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1.
Benef Microbes ; 6(5): 697-705, 2015.
Article in English | MEDLINE | ID: mdl-26192907

ABSTRACT

The efficacy of Lactobacillus paracasei V0151 (V0151), isolated from the faeces of a child, to modulate immune responses was investigated. In RAW 264.7 cells expressing an inducible nitric oxide synthase (iNOS)-directed luciferase gene, heat-inactivated V0151 stimulated iNOS expression followed by nitric oxide production. V0151 significantly elevated interferon gamma, interleukin (IL)-10, tumour necrosis factor alpha, and IL-1ß production in human peripheral blood mononuclear cells. In splenocytes isolated from ovalbumin (OVA)-sensitised BALB/c mice treated with OVA and V0151 at different bacterium-to-cell ratios (1:1, 10:1, and 20:1) for 96 h, IL-2, IL-4, IL-5, and IL-13 production was dose-dependently downregulated, whereas IL-12 was dose-dependently upregulated. Collectively, our findings indicate that V0151 might regulate pro-inflammatory factors in macrophages and splenocytes. Furthermore, the T helper 1/T helper 2 (Th1/Th2) balance was also skewed toward Th1 dominance through the elevation of Th1 cytokine production.


Subject(s)
Anti-Allergic Agents/pharmacology , Hot Temperature , Lactobacillus/immunology , Lactobacillus/radiation effects , Animals , Cells, Cultured , Cytokines/metabolism , Leukocytes, Mononuclear/immunology , Macrophages/immunology , Mice, Inbred BALB C , Nitric Oxide/metabolism , Nitric Oxide Synthase Type II/biosynthesis
2.
Mol Oral Microbiol ; 30(1): 16-26, 2015 Feb.
Article in English | MEDLINE | ID: mdl-24961744

ABSTRACT

Dental caries arises from an imbalance of metabolic activities in dental biofilms developed primarily by Streptococcus mutans. This study was conducted to isolate potential oral probiotics with antagonistic activities against S. mutans biofilm formation from Lactobacillus salivarius, frequently found in human saliva. We analysed 64 L. salivarius strains and found that two, K35 and K43, significantly inhibited S. mutans biofilm formation with inhibitory activities more pronounced than those of Lactobacillus rhamnosus GG (LGG), a prototypical probiotic that shows anti-caries activity. Scanning electron microscopy showed that co-culture of S. mutans with K35 or K43 resulted in significantly reduced amounts of attached bacteria and network-like structures, typically comprising exopolysaccharides. Spot assay for S. mutans indicated that K35 and K43 strains possessed a stronger bactericidal activity against S. mutans than LGG. Moreover, quantitative real-time polymerase chain reaction showed that the expression of genes encoding glucosyltransferases, gtfB, gtfC, and gtfD was reduced when S. mutans were co-cultured with K35 or K43. However, LGG activated the expression of gtfB and gtfC, but did not influence the expression of gtfD in the co-culture. A transwell-based biofilm assay indicated that these lactobacilli inhibited S. mutans biofilm formation in a contact-independent manner. In conclusion, we identified two L. salivarius strains with inhibitory activities on the growth and expression of S. mutans virulence genes to reduce its biofilm formation. This is not a general characteristic of the species, so presents a potential strategy for in vivo alteration of plaque biofilm and caries.


Subject(s)
Antibiosis , Biofilms/growth & development , Lactobacillus/isolation & purification , Lactobacillus/physiology , Probiotics/isolation & purification , Streptococcus mutans/growth & development , Streptococcus mutans/physiology , Bacterial Proteins/genetics , Coculture Techniques , Dental Caries/microbiology , Gene Expression Regulation, Bacterial , Glucosyltransferases/genetics , Humans , Interferon-gamma/biosynthesis , Interferon-gamma/immunology , Interleukin-10/biosynthesis , Interleukin-10/immunology , Lactobacillus/classification , Microscopy, Electron, Scanning , Polysaccharides, Bacterial/metabolism , Real-Time Polymerase Chain Reaction , Streptococcus mutans/genetics , Streptococcus mutans/ultrastructure
3.
Genet Mol Res ; 13(2): 2827-39, 2014 Jan 22.
Article in English | MEDLINE | ID: mdl-24535906

ABSTRACT

Exogenous gibberellins (GAs) are widely applied to increase crop yields, with knowledge about the physiological functioning and biochemistry mechanisms of these phytohormones improving; however, information remains limited about the effect of GAs on seed filling. In this study, the siliques (containing the seeds) of oilseed rape (Brassica napus L.) were treated with GA3 at 3 stages of seed filling. We confirmed that GA3 regulates the deposition of storage reserves in developing seeds. The percentage of crude fat in the seeds increased during the early stage, but remained stable during the middle and late stages. In comparison, the percentage of total protein decreased during the early and middle stages, but significantly increased during the late stage. In addition, Q-PCR was employed to analyze the expression level of related genes in response to GA3. It was found that the expression of WRI and ABI3 transcription factors corresponded to crude fat content and total protein content, respectively. The expression of storage reserve related genes DGAT, MCAT, SUC2, and GPT was consistent with crude fat content, whereas the expression of Napin corresponded to total protein content. The results of this study indicate that exogenous GA3 has a different effect on storage reserve deposition in seed during different stages of seed filling, and the effect might be achieved via changing the expression of related genes.


Subject(s)
Brassica napus/growth & development , Gibberellins/administration & dosage , Seeds/growth & development , Brassica napus/drug effects , Fatty Acids/metabolism , Gene Expression Regulation, Plant/drug effects , Plant Proteins/biosynthesis , Seeds/drug effects , Seeds/genetics
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