Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Lactobacillus reuteri DSM 17938 differentially modulates effector memory T cells and Foxp3 + regulatory T cells in a mouse model of necrotizing enterocolitis

doi: 10.1152/ajpgi.00038.2014

Figure Lengend Snippet: Lactobacillus reuteri DSM 17938 (LR17938) increased survival rate of mouse pups with necrotizing enterocolitis (NEC). Mice (8–10 days old) were separated from their mothers and housed in an incubator. NEC was induced by formula feeding + stress (FS) with hypoxia (5% O2-95% N2 for 10 min) and cold exposure (4°C for 5 min) twice daily for 4 days (indicated by arrows). Live animals were counted on each day for calculation of survival rate. Mice were euthanized on day 5 after NEC protocol (indicated by ×), and tissues were collected. In some cases, pups in the FS group were euthanized on day 3 or 4, if they were in pain, demonstrated labored respirations, or exhibited abdominal distension or gastrointestinal bleeding, to collect tissues for histological analysis and to calculate survival rate. LR17938 [106 colony-forming units (CFU)·g body wt−1·day−1] was added to the formula for treatment (FS + 17938). LR17938 reduced mortality rate on day 5 compared with FS without probiotic. *P = 0.027.

Article Snippet: Probiotic LR17938 Preparation Human breast milk-derived LR17938 was provided by Biogaia (Stockholm, Sweden).

Techniques:

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Lactobacillus reuteri DSM 17938 differentially modulates effector memory T cells and Foxp3 + regulatory T cells in a mouse model of necrotizing enterocolitis

doi: 10.1152/ajpgi.00038.2014

Figure Lengend Snippet: LR17938 reduced incidence and severity of NEC. A: representative intestinal histological changes in dam-fed, dam-fed + 17938, FS (NEC protocol), and FS + 17938 mice. Ileal tissues were stained with hematoxylin and eosin. Magnification ×200. Red arrows, villous core separations in animals with NEC; black arrows, normal villi in ileal tissue from dam-fed or FS + 17938 mice. B: number of animals with normal histology (NEC scores <2) and with NEC (NEC scores ≥2). n, Number of live animals from which tissue samples could be collected. Terminal ileum of each mouse was harvested at day 5 (or day 3 or 4 for some samples in the FS group). %, Incidence of NEC. *P < 0.05 vs. FS. C: FS and FS + 17938 groups with NEC scores of 2 and 3. *P < 0.05 vs. FS. LR17938 reduced NEC histological injury.

Article Snippet: Probiotic LR17938 Preparation Human breast milk-derived LR17938 was provided by Biogaia (Stockholm, Sweden).

Techniques: Staining

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Lactobacillus reuteri DSM 17938 differentially modulates effector memory T cells and Foxp3 + regulatory T cells in a mouse model of necrotizing enterocolitis

doi: 10.1152/ajpgi.00038.2014

Figure Lengend Snippet: Increased IL-1β levels in intestine of mice subjected to the NEC protocol and the modifying effect of LR17938 treatment. Cytokine levels in tissue lysates were examined using Meso Scale Discovery cytokine assay; n = 8–12 mice per group. Comparisons showed significant differences: P < 0.01 (FS vs. dam) and P < 0.05 (FS + 17938 vs. FS).

Article Snippet: Probiotic LR17938 Preparation Human breast milk-derived LR17938 was provided by Biogaia (Stockholm, Sweden).

Techniques: Cytokine Assay

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Lactobacillus reuteri DSM 17938 differentially modulates effector memory T cells and Foxp3 + regulatory T cells in a mouse model of necrotizing enterocolitis

doi: 10.1152/ajpgi.00038.2014

Figure Lengend Snippet: LR17938 reversed frequency of CD4+Foxp3+ regulatory T (Treg) cells in terminal ileum and mesenteric lymph nodes (MLNs) of mice with NEC. A: representative flow cytometric plots from terminal ileum of dam-fed mice with initial gating on lymphocyte population on the left forward scatter (FSC)-side scatter (SSC) plot (left), followed by gating on CD4+ T cells [middle; x-axis: CD4-peridinin-chlorophyll proteins (CD4-PerCP/Cy5.5); y-axis: CD–phycoerythrin (CD8-PE)], and Foxp3+ Treg cells within the group of CD8−CD4+ T cells (right; x-axis: CD4-PerCP/Cy5.5; y-axis: Foxp3-Alexa Fluor 647). B and C: frequency (%) of CD4+Foxp3+ Treg cells in terminal ileum and MLNs. n, Number of live animals from which sufficient cell numbers were obtained from tissues for analysis. FS vs. dam: P < 0.01 (B) or P < 0.001 (C); FS vs. FS + 17938: P < 0.01 (B and C); FS vs. FS + Lactobacillus acidophilus DDS (La DDS; B and C): P > 0.05. D: percentage of Treg cells in terminal ileum of dam-fed + LR17938 and dam-fed mice (P < 0.001).

