On the Infra-Red Spectra of Solutions of O-Chlorophenol and Phenol

<p><b>a–d Immunoblotting of different proteins in control and study groups of gastric adenocarcinoma.</b> (a) Representative immunoblots of different target proteins in gastric epithelium of the control and study groups (from patients 1 to 4) with GAPDH as a loading control. M, marker. Relative protein abundance of NKA α1 (b), NKA β1 (c), and E-cadherin (d) in gastric epithelium of the control and study groups. The asterisks indicate a significant difference between the control and study groups. Values were expressed as the means ± SEM. A.u., arbitrary unit. ***, P < 0.001; ****, P < 0.0001.</p

Expression Profiles of Branchial FXYD Proteins in the Brackish Medaka <em>Oryzias dancena</em>: A Potential Saltwater Fish Model for Studies of Osmoregulation

<div><p>FXYD proteins are novel regulators of Na⁺-K⁺-ATPase (NKA). In fish subjected to salinity challenges, NKA activity in osmoregulatory organs (e.g., gills) is a primary driving force for the many ion transport systems that act in concert to maintain a stable internal environment. Although teleostean FXYD proteins have been identified and investigated, previous studies focused on only a limited group of species. The purposes of the present study were to establish the brackish medaka (<em>Oryzias dancena</em>) as a potential saltwater fish model for osmoregulatory studies and to investigate the diversity of teleostean FXYD expression profiles by comparing two closely related euryhaline model teleosts, brackish medaka and Japanese medaka (<em>O. latipes</em>), upon exposure to salinity changes. Seven members of the FXYD protein family were identified in each medaka species, and the expression of most branchial <em>fxyd</em> genes was salinity-dependent. Among the cloned genes, <em>fxyd11</em> was expressed specifically in the gills and at a significantly higher level than the other <em>fxyd</em> genes. In the brackish medaka, branchial <em>fxyd11</em> expression was localized to the NKA-immunoreactive cells in gill epithelia. Furthermore, the FXYD11 protein interacted with the NKA α-subunit and was expressed at a higher level in freshwater-acclimated individuals relative to fish in other salinity groups. The protein sequences and tissue distributions of the FXYD proteins were very similar between the two medaka species, but different expression profiles were observed upon salinity challenge for most branchial <em>fxyd</em> genes. Salinity changes produced different effects on the FXYD11 and NKA α-subunit expression patterns in the gills of the brackish medaka. To our knowledge, this report is the first to focus on FXYD expression in the gills of closely related euryhaline teleosts. Given the advantages conferred by the well-developed Japanese medaka system, we propose the brackish medaka as a saltwater fish model for osmoregulatory studies.</p> </div

Salinity effects on branchial <i>fxyd11</i> mRNA abundance in the brackish medaka (Od, A) and the Japanese medaka (Ol, B).

<p>Values are means ± SEM (N = 6). Different letters indicate significant differences among salinity groups (one-way ANOVA with Tukey's comparison, <i>P</i><0.05). FW, fresh water; BW, brackish water; SW, seawater.</p

Comparisons of branchial <i>fxyd</i> mRNA abundance in the brackish medaka (Od, A) and the Japanese medaka (Ol, B).

<p>Values are means ± SEM (N = 10). Different letters indicate significant differences among <i>fxyd</i> genes, excluding <i>fxyd11</i> (one-way ANOVA with Tukey's comparison, <i>P</i><0.05).</p

Alignment of putative amino acid sequences of the FXYD proteins in the brackish medaka (Od, A) and Japanese medaka (Ol, B).

<p>Red text indicates the FXYD motif; the conserved residues in the conserved region (from the FXYD motif to the end of the transmembrane domain) are shown in blue; the predicted signal peptides are shown with a gray background; the predicted transmembrane domains are shown in green underlined text; and the predicted sites for phosphorylation are shown in pink text.</p

DataSheet1_MicroRNA-200a/200b Modulate High Glucose-Induced Endothelial Inflammation by Targeting O-linked N-Acetylglucosamine Transferase Expression.PDF

