Although nitric oxide (NO) induces neuronal cell death under some conditions, it also can prevent apoptosis resulting from growth factor withdrawal. We investigated the molecular mechanism by which NO protects undifferentiated and differentiated PC12 cells from trophic factor deprivation-induced apoptosis. PC12 cells underwent apoptotic death in association with increased caspase-3-like activity, DNA fragmentation, poly(ADP-ribose) polymerase (PARP) cleavage, and cytochrome c release after 24 hr of serum withdrawal. The apoptosis of PC12 cells was inhibited by the addition of NO-generating donor S-nitroso-N-acetylpenicillamine (SNAP) (5-100 microM) and the specific caspase-3-like protease inhibitor Ac-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-cho) but not the YVADase (or caspase-1-like protease) inhibitor N-acetyl-Tyr-Val-Ala-Asp-aldehyde (Ac-YVAD-cho). SNAP and Ac-DEVD-cho prevented the increase in DEVDase (caspase-3-like protease) activity. The SNAP-mediated suppression of DEVDase activity was only minimally reversed by the incubation of cell lysate with dithiothreitol, indicating that NO did not S-nitrosylate caspase-3-like proteases in PC12 cells. Western blot analysis showed that NO inhibited the proteolytic activation of caspase-3. The cGMP analog 8-bromo-cGMP (8-Br-cGMP) blocked apoptotic cell death, caspase-3 activity and activation, and cytochrome c release. The soluble guanylyl cyclase inhibitor 1-H-oxodiazol-[1,2,4]-[4,3-a] quinoxaline-1-one (CODQ) significantly attenuated NO-mediated, but not 8-Br-cGMP-dependent, inhibition of apoptotic cell death, PARP cleavage, cytochrome c release, and DEVDase activity. Furthermore, the protein kinase G inhibitor KT5823 reversed both SNAP- and 8-Br-cGMP-mediated anti-apoptotic events. All these apoptotic phenomena were also suppressed by NO production through neuronal NO synthase gene transfer into PC12 cells. Furthermore, similar findings were observed in differentiated PC12 cells stimulated to undergo apoptosis by NO donors and NGF deprivation. These findings indicate that NO protects against PC12 cell death by inhibiting the activation of caspase proteases through cGMP production and activation of protein kinase G.

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Animal welfare is recognized as essential for the coexistence of humans and animals. Considering the increased demand and interest in animal welfare, many methods for improving animal welfare are being devised, but which method reduces animal stress has not been scientifically verified. Therefore, reducing animal stress by providing a proper breeding environment and environmental enrichment can be the basis for animal study. In this study, stress levels were assessed based on the mouse-breeding environment. We considered that the higher the body weight and the lower the corticosterone concentration, the lower the stress. According to the results, animals in the individual ventilation cages were determined to have lower serum cortisol concentrations, while the body weight of the animals was increased when in individual ventilation cages compared with individual isolated cages and when providing environmental enrichment compared with group breeding or not providing environmental enrichment. The results provide appropriate guidelines for improving laboratory animal welfare.

High-resolution seismic survey and sediment core sampling were conducted to investigate acoustic characteristics of gas-bearing sediments in Jinhae Bay, the southeast of Korea. The sediment in Jinhae Bay is mostly homogenous mud deposited after the Holocene transgression. Along with the 410 km of chirp seismic profiling, five piston core samples were collected on the track lines.Gassy sediments are common and occur widely in the bay. Core samples were analyzed for sediment texture, physical properties (porosity, water content, bulk density, and grain density), acoustic properties (compressional wave velocity and attenuation), and electrical resistivity. X-radiograph image analysis was also performed to observe the shape of degassing cracks. There is no significant downcore variation on physical and sediment textures regardless of existence of gas bubbles. However, compressional wave velocity dramatically decreases from average 1480 to 1380$739 m=s for the cores that penetrate the gas-bearing zones. This is probably due to degassying cracks that developed by escaping gases and free gas bubbles that are still trapped in the cores. Electrical resistivity is the only geotechnical property that increases in the gas-bearing zone where compressional wave velocity abruptly decreases. This indicates the possibility of using both electrical resistivity as an index variable as well as to compressional wave velocity to identify gassy sediment microstructure because there are little changes in texture and composition of sediment.