothetical concentration of coffee melanoidins in the colon between 0.25 and 1 mg/mL, which are values comparable to the IC50 for the cytotoxic activity against colon cancer cells.
2015
- Gastro-pancreatic release of phenolic compounds incorporated in a polyphenols-enriched cheese-curd
[Articolo su rivista]
Helal, A.; Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
As functional food, enriched cheese has recently been developed. The main objectives of this study were to investigate the role of casein in the retention of polyphenol during curd formation and the release of polyphenols during in vitro gastro-pancreatic digestion of polyphenols-enriched cheese and their contribution to the antioxidant activity of digested curd. Polyphenols showed high retention coefficient in curd. The retention coefficient of polyphenol was related to the binding affinity to casein and to their hydrophilicity. The polyphenols should be added before milk coagulation since the binding decreases as casein molecules aggregate. During in vitro gastro-pancreatic digestion steps, polyphenols were released from curd due to the dilution in gastric fluid and to casein proteolysis. The addition of polyphenols to curd determined a relevant increase of antioxidant activity respect to the curd control even a part of polyphenols is degraded by alkaline pH of pancreatic fluid. Our results suggested the possibility of producing highly nutritive value cheese with high release of the polyphenols during digestion. In addition, the whey, which contains polyphenols, can be involved in different products to maximize its utilization.
2015
- Identification of ACE-inhibitory peptides from Phaseolus vulgaris after in vitro gastrointestinal digestion
[Articolo su rivista]
Tagliazucchi, Davide; Martini, Serena; Bellesia, Andrea; Conte, Angela
abstract
The objective of this study was to identify the angiotensin I-converting enzyme (ACE)-inhibitory peptides released from thermally treated Phaseolus vulgaris (pinto) whole beans after in vitro gastrointestinal digestion. The degree of hydrolysis increased during digestion reaching a value of 50% at the end of the pancreatic digestion. The<3 kDa fraction of the postpancreatic sample showed high ACE-inhibitory activity (IC50=105.6±2.1 mug of peptides/mL). Peptides responsible for the ACE-inhibitory activity were isolated by reverse-phase high-performance liquid chromatography (HPLC). Three fractions, showing the highest inhibitory activity, were selected for tandem mass spectrometry (MS/MS) experiments. Eleven of the identified sequences have previously been described as ACE-inhibitors. Most of the identified bioactive peptides have a hydrophobic amino acid, (iso)leucine or phenylalanine, or proline at the C-terminal position, which is crucial for their ACE-inhibitory activity. The sequence of some peptides allowed us to anticipate the presence of ACE-inhibitory activity.
2015
- Impact of non-starter lactobacilli on release of peptides with angiotensin-converting enzyme inhibitory and antioxidant activities during bovine milk fermentation
[Articolo su rivista]
Solieri, Lisa; Rutella, Giuseppina Sefora; Tagliazucchi, Davide
abstract
This study aimed at evaluating non-starter lactobacilli (NSLAB) isolated from cheeses for their proteolytic activity and capability to produce fermented milk enriched in angiotensin-converting enzyme (ACE)-inhibitory and antioxidant peptides. Preliminarily, 34 NSLAB from Parmigiano Reggiano (PR) and 5 from Pecorino Siciliano cheeses were screened based on their capacity to hydrolyze milk proteins. Two NSLAB strains from PR, Lactobacillus casei PRA205 and Lactobacillus rhamnosus PRA331, showed the most proteolytic phenotype and were positively selected to inoculate sterile cow milk. The fermentation process was monitored by measuring viable cell population, kinetic of acidification, consumption of lactose, and
synthesis of lactic acid. Milk fermented with Lb. casei PRA205 exhibited higher radical scavenging (1184.83 ± 40.28 umol/L trolox equivalents) and stronger ACE-inhibitory (IC50 = 54.57 ug/mL) activities than milk fermented with Lb. rhamnosus PRA331 (939.22 ± 82.68 umol/L trolox equivalents;
IC50 = 212.38 ug/mL). Similarly, Lb. casei PRA205 showed the highest production of ACE-inhibitory peptides Val-Pro-Pro and Ile-Pro-Pro, which reached concentrations of 32.88 and 7.52 mg/L after 87 and 96 h of milk fermentation, respectively. This evidence supports Lb. casei PRA205, previously
demonstrated to possess characteristics compatible with probiotic properties, as a promising functional culture able to promote health benefits in dairy foods
2015
- Melanoidins from coffee and lipid peroxidation
[Capitolo/Saggio]
Tagliazucchi, Davide
abstract
Dietary lipid oxidation products are thought to be an important risk factor in the development of cardiovascular diseases. Lipid oxidation products may be already present in oxidized food and they may be generated during digestion of highly oxidizable food such as meat. In vitro and in vivo studies are consistent for a role of coffee high molecular weight melanoidins in the prevention of oxidative damage. Since coffee melanoidins are poorly bioavailable, the gastro-intestinal tract itself might constitute their main biological site of action where they reach high concentrations following coffee consumption. In the stomach they inhibit the peroxidation of meat lipids, decreasing the synthesis of hydroperoxides and secondary lipoxidation products. The reduction in the formation of these pro-atherogenic compounds has been shown to be followed by a decrease in their absorption in the human volunteers. The ability of coffee melanoidins to inhibit lipid peroxidation may contribute to their health benefits, since dietary oxidized lipid and advanced lipid oxidation endproducts are involved in the development of atherosclerosis and other diseases.
2015
- Pomegranate ellagitannins inhibit α-glucosidase activity in vitro and reduce starch digestibility under simulated gastro-intestinal conditions
[Articolo su rivista]
Bellesia, Andrea; Verzelloni, Elena; Tagliazucchi, Davide
abstract
Pomegranate extract was tested for its ability to inhibit α-amylase and α-glucosidase activity. Pomegranate extract strongly inhibited rat intestinal α-glucosidase in vitro whereas it was a weak inhibitor of porcine α-amylase. The inhibitory activity was recovered in an ellagitannins-enriched fraction and punicalagin, punicalin and ellagic acid were identified as α-glucosidase inhibitors (IC50 of 140.2, 191.4 and 380.9 μmol/L, respectively). Kinetic analysis suggested that the pomegranate extract and ellagitannins inhibited α-glucosidase activity in a mixed mode. The inhibitory activity was demonstrated using an in vitro digestion system, mimicking the physiological gastro-intestinal condition, and potatoes as food rich in starch. Pre-incubation between ellagitannins and α-glucosidase increased the inhibitory activity, suggesting that they acted by binding to α-glucosidase. During digestion punicalin and punicalagin concentration decreased. Despite this loss, the pomegranate extract retained high inhibitory activity. This study suggests that pomegranate ellagitannins may inhibit α-glucosidase activity in vitro possibly affecting in vivo starch digestion.
2015
- Selection of Val-Pro-Pro (VPP) and Ile-Pro-Pro (IPP)–producer Lactobacillus strains for developing functional yogurt enriched of bioactive peptides
[Abstract in Atti di Convegno]
Solieri, Lisa; Rutella, Sefora; Tagliazucchi, Davide; DE VERO, Luciana; Giudici, Paolo
abstract
Accepted poster for SESSION II: The complexity of food ecosystems: physiology of single strains in pure culture vs. complex consortia
2015
- The gastro-intestinal tract as the major site of biological action of dietary melanoidins
[Articolo su rivista]
Tagliazucchi, Davide; Bellesia, Andrea
abstract
Emerging evidence from laboratory researches has highlighted the bioactivity of food melanoidins and melanoproteins. Whilst such studies have been carried out with different in vitro systems, information about melanoidins absorption and bio-availability are scarce. However, they are generally considered as poorly absorbable and bio-available compounds. Therefore, we present a review in which the gastro-intestinal tract is hypothesized to be the main site of action of food melanoidins and melanoproteins biological activity. We described recent data supporting this hypothesis both in vitro model systems and in vivo. Importantly, we focused this review only on the effect of melanoidins and melanoproteins extracted from food. Most of the studies had been carried out using water-soluble carbohydrate-based melanoidins isolated from different food sources (beer, barley coffee, coffee). In bakery products, melanoidins are protein-based structure (melanoproteins) which are largely insoluble in water. Dietary melanoidins and melanoproteins have been demonstrated to exert in vitro antioxidant and metal chelating ability in the gastro-intestinal tract reducing the formation of lipid hydroperoxides and advanced lipid oxidation end products during the digestion of meat. The reduction in the formation of these pro-atherogenic compounds has been shown to be followed by a decrease in their absorption in human volunteers. Food melanoidins have also shown in vitro anti-caries and prebiotic activities. We conclude by underlining the possible role of food melanoidins in the prevention of gastro-intestinal tract cancers. We hope this review will stimulate further research on food melanoidins and their biological activities in the gastro-intestinal tract.
