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This study investigated the antimicrobial action of oleanolic acid against Listeria monocytogenes, Enterococcus faecium, and Enterococcus faecalis. To determine the cytotoxicity of oleanolic acid, HEp-2 cells were incubated with oleanolic acid at 37°C. MICs (minimal inhibition concentrations) for L. monocytogenes, E. faecium, and E. faecalis were determined using two-fold microdilutions of oleanolic acid, and bacterial cell viability was then assessed by exposing the bacteria to oleanolic acid at 2 × MIC. To investigate the mode of antimicrobial action of oleanolic acid, we measured leakage of compounds absorbing at 280 nm, along with propidium iodide uptake. Scanning electron microscope (SEM) images were also analysed. The viability of HEp-2 cells decreased (P < 0.05) at oleanolic acid concentrations greater than 128 μg mL(-1). The MICs were 16-32 μg mL(-1) for L. monocytogenes and 32-64 μg mL(-1) for E. faecium and E. faecalis, and bacterial cell viability decreased (P < 0.05) about 3-4 log CFU mL(-1) after exposure to 2 × MIC of oleanolic acid. Leakage of 280 nm absorbing materials and propidium iodide uptake was higher in oleanolic acid -treated cells than in the control. The cell membrane was damaged in oleanolic acid-treated cells, but the control group had intact cell membrane in SEM images. The results indicate that oleanolic acid can kill L. monocytogenes, E. faecium, and E. faecalis by destroying the bacterial cell membrane.

Periodontitis has been linked to systemic inflammation, however research on its role in causing systemic diseases remains limited. Recent studies explore probiotics for microbiome modulation and enhancing natural compound bioavailability. This study investigated periodontitis-related systemic disease mechanisms, and evaluated the mitigation effects of bioconversion product using SMFM2016-RK and extracts. Four types of bioconverted milk [BM1 ( SMFM2016-RK), BM2 (BM1 +  ethanol extract), BM3 (BM1 +  hot-water extract), and BM4 (BM1+ both extracts)] were studied in a periodontitis-induced rat model. Rats were divided into six groups: normal control, skim milk with ligature, and four BM groups with ligature.   Periodontitis induction elevated trabecular resorption (0.325 ± 0.057 mm³) and histopathological symptoms. Serum ALT (55.6 ± 6.6 U/L), glucose (261.7 ± 64.3 mg/dL), insulin (1.90 ± 0.87 ng/mL), inflammation in the liver and colon, and gluconeogenesis-related enzyme expression increased. Periodontitis-induced rats showed gut dysbiosis, with decreased level and increased level. BM3 administration significantly reduced the serum glucose (190.9 ± 27.8 mg/dL), ALT (40.5 ± 5.0 U/L), inflammation, and gluconeogenesis-related enzymes, while increasing tight junction proteins expression and phylum Actinobacteria levels in the gut microbiome. The findings highlight the systemic impact of periodontitis on inflammation, glycemic control, and gut microbiome balance. BM3 effectively alleviated these effects suggesting therapeutic potential.

To decrease periodontal pathogens and increase the number of beneficial bacteria, probiotics and bioactive compounds made via microbial bioconversion are recently used. In addition, the interest regarding probiotics-mediated bioconversion with popular medicinal plants is increasing. Artemisia herba-alba, a type of wormwood, has recently been attention as a medicinal plant due to its various bioactive compounds. Therefore, we developed bioconverted milk containing A. herba-alba that effectively inhibited periodontal pathogens and α-glucosidase. To select the appropriate lactic acid bacteria for the probiotic candidate strain, 74 strains of lactic acid bacteria were screened. Among them, Lactiplantibacillus plantarum SMFM2016-RK was chosen as the probiotic due to its beneficial characteristics such as high acid and bile tolerance, antioxidant activity, and α-glucosidase inhibition. Based on the minimal bactericidal concentration against three periodontal pathogens, the following appropriate concentrations of Artemisia herba-alba extract were added to milk: 5 mg/mL of A. herba-alba ethanol extract and 25 mg/mL of A. herba-alba hot-water extract. Four bioconverted milks (BM), BM1, BM2, BM3, and BM4, were produced by combining L. plantarum SMFM2016-RK alone, L. plantarum SMFM2016-RK and ethanol extract, L. plantarum SMFM2016-RK and hot-water extract, and L. plantarum SMFM2016-RK with both extracts. As a result of antimicrobial activity, BM3 inhibited the growth of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis the most, and BM4 suppressed the growth of Fusobacterium nucleatum the most. In addition, bioconverted milk containing A. herba-alba (BM2, BM3, and BM4) inhibited α-glucosidase more effectively than BM1. The whole genome of L. plantarum SMFM2016-RK was obtained, and 3135 CDS, 67 tRNA, and 16 RNA were predicted. The genome annotation of L. plantarum SMFM2016-RK revealed 11 CDS related to proteolysis and amino acid metabolism and 2 CDS of phenolic acid-metabolizing enzymes. In conclusion, A. herba-alba-added milk bioconverted by L. plantarum SMFM2016-RK displayed both the growth inhibitory effect on periodontal pathogens and the α-glucosidase inhibitory activity; thus, it necessitates to evaluate the effects on the alleviation of periodontal diseases and glycemic control through future animal experiments.

