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Helicobacter pylori infection affects over 50% of the world's population and leads to chronic inflammation and gastric disorders, being the main pathogen correlated to gastric cancer development. Increasing antibiotic resistance levels are a major global concern and alternative treatments are needed. Soybean peptides and other compounds might be an alternative in the treatment to avoid, eradicate and/or control symptoms of H. pylori infection. This study aimed to characterize a lunasin-enriched soybean extract (LSE) using proteomics tools and to evaluate its antioxidant, anti-inflammatory and antibacterial properties against H. pylori infection. By LC-MS/MS analysis, 124 proteins were identified, with 2S albumin (lunasin and large-chain subunits) being the fourth most abundant protein (8.9%). Lunasin consists of 44 amino acid residues and an intramolecular disulfide bond. LSE at a low dose (0.0625 mg/mL) reduced ROS production in both H. pylori -infected and non-infected AGS gastric cells. This led to a significant reduction of 6.71% in the levels of pro-inflammatory interleukin (IL)-8. LSE also showed antibacterial activity against H. pylori , which can be attributed to other soybean proteins and phenolic compounds. Our findings suggest that LSE might be a promising alternative in the management of H. pylori infection and its associated symptoms., Competing Interests: The authors declare no conflicts of interest.

Isothiocyanates may have antibacterial activity against Helicobacter pylori , but there are different variables related to Brassicaceae-derived samples that could affect their efficacy. This work studied the influence of source variety, concentration, gastric digestion, and encapsulation of samples on their bioactive response against Helicobacter pylori . The antibacterial activity of raw sprouts (red cabbage and red radish) showed the highest antibacterial effect, which was consistent with a higher amount of isothiocyanates. It decreased with gastric digestion, regardless of sample encapsulation. By contrast, adult red radish leaves became antibacterial after gastric digestion. Antioxidant activity on H. pylori -infected gastric cells was similar in all samples and followed an equivalent pattern with the changes in isothiocyanates. Raw samples decreased intracellular reactive oxygen species production, but they lost this capacity after gastric digestion, regardless whether the compounds were free or encapsulated. Red cabbage sprouts, red radish sprouts, and red radish roots produced a decrease in nitric oxide production. It was consistent with a modulation of the inflammatory response and was associated to isothiocyanates concentration. Encapsulated sprout samples retained part of their anti-inflammatory activity after gastric digestion. Adult raw red radish leaves were not active, but after digestion, they became anti-inflammatory. The results obtained in this study have shown that several variables could have a significant impact on the bioactive properties of Brassicaceae-derived samples against H. pylori , providing a starting point for the design and standardization of samples with specific bioactivities (antibacterial, antioxidant, and anti-inflammatory) potentially useful for the treatment of H. pylori infection.

Franski and Beszterda-Buszczak [...].

The innate and adaptative immune systems are involved in the regulation of inflammatory and oxidative processes and mediators such as reactive oxygen species (ROS) and nitric oxide (NO). The exacerbated action of these players results in an oxidative stress status and chronic inflammation, which is responsible for the development of non-communicable diseases (NCDs). By modulating these mediators, bioactive compounds in food can exert a key role in the prevention of several NCDs. Among these compounds, soybean proteins and peptides such as lunasin have been considered to be among the most promising. The aim of this study was to obtain and characterize a soluble protein-enriched extract from a commercial soybean protein isolate and fractionate it into different fractions through ultrafiltration. Their antioxidant and immunomodulatory properties were then evaluated using biochemical and cell models. A total of 535 proteins (from 282 protein groups) were identified in the extract, in which the presence of the peptide lunasin was confirmed. The enrichment of this peptide was achieved in the 3-10 kDa fraction. The protective effects against the oxidative stress induced by LPS in the macrophage model could have been mediated by the radical scavenging capacity of the peptides present in the soybean samples. Under basal conditions, the extract and its ultrafiltered fractions activated macrophages and induced the release of NO. However, under challenged conditions, the whole extract potentiated the NO-stimulating effects of LPS, whereas the fraction containing 3-10 kDa peptides, including lunasin, counteracted the LPS-induced NO increase. Our findings suggest a promising role of soybean protein as an ingredient for functional foods and nutraceuticals aimed at promoting health and preventing oxidative stress and/or immune-alteration-associated diseases.