2014
- Bioaccessibility of polyphenols and cinnamaldehyde in cinnamon beverages subjected to in vitro gastro-pancreatic digestion
[Articolo su rivista]
A., Helal; Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
The present study was undertaken to investigate the in vitro bioaccessibility of polyphenols and cinnamaldehyde in cinnamon beverage and formulations during simulated gastro-pancreatic digestion. Cinnamon polyphenols post-pancreatic bioaccessibility was 79.8%. The decline in total polyphenols was caused by the precipitation of proanthocyanidins by pepsin. The addition of sweeteners increased the polyphenols bioaccessibility decreasing the interaction between pepsin and tannins. The addition of bovine milk had no effect on polyphenols post-pancreatic bioaccessibility. Quercetin-3-rhamnoside, syringic and coumaric acids were found to be poor bioaccessible whereas kaempferol and cinnamaldehyde were stable during digestion. The addition of sweeteners did not affect the bioaccessibility of these compounds. The final bioaccessibility was affected by milk only for cinnamaldehyde. The present study demonstrates that cinnamon beverage provides a significant source of dietary bioaccessible polyphenols and that the modality of consuming cinnamon could represent a crucial factor since the addition of sweeteners improve their bioaccessibility whereas milk has no effect on their bioaccessibility.
2014
- Cocoa brew inhibits in vitro α-glucosidase activity: The role of polyphenols and high molecular weight compounds
[Articolo su rivista]
Bellesia, Andrea; Tagliazucchi, Davide
abstract
Cocoa brew showed a dose-dependent inhibitory activity against α-glucosidase (IC50 7.87 mg/mL). The cocoa brew was fractionated by ultrafiltration in a low molecular weight fraction (LMW <10 kDa) and two melanoidin-rich fractions called high molecular weight (HMW > 30 kDa) and intermediate molecular weight (IMW 10-30 kDa) fractions. All fractions tested caused some inhibition with IMW that was the most active (IC50 2.37 mg/mL). LMW fraction was separated with Sephadex LH-20 in an unbound (containing monomeric and dimeric catechins) and a bound fraction. All the inhibitory activity was recovered in the unbound fraction. All the phenolic compounds identified with HPLC showed α-glucosidase inhibitory activity. IMW and HMW fractions were fractionated by ethanol precipitation. The fractions from IMW precipitated with 75 and 25% ethanol were found to contain power inhibitors of α-glucosidase activity (IC50 0.87 and 1.01 mg/mL, respectively). In the HMW sample, the fractions precipitated with 50 and 75% ethanol were found to be active against α-glucosidase activity. Most of the inhibitory activity against α-glucosidase of cocoa brew was due to the LMW fraction (56%) whereas IMW and HMW contributed for about 47% of the inhibitory activity. This study suggests that different components of cocoa may influence α-glucosidase activity.
2014
- Exploitation of starch industry liquid by-product to produce bioactive
peptides from rice hydrolyzed proteins
[Articolo su rivista]
L., Dei Più; A., Tassoni; D. I., Serrazanetti; M., Ferri; E., Babini; Tagliazucchi, Davide; A., Gianotti
abstract
Small peptides show higher antioxidant capacity than native proteins and may be absorbed in the intestine without further digestion. In our study, a protein by-product from rice starch industry was hydrolyzed with commercial proteolytic enzymes (Alcalase, Neutrase, Flavourzyme) and microbial whole
cells of Bacillus spp. and the released peptides were tested for antioxidant activity. Among enzymes, Alcalase was the most performing, while microbial proteolytic activity was less efficient. Conversely, the antioxidant activity was higher in the samples obtained by microbial hydrolysis and particularly with
Bacillus pumilus AG1. The sequences of low molecular weight antioxidant peptides were determined and analyzed for aminoacidic composition. The results obtained so far suggest that the hydrolytic treatment of this industrial by-product, with selected enzymes and microbial systems, can allow its exploitation for the production of functional additives and supplements rich in antioxidant peptides, to be used in new
food formulas for human consumption.
2014
- Relationship between the chemical composition and the biological activities of food melanoidins
[Articolo su rivista]
Tagliazucchi, Davide; Verzelloni, Elena
abstract
In this work the relationship between the chemical composition and the biological activities of food melanoidin-rich fractions has been investigated. Melanoidin-rich fractions were extracted by ultrafiltration (10 KDa cut-off) from coffee, barley coffee, dark beer and traditional balsamic vinegar. All the food melanoidin-rich fraction were formed mainly of carbohydrates, phenolic compounds and proteins. In dark beer, barley coffee and traditional balsamic vinegar melanoidins, glucose was the most important sugar incorporated in melanoidins. Coffee melanoidins contained the highest amount of phenolic groups, followed by traditional balsamic vinegar melanoidins. We tested radical scavenging, Fe2+-chelating and heme binding abilities of food melanoidins under gastric conditions. Melanoidin-rich fraction extracted from coffee were the most active, showing the highest radical scavenging activity, Fe2+-chelating and heme binding abilities compared to barley coffee, dark beer, and traditional balsamic vinegar. We assigned radical scavenging and Fe2+-chelating abilities to the phenolic groups present in food melanoidins.
2013
- Cocoa extract inhibits in vitro α-glucosidase activity: the role of polyphenols and melanoidins
[Abstract in Atti di Convegno]
Tagliazucchi, Davide; Bellesia, Andrea; Conte, Angela
abstract
Studies on the health benefits of cocoa have recently focused on the anti-diabetic potential of cocoa extract. The intake of cocoa has been related to a reduction in the post-prandial levels of blood glucose in diabetic rats. A possible explanation for these observations is that cocoa extract possesses the ability to inhibit enzymes involved in the digestion of carbohydrate, attenuating the post-prandial increase in glycemia. Using an in vitro model, the activity of rat intestinal α-glucosidase has been determined in the absence and presence of cocoa extract and fractions. The addition of cocoa extract inhibited α-glucosidase activity with an IC50 of 7.9 mg/mL. The cocoa extract was further fractionated by ultrafiltration in a polyphenolic–rich fraction (<10 KDa) and a melanoidins-rich fraction (>10 KDa). Both the fractions were able to inhibit α-glucosidase with an IC50 of 2.3 and 3.9 mg/mL, respectively. To identify the bioactive compounds, the polyphenolic–rich fraction was further fractionated with Sephadex LH-20 columns to separate tannins from low molecular weight flavanols. Furthermore, the melanoidins-rich fraction was separated in two fractions using ultrafiltration at 30 KDa. All the obtained fractions were characterized for their ability to inhibit α-glucosidase and for their content in polyphenolic compounds.
2013
- Effect of cinnamon beverage formulations on the bioaccessibility of polyphenols and cinnamaldehyde during in vitro gastro-pancreatic digestion
[Abstract in Atti di Convegno]
A., Helal; Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
The present study was undertaken to investigate the in vitro bioaccessibility of phenolic compounds and cinnamaldehyde in cinnamon beverage and formulations during simulated gastro-pancreatic digestion. Cinnamon polyphenol bioaccessibility was 79.8% at the end of the digestion. This decline in total polyphenols was found during the gastric step of the digestion and was caused by the precipitation of cinnamon proanthocyanidins by pepsin. The addition of sweeteners increased the polyphenol bioaccessibility to value near 90% decreasing the interaction between pepsin and tannins. The addition of bovine milk gave rise to an immediate decrease in cinnamon polyphenol concentration and negatively affect their post-pancreatic bioaccessibility. Quercetin-3-rhamnoside, syringic and coumaric acids were find to be poor bioaccessible whereas kaempferol and cinnamaldehyde were stable during digestion. The addition of sweeteners did not affect the bioaccessibility of these compounds. Milk addition caused an immediate decrease in the concentration of monomeric phenolic compounds partially due to the interaction with milk protein as probed by fluorescence spectroscopy. The final bioaccessibility was affected by milk only for cinnamaldehyde. The present study demonstrates that cinnamon beverage provide a significant source of dietary bioaccessible polyphenols and that the modality of consuming cinnamon could represent a crucial factor since the addition of sweeteners improve their bioaccessibility whereas milk negatively affect it.
2013
- Effect of grape variety on the evolution of sugars, hydroxymethylfurfural, polyphenols and antioxidant activity during grape must cooking
[Articolo su rivista]
Tagliazucchi, Davide; Verzelloni, Elena; A., Helal; Conte, Angela
abstract
Grape must cooking is a traditional practice used for the production of foodstuff worldwide such as traditional balsamic vinegar. The aim of the present work was to reveal the effect of grape variety on the evolution of the main chemical components in grape must during cooking. On this purpose, two grape must varieties (red Lambrusco and white Trebbiano grapes) were cooked and analyzed. The monosaccharides concentration decreased because cooking resulting in the formation of 5-hydroxymethylfurfural. At the end of cooking, the antioxidant activity and polyphenols concentration was higher and the 5-hydroxymethylfurfural was lower in Lambrusco than in Trebbiano must. Additional changes involved degradation of monomeric anthocyanins resulting in the formation of the corresponding phenolic acids. From a health point of view, the high antioxidant activity and polyphenols concentration and the low 5-hydroxymethylfurfural concentration make cooked red Lambrusco must a safer raw starting material for making traditional balsamic vinegar.
