Rat tissues processed with immunohistochemistry to highlight the expression of Tlr2.

<p>Panel a: spleen section used as positive control (original magnification, 40×); panel b: colon section containing an MDF (arrow) surrounded by normal mucosa; Tlr2 was weakly expressed by epithelial cells facing the gut lumen in both normal mucosa and MDF (original magnification 10×); panel c: the same MDF of panel c, shown at higher magnification (40×); panel d: colon section with normal mucosa and gut associated lymphatic tissue (GALT, indicated by an arrow) (original magnification 10×): panel e: the same GALT of panel d, shown at higher magnification (40×).</p

Representative examples of histological sections of rat colon processed with FISH (Cy3-conjugated EUB338 probe).

<p>Panel a: normal proximal colon showing a direct contact between bacteria (bright yellow signal) and the intestinal epithelium (E) (original magnification 4×). Panel b: bacteria inside a crypt in the proximal colon (original magnification 100×). Panel c: normal distal colon showing bacteria (bright yellow signal) separated from the epithelium (E) by a layer of mucous (M) (original magnification 40×). Panel d: section of a colonic tumor (T) and its adjacent normal mucosa (E) stained with DAPI (original magnification 4×); the boxed region is shown enlarged in panel e. Panel e: presence of bacteria (arrow) at the interface between tumor (T) and normal mucosa (E) (original magnification 40×). Panel f: section of an unopened colon containing an MDF (boxed) stained with DAPI (original magnification 4×); the boxed region is shown enlarged in panel g. Panel g: no bacteria are present in the MDF (original magnification 40×). Panel h: section of an unopened colon containing a tumor (T) stained with DAPI (original magnification 4×); the boxed region is shown enlarged in panel i. Panel i: no bacteria are present in the tumor (original magnification 40×).</p

Genes associated to the Tlr pathway: list of the statistically significant up and down-regulated genes in the MDF.

a<p>Genes for which the comparison between the ΔCt of the MDF with that of the corresponding normal mucosa was statistically significant (p<0.05, using t-test for paired samples).</p>b<p>For each gene, fold change between MDF and normal mucosa was calculated with the 2^−ΔΔCT method <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029918#pone.0029918-Schmittgen1" target="_blank">[26]</a>; values are means±SE (n: 7 MDF analysed).</p

Characteristic of the 75 study children, according to their final diagnosis.

<p>Note. TB: tuberculosis; BCG: Bacille Calmette-Guérin vaccine; TST = tuberculin skin test; QFT-G-IT: QuantiFERON-Gold In Tube test.</p

Characteristics of the used antigens: all the used antigens were recombinant proteins (from Hoghart et al. [14], modified).

<p>Characteristics of the used antigens: all the used antigens were recombinant proteins (from Hoghart et al. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046041#pone.0046041-Hogarth1" target="_blank">[14]</a>, modified).</p

ELISPOT assay results from 75 study children according to final diagnosis.

<p>Note.</p>*<p><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046041#s4" target="_blank">Results</a> are expressed as median and interquartile range of spot forming colonies per million PBMCs; TB:tuberculosis.</p

A receiver operator characteristic (ROC) plot is shown, illustrating sensitivity and specificity of AlaDH IFN-γ and IL-2 ELISpot results in discriminating children with latent (n = 21) and overt (n = 25) tuberculosis.

<p>Area under the ROC curve was 0.700 (95%IC: 0.547-0.853; p = 0.021 <i>vs.</i> the identity - diagonal - line) considering IFN-γ ELISpot and 0,896 (95%IC: 0.785–1.008; p<0.0001 <i>vs.</i> the identity - diagonal - line) for IL-2 ELISPOT.</p

Data_Sheet_1_Effects of the probiotic Lactiplantibacillus plantarum IMC 510® on body composition, biochemical parameters, gut microbiota composition and function, and clinical symptoms of overweight/obese subjects.docx

Background and aimIn recent decades, obesity prevalence has reached epidemic proportions and considering the pivotal role of gut microbiota (GM) in the regulation of energy balance, alternative non-pharmacological approaches involving probiotics’ administration have been proposed. The aim of the present study was to evaluate the effect of Lactiplantibacillus plantarum IMC 510® supplementation on anthropometric and biochemical parameters, GM composition and functionality, and gastrointestinal and general symptoms of overweight/obese subjects.MethodsForty overweight/obese subjects were randomly assigned to daily consume the probiotic Lactiplantibacillus plantarum IMC 510® or placebo for 3 months. Before and after the administration period, anthropometric and biochemical parameters, self-administered questionnaires, and plasma and stool samples were obtained from each participant. The GM characterization was performed with 16S rRNA sequencing, while fecal short (SCFAs) and medium (MCFAs) chain fatty acids were analyzed with a gas chromatography–mass spectrometry protocol.ResultsCompared to placebo, probiotic supplementation determined a significant decrease in body weight, BMI, waist circumference, waist-to-height ratio, and blood glucose. Moreover, probiotic administration produced a significant decrease of the genera Hafnia-Obesumbacterium and Romboutsia and an increase of Succiniclasticum spp.; conversely, placebo administration resulted in the decrease of Actinomycetaceae and an increase of both Alloprevotella spp. and of the levels of pro-inflammatory hexanoic and heptanoic acids.ConclusionThanks to its effect in increasing some beneficial gut bacteria and lowering effects on waist circumference, fasting glucose levels and gastrointestinal symptoms of obese subjects, Lactiplantibacillus plantarum IMC 510® supplementation could represent a future and encouraging strategy for the prevention or treatment of obesity.</p