Campylobacter spp. are the main cause of bacterial gastroenteritis worldwide, and broiler chicks are the main vector of transmission to humans. The high prevalence of Campylobacter in poultry meat and the increase of antibiotic resistant strains have raised the need to identify new antimicrobial agents. For this reason, the aim of the current study was to evaluate the antibacterial activity of two extracts of olive leaf against antibiotic-resistant Campylobacter strains ( C. jejuni and C. coli ) isolated from poultry food chain. The extracts of olive leaf (E1 and E2) were markedly different in their chemical compositions. While E1 was composed predominantly of highly hydrophilic compounds such as hydroxytyrosol and hydroxytyrosol glucosides (14,708 mg/100 g), E2 mainly contained moderately hydrophilic compounds, with oleuropein (20,471 mg/100 g) being prevalent. All Campylobacter strains exhibited similar antibiotic profiles, being resistant to ciprofloxacin and tetracycline. E1 showed strong antibacterial activity and reduced bacterial growth from 4.12 to 8.14 log CFU/mL, depending on the strain. Hydroxytyrosol was the main compound responsible, causing the inhibition of growth of Campylobacter strains at low concentrations (0.1-0.25 mg/mL). E2 demonstrated a lower antibacterial effect than E1, reducing growth from 0.52 to 2.49 log CFU/mL. The results of this study suggest that the optimization of the composition of olive-leaf extracts can provide improved treatment results against Campylobacter strains.

The main objective of this work is to evaluate the potential utility of an Achillea millefolium extract (yarrow extract, YE) in the control of H. pylori infection. The supercritical anti-solvent fractionation (SAF) process of YE allowed the obtaining of two different fractions: yarrow's precipitated fraction (YPF), enriched in most polar phenolic compounds (luteolin-7- O -glucoside, luteolin, and 3,5-dicaffeoylquinic acid), and yarrow's separator fraction (YSF), enriched in monoterpenes and sesquiterpenes, mainly containing camphor, artemisia ketone, and borneol. YE was effective in reducing reactive oxygen species (ROS) production in human gastric AGS cells by 16% to 29%, depending on the H. pylori strain. YPF had the highest inhibitory activity (38-40%) for ROS production. YE modulated the inflammatory response in AGS gastric cells, decreasing IL-8 production by 53% to 64%. This IL-8 inhibition also showed a strain-dependent character. YPF and YSF exhibited similar behavior, reducing IL-8 production, suggesting that both phenolic compounds and essential oils could contribute to IL-8 inhibition. YSF showed the highest antibacterial activity against H. pylori (6.3-7.1 log CFU reduction, depending on the strain) and lower MIC (0.08 mg/mL). Results obtained have shown that YE and SAF fractions (YPF and YSF) were effective as antioxidant, anti-inflammatory, and antibacterial agents regardless of the H. pylori strain characteristics., Competing Interests: The authors declare no conflicts of interest.

The aim of this work was to evaluate the influence of in vitro gastric digestion of two olive leaf extracts (E1 and E2) on their chemical composition and bioactive properties against Helicobacter pylori ( H. pylori ), one of the most successful and prevalent human pathogens. HPLC-PAD/MS analysis and anti-inflammatory, antioxidant, and antibacterial activities of both olive leaf extracts were carried out before and after their in vitro gastric digestion. The results showed that gastric digestion produced modifications of the chemical composition and bioactive properties of both olive leaf extracts. The main compounds in the extract E1 were hydroxytyrosol and its glucoside derivatives (14,556 mg/100 g), presenting all the identified compounds a more polar character than those found in the E2 extract. E2 showed a higher concentration of less polar compounds than E1 extract, with oleuropein (21,419 mg/100 g) being the major component. Gastric digestion during the fasted state (pH 2) induced an overall decrease of the most identified compounds. In the extract E1, while the anti-inflammatory capacity showed only a slight decrease (9% of IL-8 production), the antioxidant properties suffered a drastic drop (23% of ROS inhibition), as well as the antibacterial capacity. However, in the extract E2, these changes caused an increase in the anti-inflammatory (19% of IL-8 production) and antioxidant activity (9% of ROS inhibition), which could be due to the hydrolysis of oleuropein and ligustroside into their main degradation products, hydroxytyrosol and tyrosol, but the antibacterial activity was reduced. Gastric digestion during fed state (pH 5) had less influence on the composition of the extracts, affecting in a lesser degree their anti-inflammatory and antioxidant activity, although there was a decrease in the antibacterial activity in both extracts similar to that observed at pH 2.