2013
- Hydrolysis of pea and lupin proteins by microorganisms to produce bioactive peptides
[Abstract in Atti di Convegno]
L., Dei Più; Tagliazucchi, Davide; E., Babini; A., Gianotti
abstract
Small peptides and free amino acids have the advantage of being absorbed in the intestine without any digestion in the stomach. This explains their use in many formulas such as diets for nursing infants or sick adults and as stimulants for people liable for allergy development. The production of peptides through hydrolytic reactions seems to be the most promising technique to form proteinaceous antioxidants since peptides have higher antioxidant activity than raw proteins. In the present study, proteins extracted from flours of peas and lupin were hydrolyzed with microbial enzymes from whole cells of Lactobacillus fermentum MR13 and Lactobacillus rhamnosus C249. The products of the hydrolysis further separated by Size Exclusion Chromatography and the main fractions purified were analyzed for their antioxidant activity by DPPH assay. The results showed that both the total fractions, both the single fractions enhanced the antioxidant activity of the not hydrolyzed substrates. The antioxidant activities of hydrolyzed vegetal proteins seemed to be in relation to their molecular weight distribution. The peptides present in the fractions with highest antioxidant activity were identified with nanoLC-qTOF MS/MS analysis. As emerged from the present study, hydrolyzed vegetal proteins have the potential to become good ingredients and supplements in new food formulas.
2013
- Mechanism of inhibition of protein glycation by polyphenols
[Abstract in Atti di Convegno]
Bellesia, Andrea; Tagliazucchi, Davide; Conte, Angela
abstract
We have observed that protocatechuic (PC) and 3,4-dihydroxyphenylacetic (DHPA) acids, found in human plasma after the consumption of foods rich in anthocyanins and flavanols, inhibit the formation of advanced glycation endproducts (AGEs).
We have investigated the mechanisms of inhibition incubating for 7 days bovine serum albumin (BSA) (50 mg/mL) and glucose (0.8 mol/L) in the presence and absence of these polyphenols at various concentrations. Fluorescence emission at 405 nm was used to evaluate AGEs formation. Carboxymethyl-lysine (CML), Amadori products and quinoproteins were determined by ELISA, NBT assay and electrophoresis, respectively. Both PC and DHPA were able to decrease the formation of Amadori products, fluorescent AGEs and CML. These decreases were concentration dependent. Quinoproteins formation increases with the increase of polyphenol concentration. BSA was also incubated with 0.5 and 1.0 mmol/L of phenols in the absence of glucose. After 7 days of incubation the unbound polyphenols were removed by ultrafiltration. The phenols-treated BSA was incubated with glucose for 7 days. No significant differences are observed in the inhibition of AGEs formation between phenol pre-incubated and non pre-incubated BSA.
These results and the formation of quinone modified BSA suggest that polyphenols are pre-Amadori inhibitors of AGEs formation. Phenols bind BSA and decrease glucose binding. They are oxidized (pH 7.5, 37°C) in the reactive quinones intermediates and react with BSA amino groups forming quinone modified BSA and decreasing the AGEs formation.
2012
- Antioxidant activity of in vitro digested bovine milk
[Abstract in Atti di Convegno]
Verzelloni, Elena; A., Helal; Tagliazucchi, Davide; Conte, Angela
abstract
The objective of the present study was to analyze the antioxidant properties of different types of bovine milk subjected to simulated gastro-pancreatic digestion. Whole, semi-skimmed, and skimmed milk were digested in sequence with pepsin (300 U/ml; pH 2.5; 2h) then with pancreatin (0.8 g/l; pH 7.5; 2h) and bile salts (5 mg/ml). At the end of digestion, samples were subjected to ultrafiltration (3 KDa) and the high (HMW) and low (LMW) molecular weight fractions analyzed with ABTS assay. All milk types contain antioxidants with whole milk having the highest amount (622.3 ± 44.5 mg vitamin C/L). About 90% of antioxidant activity was in the HMW fraction. The degree of protein hydrolysis (DH) after digestion was different considering the different milk types (20.8 ± 0.4, 24.3 ± 0.3 and 30.7 ± 0.8 in whole, semi-skimmed and skimmed milk, respectively). Pancreatic digestion caused an increase in the antioxidant activity for all the analyzed milk (post-pancreatic ABTS value of 3374.3 ± 104.6, 2657.1 ± 39.6 and 2751.2 ± 46.9, in whole, semi-skimmed and skimmed milk, respectively). The distribution of the antioxidant activity between the LMW and HMW fractions was different considering the diverse milk types. The percentage of antioxidant activity in the LMW fraction increased to 38%, 79% and 90% in whole, semi-skimmed and skimmed milk, respectively, according to the increase in the DH.
HPLC analysis of LMW fractions revealed that all the digested milk had the same peptides pattern but the peptides concentrations in skimmed milk was higher than in semi-skimmed and whole milk. On-line HPLC-ABTS analysis of LMW fractions showed that two peptides are the major contributor to the antioxidant activity of this fraction. The two peptides were identified by MS/MS. Our data suggest that digestion of milk increased the antioxidant activity, potentially reducing the oxidative stress in the gastro-intestinal tract.
2012
- Antioxidant properties of polyphenols incorporated in casein/sodiumcaseinate films
[Articolo su rivista]
A., Helal; Tagliazucchi, Davide; Conte, Angela; S., Desobry
abstract
Radical-scavenging activity (RSA) of sodium caseinate (NaCAS) films with 0e30% added casein and antioxidants was measured. Tannic acid and catechin were added to the films as model antioxidants. RSA was measured using 2,2-diphenyl-1-picrylhydrazyl and 2,20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid); the two methods gave similar results for RSA measurement. Film casein content most influenced initial RSA, while increasing casein level resulted in an apparent decrease of RSA due to quenching. During storage, a good stability of RSA was observed. The surface antioxidant activity is of primary interest for food contact materials; a decrease in film surface RSA occurred during the first 20 days of storage, followed by an increase in surface RSA during the remainder of the 90-day storage period, especially at high relative humidity. This phenomenon was due to plasticizing of NaCAS, with possible network alteration over long storage times
2012
- Casein-antioxidants interaction, nutritional effects and technological applications
[Abstract in Atti di Convegno]
A., Helal; Tagliazucchi, Davide; Verzelloni, Elena; S., Desobry; Conte, Angela
abstract
Casein-antioxidants interaction, nutritional effects and technological applications
2012
- Manufacturing of antioxidant packaging Na-caseinate edible films
[Abstract in Atti di Convegno]
A., Helal; Tagliazucchi, Davide; Conte, Angela; S., Desobry
abstract
Manufacturing of antioxidant packaging Na-caseinate edible films
2012
- Pomegranate ellagitannins inhibit in vitro carbohydrate digestion
[Abstract in Atti di Convegno]
Tagliazucchi, Davide; Verzelloni, Elena; Bellesia, Andrea; Conte, Angela
abstract
Pomegranate is an ancient fruit studied for its anti-diabetic properties. The aim of the present work was to identify the α-glucosidase inhibitors in a pomegranate juice polyphenol-rich extract (PPRE). The PPRE, obtained using C18 solid-phase extraction, contained 5.87 ± 0.26 mmol of ellagic acid equivalent (EAE)/L and showed an inhibitor activity on rat intestinal α-glucosidase with an IC50 value of 922.2 ± 7.1 μmol of EAE/L.
The PPRE was fractioned in three different fractions using C18 columns and glucosidase inhibitory activity was recovered in an ellagitannins-enriched fraction. After HPLC separations punicalagin, punicalin and ellagic acid were identified as α-glucosidase inhibitors with IC50 values of 140.2 ± 1.1, 191.4 ± 1.3 and 380.9 ± 3.5 μmol/L, respectively.
Kinetic analysis showed that the inhibition is of mixed type with a Ki of 903.0 ± 74.7 μmol/L.
Pomegranate ellagitannins interact directly with the α-glucosidase and the non-specific binding of ellagitannins may alters the protein structure reducing the velocity of the catalysis and altering the accessibility to the active site of the substrate.
The inhibitory effect on carbohydrate hydrolysis of PPRE was tested against a real food system (potatoes) using an in vitro digestion protocol. The presence of PPRE produced a decrease in the amount of released glucose. During digestion punicalin and punicalagin concentration decreased as a consequence of their binding to salivary or potatoes proteins and of their hydrolysis. Despite this loss, the PPRE retained high inhibitory activity.
In conclusion, our data are consistent with the hypothesis that pomegranate juice can be considered a very useful food supplement to ameliorate postprandial hyperglycemia linked to type 2 diabetes and hyperglycemia-induced vascular complications.
2012
- The Type and Concentration of Milk Increase the in Vitro Bioaccessibility of Coffee Chlorogenic Acids
[Articolo su rivista]
Tagliazucchi, Davide; A., Helal; Verzelloni, Elena; Conte, Angela
abstract
Coffee with different types and concentrations of milk was digested with pepsin (2 h) and pancreatin (2 h) to
simulate gastropancreatic digestion. Chlorogenic acids (CGAs) were determined by high-performance liquid chromatography−
electrospray ionization−tandem mass spectrometry in ultrafiltrate (cutoff 3 kDa) to evaluate their bioaccessibility. After
digestion, bioaccessible CGAs decreased from 80.2 to 53.0 and 69.5 μmol/200 mL in coffee without milk and coffee-whole milk,
respectively. When whole, semiskimmed, skimmed, or diluted milk were present, the increase in bioaccessibility was dependent
on fat content (r = 0.99, p < 0.001). No relationship was observed between bioaccessibility and proteins, carbohydrates, and calcium content. The addition of milk to coffee caused an immediate decrease in the bioaccessibility due to CGAs binding to
proteins. After digestion, 86−94% of bound CGAs remained in the high molecular weight fraction. Casein bound 5-caffeoylquinic
acid with high affinity (KD of 37.9 ± 2.3 μmol/L; n = 0.88 ± 0.06).
