Development of hen egg-based protein beverages with high nutritional value.

Physical stability is a prerequisite for any emulsion-based beverage. This study aimed to optimize beverage emulsion (BE) rich in whole egg, whey protein isolate (WPI), and soyabean oil in the presence of polygalactose derivative as adsorbing hydrocolloids (AH) and partially hydrolyzed starch as non-adsorbing hydrocolloids (NAH). All formulations were optimized for stability, particle size, polydispersity index (PDI), and zeta potential (ζ). A central composite design was applied to investigate the impact of WPI concentration, hydrocolloid concentration, oil volume, and pH as independent factors on particle size, PDI, and ζ of emulsions. The optimum conditions were met upon formulating BE with 1 whole egg, 9.28 g WPI, 11.24 g AH, 5.58 mL oil at pH 7. Two optimal formulations were obtained for BE stabilized with NAH at pH 3 and 7, at which droplets had the highest electrical charge. Noting that, less amount of NAH (7.5 g) was required at pH 3 than at pH 7 (13.56 g). Oil content (3 mL) and WPI concentration (16 g) for both NAH formulations were similar. The size, polydispersity index (PDI), and zeta potential of AH-stabilized emulsion were 762 nm, 0.69, and - 32.05 mV, respectively, while for NAH emulsion, they were 150.02 nm, 0.40, and - 28.53 mV at pH 7; and 248.39 nm, 0.41, and 27.1 mV at pH 3, respectively. No conspicuous gravitational separation was observed after 20 days of storage at 4 °C. Hence, both hydrocolloids are appropriate to stabilize protein-enriched BE at neutral pH; however, physical stability was also achieved at acidic pH for NAH-stabilized emulsions.

Adaptive Radioresistance of Enterohemorrhagic Escherichia coli O157:H7 Results in Genomic Loss of Shiga Toxin-Encoding Prophages.

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a foodborne pathogen producing Shiga toxins (Stx1 and Stx2), which can cause hemorrhagic diarrhea and life-threatening infections. O157:H7 strain EDL933 carries prophages CP-933V and BP-933W, which encode Shiga toxin genes (stx1 and stx2, respectively). The aim of this work was to investigate the mechanisms of adaptive resistance of EHEC strain EDL933 to a typically lethal dose of gamma irradiation (1.5 kGy). Adaptive selection through six passages of exposure to 1.5 kGy resulted in the loss of CP-933V and BP-933W prophages from the genome and mutations within three genes: wrbA, rpoA, and Wt_02639 (molY). Three selected EHEC clones that became irradiation adapted to the 1.5-kGy dose (C1, C2, and C3) demonstrated increased resistance to oxidative stress, sensitivity to acid pH, and decreased cytotoxicity to Vero cells. To confirm that loss of prophages plays a role in increased radioresistance, clones C1 and C2 were exposed to bacteriophage-containing lysates. Although phage BP-933W could lysogenize C1, C2, and E. coli K-12 strain MG1655, it was not found to have integrated into the bacterial chromosome in C1-Φ and C2-Φ lysogens. Interestingly, for the E. coli K-12 lysogen (K-12-Φ), BP-933W DNA had integrated at the wrbA gene (K-12-Φ). Both C1-Φ and C2-Φ lysogens regained sensitivity to oxidative stress, were more effectively killed by a 1.5-kGy gamma irradiation dose, and had regained cytotoxicity and acid resistance phenotypes. Further, the K-12-Φ lysogen became cytotoxic, more sensitive to gamma irradiation and oxidative stress, and slightly more acid resistant. IMPORTANCE Gamma irradiation of food products can provide an effective means of eliminating bacterial pathogens such as enterohemorrhagic Escherichia coli (EHEC) O157:H7, a significant foodborne pathogen that can cause severe disease due to the production of Stx. To decipher the mechanisms of adaptive resistance of the O157:H7 strain EDL933, we evolved clones of this bacterium resistant to a lethal dose of gamma irradiation by repeatedly exposing bacterial cells to irradiation following a growth restoration over six successive passages. Our findings provide evidence that adaptive selection involved modifications in the bacterial genome, including deletion of the CP-933V and BP-933W prophages. These mutations in EHEC O157:H7 resulted in loss of stx1 and stx2, loss of cytotoxicity to epithelial cells, and decreased resistance to acidity, critical virulence determinants of EHEC, concomitant with increased resistance to lethal irradiation and oxidative stress. These findings demonstrate that the potential adaptation of EHEC to high doses of radiation would involve elimination of the Stx-encoding phages and likely lead to a substantial attenuation of virulence.

