Distribution of Fodrin in the Keratinocyte In Vivo and In Vitro

Distribution of fodrin in the keratinocyte, both in vivo and in vitro, was examined by immunofluorescence microscopy. In the rat epidermis in vivo, fodrin was localized in the cell periphery of the spinous layer of all the skins studied. In only the basal layer of the thick skin, however, fodrin was seen intensely in the cytoplasm. As in vitro keratinocytes, a mouse cell line (Pam 212) cultured in low (0.06 mM) as well as standard (1.87 mM) Ca2+ was examined. In low Ca2+, fodrin was observed throughout the cytoplasm without marked accumulation irrespective of the cell density. The cytoplasmic labeling in low Ca2+ looked filamentous and became aggregated when cells were treated with cytochalasin B; at least some of the aggregates coexisted with those of F-actin. In contrast, fodrin distribution was not affected with colchicine. On the other hand, in standard Ca2+, the protein became concentrated along the cell periphery and less conspicuous in the cytoplasm as the cells reached confluency. When cells were transferred from low to standard Ca2+, the distribution of fodrin changed accordingly within 180min. The present results indicate that fodrin in the keratinocyte is likely to be associated with actin filaments and that it takes two different ways of distribution both in vivo and in vitro. The peripheral and the cytoplasmic labeling of in vivo and in vitro cells are likely to correspond. It may be that fodrin changes its localization according to the cell's proliferative activity

Analysis of the protein profiles of the antibiotic-resistant Salmonella typhimurium definitive phage type (dt) 104

The emergent Salmonella typhimurium definitive phage type (DT) 104 is of particular global concern due to its frequent isolation and multiple antibiotic resistances. There is thus a need to know the kind of proteins expressed by S. typhimurium DT104 so as to provide a basis for developing an intervention. This study examined the protein profiles of a few isolates of S. typhimurium DT104 and a non-DT104 strain S. typhimurium L1388 (ST). Crude SDS-soluble whole cell and outer membrane protein (OMP) extracts revealed similar protein profiles for both phage types. A single major protein band (28.4 kDa) was observed in periplasmic fractions from both phage types. However, proteins released into growth medium was variable; one of the DT104 isolates had common proteins with the non-DT104 strain ST. Similar SDS-soluble whole cell protein profiles were observed for both phage types grown in a low-iron Medium A at 37oC; but a 38.5 kDa protein (observed in TSB-grown cells) was observed only in the temperature-tolerant DT104 isolate. The protein contents of cell-free ultracentrifuge supernatants of sonically disrupted cells of each of the DT104 isolates were significantly (P < 0.05) more than that from ST L1388, but the latter expressed a 51-kDa protein absent in the supernatants of all DT104 isolates. The higher protein content of DT104s provides possible indication of increased production of protein-like metabolites. Although the N-terminal sequence of the first twenty amino acids of the 51-kDa protein (Ala-Gln-Val-Ile-Asn-Thr-Asn-Ser-Leu-Ser-Leu-Leu-Thr-Gln-Trp-Ala-Ala-Ala-Ala-Ala) showed 14-amino acid overlap and resemblance with the flagillin, FLIC, only fourteen of its 104 trypsin digests were homologous with those of FLIC. Further work is being done to characterize this protein and to investigate its potential for use as vaccine target through antigenicity tests.African Journal of Biotechnology Vol. 4 (7), pp. 727-737, 200

Effect of TU-100 on Peyer's patches in a bacterial translocation rat model

Background: Daikenchuto (TU-100), a Japanese herbal medicine, is widely used for various gastrointestinal diseases. We have previously reported that TU-100 suppresses CPT-11-induced bacterial translocation (BT) by maintaining the diversity of the microbiome. In this study we show that TU-100 modulates the immune response during BT by inducing PD-1 expression in Peyer's patches. Methods: Eighteen male Wistar rats were divided into four groups: a control group; a control + TU-100 group, given TU-100 1000 mg/kg orally for 5 d; a BT group, given CPT-11 250 mg/kg intra-peritoneal for 2 d; and a TU-100 group, given TU-100 1000 mg/kg orally for 5 d with CPT-11 250 mg/kg intra-peritoneal on days 4 and 5. Results: The size of Peyer's patch was significantly bigger in the BT group compared to the control group (9.0 × 104 µm2 vs 29.4 × 104 µm2, P < .05), but improved in the TU-100 group (15.4 × 104 µm2, P < .005). TU-100 significantly induced PD-1 expression in Peyer's patch compared to the control group and the BT group (control vs BT vs TU-100 = 4.3 ± 4.9 vs 5.1 ± 10.3 vs 17.9 ± 17.8). The CD4+ cells were increased in the BT group (P < .05) compared to the control group but decreased in the TU-100 group. The Foxp3+ cells were increased in the BT group compared to the control group (P < .05), and further increased in the TU-100 group compared to the BT group. CPT-11 significantly increased TLR4, NF-κβ, TNF-α mRNA expressions in the BT group. TU-100 cotreatment significantly reversed these mRNA expressions. Conclusion: TU-100 may have a protective effect against BT through PD-1 expression in Peyer's patch