2012
- The first tract of alimentary canal as an extractor. Release of phytochemicals from solid food matrices during simulated digestion
[Articolo su rivista]
Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
The release of some phytochemicals (polyphenols, flavonoids, anthocyanins, carotenoids) from peach, plums, prunes, walnuts and tomatoes using an in vitro model simulating oral-gastro-hepato-pancreatic digestion has been investigated. Most of phytochemicals are extracted during the oral phase, and the amount of bioaccessible polyphenols, flavonoids and anthocyanins increased during gastric digestion in all the analyzed fruits. The transition in the hepatopancreatic environment caused a decrease in total polyphenols, whereas flavonoids (except anthocyanins) were not degraded. In most of the tested fruits, the hepatopancreatic digestion did not affectthe extraction of polyphenols and flavonoids except in walnuts, for which this phase caused an important release of both of them. Our results suggest that the first tract of alimentary canal may act as an extractor of phytochemicals where the antioxidantcompounds are progressively released from food matrix and are made available for the absorption or to exert their biological effects in this area.
2011
- Antiglycative and antioxidative properties of coffee fractions
[Articolo su rivista]
Verzelloni, Elena; Tagliazucchi, Davide; D., Del Rio; L., Calani; Conte, Angela
abstract
In this work the inhibitory activity of coffee low molecular weight compound (LMWC) and high molecular weight compound (HMWC) fractions against in vitro advanced glycation end products (AGEs) formation was investigated. The HMWC fraction was characterized for its content in total phenolic groups, proteins and carbohydrates. The chlorogenic acids of LMWC fraction were identified by liquid chromatography coupled with tandem mass spectrometry. HMWC inhibited bovine serum albumin glycation by acting as radical scavenger and Fe-chelator in the post-Amadori phase of the reaction and by inhibiting dicarbonyl reactive compounds production during glucose autoxidation. LMWC fraction was able to inhibit protein glycation and dicarbonyl reactive compounds formation more than HMWC fraction. Chlorogenic acids are the main responsible for the antiglycative activity of LMWC fraction. This study clearly shows that coffee contains molecules with in vitro antiglycative activity and in particular chlorogenic acids are of particular interest for their known bioavailability in vivo.
2011
- Antiglycative and neuroprotective activity of colon-derived polyphenol catabolites
[Articolo su rivista]
Verzelloni, Elena; C., Pellacani; Tagliazucchi, Davide; S., Tagliaferri; L., Calani; L. G., Costa; F., Brighenti; G., Borges; A., Crozier; Conte, Angela; D., Del Rio
abstract
Dietary flavonoids and allied phenolic compounds are thought to be beneficial in the control of diabetes and its complications, because of their ability to inhibit oxidative stress, protein glycation and to act as neuroprotectants. Following ingestion by humans, polyphenolic compounds entering the large intestine undergo extensive metabolism by interaction with colonic microbiota and it is metabolites and catabolites of the parent compounds that enter the circulatory system. The aim of this study was to investigate the inhibitory activity of some colonic microbiota-derived polyphenol catabolites against advanced glycation endproducts (AGEs) formation in vitro and to determine their ability, at physiological concentrations, to counteract mild oxidative stress of cultured human neuron cells. This study demonstrated that ellagitannin-derived catabolites (urolithins and pyrogallol) are the most effective antiglycative agents, whereas chlorogenic acid-derived catabolites (dihydrocaffeic acid, dihydroferulic acid and feruloylglycine) were most effective in combination in protecting neuronal cells in a conservative in vitro experimental model. In conclusion, some polyphenolic catabolites, generated in vivo in the colon, were able in vitro to counteract two key features of diabetic complications, i.e. protein glycation and neurodegeneration. These observations could lead to a better control of these events, which are usually correlated with hyperglycemia.
2011
- Effect of colon-derived polyphenol metabolites on in vitro protein glycation
[Abstract in Rivista]
Conte, Angela; Verzelloni, Elena; D., Del Rio; Tagliazucchi, Davide
abstract
Polyphenols are thought to be beneficial in the control of diabetic complications because of their ability to inhibit oxidative stress and protein glycation. Polyphenols undergo extensive metabolism during their absorption though the first-intestinal tract. Polyphenol catabolites produced by microbia of colon enter the circulatory system. The aim was to investigate the inhibitory activity of colonic-derived catabolites against protein glycation. Protein glycation was measured, after a two weeks incubation under physiological condition (pH 7.4; 37°C) of bovine serum albumin (40 mg/mL) and glucose (30 mmol/L) in presence or absence of polyphenol catabolites, with fluorescence at the excitation and emission maxima of 355 and 405 nm, respectively. A dose dependent inhibition of glycation was observed with urolithin A and B (catabolites of pomegranates and raspberries) and dihydroferulic acid (from coffee.) The IC50 of urolithin A is 12mM. Urolithin B and dihydroferulic acid are less effective. The catabolites were also tested in association, based on the dietary sources. The pomegranate-derived catabolites urolithin A, B and pyrogallol, inhibit glycation by 37 and 44% at 1 and 2µmol/L respectively, suggesting a synergic action between catabolites. The red wine-derived catabolites in association show a lower antiglycative activity (15% inhibition at 2µmol/L) while coffee catabolites are inactive. Polyphenol colon-catabolites inhibit protein glycation at concentrations observed in blood after consumption of a portion of pomegranate or raspberry. These observations suggest that some polyphenols may contribute to reduce the hyperglycemia-correlated pathologies.
2011
- Effect of cow milk on the in vitro bioaccessibility of coffee chlorogenic acids
[Relazione in Atti di Convegno]
Tagliazucchi, Davide; Verzelloni, Elena; A. M. I., Helal; Conte, Angela
abstract
To study the effect of cow milk on coffee chlorogenic acids (CQAs) bioaccessibility during simulated gastro-intestinal digestion, 50% coffee with 50% water (C), or with 50% whole milk (CM50), or with 25% whole milk-25% water (CM25), or with 10% whole milk- 40% water (CM10), or with 50% semi-skimmed milk (CSSM50), or with 50% skimmed milk (CSM50) were digested in sequence with pepsin (300 U/ml; pH 3; 2h) then with pancreatin (0.8 g/l; pH 7.5; 2h) and bile salts (5 mg/ml). At the end of the digestion periods, samples were subjected to ultrafiltration and the low molecular weight fractions were analyzed with LC-MS. The bioaccessible CQAs decreased from 91.4 ± 1.3 to 85.2 ± 5.4 and to 61.2 ± 2.0 micromol/100ml at the end of gastric and intestinal digestions respectively in sample C, suggesting CQAs degradation. After the two digestion periods higher recovery of CQAs were obtained with CM50, CM25, CM10 and CSSM50 samples respect to C. For example at the end of the digestion periods, the bioaccessible CQAs in CM50 and CSSM50 were 80.0 ± 4.6 and 67.7 ± 1.0 micromol/100 ml respectively. Similar results were obtained with 5-caffeoylquinic acid. These data suggest that the milk fats have a protective effect on CQAs degradation during digestion.
2011
- Interaction of colon-derived polyphenol metabolites with bovine serum albumin
[Abstract in Atti di Convegno]
Tagliazucchi, Davide; D., Del Rio; Verzelloni, Elena; L., Calani; A., Crozier; Conte, Angela
abstract
Following ingestion, polyphenols entering the large intestine undergo extensive metabolism by colonic microbiota and metabolites of the parent compounds enter the circulatory system. It is of limited relevance attempting to unravel the mechanism of action of intact parent polyphenols. Their colonic metabolites are more plausible candidates for investigation of putative biological activity. The aim of this work was investigating the interaction between BSA and polyphenol metabolites using tryptophan fluorescence quenching. Fluorescence spectra of BSA were recorded before and after the addition of selected metabolites, under physiological conditions (37°C and pH 7.4), in the range of 300-550 nm at an excitation wavelength of 294 nm. Calculation of KSV demonstrated that, for all the tested metabolites (with the exception of 3-hydroxyphenylacetic and 3,4-dihydroxyphenylacetic acids), the type of quenching was static, suggesting the formation of a complex between the molecules and the fluorophore (tryptophan). Urolithins had the lowest KD value and bound BSA with higher affinity with respect to the other tested compounds. The binding stoichiometry was 1:1 for all the tested metabolites, suggesting the existence of a single binding site in the protein. Competitive binding experiments with quercetin showed a displacement of urolithins from the complex, demonstrating that site I (the hydrophobic binding pocket located in the subdomain IIA of BSA, which is known to bind a variety of ligands) is likely to be the preferred binding site for these molecules. The affinity of BSA for urolithins lets hypothesise that these phenolic metabolites could circulate in vivo as protected by plasma proteins. In the presence of solid tumors there is a reduction in tumor venous blood pH below 7, as a consequence of lactate accumulation. This acidic environment may cause the release of urolithins due to pH-induced decreased affinity, and these phenolic metabolites can exert local anticancer effects.
