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•Strains grown with 5 % mangosteen extract showed increased antioxidant capacity.•Growth with 5 % mangosteen extract reduced strain biofilm formation.•Most strains were suitable for ...production of functional fermented milk products.•Mangosteen fermented milk products had increased polyphenol and antioxidant levels.
Mangosteen (Garcinia mangostana L.) is a tasty, polyphenol-rich tropical fruit. The edible part is highly appreciated by its aroma, taste and texture. The non-edible part, rich in polyphenols, has been traditionally used in Thai medicine. In this work, flavonoids and phenolic acid/derivatives were identified in mangosteen extracts (ME) from edible and non-edible portions. We first studied the effects of MEs on the growth, metabolism, antioxidant capacity, biofilm formation and antimicrobial capacity of eight bifidobacteria and lactobacilli strains from intestinal origin and two commercial probiotic strains (BB536 and GG). ME concentrations higher than 10–20 % were inhibitory for all strains. However, ME concentrations of 5 % significantly (P < 0.01) increased all strains antioxidant capacity, reduced biofilm-formation, and enhanced inhibition against Gram-positive pathogens. To apply these knowledge, bifunctional fermented milk products were elaborated with 5 % ME and individual strains, which were selected taking into account their growth with ME, and the widest range of values on antioxidant capacity, biofilm formation and antimicrobial activity (bifidobacteria INIA P2 and INIA P467, lactobacilli INIA P459 and INIA P708, and reference strain GG). Most strains survived well manufacture, refrigerated storage and an in vitro simulation of major conditions encountered in the gastrointestinal tract. As expected, products supplemented with ME showed higher polyphenol content and antioxidant capacity levels than control. After sensory evaluation, products containing strains INIA P2, INIA P708 and GG outstood as best.
There is a growing interest for healthy non-bovine functional dairy products. Therefore, in the present study, an aqueous polyphenol-rich artichoke extract (AE) and two Bifidobacterium strains of ...human origin (B. pseudolongum INIA P2 and B. longum INIA P132) were added, alone or in combination with AE, for the manufacture of six sheep milk cheeses. In general, addition of 1.25 g of lyophilized AE per 100 g of curd did not affect cheese pH, dry matter, lactoccci starter counts or organic acid levels during ripening. Bifidobacterial strains showed good viability in cheeses with AE during ripening and after simulated major gastrointestinal conditions. Cheeses with AE showed improved functional properties, with significantly (P < 0.01) higher total polyphenol content and, especially, antioxidant activity The cheese with AE and strain INIA P2 fulfil the definition of a symbiotic cheese. It contains prebiotic compounds and Bifidobacterium at adequate levels to sustain a therapeutic effect, which are able to survive major gastrointestinal conditions. This cheese could confer beneficial effects on consumers’ health by contributing to the host antioxidant capacity and increasing polyphenol bioavailability.
•Artichoke extract (AE) did not affect cheese pH, dry matter, colour or microbiota.•AE increased cheese proteolysis, polyphenol content and antioxidant activity.•INIA P2 increased AE polyphenol bioavailability and showed good viability in cheese.•AE and bifidobacteria affected cheese volatile profile.•Symbiotic cheese with AE and INIA P2 may provide polyphenol health benefits.
Enterocins are bacteriocins synthesized by
strains that show an interesting antimicrobial effectiveness against foodborne pathogens such as
. The objectives of this study were to identify and analyze ...the expression of enterocin genes of
isolated from breast-fed infants and evaluate their ability to inhibit three human isolates of virulent
, as well as some probiotic bacteria. The susceptibility of the strains of
to fifteen antibiotics was tested, detecting their resistance to cefoxitin (constitutively resistant), oxacillin, and clindamycin. The production of enterocins A, B, and P was observed in
isolates, while enterocin AS-48 was detected in an
isolate. AS-48 showed antilisterial activity by itself, while the joint action of enterocins A and B or B and P was necessary for inhibiting
, demonstrating the synergistic effect of those combinations. The presence of multiple enterocin genes does not assure the inhibition of
strains. However, the expression of multiple enterocin genes showed a good correlation with the inhibition capacity of these strains. Furthermore, the potential beneficial strains of lactobacilli and bifidobacteria examined were not inhibited by any of the enterocins produced individually or in combination, with the exception of
BB536, which was inhibited by enterocin AS-48 and the joint production of enterocins A and B or B and P. The enterocins studied here could be candidates for developing alternative treatments against antibiotic-resistant bacterial infections. Moreover, these selected enterocin-producing
strains isolated from breast-fed infants could be used as probiotic strains due to their antilisterial effect, as well as the absence of virulence factors.