Optimization of a natural antimicrobial formulation against potential meat spoilage bacteria and food-borne pathogens: Mixture design methodology and predictive modeling.

This study is about the combined antimicrobial effect of essential oils (EOs), namely Mediterranean (MN) EO, German thyme (GT) EO, Cinnamon (CN) EO, Indian (IN) EO, Asian (AN) EO, and citrus extract (CE) against spoilage bacteria (Lactobacillus sakei, Lactobacillus curvatus, Leuconostoc mesenteroides, Carnobacterium divergens, Brochothrix thermosphacta, and Pseudomonas aeruginosa) and selected pathogenic bacteria (E. coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes). Firstly, each EO and CE were screened for antibacterial activity by microdilution assay, and the most efficient antimicrobial extracts were selected based on the lowest MIC values to perform the combination assays. Afterward, a simplex-centroid mixture design was used to develop optimal antimicrobial mixtures capable of protecting meat from spoilage and pathogenic bacteria. The optimization tool allowed us to postulate models and validate them statistically as well as to create a prediction profile of the experiment. Thus, the optimal mixtures named active formulation 1 (AF1) containing MN EO/GT EO/VC EO/CE with a ratio of 1:2:2:1 and active formulation 2 (AF2) containing IN EO/AN EO/CE/VC EO with a ratio of 2:2:1:2, were developed based on the demonstration of their synergistic effect against tested bacteria. The obtained formulations at organoleptically acceptable concentrations could be applied in the preservation of meat and meat products.

Essential oils as natural antimicrobials applied in meat and meat products-a review.

Meat and meat products are highly susceptible to the growth of micro-organism and foodborne pathogens that leads to severe economic loss and health hazards. High consumption and a considerable waste of meat and meat products result in the demand for safe and efficient preservation methods. Instead of synthetic additives, the use of natural preservative materials represents an interest. Essential oils (EOs), as the all-natural and green-label trend attributing to remarkable biological potency, have been adopted for controlling the safety and quality of meat products. Some EOs, such as thyme, cinnamon, rosemary, and garlic, showed a strong antimicrobial activity individually and in combination. To eliminate or reduce the organoleptic defects of EOs in practical application, EOs encapsulation in wall materials can improve the stability and antimicrobial ability of EOs in meat products. In this review, meat deteriorations, antimicrobial capacity (components, effectiveness, and interactions), and mechanisms of EOs are reviewed, as well as the demonstration of using encapsulation for masking intense aroma and conducting control release is presented. The use of EOs individually or in combination and encapsulated applications of EOs in meat and meat products are also discussed.

Transcriptome analysis of the Clostridioides difficile response to a specific lactobacilli probiotic formulation: explanations for its mechanisms of action.

AIMS: Clostridioides difficile infections (CDI) are a major cause of morbidity and mortality in hospitalized patients. A probiotic formulation (Bio-K+) comprised of Lactobacillus acidophilus CL1285, Lacticaseibacillus casei LBC80R, and Lacti. rhamnosus CLR2 strains have been shown to reduce the incidence of CDI and antibiotic-associated diarrhea (AAD). This research aims to therefore elucidate the mechanism of action of the three probiotic strained against C. difficile R20291, independently of the acidification of the environment. . METHODS AND RESULTS: Antitoxin activity was evaluated using ELISA method and the expression of C. difficile genes was evaluated using transcriptomic analysis in co-culture assays conducted in a bioreactor allowing precise control of the pH. The fermentation results demonstrated a decrease for toxin A and many genes directly related to C. difficile virulence were underexpressed in the co-cultures. CONCLUSIONS: The lactobacilli tested could have a role in the motility, the quorum sensing (QS), the survival of the spores, and the germination potential of the spores, which are essential elements for the virulence of C. difficile. .

Influence of growth parameters on bacteriocin-like inhibitory substances (BLIS) production by lactic acid bacteria.

