An experimental investigation evaluated the possibility of increasing the nutritional value of fermented table olives by adding olive leaf extract (OLE). OLE was added to table olives fermented using indigenous bacteria and yeasts, and a commercial starter (Lactobacillus plantarum strain). Microbiological, physico-chemical, and sensory analyses showed that OLE addition resulted in fermented olives with higher levels of antioxidant, anti-inflammatory, and antimicrobial substances, but did not adversely affect their qualities. Moreover, OLE and the commercial starter functioned synergically against spoilage microorganisms. In addition, fermented olives had higher values of hardness, total phenols, antioxidant activity, hydroxytyrosol, and verbascoside. Nonanal and ethanol contents were lower in fermented olives when Lactobacillus plantarum and OLE were used, indicating lower degrees of oxidation and fermentation. Finally, olives fermented with OLE had a less bitter taste.

Proceedings of the 1th International conference on "Microbial Diversity 2011 - Environmental Stress and Adaptation, October 26 – 28, Milan (Italy). Pages 102-105.

SelectedLactobacillus paracasei FC2-5, Lactobacillus casei LC01 and Lactobacillus curvatus 2770 were used for cheese making as adjunct cultures (AC) or attenuatedadjunct cultures (AAC). AAC were obtained by sonication treatment. ItalianCaciotta-typecheeses were manufactured on an industrial plant scale, including control cheese (CC) without AC, and ripening was lasted 60 days at 10 °C. AAC did not increase the production of lactic acid compared with the CC. AC acidified during manufacture and throughout ripening, affected the moisture and texture of the cheese. As shown by the plate count and confirmed by RAPD-PCR, the cell numbers of non-starterlactobacilli varied between cheeses manufactured with AC or AAC. The major differences between cheeses were the accumulation of free amino acids and the synthesis of some key volatile components. As shown by PT-GC/MS analysis, the levels of ketones, secondary alcohols and sulphur compounds were highest in the cheese manufactured with AAC.

Aims:  To screen the glutamate dehydrogenase (GDH) activity of nonstarter lactic acid bacteria (NSLAB) and to determine the effects of temperature, pH and NaCl values used for cheese ripening on enzyme activity and expression of GDH gene. Methods and Results:  A subcellular fractionation protocol and specific enzyme assays were used. The effect of temperature, pH and NaCl on enzyme activity was evaluated. The expression of GDH gene was monitored by real-time PCR. One selected strain was also used as adjunct starter for cheese making to evaluate the catabolism of free amino acids and the production of volatile organic compounds (VOC) during cheese ripening. The cytoplasm fraction of all strains showed in vitro NADP-dependent GDH activity. NADP-GDH activity was markedly strain dependent and varied according to the interactions between temperature, pH and NaCl. Lactobacillus plantarum DPPMA49 showed the highest NADP-GDH activity under temperature, pH and NaCl values found during cheese ripening. RT-PCR analysis revealed that GDH expression of Lact. plantarum DPPMA49 was down-expressed by low temperature (<13°C) and over-expressed by NaCl (1·87–5·62%). According to NADP-GDH activity, the highest level of VOC (alcohols, aldehydes, miscellaneous and carboxylic acids) was found in cheeses made with DPPMA49. Conclusions:  The results of this study may be considered as an example of the influence of temperature, pH and NaCl on enzyme activity and expression of functional genes, such as GDH, in cheese-related bacteria. Significance and Impact of the Study:  It focuses on the phenotypic and molecular characterization of the NADP-GDH in lactobacilli under cheese-ripening conditions. The findings of this study contribute to the knowledge about enzymes involved in the catabolism of amino acids, to be used as an important selection trait for cheese strains.