Article Snippet: Probiotic LR17938 Preparation Human breast milk-derived LR17938 was provided by Biogaia (Stockholm, Sweden).

Techniques:

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Lactobacillus reuteri DSM 17938 differentially modulates effector memory T cells and Foxp3 + regulatory T cells in a mouse model of necrotizing enterocolitis

doi: 10.1152/ajpgi.00038.2014

Figure Lengend Snippet: Increased Treg cells induced by LR17938 during the NEC protocol in terminal ileum of mice were predominantly thymus-derived Treg cells with Helios expression. Percentage of Helios+ cells within CD4+Foxp3+ Treg cells is shown. *Undetectable due to low numbers of Treg cells in the ileum. There were no differences between FS + 17938 and dam-fed mice. All groups demonstrated high Helios expression in ∼75% of Treg cells.

Article Snippet: Probiotic LR17938 Preparation Human breast milk-derived LR17938 was provided by Biogaia (Stockholm, Sweden).

Techniques: Derivative Assay, Expressing

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Lactobacillus reuteri DSM 17938 differentially modulates effector memory T cells and Foxp3 + regulatory T cells in a mouse model of necrotizing enterocolitis

doi: 10.1152/ajpgi.00038.2014

Figure Lengend Snippet: LR17938 reduced the increased activated effector/memory T (Tem) cells in intestinal mucosa of mice with NEC. A: representative flow cytometric plots from terminal ileum of mice. Population of CD4+Foxp3− non-Treg cells was initially defined (dotted rectangle in 1st left plot) according to expression of CD4 and Foxp3 (x-axis: CD4-PerCP/Cy5.5; y-axis: Foxp3-Alexa Fluor 647). Two different effector T cells were further characterized in the non-Treg cell population on the basis of expression of CD44 and CD45RB (x-axis: CD44-FITC; y-axis: CD45RB-PE). Percentages of T cells with phenotypes of CD44+CD45RBlo (Tem) and CD44+CD45RBhi (transitional effector T) (rectangle in the 2nd left plot) are indicated. Representative plots show percentages of Tem and transitional effector T cells in dam-fed, dam-fed + 17938, FS, FS + 17938, and FS + La DDS mice. B and C: frequency (%) of Tem cells and transitional effector T cells in ileum of mice among the different groups. n, Number of live animals from which sufficient cell numbers were successfully obtained from tissues for analysis. Significant P values for multigroup comparisons are shown. *P < 0.05 FS + 17938 vs. dam.

Article Snippet: Probiotic LR17938 Preparation Human breast milk-derived LR17938 was provided by Biogaia (Stockholm, Sweden).

Techniques: Expressing

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Acerola exosome-like nanovesicles to systemically deliver nucleic acid medicine via oral administration

doi: 10.1016/j.omtm.2021.03.006

Figure Lengend Snippet: Protective effect of AELNs on miRNA (A) The procedure: (1) mixing of nucleic acids and AELNs on ice for 30 min, (2) collecting the supernatant upon ultracentrifugation, (3) treatment with RNase, acid (HCl; pH 2.0), or base (NaOH; pH 10.0), and (4) measurement of miRNA by quantitative real-time PCR. (B) Comparison of the miRNA protective effect is shown for miRNA alone (white bar), miRNA and AELN mixture (black bar), and miRNA and milk exosome mixture (gray bar). The vertical axis represents the expression level of miR-340, which was standardized with ath-miR-159. ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: As a control experiment, human breast milk exosome (Cosmo Bio, Tokyo, Japan) was used as a substitute for AELNs to form a complex with nucleic acids in the same procedure.

Techniques: Real-time Polymerase Chain Reaction, Expressing