This study aimed to investigate the possible production of bacteriocin-like inhibitory substances (BLIS) and optimize it by Lactobacillus sakei and combined Lactobacillus curvatus/Pediococcus acidilactici, lactic acid bacteria used for the meat fermentation. The effect of growth parameters on BLIS production was evaluated by measuring the antibacterial activity of the cell-free supernatant of these bacteria. The stability of the cell-free supernatant of combined L.curvatus/P.acidilactici bacteria was also examined after heat treatment and pH variations. Results showed highest BLIS production by L.curvatus/P.acidilactici was at growth temperatures of 28°C and pH 5, while the optimum condition for production of BLIS by L.sakei was 24°C and pH 6.5. The BLIS production by L. curvatus/P. acidilactici bacteria was greatly influenced by carbon and nitrogen sources. The antimicrobial activity of BLIS in supplemented De Man, Rosa, and Sharpe broth (MRS) was 2044.44 AU/mL, 4.84-fold greater than unsupplemented MRS. None of the carbon sources used in this study affected the BLIS production by the L.sakei, while pea peptone enhanced the antimicrobial activity and showed a value of 188.89 AU/mL, 1.47-fold greater than unsupplemented MRS. The characterization of the cell-free supernatant of L. curvatus/P. acidilactici bacteria showed the heat stability and activity of BLIS at pH 2-10.

Food grade nanoemulsion development to control food spoilage microorganisms on bread surface.

In this study, the effect of emulsifier mixture and their concentrations on the development of nanoemulsion was studied. The impact of sonication and microfluidization processing conditions on the physicochemical properties and in vitro antimicrobial activity was also evaluated. The optimal nanoemulsion formulation was then evaluated on bread surface against B. subtilis. Results showed that a hydrophilic-lipophilic balance HLB = 12 and emulsifier: oil ratio of 1:1 allowed the formation of stable nanoemulsion. Also, both microfluidization and sonication allowed the formation of nanoscale-emulsion. Sonication treatment for 10 min allowed a maintain the total flavonoid content and a slight reduction of total phenol content. Furthermore, employing sonication resulted to the lowest polydispersity index suggesting more stable nanoemulsion. Nanoscale-emulsion showed a good in vitro antimicrobial activity against L. monocytogenes and E. coli. The application of nanoemulsion on bread surface inoculated with B. subtilis showed a delay of the decay. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05660-5.

Silver nanoparticles-essential oils combined treatments to enhance the antibacterial and antifungal properties against foodborne pathogens and spoilage microorganisms.

Plant-derived essential oils (EOs) and commercial silver nanoparticles (AgNPs) were tested to evaluate their antibacterial and antifungal efficiency against two pathogenic bacteria (Escherichia coli O157:H7 and Salmonella Typhimurium) and three spoilage fungi (Aspergillus niger, Penicillium chrysogenum, and Mucor circinelloides). A broth microdilution assay was used to determine the minimal inhibitory concentration (MIC) of EOs and AgNPs. In the MIC assay, the cinnamon EO, Mediterranean formulation, citrus EO and spherical-shaped silver nanoparticles (AgNPs) (AGC 1, AGC 0.5, AGPP and AGPPH) showed moderate to high antibacterial and antifungal properties, with MIC ranging from 7.8 to 62.5 ppm for AgNPs and 312.5-1250 ppm for EOs against the tested bacteria and fungi. The possible interaction between the EOs and the AgNPs was determined using a checkerboard method by evaluating fractional inhibitory concentration (FIC) values. The combination of two or more EOs and AgNPs (Active combination 1: AGPPH+cinnamon EO, Active combination 2: AGC 0.5+Mediterranean formulation+citrus EO, Active combination 3: AGPP+cinnamon EO+Asian formulation+lavang EO) showed synergistic effects (FIC <1.0) against all tested bacteria and fungi. A modified Gompertz model was used to evaluate growth parameters including maximum colony diameter (A), maximum growth rate (Vm), and lag phase (λ), under the three active combinations suggested by the checkerboard method using a vapor assay. The three active combinations 1, 2 and 3 reduced the growth rate and maximum colony diameter of E. coli, S. Typhimurium, A. niger, P. chrysogenum, and M. circinelloides, and extended their lag phase from 1 to 5 days. In in situ tests with inoculated rice, the three active combinations showed a significant reduction of all tested bacteria and fungi at 27 °C for 28 days.