Novel bioactive metabolites have been developed through a bioconversion of dairy products or other foods using probiotics isolated from dairy products or other fermented foods. These probiotics-mediated bioconversion (PMB) metabolites show antioxidant, anti-inflammatory, antimicrobial, epithelial barrier, and anticancer activities. In addition, the effect of PMB metabolites in periodontitis is recently reported in several studies. Periodontitis is a chronic inflammatory disease caused by infections, and the tooth support tissue is destroyed. Common treatments for periodontitis include scaling and root planning with systemic antibiotics. However, the overuse of antibiotics has led to the emergence of drug-resistant microorganisms and disturbs the beneficial bacteria, including lactobacilli in the oral cavity. For this reason, PMB metabolites, such as fermented milk, have been suggested as substitutes for antibiotics to reduce periodontitis. This paper reviews the recent studies on the correlation between periodontitis and PMB metabolites and classifies the efficacy of major PMB metabolites for periodontitis. The review suggests that PMB is effective for periodontitis, and further studies are needed to confirm the therapeutic effect of PMB metabolites on periodontitis.

Pseudomonas aeruginosa is a pathogen that can cause serious infections and usually coexists with other pathogens, such as Staphylococcus aureus. Virulence factors are important for maintaining a presence of the organisms in these multispecies environments, and DesB plays an important role in P. aeruginosa virulence. Therefore, we investigated the effect of DesB on S. aureus reduction under competitive situation. Liquid cultures of P. aeruginosa wild type (WT) and its desB mutant were spotted on agar plates containing S. aureus, and the size of the clear zones was compared. In addition, interbacterial competition between P. aeruginosa and S. aureus was observed over time during planktonic coculture. The transcriptional profiles of the WT and desB mutant were compared by qRT-PCR and microarray to determine the role of DesB in S. aureus reduction at the molecular level. As a result, the clear zone was smaller for the desB mutant than for P. aeruginosa PAO1 (WT), and in planktonic coculture, the number of S. aureus cells was reduced in the desB mutant. qRT-PCR and microarray revealed that the expression of MvfR-controlled pqsA-E and phnAB operons was significantly decreased, but the mexEF-oprN operon was highly expressed. The results indicate that intracellular levels of 4-hydroxy-2-heptylquinoline (HHQ), a ligand of MvfR, are reduced due to MexEF-OprN-mediated efflux in desB mutant, resulting in the decrease of MvfR binding to pqsA-E promoter and the reduction of 4-hydroxy-2-alkylquinolines (HAQs) synthesis. Overexpression of mexEF-oprN operon in desB mutant was phenotypically confirmed by observing significantly increased resistance to chloramphenicol. In conclusion, these results suggest that DesB plays a role in the inhibition of S. aureus growth by controlling HAQ synthesis.

The objective of this study was to elucidate the role of the icaA gene in biofilm formation of Staphylococcus aureus exposed to NaCl. The icaA-deletion mutant of S. aureus ATCC 13565 was constructed with the temperature-sensitive plasmid pIMAY. Microtiter plate assays were performed to confirm biofilm formation for both the wild type and the mutant at 0% (control), 2, 4, and 6% NaCl. The microtiter plate assay revealed that biofilm formation by the wild type increased ( P < 0.05) as NaCl concentration increased, but biofilm formation of the mutant was not affected by NaCl concentration. Biofilm formation by the mutant was lower ( P < 0.05) than that by the wild type. These results indicate that icaA plays an important role in biofilm formation by S. aureus when the pathogen is exposed to NaCl.