Helicobacter pylori ( H. pylori ) is one of the major human pathogens and the main cause of pathological damages that can progress from chronic gastritis to gastric cancer. During the colonization of gastric mucosa, this bacterium provokes a strong inflammatory response and subsequent oxidative process, which are associated with tissue damage. Therefore, the objective of this research was to evaluate the ability of two olive-leaf extracts (E1 and E2) to modulate the inflammatory response and oxidative stress in H. pylori -infected human gastric AGS cells. The obtained results showed that both extracts significantly decreased interleukin-8 (IL-8) secretion and reactive oxygen species (ROS) production in human gastric AGS cells. Both extracts also showed antibacterial activity against different H. pylori strains. HPLC-PAD-MS characterization demonstrated that extract E1 was mainly composed of highly hydrophilic compounds, such as hydroxytyrosol (HT) and its glucosides, and it was the most effective extract as an antibacterial agent. In contrast, extract E2 was composed mostly of moderately hydrophilic compounds, such as oleuropein (OLE), and it was more effective than extract E1 as an anti-inflammatory agent. Both extracts exhibited similar potential to decrease ROS production. These results show the importance of standardizing the extract composition according to the bioactive properties that should be potentiated.

Helicobacter pylori ( H. pylori ) is a pathogenic bacteria identified as a potential risk factor for gastritis, gastric ulcers and gastric cancer. During the stomach colonization, H. pylori triggers a strong inflammatory response and subsequent oxidative stress, which are associated with tissue damage. For this reason, it is of particular interest to develop alternative natural tools that enable modulation of the associated damaging immune response. With this purpose, we obtained grape seed extract (GSE) from sweet (not fermented) food grade seeds. The aim of our study was to investigate the effect of GSE and its two enriched procyanidins fractions (OPC and PPC) on the inflammatory process and oxidative stress produced by different H. pylori strains in human gastric epithelial cells (AGS). Anti-inflammatory activity was evaluated by measuring the level of interleukin-8 (IL-8) secretion. IL-8 production was significantly reduced in H. pylori -infected human gastric epithelial cells pre-treated with GSE or its enriched fractions when compared with non-pre-treated infected cells (from 21.6% to 87.8%). Pre-treatment with GSE or its fractions significantly decreased intracellular reactive oxygen species (ROS) production in AGS cells after infection, depending on the H. pylori strain. Our results also showed that GSE and its fractions demonstrate antibacterial activity against all strains of H. pylori used in the study. This work demonstrates the effectiveness of GSE enriched in procyanidins against the main events associated with H. pylori infection.

Strains of Helicobacter pylori (H. pylori) resistant to various antibiotics have increased in recent years. In this context, the search for new therapeutic approaches is crucial. The aim of the present study was to demonstrate the antibacterial activity of a procyanidin-rich extract obtained from food-grade winery grape seeds against 14 H. pylori strains and elucidate its phenolic composition. Ten strains (71.4%) showed resistance to at least some of the tested antibiotics, while four isolates (28.6%) were susceptible to all antibiotics. Resistance to more than one class of antibiotics was observed in six strains (42.9%). The extract was able to inhibit the growth of all H. pylori strains in a range of a minimum inhibitory concentration (MIC) from 0.015 mg/mL to 0.125 mg/mL, confirming also the existence of a strain-dependent effect. The phenolic composition determined by reverse phase high pressure liquid chromatography, photodiode array, and mass spectrometry detection (RP-HPLC-PAD-MS) analysis revealed the presence of 43 individual compounds and allowed the quantification of 41 of them, including seven procyanidin tetramers, seven procyanidin pentamers, and six galloylated procyanidin dimers, trimers, and tetramers. The extract was composed mainly by catechin and procyanidin oligomers with a total amount of 5.801 mg/100 g, which represent 92% of the total individual phenolic content. Among them, the most abundant were catechins (2.047 mg/100 g), followed by procyanidin dimers (1.550 mg/100 g), trimers (1.176 mg/100 g), tetramers (436 mg/100 g), and pentamers (296 mg/100 g) that represent 35, 27, 20, 8, and 5%, respectively of the total flavanol constituents. The composition profile information may help to improve the production process of useful antibacterial extracts against H. pylori .