Norovirus elimination on the surface of fresh foods.

Fresh foods like fruits, vegetables and shellfish are potential sources for viral infections such as human norovirus (NoV). Chemical treatment like chlorination is a well-known process for food pathogens and virus elimination. However, with the increase of the consumer demand for less toxic treatment, the use of natural antimicrobials like essential oils from spice or plants, fruit extracts, and cold pasteurization treatments (fermentation, irradiation, ozonation and high pressure) could be considered. The aim of this review is to discuss these technologies and their efficacy to eliminate NoV on the surface of fresh food.

Physico-chemical Properties and Sensorial Appreciation of a New Fermented Probiotic Beverage Enriched with Pea and Rice Proteins.

OBJECTIVES: The purpose of this study was to evaluate the physico-chemical stability, the sensorial properties, and the microbial quality of a fermented beverage enriched with pea and rice proteins (PRF) during storage at 4 °C. To investigate the effect of the protein enrichment and fermentation, the PRF beverage quality was compared with non-fermented and non-enriched beverages. METHODS: The beverage was supplemented with a 50/50 mixture of pea and rice protein concentrate to 13% concentration. Following inoculation with 108 CFU/mL of lactic acid bacteria, it was incubated at 37 °C for 14 h. RESULTS: Results showed that the enrichment with protein induced an increase in pH, titratable acidity and viscosity of the PR products, while the fermentation led to a decrease of pH and viscosity. However, a significant increase of the viscosity of PRF from 39 to 57 cP was observed during the 143 days of storage (P ≤ 0.05). The PRF beverage contained significantly more peptides < 200 Da than the non-fermented one (PRNF) and these small peptides were also released during the storage. Despite the physico-chemical modifications, the sensorial properties of the PRF product were appreciated over the storage, particularly for the texture. Furthermore, the beverage maintained a high concentration of viable probiotics during the entire storage with 8.4 log colony form unit (CFU)/mL after 143 days. CONCLUSION: Applying probiotics and the mixture of rice and pea proteins in the fermented beverage can enhance nutritional and nutraceutical value of the product.

A rapid way of formulation development revealing potential synergic effects on numerous antimicrobial combinations against foodborne pathogens.

The interactions between various essential oils (EOs) were evaluated for the development of antimicrobial formulations. A full factorial design was applied for testing eight EOs (Mustard, Thyme, Garlic, Oregano, Chinese cinnamon, Cinnamon bark, Red bergamot, Winter savory) against nine bacteria (E.coli O157:H7 RM1239, E.coli O157:H7 RM 1931, E.coli O157:H7 RM 1933, E.coli O157:H7 RM 1934, E.coli O157:H7 380-94, Listeria monocytogenes LM 1045, Listeria innocua ATCC 51742, Salmonella Typhimurium SL 1344, Salmonella enterica Newport ATCC 6962) and two molds (Penicillium chrysogenum ATCC 10106, Aspergillus niger ATCC 1015). Results showed that combinations of Thyme + Oregano, Oregano + Cinnamon bark, Chinese cinnamon + Cinnamon bark have shown high interactions in Factorial design and validated to be mostly additive effects against tested bacteria. The combination of Mustard + Thyme, Mustard + Garlic, and Thyme + Garlic EOs showed high interactions and also all additive effects against tested molds. The corresponding results of Factorial design and checkerboard render the designation to demonstrate the highly efficient formulations and interactions rapidly among abundant mixtures.

Potential role of probiotics in reducing Clostridioides difficile virulence: Interference with quorum sensing systems.

Opportunistic pathogenic bacteria may cause disease after the normally protective microbiome is disrupted (typically by antibiotic exposure). Clostridioides difficile is one such pathogen having a severe impact on healthcare facilities and increasing costs of medical care. The search for new therapeutic strategies that are not reliant on additional antibiotic exposures are currently being explored. One such strategy is to disrupt the production of C. difficile virulence factors by interfering with quorum sensing (QS) systems. QS has been well studied in other bacteria, but our understanding in C. difficile is not so well understood. Some probiotic strains or combinations of strains have been shown to be effective in the treatment or primary prevention of C. difficile infections and may possess multiple mechanisms of action. One mechanism of probiotics might be the inhibition of QS, but their role has not been clearly defined yet. A literature search was conducted using standard databases (PubMed, Google Scholar) from database inception to August 2020. The objective of this paper is to update our understanding of how QS leads to toxin production by C. difficile, which is important in pathogenesis, and how QS inhibitors or probiotics may disrupt this pathway. We found two main QS systems for C. difficile (Agr and Lux systems) that are involved in C. difficile pathogenesis by regulating toxin production, motility and adherence. Probiotics and other QS inhibitors targeting QS systems may represent important new directions of therapy and prevention of CDI.