Nitrite plays a major role in inhibiting the growth of foodborne pathogens, including Clostridium botulinum (C. botulinum) that causes botulism, a life-threatening disease. Nitrite serves as a color-fixing agent in processed meat products. However, N-nitroso compounds can be produced from nitrite, which are considered as carcinogens. Thus, consumers desire processed meat products that contain lower concentrations (below conventional concentrations of products) of nitrite or no nitrite at all, although the portion of nitrite intake by processed meat consumption in total nitrite intake is very low. However, lower nitrite levels might expose consumers to risk of botulism poisoning due to C. botulinum or illness caused by other foodborne pathogens. Hence, lower nitrite concentrations in combination with other factors such as low pH, high sodium chloride level, and others have been recommended to decrease the risk of food poisoning. In addition, natural compounds that can inhibit bacterial growth and function as color-fixing agents have been developed to replace nitrite in processed meat products. However, their antibotulinal effects have not been fully clarified. Therefore, to have processed meat products with lower nitrite concentrations, low pH, high sodium chloride concentration, and others should also be applied together. Before using natural compounds as replacement of nitrite, their antibotulinal activities should be examined. KCI Citation Count: 24

The survival of Escherichia coli and Salmonella strains during diced white radish kimchi fermentation was studied. Kimchi batches inoculated with the pathogens were fermented at 4, 15, and 25°C for 42 to 384 h. Cell counts of E. coli and Salmonella were enumerated on E. coli-coliform count plates and xylose lysine deoxycholate agar, respectively. Baranyi (primary model) and polynomial (secondary model) models, validated by root mean square error, were used to describe the kinetic behavior of the pathogens. In the primary model, both the death phase shoulder ( E. coli: 208.18 to 8.25 h, 4 to 25°C; Salmonella: 79.91 to 0.97 h, 4 to 25°C) and bacterial cell counts (log CFU per gram per hour) decreased with increasing temperature ( P < 0.05) (death rate: E. coli: -0.02 to -0.09, 4 to 25°C; Salmonella: -0.01 to -0.10, 4 to 25°C), the results being equally significant in the secondary model. The root mean square error (0.480 to 0.485) showed that the model performance was good. The fermentation temperature and time are the critical factors that control pathogenic E. coli and Salmonella in kimchi.

This study was conducted to examine the kinetic behavior of Campylobacter jejuni in raw beef tartare by using mathematical models and to identify genes related to C. jejuni survival at cold temperatures. C. jejuni was inoculated onto beef tartare samples, stored at 4, 10, 15, 25, and 30°C, plated on modified charcoal-cefoperazone-deoxycholate agar, and enumerated. The survival data was fitted to the Weibull model to calculate delta (δ), which is the time required for the first 1-log reduction of the cells. The Davey model was then fitted to the δ to evaluate the effect of temperature. To evaluate the performance of the developed model, the root mean square error (RMSE) was calculated by comparing the observed data with the predicted data. The mRNA was extracted from samples stored at 4 and 30°C under aerobic and anaerobic conditions, and the expression of oxidative stress response and stress response genes was evaluated. C. jejuni survived in beef tartare longer at 4°C (δ = 657.1 ± 79.6 min) than at other temperatures (9.7 ± 11.2 to 465.7 ± 139.3°C) even under aerobic conditions. The RMSE (0.475) suggested that the developed model was appropriate to describe the kinetic behavior of C. jejuni. Quantitative real-time PCR results revealed that oxidative stress and stress response genes were related to C. jejuni survival at cold temperatures, even under aerobic conditions. These results indicate that the model will be useful for describing the kinetic behavior of C. jejuni in beef tartare and that this pathogen can survive at cold temperatures because of the expression of the sodB, katA, and clpP genes.

Memory T cells contribute to rapid viral clearance during re-infection, but the longevity and differentiation of SARS-CoV-2-specific memory T cells remain unclear. Here we conduct ex vivo assays to evaluate SARS-CoV-2-specific CD4 + and CD8 + T cell responses in COVID-19 convalescent patients up to 317 days post-symptom onset (DPSO), and find that memory T cell responses are maintained during the study period regardless of the severity of COVID-19. In particular, we observe sustained polyfunctionality and proliferation capacity of SARS-CoV-2-specific T cells. Among SARS-CoV-2-specific CD4 + and CD8 + T cells detected by activation-induced markers, the proportion of stem cell-like memory T (T SCM ) cells is increased, peaking at approximately 120 DPSO. Development of T SCM cells is confirmed by SARS-CoV-2-specific MHC-I multimer staining. Considering the self-renewal capacity and multipotency of T SCM cells, our data suggest that SARS-CoV-2-specific T cells are long-lasting after recovery from COVID-19, thus support the feasibility of effective vaccination programs as a measure for COVID-19 control. T cells are instrumental to protective immune responses against SARS-CoV-2, the pathogen responsible for the COVID-19 pandemic. Here the authors show that, in convalescent COVID-19 patients, memory T cell responses are detectable up to 317 days post-symptom onset, in which the presence of stem cell-like memory T cells further hints long-lasting immunity.