18 research outputs found
Epithelium-Intrinsic MicroRNAs Contribute to Mucosal Immune Homeostasis by Promoting M-Cell Maturation.
M cells in the follicle-associated epithelium (FAE) of Peyer's patches (PPs) serve as a main portal for external antigens and function as a sentinel in mucosal immune responses. The scarcity of these cells has hampered identification of M cell-specific molecules. Recent efforts have begun to provide insight into antigen transcytosis and differentiation of M cells; however, the molecular mechanisms underlying these processes are not fully elucidated. Small non-coding RNAs including microRNA (miRNA) have been reported to regulate gene expression and control various biological processes such as cellular differentiation and function. To evaluate the expression of miRNAs in FAE, including M cells, we previously performed microarray analysis comparing intestinal villous epithelium (VE) and PP FAE. Here we confirmed FAE specific miRNA expression levels by quantitative PCR. To gain insight into miRNA function, we generated mice with intestinal epithelial cell-specific deletion of Dicer1 (DicerÎIEC) and analyzed intestinal phenotypes, including M-cell differentiation, morphology and function. DicerÎIEC mice had a marked decrease in M cells compared to control floxed Dicer mice, suggesting an essential role of miRNAs in maturation of these cells. Furthermore, transmission electron microscopic analysis revealed that depletion of miRNA caused the loss of endosomal structures in M cells. In addition, antigen uptake by M cells was impaired in DicerÎIEC mice. These results suggest that miRNAs play a significant role in M cell differentiation and help secure mucosal immune homeostasis
Novel ABCA12 mutations identified in two cases of non-bullous congenital ichthyosiform erythroderma associated with multiple skin malignant neoplasia
Efficacy and Safety of Daikenchuto for Constipation and Dose-Dependent Differences in Clinical Effects
Background. Daikenchuto (DKT) is a Kampo medicine used for the treatment of constipation. In this study, we evaluated the effectiveness of DKT against constipation. Patients and Methods. Thirty-three patients administered DKT for constipation were selected and divided into low-dose (7.5âg DKT; n=22) and high-dose (15âg DKT; n=11) groups. We retrospectively evaluated weekly defaecation frequency, side effects, and clinical laboratory data. Results. Median defaecation frequencies after DKT administration (5, 5.5, 5, and 8 for the first, second, third, and fourth weeks, resp.) were significantly higher than that before DKT administration (2) in all 33 cases (P<0.01). One case (3%) of watery stool, one case of loose stools (3%), and no cases of abdominal pain (0%) were observed. Median defaecation frequencies in the high-dose group (7 and 9) were significantly higher than those in the low-dose group (4 and 3) in the first (P=0.0133) and second (P=0.0101) weeks, respectively. There was no significant change in clinical laboratory values. Conclusion. We suggest that DKT increases defaecation frequency and is safe for treating constipation
Evaluation and Comparison of Daiokanzoto and Lubiprostone for Constipation: A Retrospective Cohort Study
Morphology of M cells in Dicer<sup>ÎIEC</sup>.
<p>Electron micrographs of Dicer<sup>F/F</sup> and Dicer<sup>ÎIEC</sup> PP. (A) Surface of FAE by scanning electron microscopy. Arrowheads indicate M cells. Scale bars: 10 ÎŒm (B) Transmission electron micrographs of an M cell in FAE and enterocyte in VE. Scale bars: 1.8 ÎŒm (C) High magnification image of (B). Arrow indicate the endosomes in M cell. Scale bars: 500 nm.</p
miRNA expression profiles in intestinal epithelium.
<p>Q-PCR analysis was performed for miRNA expression in FAE and VE. The relative levels of each miRNA relative to the small nucleolar RNA <i>Sno202</i> are shown. Values are mean ± SE of three samples from different mice. *P<0.05.</p
FAE miRNAs involved in M cell maturation.
<p>(A) Flowchart of M cell maturation. (B) Q-PCR analysis was performed for <i>Marcksl1</i>, <i>SpiB</i>, <i>Ccl9 and Gp2</i> mRNA expression in Dicer<sup>ÎIEC</sup> FAE and Dicer<sup>F/F</sup> FAE. The relative expression levels of each gene to <i>Gapdh</i> are shown. Values represent the mean ± SD of three samples from different mice. *<i>P</i> < 0.05 **<i>P</i> < 0.01.</p
Total number of M cells in Peyerâs patches is decreased in Dicer<sup>ÎIEC</sup> mice.
<p>(A) H&E staining of small intestines VE region of Dicer<sup>ÎIEC</sup> and Dicer<sup>F/F</sup>. (B) H&E staining in PPs of Dicer<sup>ÎIEC</sup> and Dicer<sup>F/F</sup>. (C) The total number of follicles and surface area in Dicer<sup>ÎIEC</sup> and Dicer<sup>F/F</sup>. Data are means± SE (n = 3). *<i>P</i> < 0.05. (D) Whole mount immunostaining of PPs with anti-GP2 (red) and F-actin (green) analyzed using a confocal microscope. Scale bars: 100 ÎŒm (E) M cell number/mm<sup>2</sup> in FAE of each mouse strain. Data are means and SE. *<i>P</i> < 0.05.</p
Impaired antigen uptake by Dicer<sup>ÎIEC</sup> M cells.
<p>(A) Dicer<sup>ÎIEC</sup> and Dicer<sup>F/F</sup> mice were inoculated by gavage with 1 x 10<sup>11</sup> FluoSpheres. After 4 hours, frozen sections were prepared to examine translocated beads in PPs. Scale bars: 100 ÎŒm (B) Count data of beads taken up in PP each mouse strain. Data are expressed as the mean ± SE of four different samples for each group. **<i>P</i> < 0.01. (C) Dicer<sup>ÎIEC</sup> and Dicer<sup>F/F</sup> mice were inoculated intragastrically by gavage with 1 x 10<sup>8</sup> CFU of <i>Yersinia enterocolitica</i>. After 24 hours, the bacterial translocation to Peyerâs patches was examined by plating PP homogenates. Data are expressed as the mean ± SE of five different mice/each group. *<i>P</i> < 0.05.</p