Chemopreventive Properties of Extracts Obtained from Blueberry (Vaccinium myrtillus L.) and Jabuticaba (Myrciaria cauliflora Berg.) in Combination with Probiotics.

The association of probiotics and fruit extracts may influence the chemopreventive effect of colorectal cancer. In this context, antiproliferative activity was evaluated to select the best extracts that would be added probiotics, after addition of Bifidobacterium or Lactobacillus in the extracts the antiradical and antioxidant activity, quinone reductase (QR) assay and apoptosis assay were evaluated. Four extracts were isolated: E1: rich in total phenolic compounds; E2: rich in water-soluble phenolic compounds; E3: rich in most apolar phenolic compounds and E4: rich in anthocyanins. The antiproliferative results showed that the best extracts for blueberry and jabuticaba were, respectively the extract E4 and E2. After addition of the probiotic bacteria in these best extracts, it was observed that E2 from jabuticaba presented significantly higher antiradical and antioxidant activity values compared to E4 from blueberry before and after addition of probiotics. There was also a 9-fold increase in activity of QR by the E2 from jabuticaba with Lactobacillus (JL). Likewise, this same extract showed a significant increase both in apoptotic and necrotic cells for both cells. In conclusion, extract rich in water-soluble phenolic compounds (E2) from jabuticaba presented a greater chemopreventive effect compared to the others.

Microbial radiosensitization using combined treatments of essential oils and irradiation- part B: Comparison between gamma-ray and X-ray at different dose rates.

Stored rice and rice products are prone to contamination by pathogenic fungi and bacteria such as Aspergillus niger, Bacillus cereus, and Paenibacillus amylolyticus. Treatment with antimicrobial essential oils (EOs) and irradiation are options to control spoilage organisms. Microbial samples with or without fumigation with an oregano/thyme EO mixture were irradiated at 0.5, 0.75, 1.0, 1.5, 2.0, 2.5, 3.0 and 3.5 kGy for calculation of a D10 value. The relative sensitivity was calculated as the ratio of D10 values for the irradiation plus oregano and thyme EO combination and irradiation alone treatments. In all cases, irradiation plus fumigation with the oregano and thyme EO mixture showed increased efficacy compared with irradiation alone. The relative sensitivity of γ-ray irradiation against A. niger was 1.22, 1.33, and 1.24 for radiation dose rates of 10.445, 4.558, and 0.085 kGy/h, respectively, however against B. cereus it was 1.28, 1.45, and 1.49, and against P. amylolyticus it was 1.35, 1.33, and 1.38, for respective γ-ray irradiation dose rates. The relative sensitivity of X-ray irradiation against A. niger, B. cereus, and P. amylolyticus was 1.63, 1.21, and 1.31, respectively, at the X-ray dose rate of 0.76 kGy/h. The results showed that the relative sensitivity of γ-ray irradiation was higher against the two bacteria than the fungus, whereas X-ray showed higher sensitivity against the fungus than the two bacteria. There was no consistent positive or negative relationship between dose rate and relative sensitivity. The results demonstrated the potential of an oregano and thyme EOs mixture as an antimicrobial agent and its efficacy to increase the radiosensitization of A. niger, B. cereus, and P. amylolyticus during γ-ray or X-ray irradiation treatments.

Correlation between chemical composition and antimicrobial properties of essential oils against most common food pathogens and spoilers: In-vitro efficacy and predictive modelling.