The effectiveness of a preparative integrated ultrafiltration/solid-phase extraction (UF/SPE) process for purification of oligomeric procyanidins (OPCs) from a crude grape seed extract (GSE) was studied for the first time. The separation of OPCs from polymeric procyanidins (PPCs) by UF was very efficient. The membrane showed an acceptable filtration flux of 6 to 3.5 L/h·m 2 at 0.5 bar of transmembrane pressure and 95% recovery of its water flux after chemical cleaning. The process was scalable to a pilot scale. The separation of very polar and ionic species from OPCs by SPE (XAD7HP and XAD16 resins) was also very good, but both adsorbents lost their retention capacities quickly, due probably to irreversible retention of OPCs/PPCs. Even though the global purification of OPCs by the integrated UF/SPE process allowed the recovery of 24.2 g of highly purified OPCs (83% purity) from 14.4 L of crude grape seed extract, the use of these adsorbents for further purification of the OPCs was very limited.

The consumption of plums in a fresh form is seasonal, therefore the transformation of plum juice extracts into powdered form is a good alternative for its longer availability throughout the year. The drying process can moderate the physical and chemical properties of the plum extracts, thus, this study examined the changes in biological activity, i.e., antibacterial, antioxidant, and anti-inflammatory properties moderated by freeze, vacuum, and spray drying. It was suggested that the drying processes and the applied parameters might moderate the content of polyphenolic compounds in the powders, which influence the different levels of growth inhibition against the foodborne pathogens (17% to 58% of inhibition), demonstrating a strain-dependent effect. These powders could also induce cellular protection against oxidative stress by preventing intracellular reactive oxygen species (ROS) accumulation (23% to 37% of reduction), but the level of antioxidant capacity may be determined by the conditions applied during the drying process. Moreover, plum extract powders exhibited a greater anti-inflammatory capacity (24% to 39% of inhibition), which would be influenced both, by the type of treatment used and by the temperature used in each treatment. The results demonstrate that the selection of the drying method can be an effective tool for modulating the composition, physical, and bioactive properties of plum extracts powders., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Campylobacter is the leading cause of bacterial diarrheal disease worldwide. Although most episodes of campylobacteriosis are self-limiting, antibiotic treatment is usually needed in patients with serious enteritis, and especially in childrens or the elderly. In the last years, antibiotic resistance in Campylobacter has become a major public health concern and a great interest exists in developing new antimicrobial strategies for reducing the impact of this food-borne pathogen on human health. Among them, the use of silver nanoparticles as antibacterial agents has taken on increased importance in the field of medicine. The aim of the present study was to evaluate the antimicrobial effectiveness of glutathione-stabilized silver nanoparticles (GSH-Ag NPs) against multidrug resistant (MDR) Campylobacter strains isolated from the chicken food chain (FC) and clinical patients (C). The results obtained showed that GSH-Ag NPs were highly effective against all MDR Campylobacter strains tested. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were in a range from 4.92 to 39.4 μg/mL and 9.85 to 39.4 μg/mL, respectively. Cytotoxicity assays were also assessed using human intestinal HT-29, Caco-2, and CCD-18 epithelial cells. Exposure of GSH-Ag NPs to intestinal cells showed a dose-dependent cytotoxic effect in all cell lines between 9.85 and 39.4 μg/mL. More than 60% of the tested Campylobacter strains were susceptible to GSH-Ag NPs concentrations ≤ 9.85 μg/mL, suggesting that practical inhibitory levels could be reached at low GSH-Ag NPs concentrations. Further works are needed with the purpose to evaluate the practical implications of the toxicity studies and to know more about other attributes linked to the biological compatibility. This behavior makes GSH-Ag NPs as a promising tool for the design of novel antibacterial agents for controlling Campylobacter .

