Yeast strains used in the investigation.

<p>Yeast strains used in the investigation.</p

Immunofluorescence staining of <i>C. albicans</i> ghost cells and zymosan with various fluorescent probes specific for yeast cell wall glycans.

<p><i>C. albicans</i> ghost cells and zymosan were stained with an anti-chitin WGA monoclonal antibody (mAb) (a and b), anti-β-1,3 glucan (c and d), GNL (e and f), ConA (g and h) and DAPI (i and j), respectively. <i>C. albicans</i> ghost cells (b and d) and zymosan (a and c) were strongly stained with both anti-chitin WGA and anti-β-1,3 glucan mAb. Strong immunofluorescent staining was detected in zymosan using GNL and ConA (e and g), whereas no immunofluorescent signals were observed in <i>C. albicans</i> ghost cells (f and h). While zymosan was strongly stained with nuclear DAPI (i), only a weak signal was observed in <i>C. albicans</i> ghost cells (j).</p

Structural analysis of glycan fraction.

<p>(<b>A</b>)<b> MALDI-MS analysis of the native fraction</b>. Native glycan consisted of a large hexose polymer. (<b>B</b>)<b> Zymolyase digestion of the polymer</b>. This produced small hexose oligomers, 2–5 residues in size, which were identified by ¹H-¹³C HSQC-NMR (<b>C</b>) as β-1,3-linked glucans partially substituted by single β-glucose residues linked to C-6 positions.</p

Effect of different strains and cell wall extracts on <i>C. albicans</i> colonization and inflammation in the curative <i>C. albicans</i> DSS model.

<p>Big improvement (+++), improvement (++), unchanged (+) and worsening symptoms (−).</p

Schematic diagram of glycan preparation from <i>S. cerevisiae</i>.

<p>Schematic diagram of glycan preparation from <i>S. cerevisiae</i>.</p

Summary of the effects of <i>S. cerevisiae</i> strains or glycan fractions on <i>Candida</i> DSS-treated mice.

<p>Radar blots showed the effect of each <i>S. cerevisiae</i> strain or glycan fraction on: (1) mortality; (2) body weight loss; (3) clinical score; (4) histological score; and (5) <i>C. albicans</i> colonization. Sb, Sc1-1 and Sc3 strains improved the intestinal damage due to DSS-induced colitis and <i>C. albicans</i> colonization. Sc1-2, Sc2 and Sc4 strains had a dramatic effect on all inflammation signs including <i>Candida</i> colonization. Regarding the glycan fractions, both β-glucans and F2 reduced all colitis parameters and eliminated <i>C. albicans</i> colonization, while MP was less effective at controlling these parameters.</p

Schematic diagram of β-glucan preparation from <i>C. albicans</i>.

<p>Schematic diagram of β-glucan preparation from <i>C. albicans</i>.</p

Comparison of the probiotic potential of <i>S. cerevisiae</i> var. <i>boulardii</i> and <i>S. cerevisiae</i> 1-1 strain.

<p>(<b>A</b>)<b> Experimental design</b>. Low doses of DSS were used for 2 weeks followed by a 3-day restitution period. Either <i>S. cerevisiae</i> strains or β-glucan fractions were inoculated daily by oral gavage starting 3 days after <i>C. albicans</i> challenge. Three days post-<i>C. albicans</i> challenge was chosen to start <i>S. cerevisiae</i> treatment as both <i>Candida</i> colonization and colonic inflammation are well established at this time point. (<b>B</b>)<b> Percentage survival of mice</b>. The results are shown as percent survival from the time of <i>C. albicans</i> challenge and DSS treatment. The survival data were significantly different by the log-rank test. A total of 140 mice were divided into five control groups: water (n = 16 mice), Ca (n = 16 mice), Sb (n = 16 mice), Sc1-1 (n = 16 mice) and DSS alone (n = 16 mice), and three experimental groups: CaD (n = 20 mice), CaDSb (n = 20 mice), and CaDSc1-1 (n = 20 mice). (*<i>P</i><0.05 for CaDSb mice vs. CaD mice; and ‡<i>P</i><0.05 for CaDSc1-1 mice vs. CaD mice.) (<b>C</b>)<b> Number of </b><b><i>C. albicans</i></b><b> colony forming units recovered from stools</b>. Each data set represents the mean values of 10 mice/group. (*<i>P</i><0.05 for CaDSb mice vs. CaD mice; and ‡<i>P</i><0.05 for CaDSc1-1 mice vs. CaD mice.) (<b>D</b>)<b> Clinical analysis of DSS-induced colitis in mice</b>. Both <i>S. cerevisiae</i> var. <i>boulardii</i> and <i>S. cerevisiae</i> Sc1-1 significantly reduced the clinical score. (*<i>P</i><0.05 for CaDSb mice vs. D mice; ‡<i>P</i><0.05 for CaDSb mice vs. CaD mice; †<i>P</i><0.05 for CaDSc1-1 mice vs. D mice; and **<i>P</i><0.05 for CaDSc1-1 mice vs. CaD mice.) (<b>E</b>)<b> Histological score</b>. The histological score was determined by two independent, blinded examiners (degree of inflammation: 0, no changes, to 6, extensive cell infiltration and tissue damage). The histological score increased in both DSS and CaDSS groups. The histological score was significantly lower in both CaDSb and CaDSc1-1 groups when compared to CaDSS and DSS-treated groups. (*<i>P</i><0.05 for CaDSb mice vs. D mice; ‡<i>P</i><0.05 for CaDSb mice vs. CaD mice; †<i>P</i><0.05 for CaDSc1-1 mice vs. D mice; and **<i>P</i><0.05 for CaDSc1-1 mice vs. CaD mice.) (<b>F and G</b>)<b> Relative expression levels, determined by real-time quantitative PCR, of TNF-</b><b><i>α</i></b><b> and IL-10 mRNA in the colon</b>. Data are expressed as the mean ± SE of five mice in each group. (*<i>P</i><0.05 for CaDSb mice vs. D mice; †<i>P</i><0.05 for CaDSb mice vs. CaD mice; ‡<i>P</i><0.05 for CaDSc1-1 mice vs. D mice; and **<i>P</i><0.05 for CaDSc1-1 mice vs. CaD mice.)</p

Effect of β-oligoglucoside fractions derived from <i>C. albicans</i> on <i>Candida</i> DSS-treated mice.