2010
- Changes in major antioxidant compounds during aging of traditional balsamic vinegar
[Articolo su rivista]
Verzelloni, Elena; Tagliazucchi, Davide; Conte, Angela
abstract
Traditional balsamic vinegar (TBV) shows antioxidant capacity that increases passing from cask 5, containing the youngest vinegar, to cask 1 containing the oldest vinegar. This increase in antioxidant capacity is a consequence of both the concentration of compounds already present and of new antioxidants formation and is positively related to the increase in the polyphenolic content and in the Maillard reaction/caramellisation products. During TBV ageing, some reactions involving flavonoids and tannins take place. Tannins can undergo acid-catalyzed cleavage of their interflavonoid bonds with subsequent condensation of other flavonoid molecules. In addition, the low pH, the decrease of the water content and the presence of aldehydes, may promote flavonoids polycondensation. These reactions explain the observed increase in polymeric phenolic compounds and the decrease in monomeric flavonoids. During TBV ageing an increase in the browning index is observed as a consequence of the polycondensation reactions of flavonoids and of brown melanoidins accumulation.
2010
- Contribution of melanoidins to the antioxidant activity of traditional balsamic vinegar during ageing
[Articolo su rivista]
Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
The contribution of high molecular weight melanoidins to the overall antioxidant activity of traditional balsamic vinegar has been determined. High molecular weight melanoidins are gradually synthesized and accumulated during traditional balsamic vinegar ageing contributing significantly to the browning of the vinegar. The increase in antioxidant activity of traditional balsamic vinegar during ageing is largely due to the formation of high molecular weight melanoidins and only in lower lesser part to the concentration of polyphenols deriving from the grape or to the formation of low molecular weight Maillard reaction or caramelisation products during must cooking and vinegar ageing. During ageing, low molecular weight compounds are progressively incorporated into the melanoidins skeleton and may contribute significantly to the antioxidant activity of high molecular weight melanoidins. Among these compounds, it has been shown that antioxidant phenolic compounds are progressively incorporated into the melanoidins skeleton during traditional balsamic vinegar ageing.
2010
- Effect of dietary melanoidins on lipid peroxidation during simulated gastric digestion: their possible role in the prevention of oxidative damage
[Articolo su rivista]
Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
The ability of high molecular weight melanoidins extracted from coffee, barley coffee and dark beer to inhibit lipid peroxidation during simulated gastric digestion of turkey meat has been investigated. Results showed that melanoidins decrease the synthesis of lipid hydroperoxides and secondary lipoxidation products. Coffee melanoidins at 3 mg/ml reversed the reaction and broke down hydroperoxides to concentration lower than the initial value. Barley coffee and dark beer melanoidins were less effective and even at 12 mg/ml did not reverse the reaction. The proposed mechanism of action involved Fe2+ chelating capacity, heme binding ability and radical scavenging activity. Melanoidins were characterized for their content in total proteins, carbohydrates and phenolics and the relationship between the chemical composition and the antioxidant activity of dietary melanoidins was investigated. Coffee melanoidins that contain more phenolics and proteins respect to the other melanoidins, showed greater antioxidant activity respect to the other melanoidins tested.
2010
- From balsamic to healthy: traditional balsamic vinegar melanoidins inhibit lipid peroxidation during simulated gastric digestion of meat
[Articolo su rivista]
Verzelloni, Elena; Tagliazucchi, Davide; Conte, Angela
abstract
In this work traditional balsamic vinegar (TBV) melanoidins were characterized for chemical composition and antioxidant activity and their antiperoxidative effect during an in vitro gastric digestion of turkey meat was studied. The most important constituents of TBV melanoidins were carbohydrates (51% w/w) of which glucose (35% w/w) and fructose (10% w/w) are the main representatives, hydroxymethylfurfural (HMF) (7.2% w/w), phenolic groups (4.6% w/w) and proteins (1.2% w/w). The antioxidant capacity of melanoidins was studied, measuring lipid hydroperoxides and secondary lipoxidation products formed during in vitro gastric digestion of turkey meat. The most important mechanisms in their antioxidant activity resulted radical scavenging and Fe2+-chelating activities. Pepsin inhibiting ability has been excluded.. TBV melanoidins were also able to bind heme under gastric conditions potentially preventing its absorption and prooxidant and cytotoxic effects. Our results support the idea that TBV melanoidins may have a role in oxidative damage prevention. Fe2+-chelating and heme-binding activities as well as mechanisms of antioxidant activity of TBV melanoidins were also compared with coffee, barley coffee and dark beer melanoidins.
2010
- In vitro bio-accessibility and antioxidant activity of grape polyphenols
[Articolo su rivista]
Tagliazucchi, Davide; Verzelloni, Elena; D., Bertolini; Conte, Angela
abstract
The bio-accessibility (the release of compounds from solid food matrices) of grape polyphenols using an in vitro model simulating gastro-intestinal conditions has been investigated. In vitro studies are needed to unravel the factors affecting the release of antioxidants during digestion. The amount of bio-accessible polyphenols, flavonoids and anthocyanins increases during gastric digestion. The transition in the intestinal environment causes a decrease in all the analyzed classes of polyphenols followed by a renewal in the extraction of polyphenols and flavonoids but not of anthocyanins. The stability under gastro-intestinal conditions of pure phenolic acids, flavonoids and resveratrol has been analyzed and discussed. Changes in antioxidant activity during digestion were correlated to the changes in polyphenols concentration as well as to the pH. Our results suggest that the gastro-intestinal tract may act as an extractor where polyphenols are progressively released from solid food matrix and made available for the absorption or to exert their biological effects in the gastro-intestinal tract.
2010
- Sugar Conversion Induced by the Application of Heat to Grape Must
[Articolo su rivista]
Falcone, Pasquale Massimiliano; Tagliazucchi, Davide; Verzelloni, Elena; Giudici, Paolo
abstract
Two lots of the grape Trebbiano cultivar were harvested from the same vineyard 15 days apart, and their musts were cooked in an open stainless steel pan directly heated by fire; the kinetics of formation or disappearance of key constituents was then monitored for at least 16 h. From an engineering standpoint, the vessel behaved like a nonisothermal batch reactor in which the volume of the grape must necessarily decreased while its composition changed profoundly as a result of chemical reactions. Brix, total titratable acids, acetic acid, malic acid, lactic acid (D and L), pH, water activity, 5-HMF, and phenolic and radicalscavenging compounds were proposed as markers of the extent of cooking for which water vaporization and sugar degradation were identified as the two main driving factors. Acid-catalyzed dehydration was hypothesized as the predominant mechanism for sugar degradation, assuming a direct role of water vaporization; however, contributions of Maillard degradation pathways and other parallel reversible reactions were also hypothesized. Fractional conversion of 5-HMF and radical-scavenging compounds were proposed as quantitative markers for the extent of sugar degradation at, respectively, the early and advanced stages of cooking. Selectivity indices were also proposed as a performance criterion to design cooking processes in relation to sugar degradation.
2008
- A rheological approach to the quantitative assessment of traditional balsamic vinegar quality
[Articolo su rivista]
Falcone, Pasquale Massimiliano; Verzelloni, Elena; Tagliazucchi, Davide; Giudici, Paolo
abstract
This work aimed at evaluating the ability of rheological measurements to describe the quality of Traditional Balsamic Vinegars (TBV) with respect to their composition and sugar crystallization. With this aim, shearing experiments were conducted on 100 samples of TBV using a controlled-strain rotational rheometer, whereas the concentration of glucose, fructose, acetic acid, gluconic acid, malic acid as well as refractive index and titrable acidity were determined according to the normal procedure for quality assessment of TBV. A broadrange for the rheological response was observed including Newtonian and shear-thinning behavior. A Joining Tree Cluster Analysis (JTCA) was able to classify the investigated vinegars into meaningful classes based on the stress data alone. Moreover, Principal Component Analysis (PCA) was performed to link composition data to rheological properties, and finally, Multivariate Regression Analysis (MRA) evaluated the contribution of the main constituents to vinegar viscosity. The results suggested that rheological analysis must be carried out in order to spot some fundamental aspects of the vinegar texture, which cannot be detected otherwise. In particular, two rheological parameters were proposed as objective and quantitative descriptors of TBV quality: the shear viscosity measured at500 s-1 accounting for the vinegar composition, and the flow behavior index accounting for sugar crystallization.
2008
- Antioxidant properties of traditional balsamic vinegar and boiled must model systems
[Articolo su rivista]
Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
Traditional balsamic vinegar (TBV) is a natural product prepared with cooked and concentrated locally grown grape must. It has been demonstrated that TBV contains phenols and shows antioxidant activity. In this study we investigated the antioxidant properties of TBV in relation to its content in phenolic compounds, polymeric tannins and Maillard reaction products (MRPs). Results show that TBV has a high antioxidant activity measured with both FRAP and ABTS assay that is higher or equal to those obtained in some red wine. About 45% of the antioxidant activity of TBV is due to the total polyphenolic fraction. Among polyphenols, tannins contribute to about 50% of the antioxidant activity of the total polyphenolic fraction. The residual antioxidant activity of TBV is due to the melanoidins (about 45%) synthesized during the boiling of the must and the ageing of TBV and to other compounds such as low molecular weight MRPs. When we investigated the effect of heating on the browning and on the formation of antioxidant MRPs in must model systems we observed a major formation of antioxidant MRPs for the model system containing both amino acids and sugars with respect to the model system containing only sugars. We also tested the effect of some representative phenolic compounds present in must. Only polyphenols were stable in the model solution, however, in our experimental conditions they did not influence the browning and the formation of MRPs. Indipendently of their bioavailability, dietary antioxidants play an important role in protecting the gastrointestinal tract itself from oxidative damage and could protect against a built up of peroxides and their assimilitation in the digestive tract.
