Ultraviolet Action Spectrum and Effect of EPC-K1 on Ultraviolet Radiation-induced Injury in Cultured Normal Human Epidermal Keratinocytes

This study was aimed to determine the ultraviolet (UV: 235-310nm) action spectrum for killing normal human epidermal keratinocytes (NHEK) and to investigate the preventive effect of EPC-K1, a phosphate diester of vitamin C and vitamin E on UV radiation-induced NHEK injury. NHEK were cultured in EpiLife medium supplemented with Human Keratinocyte Growth Supplement Kit. NHEK viability was determined by crystal violet (CV) staining 48 h after the UV irradiation. The mRNA expressions of the C/EBP homologous protein (Chop) transcription factor and endoplasmic reticulum-resident molecular chaperone, Bip, were determined by RT-PCR analyses. UV was especially effective in killing NHEK when applied in the wavelength region of 250-280nm. The minimum exposure dose required to kill 50% of cells (LD50) was 1.64mJ/cm2 at 269nm. At 235 and 310nm, the LD50 for NHEK was 6.62 and 293mJ/cm2, respectively. Irradiation of 660-mJ/cm2 at 310nm significantly decreased the cell viability to 30% of control (without irradiation). The addition of 0.1mM EPC-K1 after irradiation returned the cell viability to 118%. Six hours after the 660-mJ/cm2 irradiation at 310nm, Chop and Bip mRNA levels in NHEK were increased to 487% and 283%, respectively, and were not significantly affected by EPC-K1. Chop and Bip are responsive to ER stress. These results suggested that EPC-K1 exerts a protective effect against UV-induced NHEK injury, and further studies should investigate the molecular mechanism underlying this effect

Effect of Ethylene Diurea on Oxygen-induced Ischemic Retinopathy in the Neonatal Rat

This study investigated the effect of N-[2- (2-oxo-1-imidazolidinyl) ethyl]-N-phenylurea (ethylene diurea, EDU) on oxygen-induced ischemic retinopathy (OIR) in a neonatal rat model. OIR was induced by maintaining daily cycles of 80% oxygen (20.5h), ambient air (0.5h), and a progressive return to 80% oxygen (3h) for 12 days (postnatal day: P12). The rats were treated intraperitoneally with EDU (30mg/kg body weight) or distilled water (DW) from P6 to P17. At P18, the percentage of avascular areas in the total retinal area (%AVA) was measured, and retinal neovascularization (NV) was scored in ADPase-stained retinas. Retinal superoxide dismutase (SOD) activity in the retina was also determined by a chemiluminescence method. The mean %AVA in the EDU-treated group (9.3 ± 1.7%, n = 16) was lower than in the DW group (18.2 ± 4.7%, n = 17). EDU did not significantly affect NV, but significantly increased SOD activity (1.36 ± 0.13 units/mg protein, n = 4) compared to DW treatment (1.04 ± 0.01 units/mg protein, n = 4, P = 0.032) at P18. These results suggest that EDU treatment decreased the %AVA, accompanied by an increase in normal retinal vascular growth and/or a decrease in vessel proliferation. The increased SOD activity observed in the present study is likely to involve the EDU-mediated effects

Effects of Green Tea Fractions on Oxygen-Induced Retinal Neovascularization in the Neonatal Rat

This study aimed to investigate the preventive effects of green tea fractions (GTFs) on rat model of oxygen-induced retinopathy (OIR). Neonatal Sprague-Dawley rats were exposed to daily cycles of 80% oxygen (20.5 h), ambient air (0.5 h), and progressive return to 80% oxygen (3 h) until postnatal day 12 (P12), then the rats were placed in ambient air until P18. The green tea was fractionated by DM-A50, DM-W, M-B, and M-W. The rats were treated once daily from P6 to P17 by gastric gavage of GTFs (0.05 or 0.01 g/ml) or distilled water (DW) at 50 µl/10 g body weight. On P18, the rats were sacrificed and the retinal samples were collected. The retinal neovascularization (NV) was scored and avascular areas (AVAs) were measured as a % of total retinal area (%AVAs) in ADPase stained retinas. The NV scores in 0.01 g/ml M-W were significantly lower than those in DW. The %AVAs in 0.05 g/ml DM-A50 and in 0.05 g/ml and 0.01 g/ml M-W were significantly lower than those in DW. There were less catechins, and less caffeine in M-W fraction compared with other GTFs, suggesting components of green tea except for catechins and caffeine might suppress the neovascularization in rat model of OIR

Porphyrin‐uptake in liposomes and living cells using an exchange method with cyclodextrin

The water‐solubilisation of porphyrin derivatives is very important for biological applications. Although liposomal drug carriers for porphyrin derivatives have shown significant promise in the field of medicinal chemistry (e.g., as sensitisers for photodynamic therapy), it is currently not possible to prepare lipid‐membrane‐incorporated tetraphenylporphyrin (TPP) with a high concentration of TPP using conventional methods. In this study, we have succeeded in preparing lipid-membrane‐incorporated TPP and zinc(II) tetraphenylporphyrin (ZnTPP) from the corresponding TPP or ZnTPP•cyclodextrin complex using the exchange method in lipid‐membranes composed of liposomes. Furthermore, the exchange method allowed for the incorporation of TPP or ZnTPP into the plasma membranes of HeLa cells. However, it was not possible to prepare lipid‐membrane‐incorporated porphyrin derivatives with polar and hydrophilic groups in the meso positions using this exchange reaction.Electronic supplementary information (ESI) available: Experimental procedures, 1H NMR spectra, DLS measurements, cryo-TEM images, phase contrast and fluorescence images. See DOI: 10.1039/c5ra24985This work was supported by JSPS KAKENHI a Grant‐in‐Aid for Scientific Research (B) (Grant No. 25288037) and a Grant‐in‐Aid for Young Scientists (A) (Grant No. 24681028)

Design Methodology and Performance Evaluation of High Energy Absorbing Rockfall Protection Net by Full-Scale Experiments and Dynamic Analysis

It is obligatory to verify the performance of high-energy-absorbing pocket-type rockfall protection nets by full-scale experiments. While the performance verification by full-scale experiments can provide a practical evaluation, it requires a lot of cost and time because it is necessary to conduct the experiments under several conditions depending on the structure of the rockfall protection net. Therefore, the performance verification by combining full-scale experiments and numerical analysis may be a useful tool. In this study, a full-scale experiment of a high-energy-absorbing pocket-type rockfall protection net was conducted to evaluate the net performance with respect to the required capability, and reusability and repairability of the components by understanding the behavior of the net under the action of heavy weights. Throughout the experiment, the deformation of the net, the change of the wire rope tension with time, and the displacement of the top of the net and the top of the pillar were measured in detail. Moreover, it was confirmed that it is possible to evaluate the performance verification based on the performance requirements. Then, the full-scale experiment was replicated by numerical simulations to examine the effects of the rock shape on the behavior and performance of the simulated net. Through the thorough investigations on the deformation of the net, the relation between the maximum tension of the wire rope and the slip length of the shock absorber, the temporal change of the tension of each wire rope, and the absorbed energy of each structural member, it was judged that the reproducibility of the model predictions is adequately high. Specifically, when the impact energy is constant, the effect of the rock shape on the behavior and performance is considered to be small