Besides the variety of colours and flavours, microgreens show interesting nutritional properties, mainly regarding their contents of mineral nutrients and bioactive compounds. To date, the literature has prevalently focused on the individual nutritional features of microgreens usually belonging to Brassicaceae. The present study reports an articulated nutritional profile of six genotypes of microgreens, belonging to three species and two families: chicory (Cichorium intybus L., Puglia's local variety 'Molfetta', CM, and cultivar 'Italico a costa rossa', CR) and lettuce (Lactuca sativa L. Group crispa, cultivar 'Bionda da taglio', LB, and 'Trocadero', LT), from Asteraceae; and broccoli (Brassica oleracea L. Group italica Plenk, Puglia's local variety 'Mugnuli', BM, and cultivar 'Natalino', BN) from Brassicaceae. All the microgreens, except LB, can be considered good sources of Ca, whilst LT and CM also showed considerable amounts of K. As regards bioactive compounds, Brassica microgreens were the richest in phenolic antioxidants. The microgreens also presented higher amounts of α-tocopherol and carotenoids compared to mature vegetables. In particular, broccoli microgreens and LB showed the highest amounts of vitamin E, while Asteraceae microgreens presented the highest levels of carotenoids. Due to their delicate tissues, fresh cut microgreens showed a shelf life not exceeding ten days at 5 °C. The results obtained highlight the possibility to exploit genetic biodiversity in order to obtain tailored microgreens with the desired nutritional profiles, with particular regard to mineral nutrients and bioactive compounds. Appropriate pre- and post-harvest strategies should be developed, so as to allow microgreens to retain as long as possible their nutritional value.

This work showed the effect of pheromone plantaricinA (PlnA) on the proliferation and migration of the humankeratinocytes NCTC 2544. PlnA was chemically synthesized and used as pure peptide or biologically synthesized during co-cultivation of Lactobacillusplantarum DC400 and Lactobacillus sanfranciscensis DPPMA174. The cell-free supernatant (CFS) was used as the crude preparation containing PlnA. The inductive effect of PlnA on the proliferation of NCTC 2544 cells was higher than that found for hyaluronic acid, a well known skin protective compound. As shown by scratch assay and image analyses, PlnA enhanced the migration of NCTC 2544 cells. Compared to the basal serum free medium (control), the highest inductive effect was found using 10 μg/ml of chemically synthesized PlnA. Similar results (P > 0.05) were found for CFS. In agreement, the percentage of the starting scratch area was decreased after treatment (24 h) with PlnA. The expression of transforming growth factor-β1 (TGF-β1), keratinocyte growth factor 7 (FGF7), vascular endothelial growth factor (VEGF-A), and interleukin-8 (IL-8) genes was affected by PlnA. Compared to control, TGF-β1gene was under expressed in the first 4 h of treatments and up-regulated after 8–24 h. On the contrary, FGF7gene was strongly up-regulated in the first 4 h of treatments. Compared to control, VEGF-A and IL-8genes were always up-regulated during the 4–24 h from scratching. Since capable of promoting the proliferation and migration of the humankeratinocytes and of stimulating IL-8 cytokine, the use of PlnA for dermatological purposes should be considered.

This work aimed at showing the effect of pheromone plantaricin A (PlnA) by Lactobacillus plantarum DC400 towards other sourdough lactic acid bacteria and the potential of PlnA to protect the function of the human intestinal barrier. Growth and survival of sourdough lactic acid bacteria were differently affected by co-cultivation with L. plantarum DC400. Compared to mono-cultures, Lactobacillus sanfranciscensis DPPMA174 and Pediococcus pentosaceus 2XA3 showed growth inhibition and decreased viability when co-cultured with L. plantarum DC400. L. sanfranciscensis DPPMA174 induced the highest synthesis of PlnA. Survival of strain DPPMA174 only slightly varied by comparing the addition of PlnA to the culture medium and the co-cultivation with L. plantarum DC400. Compared to mono-culture, the proteome of L. sanfranciscensis DPPMA174 grown in co-culture with L. plantarum DC400 showed the variation of expression of 58 proteins (47 over expressed and 11 repressed). Thirty-four of them were also over expressed or repressed during growth of DPPMA174 with PlnA. Fifty-one of the above 58 proteins were identified. They had a central role in stress response, amino acid, energy and nucleotide metabolisms, membrane transport, regulation of transcription, and cell redox homeostasis. PlnA markedly increased the viability of human Caco-2/TC7 cells and the transepithelial electrical resistance.