In Albariño white wines, aging of wines on lees is a technique not used or only used empirically by some producers to obtain a distinctive character in the final wine. This study analyzes the influence of a short aging on lees on the chemical and sensorial parameters of this young white wine. Albariño grape must was inoculated with a locally selected yeast (Saccharomyces cerevisiae 1) and the effect of a short aging on lees was studied during different times (10, 20, 30, 40, and 50 d). Mannoprotein content and the aromatic profile were determined and a sensorial analysis of the wines was conducted. Results showed that aging time was correlated with the concentration of some key aroma compounds and mannoproteins in Albariño wines. The best sensorial character was obtained in wines aged 20 d on lees. Further aging times decreased the sensorial quality of Albariño wine and modified its volatile profile and mannoprotein concentration., (© 2015 Institute of Food Technologists®)

In this study, a recombinant Saccharomyces cerevisiae strain EKD13 overproducing mannoproteins has been used to obtain Albariño white wines. The inoculated strain prevailed and produced complete fermentation of the must, as also occurred in the case of spontaneous (non-inoculated) fermentation and in the must inoculated with the S. cerevisiae EC1118 strain. The analytical study of the wines obtained showed that the most important chemical differences among the wines produced with EKD-13, corresponded to the high concentration of mannoproteins, 2-phenyl ethanol and tyrosol. These differences were attributed to the expression, during must fermentation, of genes modified in the recombinant EKD-13 strain. The results obtained imply that this strain could be potentially useful to produce wines rich in mannoproteins that have distinctive characteristics compared to other similar wines, modifying the sensorial and technological parameters of the wines obtained., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Aim: This study was aimed to investigate the influence of malolactic fermentation (MLF) on sensory profile and organoleptic characteristics of Albariño and Caiño white wines., Methods and Results: Autochthonous bacteria were isolated from wines after alcoholic fermentation (AF) and further identified as Pediococcus damnosus by 16S rRNA gene sequence analysis. When a commercial Oenococcus oeni starter was inoculated into Albariño and Caiño white wines to perform MLF, which was checked by HPLC quantification of malic and lactic acids, it was shown that autochthonous Ped. damnosus strains were able to predominate over the commercial O. oeni starter and perform MLF in Caiño wine. By contrast, neither commercial strain nor indigenous Pediococcus carried out MLF in Albariño wine. However, MLF was achieved when autochthonous strains that predominated in Caiño were inoculated into Albariño. Sensory analysis showed that after the MLF Albariño increased its body and softness, while Caiño result a more mature wine., Conclusions: MLF can positively affect Albariño and Caiño wines giving them new attributes. Pediococci isolated and characterized in this work can successfully perform MLF without negative effects on the wine, because no production of biogenic amines or exopolysaccharides by the selected pediococcus strains was detected., Significance and Impact of the Study: The effect of MLF in the sensory profile of Albariño and Caiño wines has never been studied before. Results obtained in this work showed that Ped. damnosus strains can be considered as a new topic of investigation on malolactic starter., (© 2013 The Society for Applied Microbiology.)

Campylobacter is the most common cause of bacterial food-borne diarrhoeal disease throughout the world. The principal risk of human contamination is handling and consumption of contaminated poultry meat. To colonize poultry, Campylobacter adheres to and persists in the mucus layer that covers the intestinal epithelium. Inhibiting adhesion to the mucus could prevent colonization of the intestine. The aim of this study was to investigate in vitro the protective effect of defined commercial human probiotic strains on the adhesion of Campylobacter spp. to chicken intestinal mucus, in a search for alternatives to antibiotics to control this food-borne pathogen. The probiotic strains Lactobacillus rhamnosus GG and Propionibacterium freudenreichii ssp. shermanii JS and a starter culture strain Lactococcus lactis ssp. lactis adhered well to chicken intestinal mucus and were able to reduce the binding of Campylobacter spp. when the mucus was colonized with the probiotic strain before contacting the pathogen. Human-intended probiotics could be useful as prophylactics in poultry feeding for controlling Campylobacter spp. colonization., (© 2012 Blackwell Verlag GmbH.)

