RESUMEN
The present study focuses on the investigation of the oxidized cell-free DNA (cfDNA) properties in several experimental models, including cultured cerebellum cells, peripheral blood lymphocytes (PBL), plasma, and hippocampus under an acute and chronic unpredictable stress model in rats. Firstly, our study shows that Spectrum Green fluorescence-labeled oxidized cfDNA fragments were transferred into the cytoplasm of 80% of the cerebellum culture cells; meanwhile, the nonoxidized cfDNA fragments do not pass into the cells. Oxidized cfDNA stimulates the antioxidant mechanisms and induction of transcription factor NRF2 expression, followed by an activation of NRF2 signaling pathway genes-rise of Nrf2 and Hmox1 gene expression and consequently NRF2 protein synthesis. Secondly, we showed that stress increases plasma cfDNA concentration in rats corresponding with the duration of the stress exposure. At the same time, our study did not reveal any significant changes of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) level in PBL of rats under acute or chronic stress, probably due to the significantly increased Nrf2 expression, that we found in such conditions. 8-oxodG is one of the most reliable markers of DNA oxidation. We also found an increased level of 8-oxodG in the hippocampal homogenates and hippocampal dentate gyrus in rats subjected to acute and chronic stress. Taken together, our data shows that oxidized cfDNA may play a significant role in systemic and neuronal physiological mechanisms of stress and adaptation.
Asunto(s)
Antioxidantes/metabolismo , Ácidos Nucleicos Libres de Células/metabolismo , Estrés Oxidativo , 8-Hidroxi-2'-Desoxicoguanosina/análisis , Animales , Ácidos Nucleicos Libres de Células/sangre , Ácidos Nucleicos Libres de Células/química , Células Cultivadas , Cerebelo/citología , Cerebelo/metabolismo , Citoplasma/metabolismo , Regulación de la Expresión Génica , Hemo Oxigenasa (Desciclizante)/genética , Hemo Oxigenasa (Desciclizante)/metabolismo , Hipocampo/metabolismo , Linfocitos/metabolismo , Masculino , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , Ratas , Ratas Wistar , Transducción de SeñalRESUMEN
A 263 GHz continuous-wave (CW) gyrotron was developed at the IAP RAS for future applications as a microwave power source in Dynamic Nuclear Polarization / Nuclear magnetic resonance (DNP/NMR) spectrometers. A new experimental facility with a computerized control was built to test this and subsequent gyrotrons. We obtained the maximum CW power up to 1 kW in the 15 kV/0.4 A operation regime. The power about 10 W, which is sufficient for many spectroscopic applications, was realized in the low current 14 kV/0.02 A regime. The possibility of frequency tuning by variation of the coolant temperature about 4 MHz/1 °C was demonstrated. The spectral width of the gyrotron radiation was about 10(-6).