Plantaricin A (PlnA) is a peptide with antimicrobial and pheromone activities. PlnA was synthesized chemically and used as a pure peptide or synthesized biologically using Lactobacillus plantarum DC400 co-cultured with Lactobacillus sanfranciscensis DPPMA174. Cell-free supernatant (CFS) was used as a crude PlnA preparation. As estimated using the 3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide and the 2',7'-dichlorofluorescein diacetate assays, both PlnA preparations increased the antioxidant defenses of human NCTC 2544 keratinocytes. PlnA (10 ?g/ml) had a higher activity than hyaluronic acid or 125 ?g/ml ?-tocopherol. Effects on the transcriptional regulation of filaggrin (FLG), involucrin (IVL), hyaluronan synthase (HAS2), human ?-defensin-2 (HBD-2) and tumor necrosis factor-alpha (TNF-?) genes were assayed. Compared with the control, expression of the FLG gene in NCTC 2544 cells increased in cells treated with hyaluronic acid, 1 or 10 ?g/ml PlnA. Compared with the control, the level of IVL gene expression increased in NCTC 2544 cells treated with 10 ?g/ml PlnA. No significant difference was found between the level of the HAS2 gene expressed by control cells and cells treated with PlnA. Compared with chemically synthesized PlnA, the up-regulation of the HBD-2 gene by CFS was higher. Compared with the control, expression of TNF-? decreased in NCTC 2544 cells after treatment with 1 or 10 ?g/ml of chemically synthesized PlnA. In contrast, the level of TNF-? was highest in the presence of 10 ?g/ml CFS-PlnA. These findings suggest that the PlnA was positively sensed by human keratinocytes, promoting antioxidant defenses, barrier functions and antimicrobial activity of the skin.

This study aimed at investigating the effect of corn, rice and amaranth gluten-free (GF) sourdoughs on the release of nitric oxide (NO) and synthesis of pro-inflammatory cytokines by duodenal mucosa biopsies of eight coeliac disease (CD) patients.

This study aimed at investigating the addition of legume (chickpea, lentil and bean) flours to wheat flour bread. Type I sourdough containing legumes or wheat–legume flours were prepared and propagated (back slopped) in laboratory, according to traditional protocols that are routinely used for making typical Italian breads. Based on kinetic of acidification and culture-dependent data, the wheat–legume sourdough was further characterized and selected for bread making. As determined by RAPD-PCR and partial sequencing of 16S rDNA gene analyses, lactic acid bacteria in wheat–legume sourdough included Lactobacillus plantarum, Lactobacillus sanfranciscensis, Leuconostoc mesenteroides, Lactobacillus fermentum, Weissella cibaria, Lactobacillus pentosus, Lactobacillus coryneformis, Lactobacillus rossiae, Lactobacillus brevis, Lactobacillus parabuchneri and Lactobacillus paraplantarum. Two breads containing 15% (w/w) of legume (chickpea, lentil and bean) flours were produced using selected wheat–legume sourdough (WLSB) and traditional wheat sourdough (WSB). Compared to wheat yeasted bread (WYB), the level of total free amino acids (FAA) was higher in WSB and WLSB. Phytase and antioxidant activities were the highest in WLSB. Compared to bread WYB, the addition of legume flours decreased the in vitro protein digestibility (IVPD) (WYB versus WSB). However, the dough fermentation with WSLB favored an increase of IVPD. According to the levels of carbohydrates, dietary fibers and resistant starch, WSB and WLSB showed lower values of hydrolysis index (HI) compared to WYB. As showed by texture and image analyses and sensory evaluation of breads, a good acceptability was found for WSB and, especially, WLSB breads.