2007
- Occurence of two NOS isoforms in the developing gut of sea bass Dicentrarchus labrax (L.)
[Articolo su rivista]
Pederzoli, A.; Conte, A.; Tagliazucchi, D.; Gambarelli, A.; Mola, L.
abstract
In this work we have examined the appearance and distribution of nitric oxide synthase (NOS), with histochemical, immunohistochemical and biochemical methods, during development of the sea bass (Dicentrarchus labrax) gut. The data showed that both the calcium-calmodulin dependent neuronal isoform (nNOS) and calcium-independent inducible isoform (iNOS) are present in the larval gut of sea bass. The nNOS-immunoreactivity was present in the epithelial cells and enteric nerve cells of gut both in the 8-day-old specimens and in the 24-day-old-larvae. In the adult nNOS-immunoreactivity disappeared from epithelial cells, remaining in the wall intramural neurons and fibers. The iNOS-immunoreactivity was present in the epithelial cells of 24-day-old-larvae and was not detectable in the adult gut. Western blot analysis and determination of NOS activity also demonstrated the presence of the two NOS isoforms, nNOS and iNOS, in the gut of 24-day-old specimens. The presumably different roles played by the two isoforms of enzyme are discussed. The presence of nNOS isoform in the gut enteric neurons of the same larval stages of D. labrax in which we previously demonstrated the presence of substance P and Vasoactive Intestinal Polypeptide (VIP), may suggest that all these three components of the motility control system are already present in the larval phase. Nitric oxide (NO) may be also involved in the early immune response. The present results on the occurrence of iNOS isoform in epithelial gut cells of the same regions in which the gut-associated lymphoid tissue (GALT) will differentiate, may suggest for NO a role in early defence mechanisms, before the establishment of immune responses in GALT. Finally, the developmental and regional differences in nNOS and iNOS expression also suggest a regulatory role in development and differentiation of the sea bass gut.
2007
- Relationship between the antioxidant properties and the phenolic and flavonoid content in traditional balsamic vinegar
[Articolo su rivista]
Verzelloni, Elena; Tagliazucchi, Davide; Conte, Angela
abstract
The antioxidant properties of traditional balsamic vinegar as regards its phenolic and flavonoid content comparing it to selected vinegars and red wines have been investigated. The polyphenols were separated from interfering compounds utilizing C18 columns. The polyphenolic content was determined utilizing both Folin–Ciocalteu and peroxidase assays. The antioxidant capacity was quantified using both ABTS and FRAP assays. The results show that traditional balsamic vinegar has lower antioxidant activity and phenolic and flavonoid content than Nero d’Avola but higher than the other tested products. The antioxidant capacity of wines and vinegars is highly correlated with their phenolic content, measured by peroxidase assay and it is also highly correlated with their flavonoid content while in traditional balsamic vinegar and balsamic vinegar this correlation diminishes. The study describes a simple and fast method of separating from other compounds and of measuring polyphenols in the analysis of red wines and vinegars with complex composition such as traditional balsamic vinegar.
2006
- Confronto fra le proprietà antiossidanti e il contenuto fenolico di vini rossi e aceto balsamico tradizionale
[Relazione in Atti di Convegno]
Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
Negli ultimi anni i composti polifenolici sono stati studiati per le loro proprietà antiossidanti e i potenziali effetti benefici sulla salute umana. Nel presente lavoro sono state studiate le proprietà antiossidanti e il contenuto polifenolico totale di alcuni campioni di vino rosso e di aceto balsamico tradizionale (ABT). Per determinare il contenuto polifenolico totale sono stati utilizzati il saggio di Folin-Ciocalteu ed il saggio della perossidasi. Sulla base dei nostri risultati, il saggio della perossidasi sembra essere il metodo più preciso per determinare il contenuto polifenolico in matrici estremamente complesse come l’ABT. Considerando i dati ottenuti con il saggio della perossidasi, l’ABT contiene meno polifenoli del Nero d’Avola ma più polifenoli degli altri vini analizzati. Le proprietà antiossidanti dei campioni sono state analizzate in funzione di quello che era il loro contenuto in polifenoli e analizzando quelle che erano le capacità di ciascun campione di inattivare i radicali liberi generati in fase acquosa e di inibire la perossidazione lipidica durante una digestione simulata di carne suina. I risultati mostrano come la capacità antiossidante totale dei campioni era: Nero d’Avola > ABT > Dolcetto d’Alba > Lambrusco. Inoltre, sia il vino rosso sia l’ABT sono in grado di inibire in maniera significativa la formazione di perossidi lipidici durante una digestione simulata di carne suina. In conclusione, l’ABT rappresenta un ottimo condimento che contribuisce positivamente sia sulla qualità dei pasti che sulla quantità totale di antiossidanti introdotti durante i pasti.
2006
- Effect of chain length and aldehydic function on some biological properties of parropolyenes
[Articolo su rivista]
Tagliazucchi, Davide; Verzelloni, Elena; E., Pini; G., Ronca; Conte, Angela
abstract
Parropolyenes, also called parroenes, parrodienes and psittacofulvines, are aldehydes costituted of an unsaturated chain ranging from 8 to 18 carbon atoms. These compounds have been isolated from the plumage of parrots and are also synthesized in the laboratory. Parropolyenes have antiproliferative and antioxidant activities. We investigated the effect of chain length and the role of aldehydic function of some parropolyenes by comparing their biological activities with those of their corresponding alcohols. Trans, trans-Δ2,4-hexadienal at a concentration of 100 μM significantly inhibited the growth of SH-SY5Y cells after 72 h of incubation. The corresponding alcohol was less effective. All-trans-Δ2,4,6-octatrienal and all-trans-Δ2,4,6,8,10-dodecapentaenal were more effective since significant inhibition of growth was observed at a concentration of 10 μM. Again, the corresponding alcohols were less effective. All-trans-Δ2,4,6-octatrienal and all-trans-Δ2,4,6,8,10-dodecapentaenal at 1 μM concentration protected 2-deoxyribose from degradation by ferrous ions. Trans, trans-Δ2,4-hexadienal at the same concentration does not show this protective effect. The protection increased with increasing parropolyene chain length. No significant difference was observed between aldehydes and their corresponding alcohols. Parropolyenes increased the rate of lysis of erythrocyte membrane in a lactoperoxidase-potassium iodide-hydrogen peroxide assay. The cytolytic effect increased with increasing parropolyene chain length. In this assay no difference was observed between aldehydes and alcohols. In all assays the effects were concentration dependent.
2006
- I Microrganismi dell'aceto balsamico. Atti del convegno: "Anche la tradizione va studiata" Risultati di un biennio di indagini per l'individuazione e il possibile impiego di starter per Aceto Balsamico Tradizionale.
[Curatela]
Licciardello, Fabio; Falcone, Pasquale Massimiliano; Tagliazucchi, Davide; Giudici, Paolo; Solieri, Lisa; Gullo, Maria; DE VERO, Luciana
abstract
Il ruolo e l'importanza dei microrganismi sulla qualità dell'aceto balsamico tradizionale.
2006
- L’approccio reologico nella valutazione della qualità degli Aceti Balsamici Tradizionali
[Relazione in Atti di Convegno]
P. M., Falcone; S., Chillo; Verzelloni, Elena; Tagliazucchi, Davide; Giudici, Paolo; M. A., Del Nobile
abstract
In questa ricerca è stata valutata la possibilità di studiare in modo oggettivo la qualità degli Aceti Balsamici Tradizionali (ABT) e come questa dipende dalla composizione e stato fisico dei soluti, attraverso la determinazione della viscosità dinamica. Considerata la complessità e la naturale variabilità delle materie prime impiegate nella produzione di tali aceti, questo studio è stato condotto in due fasi. Nella prima fase è stato utilizzato un sistema modello contenente i principali costituenti degli ABT quali glucosio, fruttosio, acido acetico e acqua. Allo scopo di esplorare un ampio intervallo di composizione, la concentrazione dei singoli soluti è stata modulata in accordo ad un piano sperimentale frazionario a cinque livelli e tre variabili. La viscosità dinamica è stata determinata utilizzando un reometro rotazionale. L’influenza della composizione sulla viscosità dinamica è stata studiata attraverso l’analisi statistica basata sulla Metodologia delle Superfici di Iso-Risposta. NelIa seconda fase è stato possibile verificare le correlazioni osservate tra composizione e viscosità direttamente su sistema reale, attraverso l’analisi reologica di alcuni ABT con diversa composizione. Sono stati analizzati anche campioni di ABT contenenti soluti in forma cristallina. I risultati dimostrano che la frazione ponderale di glucosio e il rapporto glucosio/fruttosio influiscono in modo complesso e determinante sulla viscosità delle soluzioni modello. Nel caso degli ABT il maggiore contributo alla viscosità dinamica è dato dai soluti minori i quali nel loro insieme determinano livelli di viscosità mediamente due ordini di grandezza superiori a quelli delle soluzioni modello. L’analisi reologica può risultare utile nel fornire informazioni oggettive su alcuni aspetti della qualità degli ABT legati alla concentrazione e stato fisico dei soluti.