Using disk diffusion assay and broth microdilution, we evaluated the antimicrobial activity of 38 commercially available essential oils (EOs) against 24 food pathogens and spoilers. These including E. coli O157: H7 (3 types), Listeria (3 types), Bacillus (2 types), Salmonella enterica (2 types), Staphylococcus aureus (3 types), Clostridium tyrobutiricum, Pseudomonas aeruginosa, Brochotrix thermosphacta, Campylobacter jejuni, Carnobacterium divergens, Aspergillus (2 types), and Penicillium (4 types). Correlation between EOs' chemical composition and antimicrobial properties was studied using R software. Moreover, statistical models representing the relationship were generated using Design Expert®. The predictive models identified the chemical attributes of EOs that drive their antimicrobial properties while providing an understanding of their interactions. Thyme (Aldrich, Novotaste), cinnamon (Aliksir, BSA), garlic (Novotaste), Mexican garlic blend N & A (Novotaste), and oregano (BSA) were the strongest antimicrobial. The most sensitive pathogens were P. solitum (MIC of 19.53 ppm) and L. monocytogenes (MIC of 39 ppm). The correlation analysis showed that phenols and aldehydes had the strongest positive effects on the antimicrobial properties followed by the sulfur containing compounds and the esters; while the effects of monoterpenes and ketones were negative. Different sensitivity of food pathogens to chemical families was observed. For instance, phenols and aldehydes exhibited a linear inhibitory effect on L. monocytogenes (LM1045, MIC), while sesquiterpene and ester showed a significant effect on S. aureus (ATCC 6538, MIC). The developed predictive models are expected to predict the antimicrobial properties based on the chemical families of essential oils.

Gamma irradiation triggers a global stress response in Escherichia coli O157:H7 including base and nucleotides excision repair pathways.

Shiga toxin-producing Escherichia coli O157:H7, one of the most severe human foodborne pathogens, can withstand several stresses, including some levels of γ-irradiation. In this study, the response of E. coli O157:H7 to a sensitization irradiation dose of 0.4 kGy was assessed using RNA-seq transcriptomic at 10 (t10) and 60 (t60) min post-irradiation, combined with an isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis at 60 min post-irradiation. Several functions were induced by the treatment, such as base excision repair and nucleotide excision repair pathways; sulfur and histidine metabolism, and virulence mechanisms. Additionally, the sulA gene, coding for the cell division repressor, together with other genes involved in SOS response and repair mechanism (including recA, recN, recJ, recQ, mutM and uvrB) were up-regulated at t60. As the early response to irradiation stress (t10), dnaK, groEL, ibpA, sulfur metabolism genes, as well as those related to oxidative stress were up-regulated, while histidine biosynthesis genes were down-regulated. Acid stress, heat shock, UV resistance and several virulence genes, especially stx2A/stx2b which code for the Shiga toxins characteristic of O157:H7, were upregulated at 60 min post-irradiation. The treatment was also found to increase the levels of CysN, MutM, DinG and DnaC in the cells, proteins involved respectively in sulfur metabolism, base excision repair, recombinational DNA repair and chromosome replication. Our results provide insights into the resistance response of E. coli O157:H7 to a non-lethal irradiation dose. Our findings indicate that E. coli O157:H7 can resist to γ-irradiation through important modifications in genes expression and proteins profiles.

Potential synergistic antimicrobial efficiency of binary combinations of essential oils against Bacillus cereus and Paenibacillus amylolyticus-Part A.

The checkerboard method was used to study the potential interactions between eight essential oils (Basil, Cinnamon, Eucalyptus, Mandarin, Oregano, Peppermint, Tea tree, and Thyme) when used as antibacterial agents against Bacillus cereus LSPQ 2872 and Paenibacillus amylolyticus ATCC 9995. The minimum inhibitory concentration (MIC) of each essential oil (EO) and the fractional inhibitory concentration (FIC) index for the binary combinations of essential oils (EOs) were determined. According to FIC index values, some of the compound binary combinations showed an additive effect; however, Thyme/Tea tree and Cinnamon/Thyme EOs exhibited a synergistic effect against P. amylolyticus and B. cereus, respectively. Cinnamon/Thyme EOs mixture exhibited no interactive effect against P. amylolyticus, but a synergistic effect against B. cereus. The combination of Oregano/Thyme EOs displayed the best antibacterial activity and showed a synergistic effect against B. cereus and P. amylolyticus bacteria. The Oregano/Thyme EOs mixture has potential application in food preservation to reduce the contamination of B. cereus and P. amylolyticus.