Lck inhibition attenuate lung fibrosis by suppressing Treg activity

Background Lymphocyte-specific protein tyrosine kinase (Lck) is a member of the Src family of tyrosine kinases. The significance of Lck inhibition in lung fibrosis has not yet been fully elucidated, even though lung fibrosis is commonly preceded by inflammation caused by infiltration of Tcells expressing Lck. In this study, we examined the effect of Lck inhibition in an experimental mouse model of lung fibrosis. We also evaluated the effect of Lck inhibition on the expression of TGF-β1, an inhibitory cytokine regulating the immune function, in regulatory T-cells (Tregs). Methods Lung fibrosis was induced in mice by intratracheal administration of bleomycin. A-770041, a Lck-specific inhibitor, was administrated daily by gavage. Tregs were isolated from the lung using a CD4+CD25+ Regulatory T-cell Isolation Kit. The expression of Tgfb on Tregs was examined by flow cytometry and quantitative polymerase chain reaction. The concentration of TGF-β in bronchoalveolar lavage fluid (BALF) and cell culture supernatant from Tregs was quantified by an enzyme-linked immunosorbent assay. Results A-770041 inhibited the phosphorylation of Lck in murine lymphocytes to the same degree as nintedanib. A-770041 attenuated lung fibrosis in bleomycin-treated mice and reduced the concentration of TGF-β in BALF. A flow-cytometry analysis showed that A-770041 reduced the number of Tregs producing TGF-β1 in the lung. In isolated Tregs, Lck inhibition by A-770041 decreased the Tgfb mRNA level as well as the concentration of TGF-β in the supernatant. Conclusions These results suggest that Lck inhibition attenuated lung fibrosis by suppressing TGF-β production in Tregs and support the role of Tregs in the pathogenesis of lung fibrosis

Photo-excitation band-structure engineering of 2H-NbSe2_2 probed by time- and angle-resolved photoemission spectroscopy

We investigated the nonequilibrium electronic structure of 2H-NbSe$_2$ by time- and angle-resolved photoemission spectroscopy. We find that the band structure is distinctively modulated by strong photo-excitation, as indicated by the unusual increase in the photoelectron intensities around E$_F$. In order to gain insight into the observed photo-induced electronic state, we performed DFT calculations with modulated lattice structures, and found that the variation of the Se height from the Nb layer results in a significant change in the effective mass and band gap energy. We further study the momentum-dependent carrier dynamics. The results suggest that the relaxation is faster at the K-centered Fermi surface than at the $\Gamma$-centered Fermi surface, which can be attributed to the stronger electron-lattice coupling at the K-centered Fermi surface. Our demonstration of band structure engineering suggests a new role for light as a tool for controlling the functionalities of solid-state materials.Comment: 7 pages, 5 figure

Dual effects of FGFR inhibition in lung fibrosis

[Rationale] Fibroblast growth factors (FGF) are major factors associated with the pathogenesis of pulmonary fibrosis. Nintedanib, a tyrosine kinase inhibitor targeting several growth factor receptors including the FGF receptor (FGFR), has been approved for the treatment of idiopathic pulmonary fibrosis (IPF). On the other hand, recent reports suggest that FGF are required for epithelial recovery. In this study, we focused on FGF signaling to both fibroblasts and alveolar epithelial cells (AECs), and examined the effect of a pan-FGFR blocker on experimental pulmonary fibrosis in mice. [Methods] The effects of BGJ398, a pan-FGFR inhibitor, on the migration and proliferation of fibroblasts and AECs were assessed using transwell migration or 3H-thymidine incorporation assays. The expression of FGFR was analyzed using immunoblot or flow cytometry. We also investigated the effect of BGJ398 on the pulmonary fibrosis induced by bleomycin in mice. [Results] Both lung fibroblasts and AECs expressed FGFRs. BGJ398 significantly inhibited the proliferation and migration of lung fibroblasts stimulated with FGF2. BGJ398 also reduced the proliferation of AECs in response to FGF2. Although the administration of BGJ398 ameliorated pulmonary fibrosis in bleomycin-treated mice, it increased mortality due to alveolar injury and inhibition of AEC regeneration. [Conclusions] These data suggest that the total inhibition of FGFR signaling can suppress lung fibrosis by inhibiting fibroblast activities, although alveolar injury is simultaneously caused