2006
- RELATIONSHIP BETWEEN THE ANTIOXIDANT PROPERTIES AND THE PHENOLIC AND FLAVONOID CONTENT IN TRADITIONAL BALSAMIC VINEGAR
[Relazione in Atti di Convegno]
Verzelloni, Elena; Tagliazucchi, Davide; E., Panicucci; Conte, Angela
abstract
The antioxidant properties of traditional balsamic vinegar as regards its phenolic and flavonoidcontent comparing it to selected vinegars and red wines have been investigated. Thepolyphenols were separated from interfering compounds utilizing C18 columns. Thepolyphenolic content was determined utilizing both Folin-Ciocalteu and peroxidase assays. Theantioxidant capacity was quantified using both ABTS and FRAP assays. The results show thattraditional balsamic vinegar has lower antioxidant activity and phenolic and flavonoid contentthan Nero d’Avola but higher than the other tested products. The antioxidant capacity of winesand vinegars is highly correlated with their phenolic content, measured by peroxidase assay andit is also highly correlated with their flavonoid content while in traditional balsamic vinegar andbalsamic vinegar this correlation diminishes suggesting that other antioxidant compounds arepresent. The study describes a simple and fast method of separating from other compounds andof measuring polyphenols in the analysis of red wines and vinegars with complex compositionsuch as traditional balsamic vinegar.
2006
- Vinegar polyphenols increase the proteolysis and decrease the oxidative stress during digestion by pepsin
[Abstract in Atti di Convegno]
Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
Vinegar polyphenols increase the proteolysis and decrease the oxidative stress during digestion by pepsin
2005
- Calcium/calmodulin dependence of nitric oxide synthase from Viviparus aeter immunocytes
[Articolo su rivista]
Tagliazucchi, Davide; Conte, Angela
abstract
The calcium ion dependence of soluble and particulate NOS activity from Viviparus ater hemocytes has been investigated. At 2 nM calcium ion concentration the NOS activity measured by citrulline formation is 27.1 ± 2.2 and 9.3 ± 0.8 pmol.min-1.106cell-1 for soluble and particulate NOS respectively. The increase of free calcium ion concentration up to 300 nM increases the enzyme activity to 57.5 ± 4.1 and 23.5 ± 1.2 respectively. The 50% activation of the calcium dependent activity is 91 and 97 nM Ca2+ for soluble and particulate enzyme. Trifluoperazine, an inhibitor of calmodulin dependent enzyme, partially inhibits both activities. The soluble NOS is five time more sensible than particulate NOS. The behaviour of both activities to three NOS inhibitors (7-nitroindazole, S-methylisothiourea sulfate, diphenyleneiodonium) is very similar with not significant differences in IC50 values. The calcium ion dependence of NOS activities in a range of free calcium ion variations which are transiently observed in receptor stimulated cells suggests that NO in V. ater hemocytes has not only a defensive role but also a signalling relevance in cross-talking between hemocytes and other cells.
2005
- Differente tolleranza all’ossigeno e dipendenza dal Ca2+ della NOS nel cervello di tre diverse specie di pesci
[Abstract in Atti di Convegno]
Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
nd
2005
- Effect of some phenolic compounds and beverages on pepsin activity during simulated gastric digestion
[Articolo su rivista]
Tagliazucchi, Davide; Verzelloni, Elena; Conte, Angela
abstract
The effect of some polyphenols (resveratrol, catechin, epigallocatechin-3-gallate, and quercetin) and beverages (red wine and green tea) on the enzymatic activity of pepsin during the digestion of three different substrates (pork meat, insoluble azocasein, and denatured hemoglobin) has been investigated. The tested polyphenols and beverages increase the initial velocity of the reaction, and the activating effect is concentration dependent. The order of effectiveness of polyphenols in increasing the initial velocity of the reaction is resveratrol >= quercetin > epigallocatechin-3-gallate > catechin. The kinetic data obtained with soluble denatured hemoglobin show that the Km for the substrate is not changed, whereas the V-max of the reaction is increased. Pepsin activity follows a simple Michaelis-Menten kinetic suggesting that k(3) is increased by polyphenols. To the authors' knowledge, the present study is the first demonstration that some polyphenols and related beverages are able to enhance the enzymatic activity of pepsin.
2005
- Occurrence of nitric oxide synthase in Megoura viciae Buckton (Homoptera, Aphididae): an histochemical and immunohistochemical localisation
[Articolo su rivista]
S., Ganassi; Tagliazucchi, Davide; Mola, Lucrezia
abstract
Nitric oxide (NO) is known to be involved in many physiological reactions of insects. We analysed NOS localisation in aphids of the species Megoura viciae by means of histochemical reaction for the NADPH-diaphorase activity and immunohistochemical methods for uNOS, nNOS and iNOS. The obtained data provided a complex and peculiar pattern of NOS distribution in cells and tissue of M. viciae.The histochemical reaction for NADPH-diaphorase was an indicative, but not exact marker of NOS localisation in aphids. The use of anti uNOS antiserum (frequently applied in insects) was of limited value in our specimens, whereas more satisfactory results were obtained with anti nNOS and iNOS antisera of human origin. The results of Western blot analysis confirmed the immunohistochemical ones, showing an aphid protein that reacted strongly with the polyclonal antibody anti-iNOS and anti-nNOS while a similar protein band was weakly immunoreactive with the polyclonal antibody anti-uNOS. Our results suggest that NO, prevalently synthesised by calcium/calmodulin-dependent isoform, plays important physiological roles both in adult and embryological stages of aphids. The data of principal interest was NOS presence in bacteriocytes, cells that host symbiotic prokaryotes belonging to the species Buchnera aphidicola, and in nuclei of adipocytes and gut cells.
2005
- Variazione plasmatica delle HSPs 70 in pazienti gonartrosici sottoposti a terapia termale con applicazione di fanghi caldi delle Terme di Castrocaro
[Relazione in Atti di Convegno]
G., Ronca; G., Agostini; F., Russo; M., Conti; Tagliazucchi, Davide; Conte, Angela
abstract
Le proteine dello shock termico o heat shock protein (HSP) sono proteine che aumentano nelle cellule in seguito a stress di varia natura, il primo dei quali ad essere stato identificato è stato quello termico. In questa ricerca abbiamo valutato il livello di HSP70 e HSP60 nel plasma di 20 pazienti maschi gonartrosici trattati in ambiente termale con 9 applicazioni di fanghi caldi (50°C) per 20 minuti sul 50-70% della superficie corporea. Le HSP70 aumentavano già alla terza applicazione, l'aumento diventa significativo alla nona applicazione e permane anche dopo 30-35 giorni. L'aumento si correla al miglioramento degli indici patologici della gonartrosi. Le HSP 60 invece non subiscono modificazione. L'applicazione dello stimolo termico in ambiente termale determina un aumento delle difese intracellulari, modula la risposta infiammatoria sia locale che generale diminuendo il dolore e migliorando la funzione articolare.
2005
- Variazione plasmatica delle HSPs70 in pazienti gonartrosici sottoposti a terapia termale con applicazione di fanghi caldi delle Terme di Castrocaro(FC)
[Articolo su rivista]
G., Ronca; G., Agostini; F., Russo; M., Conti; Tagliazucchi, Davide; Conte, Angela
abstract
Le proteine dello shock termico o heat shock protein (HSP) sono proteine che aumentano nelle cellule in seguito a stress di varia natura, il primo dei quali ad essere stato identificato è stato quello termico. In questa ricerca abbiamo valutato il livello di HSP70 e HSP60 nel plasma di 20 pazienti maschi gonartrosici trattati in ambiente termale con 9 applicazioni di fanghi caldi (50°C) per 20 minuti sul 50-70% della superficie corporea. Le HSP70 aumentavano già alla terza applicazione, l'aumento diventa significativo alla nona applicazione e permane anche dopo 30-35 giorni. L'aumento si correla al miglioramento degli indici patologici della gonartrosi. Le HSP 60 invece non subiscono modificazione. L'applicazione dello stimolo termico in ambiente termale determina un aumento delle difese intracellulari, modula la risposta infiammatoria sia locale che generale diminuendo il dolore e migliorando la funzione articolare.
2004
- NO in the developing gut of Dicentrarchus labrax: an early immune role ?
