Starch-based orodispersible film for diclofenac release

The form of drug administration affects the success of treatment, since it can influence adherence of the patient to the therapy. The use of orodispersible films has emerged as a way to overcome some drawbacks of conventional methods of drug delivery, especially for patients experiencing difficulty in swallowing. These films are prepared using a matrix that incorporates the drug and contains other substances that confer the properties of the system. The present work describes the use of thermoplastic starch as a carrier for the model drug diclofenac, including film preparation and testing of its orodispersible potential. Preparation of the film employed a microwave oven to gelatinize and plasticize corn starch, with incorporation of the model drug, followed by solvent-casting. The samples were characterized using mechanical tests, analyses of water uptake and water content, and Fourier transform infrared spectroscopy. The results indicated that the film presented promising properties as an alternative system for oral drug administration, with good incorporation and distribution of the drug in the matrix. The material displayed satisfactory mechanical properties, which are crucial for this type of material, due to the need for oral administration and handling before use

“Transmantle sign”を示す限局性皮質異形成における神経細胞の成熟と分化の未熟性：層特異的マーカー発現による解析

Transmantle dysplasia is a rare type of focal cortical dysplasia (FCD) characterized by expansion of the cortex from the deep white matter to the surface and in which there is a FCD IIA or IIB pathologic pattern. To characterize possible mechanisms underlying this regional disorder of radial migrating cells, we studied the expression patterns of neocortical layer-specific markers using immunohistochemistry in surgical specimens from 5 FCD IIA and 4 FCD IIB cases in children. All neuronal cells expressed the mature neuron marker MAP2/2B but not the microglia markers Iba-1 and CD68. Some layer-specific markers showed distinct expression patterns. TBR1-positive, SATB2-positive, and FOXP1-positive cells were diffusely distributed in the cortex and/or the white matter. TBR1-positive and FOXP1-positive cells were generally more numerous in FCD IIB than in FCD IIA and were mostly in the cortical molecular and upper layers. FOXP1-, FOXP2-, and CUTL1-positive cells also expressed the immature neuron marker, Nestin/PROX1, whereas TBR1-, CTIP2-, and SATB2-positive cells only expressed MAP2/2B. These data highlight differences between FCD IIB and FCD IIA with more cells having the immature marker in upper layer markers in the former. By analyzing layer-specific marker expression patterns, we identified apparent neuronal maturation differences between FCD IIA and FCD IIB in cases of transmantle dysplasia.博士（医学）・乙第1312号・平成25年5月29

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 (DicerΔIEC) and analyzed intestinal phenotypes, including M-cell differentiation, morphology and function. DicerΔIEC 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 DicerΔIEC mice. These results suggest that miRNAs play a significant role in M cell differentiation and help secure mucosal immune homeostasis

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

Morphology of M cells in Dicer^ΔIEC.

<p>Electron micrographs of Dicer^F/F and Dicer^ΔIEC PP. (A) Surface of FAE by scanning electron microscopy. Arrowheads indicate M cells. Scale bars: 10 μm (B) Transmission electron micrographs of an M cell in FAE and enterocyte in VE. Scale bars: 1.8 μm (C) High magnification image of (B). Arrow indicate the endosomes in M cell. Scale bars: 500 nm.</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^ΔIEC FAE and Dicer^F/F 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

Delayed maturation and differentiation of neurons in focal cortical dysplasia with the transmantle sign: analysis of layer-specific marker expression

Transmantle dysplasia is a rare type of focal cortical dysplasia (FCD) characterized by expansion of the cortex from the deep white matter to the surface and in which there is a FCD IIA or IIB pathologic pattern. To characterize possible mechanisms underlying this regional disorder of radial migrating cells, we studied the expression patterns of neocortical layer-specific markers using immunohistochemistry in surgical specimens from 5 FCD IIA and 4 FCD IIB cases in children. All neuronal cells expressed the mature neuron marker MAP2/2B but not the microglia markers Iba-1 and CD68. Some layer-specific markers showed distinct expression patterns. TBR1-positive, SATB2-positive, and FOXP1-positive cells were diffusely distributed in the cortex and/or the white matter. TBR1-positive and FOXP1-positive cells were generally more numerous in FCD IIB than in FCD IIA and were mostly in the cortical molecular and upper layers. FOXP1-, FOXP2-, and CUTL1-positive cells also expressed the immature neuron marker, Nestin/PROX1, whereas TBR1-, CTIP2-, and SATB2-positive cells only expressed MAP2/2B. These data highlight differences between FCD IIB and FCD IIA with more cells having the immature marker in upper layer markers in the former. By analyzing layer-specific marker expression patterns, we identified apparent neuronal maturation differences between FCD IIA and FCD IIB in cases of transmantle dysplasia.博士（医学）・乙第1312号・平成25年5月29日identifier:Journal of neuropathology and experimental neurology Vol.71 No.8 p.741-749identifier:00223069identifier:http://ginmu.naramed-u.ac.jp/dspace/handle/10564/2590identifier:Journal of neuropathology and experimental neurology, 71(8): 741-74

Primer sequence of q-PCR.

<p>Primer sequence of q-PCR.</p

Total number of M cells in Peyer’s patches is decreased in Dicer^ΔIEC mice.

<p>(A) H&E staining of small intestines VE region of Dicer^ΔIEC and Dicer^F/F. (B) H&E staining in PPs of Dicer^ΔIEC and Dicer^F/F. (C) The total number of follicles and surface area in Dicer^ΔIEC and Dicer^F/F. 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 μm (E) M cell number/mm² in FAE of each mouse strain. Data are means and SE. *<i>P</i> < 0.05.</p

Impaired antigen uptake by Dicer^ΔIEC M cells.

<p>(A) Dicer^ΔIEC and Dicer^F/F mice were inoculated by gavage with 1 x 10¹¹ FluoSpheres. After 4 hours, frozen sections were prepared to examine translocated beads in PPs. Scale bars: 100 μm (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^ΔIEC and Dicer^F/F mice were inoculated intragastrically by gavage with 1 x 10⁸ CFU of <i>Yersinia enterocolitica</i>. After 24 hours, the bacterial translocation to Peyer’s 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