<p>(<b>A</b>)<b> Percentage survival of mice</b>. The results are shown as percent survival from the time of <i>C. albicans</i> challenge and DSS treatment. Three days after <i>C. albicans</i> challenge, mice were given <i>C. albicans</i> β-glucans (F2) for 2 weeks. The survival data were significantly different by the log-rank test. A total of 80 mice were divided into four experimental groups: D (n = 20 mice), CaD (n = 20 mice), DF2 (n = 20 mice), and CaDF2 (n = 20 mice). (*<i>P</i><0.05 for DF2 mice vs. D mice; and ‡<i>P</i><0.05 for CaDF2 mice vs. CaD mice.) (<b>B</b>)<b> Clinical analysis of DSS-induced colitis in mice</b>. Each data set represents the mean value for each body weight. (*<i>P</i><0.05 for DF2 mice vs. D mice; and ‡<i>P</i><0.05 for CaDF2 mice vs. CaD mice.) (<b>C</b>)<b> Histological analysis of DSS-induced colitis in mice</b>. Panels (a) and (b) correspond to colon sections from DF2 and CaDF2 mice, respectively. No histological signs of colonic inflammation were seen in either DF2 or CaDF2 mice. (<b>D</b>)<b> Histological score</b>. The histological score decreased significantly in both DF2 and CaDF2 mice. (†<i>P</i><0.05 for DF2 mice vs. D mice; ‡<i>P</i><0.05 for DF2 mice vs. CaD mice; *<i>P</i><0.05 for CaDF2 mice vs. D mice; and ¶<i>P</i><0.05 for CaDF2 mice vs. CaD mice.) (<b>E</b>)<b> Number of </b><b><i>C. albicans</i></b><b> colony forming units recovered from stools</b>. Each data set represents the mean values of eight mice/group. (*<i>P</i><0.05 for CaDF2 mice vs. CaD mice.) (<b>F</b>)<b> Number of </b><b><i>C. albicans</i></b><b> colony forming units (CFU) recovered from different gut compartments</b>. Each data set represents the mean count of 16 mice/group. In mice receiving F2, numbers of <i>C. albicans</i> wild-type CFUs from the stomach, ileum and colon were significantly lower than those of <i>CaD</i> mice. (*, †, ‡ <i>P</i><0.05 for CaDF2 mice vs. CaD mice.)</p

Effect of glycan fractions derived from <i>S. cerevisiae</i> on <i>Candida</i> DSS-treated mice.

<p>(<b>A</b>)<b> Percentage survival of mice</b>. The results are displayed as percent survival from the time of <i>C. albicans</i> challenge and DSS treatment. Three days after <i>C. albicans</i> challenge, mice were given either <i>S. cerevisiae</i> (Sc2), mannoproteins (Sc2MP) or β-glucan fractions (Sc2GP) for 2 weeks. The survival data were significantly different by the log-rank test. A total of 40 mice were divided into four experimental groups. (*<i>P</i><0.05 for CaDSc2GP mice vs. CaD mice; and ‡<i>P</i><0.05 for CaDSc2MP mice vs. CaD mice.) (<b>B</b>)<b> Difference in body weight loss in mice</b>. Each data set represents the mean value for each body weight. (*<i>P</i><0.05 for CaDSc2GP mice vs. CaD mice, and †<i>P</i><0.05 for CaDSc2MP mice vs. CaD mice.) (<b>C</b>)<b> Number of </b><b><i>C. albicans</i></b><b> colony forming units recovered from stools</b>. Each data set represents the mean value of 10 mice/group. (*<i>P</i><0.05 for CaDSc2GP mice vs. CaD mice.) (<b>D</b>)<b> Histological analysis of DSS-induced colitis in mice</b>. Panel (a) shows a colon section from a <i>C. albicans</i>+mannoprotein DSS-treated mouse; panel (b) shows a colon section from a <i>C. albicans</i>+β-glucan DSS-treated mouse. Inflammatory cell infiltrates were insignificant in both CaDSc2MP and CaDSc2GP mice. (<b>E</b>)<b> Clinical analysis of DSS-induced colitis in mice</b>. Clinical score was determined by assessing weight loss, change in stool consistency and the presence of gross bleeding. The clinical score ranged from 0 to 8. (*<i>P</i><0.05 for CaDSc2 mice vs. CaD mice; †<i>P</i><0.05 for CaDSc2MP mice vs. CaD mice; and ‡<i>P</i><0.05 for CaDSc2GP mice vs. CaD mice.) (<b>F</b>)<b> Histological score</b>. The histological score decreased significantly in CaDSc2GP mice. (*<i>P</i><0.05 for CaDSc2 mice vs. CaD mice; †<i>P</i><0.05 for CaDSc2MP mice vs. CaD mice; and ‡<i>P</i><0.05 for CaDSc2GP mice vs. CaD mice.)</p