[Abstract in Rivista]
Mola, Lucrezia; Pederzoli, Aurora; Gambarelli, Andrea; Tagliazucchi, Davide; Conte, Angela
abstract
It is known that nitric oxide (NO) plays an important role in the immune-neuroendocrine communications. In this issue, we have examined the appearance and distribution of nitric oxide sinthase (NOS) histochemically, immunohistochemically and biochemically during development of the sea bass gut. In 4 and 24-day-old larvae the NOS activity evaluated by NADPH-diaphorase was strong in all epithelial cells and in cell and fibers of intestinal wall, at all gut levels. In the same larval stages and localizations immunoreactive material to antibodies against nNOS and iNOS was present. In the 5-month-old adult gut both enzimatic activity for NADPH-diaphorase and immunoreactivity (IR) to anti-nNOS disappeared from epithelium remaining in the gastro-enteric nervous system. The IR to anti-iNOS completely disappeared. Western blot analysis showed that neuronal (about 150,000 mol. wt band) and inducible (about 135,000 mol wt band) NOS-immunoreactive proteins are present in 24 day-old larvae gut. In the 5 month-old adult gut nNOS and iNOS IR disappeared in the soluble fraction of crude gut homogenates. A small amount of nNOS IR was present in particulate gut fraction. Our data show that both the calcium-calmodulin dependent nNOS and calcium-independent iNOS are present in the larval gut of sea bass. In this species the maturation of cell-mediated immune responses and humoral immune system takes place respectively around the first and second month post hatching (Scapigliati et al., 2002). The presence of inducible NOS in the same regions of the sea bass gut in which the GALT will differentiate, may suggest for NO a role in early defence mechanisms, before of establishment of immune responses in GALT.
2003
- Effect of resveratrol and catechin on PC12 tyrosine kinase activities and their synergistic protection from beta-amyloid toxicity
[Articolo su rivista]
Conte, Angela; S., Pellegrini; Tagliazucchi, Davide
abstract
Beta-Amyloid peptide (beta-AP) is the main component of amyloid deposits around the cerebral vessel and in the brain parenchyma in Alzheimer's disease and Down's syndrome. In vitro studies in neuronal cells or in PC12 and Hela cell lines have shown that the aggregate form of beta-AP is toxic. Many genetic and environmental factors including metal ions, proteoglycans, plasma proteins and antioxidants modify beta-AP toxicity. We investigated the effect of two plant polyphenols-resveratrol and catechin-on soluble and particulate tyrosine kinase activity from PC12 cells and the protective action of these compounds against beta-AP (1-41) toxicity. beta-AP (1-41) decreased PC12 viability with an IC50 value of 1.1 +/- 0.14 x 10(-8) M. Resveratrol and catechin protected PC12 cells from beta-AP (1-41) toxicity. With 25 muM resveratrol the IC50 value increased to 2.2 +/- 0.19 x 10(-7) M. In the presence of beta-AP (1-41) resveratrol showed a concentration-dependent biphasic effect, and at a concentration of up to 40 muM it protected PC12 cells from beta-AP (1-41) toxicity. At concentrations higher than 40 muM, an inhibitory activity on cell proliferation appeared. This antiproliferative effect was also seen in the absence of beta-AP (1-41). With 100 muM catechin the IC50 value increased from 1.1 +/- 0.14 x 10(-8) M to 3.2 +/- 0.25 x 10(-7) M beta-AP (1-41). The protective effect was concentration dependent. Resveratrol and catechin had a synergistic protective action. In the presence of 40 muM catechin and 10 muM resveratrol or 20 muM resveratrol and 10 muM catechin, the toxicity determined by 10(-7) M beta-AP (1-41) was almost completely removed. Resveratrol and catechin had different effects on PC12 tyrosine kinase activity. With peptide 1-17 of gastrin as substrate, resveratrol inhibited particulate tyrosine kinases while it had no effect on soluble activity. With the same substrate, catechin increased the activity of soluble fraction while it inhibited particulate activity When peptide 6-20 of cell division kinase p34(cdc2) was utilized, catechin showed an opposite effect, inhibiting soluble tyrosine kinase activity and increasing particulate activity With peptide 6-20, resveratrol inhibited both soluble and particulate activities. These results demonstrate that resveratrol and catechin have different activities on the signal transduction pathway involving protein phosphorylation. These differences may contribute not only to the different effects of these compounds on PC12 growth but also to the synergistic effect against beta-AP (1-41) toxicity The different activity of resveratrol and catechin on signal transduction pathways, as well as the differences in metal chelation, partition coefficient between water and lipids, hydrogen donation redox potential and enzyme inhibition may be at least in part based on synergistic protection against beta-AP (1-41) toxicity.
2003
- Relationship between iron and protein content of dishes and polyphenol content in accompanying wines
[Articolo su rivista]
G., Ronca; L., Palmieri; S., Maltinti; Tagliazucchi, Davide; Conte, Angela
abstract
The traditional combination of wines and dishes is highly complex, elaborated and refined. The aim of this study was to investigate the possible relationship between the chemical composition of wines and dishes that determines their combination. We determined the content of total polyphenols in 56 wines. The content of total proteins, total lipids, kilocalories, sodium, potassium, calcium, copper and zinc were determined in 44 raw foods and 44 dishes. Nine gourmets independently chose three wines for each food. We correlated the content of the chemical constituents of foods with the phenol content of wines combined with each food by the gourmets. A significant positive correlation was obtained between the phenol content of wines and iron (r = 0.81, p < 0.0001) and total protein content (r = 0.66, p < 0.0001) of foods. Nine gourmets composing a second panel chose three wines for each dish. A significant positive correlation was also obtained between the phenol content of wines and iron (r = 0. 69, p < 0. 000 1), total protein (r = 0. 50, p < 0. 0006) and potassium (r = 0. 45, p < 0. 002) in dishes combined with wines by the second panel of gourmets. Plant phenols decrease the intestinal absorption of iron and have antioxidant activity in the intestinal tract and elsewhere in the body These positive effects compensate the negative antinutritional activity toward protein digestion. The traditional combination of wines and dishes appears to be very favorable since wines poor in phenols are combined with dishes poor in iron and/or proteins to minimize their possible antinutritional effects, while phenol-rich wines are combined with dishes rich in iron to decrease iron absorption and prandial peroxidative stress.
2003
- Synergistic protection of PC12 cells from β-amyloid toxicity by resveratrol and catechin
[Articolo su rivista]
Conte, Angela; S., Pellegrini; Tagliazucchi, Davide
abstract
Beta-Amyloid peptide (beta-AP) elicits a toxic effect on neurons in vitro and in vivo. Many environmental factors including antioxidants, metal ions and proteoglycans modify beta-AP toxicity. We have investigated on PC12 cells, the protective effect from beta-AP (1-41) of two plant polyphenols, resveratrol and catechin. PC12 cells treated with 10(-6) M beta-AP (1-41) for 16 h decrease the cell viability at 24% of the control with an IC50 value of 1.1 +/- 0.14 x 10(-8) M. Twenty-five micromolar resveratrol and 50 muM catechin protect PC12 cells from beta-AP (1-41) toxicity with the IC50 value increased at 2.2 +/- 0.19 x 10(-7) M and at 8.9 +/- 0.7 x 10(-8) M, respectively. While the protective effect is concentration dependent for catechin, resveratrol shows a concentration-dependent biphasic effect. Up to 50 muM concentration, resveratrol protects PC12 cells from beta-AP (1-41) toxicity. At concentration higher than 50 muM, an inhibitory activity on cell proliferation appears. This antiproliferative effect is shown also in the absence of beta-AP (1-41). Resveratrol and catechin have a synergistic protective action. In the presence of 50 muM catechin and 10 muM resveratrol or 25 muM resveratrol and 10 muM catechin, the toxicity determined by 10(-7) M beta-AP (1-41) is almost completely abolished. Catechin is more effective than resveratrol in protecting PC12 cells from the toxicity of hydrogen peroxide. The protection from Oxygen Reactive Species (ROS) toxicity is concentration dependent for both resveratrol and catechin. In this case the protection is merely additive and the synergistic effect is not observed. These results demonstrate that resveratrol and catechin protect PC12 cells from beta-AP (1-41) toxicity and that their protective effect is synergistic. Such a protective effect probably is not due only to their antioxidant activity. The different chemical and biological activity shown by these compounds on several cell types and the complexity of the beta-AP (1-41) toxicity may explain the synergistic protective effect and suggest that the utilization of different compounds with synergistic activity may protect more effectively from complex mechanisms of toxicity.
2002
- 50 Hz magnetic fields of varying flux intensity affect cell shape changes in invertebrate immunocytes: the role of potassium ion channels
[Articolo su rivista]
Ottaviani, Enzo; Malagoli, Davide; A., Ferrari; Tagliazucchi, Davide; Conte, Angela; Gobba, Fabriziomaria
abstract
The effect induced by exposure to 50 Hz magnetic fields (MFs) in immunocytes from the mussel Mytilus galloprovincialis is evaluated. The whole animal was exposed for 15 and 30 min to MF intensities ranging from 200 to 1,000 microT. The changes in the cellular shape of immunocytes, expressed as shape factor (SF), were studied at different times after addition of the chemotacting substance N-formyl-Meth-Leu-Phe (fMLP). Results show that MFs provoke differing delays in fMLP-induced cellular shape changes: 200 microT are ineffective, while levels from 300 microT upwards cause a significant increase in immunocyte SF values compared to controls. Reactivation of the cells is possible up to an intensity of 600 microT. The use of PCO 400, an opener of ATP-sensitive K+ channels, shows that potassium channels are involved in the effect of MFs on M. galloprovincialis immunocytes.