Soy beverages can be a source of bioactive isoflavones, with potential human health benefits. In this work, the suitability of three Lacticaseibacillus and three Bifidobacterium probiotic strains as ...functional starters for soy beverage fermentation were evaluated, alongside with the effect of refrigerated storage on the viability of the strains and the isoflavone composition of the fermented beverages. The three bifidobacteria strains suffered a decrease in their viability during refrigeration and only Bifidobacterium breve INIA P734 produced high concentrations of bioactive isoflavones. Meanwhile, L. rhamnosus GG and L. rhamnosus INIA P344 produced high levels of aglycones and, with L. paracasei INIA P272, maintained their viability during the refrigeration period, constituting promising starters to obtain functional soy beverages that could gather the benefits of the bioactive isoflavone aglycones and the probiotic strains. Moreover, the three lactobacilli caused an increase in the antioxidant capacity of the fermented beverages, which was maintained over the refrigerated storage.
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•L. paracasei INIA P272 and L. rhamnosus INIA P344 draft genomes were analysed.•Probiotic potential disclosed by both strains was complemented by genome analysis.•Genomic safety ...assessment confirmed both strains suitable as food grade ingredients.•A validated strategy for in silico adhesion capacity determination was established.
Lacticaseibacillus paracasei INIA P272 and Lacticaseibacillus rhamnosus INIA P344, isolated from breast-fed infants, are two promising bacterial strains for their use in functional foods according to their demonstrated probiotic and technological characteristics. To better understand their probiotic characteristics and evaluate their safety, here we report the draft genome sequences of both strains as well as the analysis of their genetical content. The draft genomes of L. paracasei INIA P272 and L. rhamnosus INIA P344 comprise 3.01 and 3.26 Mb, a total of 2994 and 3166 genes and a GC content of 46.27 % and 46.56 %, respectively. Genomic safety was assessed following the EFSA guidelines: the identification of both strains was confirmed through Average Nucleotide Identity, and the absence of virulence, pathogenic and antibiotic resistance genes was demonstrated. The genome stability analysis revealed the presence of plasmids and phage regions in both genomes, however, CRISPR sequences and other mechanisms to fight against phage infections were encoded. The probiotic abilities of both strains were supported by the presence of genes for the synthesis of SCFA, genes involved in resistance to acid and bile salts or a thiamine production cluster. Moreover, the encoded exopolysaccharide biosynthesis genes could provide additional protection against the deleterious gastrointestinal conditions, besides which, playing a key role in adherence and coaggregation of pathogenic bacteria together with the high number of adhesion proteins and domains encoded by both genomes. Additionally, the bacteriocin cluster genes found in both strains, could provide an advantageous ability to compete against pathogenic bacteria. This genomic study supports the probiotic characteristics described previously for these two strains and satisfies the safety requirements to be used in food products.
Limosilactobacillus reuteri is a beneficial bacterium that inhabits the gastrointestinal tract of different mammals. Diverse beneficial effects have been attributed to specific strains, in part ...mediated by the production of reuterin. Here, we report the draft genome sequence of L. reuteri INIA P572, a reuterin-producing strain isolated from pig feces.
The objective of this study is to explore the possibility of applying a point system as a guide to the management of waiting lists in National Health Systems. Following recent contributions in the ...axiomatic theory of justice, the ethical properties of a point system are illustrated. In addition, we present the results of an experiment whose objective was to develop a point system for cataract extraction, based on social preferences. The results of the experiment have shown that the analytic methods used here, of focus groups, interview-administered questionnaires, conjoint analysis, and rank-ordered logit, can be usefully combined to determine the total priority score for each patient.
The objective of this study is to explore the possibility of applying a point system as a guide to the management of waiting lists in National Health Systems. Following recent contributions in the ...axiomatic theory of justice, the ethical properties of a point system are illustrated. In addition, we present the results of an experiment whose objective was to develop a point system for cataract extraction, based on social preferences. The results of the experiment have shown that the analytic methods used here, of focus groups, interview-administered questionnaires, conjoint analysis, and rank-ordered logit, can be usefully combined to determine the total priority score for each patient.