Cancer Chemopreventive, Antiproliferative, and Superoxide Anion Scavenging Properties of Kluyveromyces marxianus and Saccharomyces cerevisiae var. boulardii Cell Wall Components.

This study investigated the cancer chemopreventive, the antiradical, and the antiproliferative properties of polysaccharides extracts from cell wall of Saccharomyces boulardii and Kluyveromyces marxianus. ß-glucan, mannan, and chitin were also quantified to identify the most important extract responsible for these biological properties. Soluble and insoluble glucans as well as mannoprotein were extracted from cell wall using single hot-alkaline method. Superoxide anion scavenging (antiradical capacity), NAD(P)H: quinone reductase (QR) (EC 1.6.99.2) induction, and antiproliferative assays were done for the evaluation of biological properties of those extracts. The insoluble glucan from S. boulardii revealed the most relevant biological properties by increasing QR activity and exhibiting the highest growth inhibition against colorectal cancer cells. Moreover, high amount of glucan, high glucan/total sugars ratios, and low chitin/glucan ratios were shown to have an impact on enhancing cancer chemopreventive and antiproliferative properties. To our knowledge, this is the first study that demonstrates QR activity by yeast cell wall components in a dose-dependent manner.

Effect of Saccharomyces Boulardii Cell Wall Extracts on Colon Cancer Prevention in Male F344 Rats Treated with 1,2-Dimethylhydrazine.

The effect of Saccharomyces boulardii cell wall extracts on colon cancer prevention in rats treated with 1,2-dimethylhydrazine was investigated. A crude insoluble glucan (0.5 and 1.0 mg/kg/day) and a crude mannoprotein extract (0.3 and 3.0 mg/kg/day) were administered in rats by gavage for 12 weeks along with a high fat low fiber diet whereupon rats were sacrificed and aberrant crypt foci (ACF) were counted in the colon. Moreover, NAD(P)H: quinone reductase (QR) and harmful fecal enzymes (ß-glucosidase and ß-glucuronidase) were quantified in the liver and in the caecum, respectively. Results showed a reduction in ACF counts, a decreased ß-glucuronidase activity and an increased QR activity when rats were treated only with insoluble glucan. While these enzymatic modulations may be constituted one of the mechanisms that is responsible for the reduction of ACF counts observed, the reduction of ACF counts caused by insoluble glucan should be addressed, at least, as a biomarker of their cancer-prevention properties. To our knowledge, this is the first study demonstrated that crude cell wall extract obtained from S. boulardii could have a potential role in colon cancer prevention in vivo by revealing the potential implication of QR and ß-glucuronidase modulation.

Human L-ficolin recognizes phosphocholine moieties of pneumococcal teichoic acid.

Human L-ficolin is a soluble protein of the innate immune system able to sense pathogens through its fibrinogen (FBG) recognition domains and to trigger activation of the lectin complement pathway through associated serine proteases. L-Ficolin has been previously shown to recognize pneumococcal clinical isolates, but its ligands and especially its molecular specificity remain to be identified. Using solid-phase binding assays, serum and recombinant L-ficolins were shown to interact with serotype 2 pneumococcal strain D39 and its unencapsulated R6 derivative. Incubation of both strains with serum triggered complement activation, as measured by C4b and C3b deposition, which was decreased by using ficolin-depleted serum. Recombinant L-ficolin and its FBG-like recognition domain bound to isolated pneumococcal cell wall extracts, whereas binding to cell walls depleted of teichoic acid (TA) was decreased. Both proteins were also shown to interact with two synthetic TA compounds, each comprising part structures of the complete lipoteichoic acid molecule with two PCho residues. Competition studies and direct interaction measurements by surface plasmon resonance identified PCho as a novel L-ficolin ligand. Structural analysis of complexes of the FBG domain of L-ficolin and PCho revealed that the phosphate moiety interacts with amino acids previously shown to define an acetyl binding site. Consequently, binding of L-ficolin to immobilized acetylated BSA was inhibited by PCho and synthetic TA. Binding of serum L-ficolin to immobilized synthetic TA and PCho-conjugated BSA triggered activation of the lectin complement pathway, thus further supporting the hypothesis of L-ficolin involvement in host antipneumococcal defense.