<p>Background and Aims: Increased O-linked N-acetylglucosamine (O-GlcNAc) modification of proteins by O-GlcNAc transferase (OGT) is associated with diabetic complications. Furthermore, oxidative stress promotes endothelial inflammation during diabetes. A previous study reported that microRNA-200 (miR-200) family members are sensitive to oxidative stress. In this study, we examined whether miR-200a and miR-200b regulate high-glucose (HG)-induced OGT expression in human aortic endothelial cells (HAECs) and whether miRNA-200a/200b downregulate OGT expression to control HG-induced endothelial inflammation.</p><p>Methods: HAECs were stimulated with high glucose (25 mM) for 12 and 24 h. Real-time polymerase chain reaction (PCR), western blotting, THP-1 adhesion assay, bioinformatics predication, transfection of miR-200a/200b mimic or inhibitor, luciferase reporter assay, and transfection of siRNA OGT were performed. The aortic endothelium of db/db diabetic mice was evaluated by immunohistochemistry staining.</p><p>Results: HG upregulated OGT mRNA and protein expression and protein O-GlcNAcylation levels (RL2 antibody) in HAECs, and showed increased intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin gene expression; ICAM-1 expression; and THP-1 adhesion. Bioinformatics analysis revealed homologous sequences between members of the miR-200 family and the 3′-untranslated region (3′-UTR) of OGT mRNA, and real-time PCR analysis confirmed that members of miR-200 family were significantly decreased in HG-stimulated HAECs. This suggests the presence of an impaired feedback restraint on HG-induced endothelial protein O-GlcNAcylation levels because of OGT upregulation. A luciferase reporter assay demonstrated that miR-200a/200b mimics bind to the 3′-UTR of OGT mRNA. Transfection with miR-200a/200b mimics significantly inhibited HG-induced OGT mRNA expression, OGT protein expression; protein O-GlcNAcylation levels; ICAM-1, VCAM-1, and E-selectin gene expression; ICAM-1 expression; and THP-1 adhesion. Additionally, siRNA-mediated OGT depletion reduced HG-induced protein O-GlcNAcylation; ICAM-1, VCAM-1, and E-selectin gene expression; ICAM-1 expression; and THP-1 adhesion, confirming that HG-induced endothelial inflammation is partially mediated via OGT-induced protein O-GlcNAcylation. These results were validated in vivo: tail-vein injection of miR-200a/200b mimics downregulated endothelial OGT and ICAM-1 expression in db/db mice.</p><p>Conclusion: miR-200a/200b are involved in modulating HG-induced endothelial inflammation by regulating OGT-mediated protein O-GlcNAcylation, suggesting the therapeutic role of miR-200a/200b on vascular complications in diabetes.</p

Intestinal FXYD12 and sodium-potassium ATPase: A comparative study on two euryhaline medakas in response to salinity changes - Fig 8

<p><b>Effects of salinity on intestinal NKA activity in the Indian medaka (Od; A) and the Japanese medaka (Ol; B).</b> The values are means ± SEM (N = 6). Dissimilar letters indicate significant differences among various salinity groups (<i>P < 0</i>.<i>05</i>). FW, fresh water; BW, brackish water; SW, seawater.</p

Intestinal FXYD12 and sodium-potassium ATPase: A comparative study on two euryhaline medakas in response to salinity changes - Fig 2

<p><b>Immunohistochemical localization of NKA α-subunit (NKA) and FXYD12 in paraffin cross sections of intestines of the brackish water-acclimated Indian medaka (Od; A-C) and fresh water-acclimated Japanese medaka (Ol; D-E).</b> Immunosignals of NKA (<b>B</b>, <b>E</b>) and FXYD12 (<b>C</b>, <b>F</b>) were both detected in the basolateral membrane of intestinal epithelium, compared with the negative control (<b>A</b>, <b>D</b>). V, villus; *, lumen. Scale bar: 20 μm.</p

Co-immunoprecipitation of OdNKA with OdFXYD11 in freshwater-acclimated brackish medaka.

<p>FXYD11 was immunoprecipitated from gill total lysates with a primary antibody, and then the immune complexes were analyzed by SDS-PAGE and immunoblotted for the NKA protein. Immunoreactive bands for NKA were detected at 100 kDa. F11, western blot detection of the FXYD11 antibody (experimental group); Pre, negative immunoblot control using pre-immune serum for the immunoprecipitation; α5, positive immunoblot control using the same antibody (NKA, α5) for the immunoprecipitation; G, positive immunoblot control using gill total lysates without the immunoprecipitation.</p

Intestinal FXYD12 and sodium-potassium ATPase: A comparative study on two euryhaline medakas in response to salinity changes - Fig 4

<p><b>Double immunofluorescence staining of NKA α-subunit (NKA; green; A-C) and FXYD12 (red; D-F) in intestinal cross-cryosections of the Japanese medaka (Ol).</b> The merged images (yellow; <b>G</b>, <b>H</b>, <b>I</b>) revealed that FXYD12 colocalised to the basolateral membrane of NKA-immunoreactive cells in the fresh water- (FW; <b>A</b>, <b>D</b>, <b>G</b>), brackish water- (BW; <b>B</b>, <b>E</b>, <b>H</b>), and seawater- (SW; <b>C</b>, <b>F</b>, <b>I</b>) acclimated fish. V, villus; *, lumen. Scale bar: 20 μm.</p