Biomarkers for cancer immunotherapy are an unmet medical need. The group of Daniela Thommen at the NKI recently reported on novel methodologies based on short-term cultures of patient-derived tumor ...fragments whose cytokine concentrations in the supernatants and activation markers on infiltrating T cells were associated with clinical response to PD-1 blockade. We set up a similar culture technology with tumor-derived fragments using mouse tumors transplanted into syngeneic immunocompetent mice to test an agonist anti-CD137 mAb and its combinations with anti-PD-1 and/or anti-TGF-β. Increases in IFNγ concentrations in the tissue culture supernatants were detected upon in-culture activation with the anti-CD137 and anti-PD-1 mAb combinations or concanavalin A as a positive control. No other cytokine from a wide array was informative of stimulation with these mAbs. Interestingly, increases in Ki67 and other activation markers were substantiated in lymphocytes from cell suspensions gathered at the end of 72 h cultures. In mice bearing bilateral tumors in which one was excised prior to in vivo anti-CD137 + anti-PD-1 treatment to perform the fragment culture evaluation, no association was found between IFNγ production from the fragments and the in vivo therapeutic outcome in the non-resected contralateral tumors. The experimental system permitted freezing and thawing of the fragments with similar functional outcomes. Using a series of patient-derived tumor fragments from excised solid malignancies, we showed IFNγ production in a fraction of the studied cases, that was conserved in frozen/thawed fragments. The small tumor fragment culture technique seems suitable to preclinically explore immunotherapy combinations.Biomarkers for cancer immunotherapy are an unmet medical need. The group of Daniela Thommen at the NKI recently reported on novel methodologies based on short-term cultures of patient-derived tumor fragments whose cytokine concentrations in the supernatants and activation markers on infiltrating T cells were associated with clinical response to PD-1 blockade. We set up a similar culture technology with tumor-derived fragments using mouse tumors transplanted into syngeneic immunocompetent mice to test an agonist anti-CD137 mAb and its combinations with anti-PD-1 and/or anti-TGF-β. Increases in IFNγ concentrations in the tissue culture supernatants were detected upon in-culture activation with the anti-CD137 and anti-PD-1 mAb combinations or concanavalin A as a positive control. No other cytokine from a wide array was informative of stimulation with these mAbs. Interestingly, increases in Ki67 and other activation markers were substantiated in lymphocytes from cell suspensions gathered at the end of 72 h cultures. In mice bearing bilateral tumors in which one was excised prior to in vivo anti-CD137 + anti-PD-1 treatment to perform the fragment culture evaluation, no association was found between IFNγ production from the fragments and the in vivo therapeutic outcome in the non-resected contralateral tumors. The experimental system permitted freezing and thawing of the fragments with similar functional outcomes. Using a series of patient-derived tumor fragments from excised solid malignancies, we showed IFNγ production in a fraction of the studied cases, that was conserved in frozen/thawed fragments. The small tumor fragment culture technique seems suitable to preclinically explore immunotherapy combinations.
Objective
We aimed to investigate the prevalence of TOR1A, GNAL and THAP1 variants as the cause of dystonia in a cohort of Spanish patients with isolated dystonia and in the literature.
Methods
A ...population of 2028 subjects (including 1053 patients with different subtypes of isolated dystonia and 975 healthy controls) from southern and central Spain was included. The genes TOR1A, THAP1 and GNAL were screened using a combination of high‐resolution melting analysis and direct DNA resequencing. In addition, an extensive literature search to identify original articles (published before 10 August 2020) reporting mutations in TOR1A, THAP1 or GNAL associated to dystonia was performed.
Results
Pathogenic or likely pathogenic variants in TOR1A, THAP1 and GNAL were identified in 0.48%, 0.57% and 0.29% of our patients, respectively. Five patients carried the variation p.Glu303del in TOR1A. A very rare variant in GNAL (p.Ser238Asn) was found as a putative risk factor for dystonia.
In the literature, variations in TOR1A, THAP1 and GNAL accounted for about 6%, 1.8% and 1.1% of published dystonia patients, respectively.
Conclusions
There is a different genetic contribution to dystonia of these three genes in our patients (about 1.3% of patients) and in the literature (about 3.6% of patients), probably due the high proportion of adult‐onset cases in our cohort. As regards age at onset, site of dystonia onset, and final distribution, in our population there is a clear differentiation between DYT‐TOR1A and DYT‐GNAL, with DYT‐THAP1 likely to be an intermediate phenotype.
Although the molecular mechanisms underlying isolated dystonia are largely unknown, it is known that its pathogenesis is genetically heterogeneous. Causative mutations in three genes (TOR1A, THAP1, and GNAL) have been identified in patients from different populations. This study is the largest and most complete study of TOR1A, THAP1 and GNAL genes in the Spanish population and a review of the recent genetic findings in these genes. The incidence of pathogenic or likely pathogenic variants in these three genes in our population is low. As regards age at onset, site of dystonia onset, and final distribution, in our population there is a clear differentiation between DYT‐TOR1A and DYT‐GNAL, with DYT‐THAP1 likely to be an intermediate phenotype.
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