Unlabelled: In the present work, 3 different yeast strains (1, 2, and 3) were used to elaborate white wines using Albariño must. The concentration of polymeric mannose was determined using a method based on the mannoprotein precipitation, hydrolysis and analysis of sylylated mannose derivatives by gas chromatography. Wines elaborated with the strain 1 (W1) presented a higher mannoprotein concentration than the other wines. The analysis of the volatile composition of wines showed significant differences (P < 0.05) among them, being W1 which presented the highest concentration of aroma compounds, mainly terpens and norisoprenoids. The sensorial analysis of wines also showed that W1 had the best quality. The results obtained from this work demonstrate that mannoproteins could be involved in the behavior observed. Some evidences were obtained using a model wine, where 2 major terpens in W1 were preferentially retained by the colloids rich in mannoproteins released by strain 1., Practical Application: White wines elaborated with yeast strains overproducing mannoproteins could have better quality than others. Mannoproteins could contribute to aroma enhancement of Albariño white wines., (Journal of Food Science © 2012 Institute of Food Technologists® No claim to original US government works.)

Yeast cell wall (YCW) preparations and yeast mannoprotein extracts have been effective against some enteropathogenic bacteria as Campylobacter jejuni, Escherichia coli, and Salmonella, and they can affect the population of beneficial lactic acid bacteria (LAB). In this work, we studied the effect of a mannoprotein extract on five strains of LAB. This extract was metabolised by the bacteria, enhancing their survival in simulated gastrointestinal juice, and increasing the adherence of Lactobacillus plantarum, L. salivarius, and Enterococcus faecium to Caco-2 cells. Yeast mannoproteins are promising naturally occurring compounds that could be used to enhance LAB intestinal populations and control pathogens., (© 2012 Institute of Food Technologists®)

The effect of growth phase on the adherence to and invasion of Caco-2 epithelial cells by five strains of Campylobacter was studied. No significant differences were observed between the behaviors in the exponential or stationary phases for the most stationary-phase tolerant strains (C. jejuni 118 and C. coli LP2), while the strains that produced a greater reduction in the viability in the stationary phase (C. jejuni 11351, C. jejuni 11168 and C. jejuni LP1), also presented reduced adherence to and invasion of Caco-2 cells. In order to find a possible explanation for the observed differences, the presence of putative virulence factors was studied in the analyzed strains. In spite of the fact that C. jejuni 118 and C. jejuni 11168 strains showed a different adherence to and invasion of Caco-2 cells behavior, they posses identical alleles for ciaB, cadF, and pldA loci. From the virulence factors analyzed, only the flaA locus was different among both strains., ((c) 2010 Elsevier B.V. All rights reserved.)

The ability of pectic oligosaccharides (POS) to inhibit adherence to and invasion of undifferentiated (UC) and differentiated (DC) Caco-2 cells by Campylobacter jejuni (C. jejuni) was investigated. It was observed that both adherence and invasion were significantly higher in UC than in DC. POS (2.5mg/ml) had no significant effect on the number of bacteria which can adhere to cells, but they significantly inhibited cell invasion. The extent of the anti-invasive effect of POS was dependent on the concentration, although the entire range tested (from 2.5mg/ml to 0.05 mg/ml) was capable of inhibiting the invasion of Caco-2 cells by Campylobacter to some degree. The pre-incubation or not of C jejuni with POS did not influence the behaviour observed. The results obtained in this work suggest that POS could be potentially useful as alternatives to antibiotics in the control of C. jejuni., ((c) 2009 Elsevier B.V. All rights reserved.)

The main objective of the present work was to study the influence of yeast-derived mannoproteins on the adherence to and invasion of Caco-2 cells by Campylobacter jejuni. Mannoprotein fractions were prepared by enzymatic and thermal extraction methods. The method used to prepare the mannoprotein extracts influenced their composition and determined the efficacy of the extract against C. jejuni adherence and/or invasion. The availability of mannose in the mannoprotein fraction seemed to be important for inhibiting effective adherence and invasion of Caco-2-cells by C. jejuni, although protein moieties also played a role in the process. The study of the mechanisms involved in the inhibition of C. jejuni adherence and invasion by mannoproteins may have further implications in the control of this foodborne pathogen.

A selection method, based on a temperature-sensitive autolytic phenotype, has been used to genetically improve a second fermentation Saccharomyces cerevisiae yeast strain by UV mutagenesis. The mutations carried by the resulting strains affected cell morphology, growth kinetics, sporulation and the release of nitrogenous compounds in an accelerated autolysis experimental model. Their fermentation power was not severely impaired.