Newcastle disease is one of the most important pathogens of domestic poultry including chickens. The hemagglutinin-neuraminidase (HN) of Newcastle disease virus is the principal target of neutralizing and protective antibodies against Newcastle disease. In this paper, we transformed tobacco plants with Agrobacterium tumefaciens EHA105 to generate the plants expressing HN of Newcastle disease virus (NDV). The insertion and copy numbers of HN gene in the genomic DNA of phosphinothricin-resistant plants were confirmed by PCR and Southern blot, respectively. The presence of the HN-specific transcript in the total RNAs of the leaves of transgenic plants was verified by Northern analysis. The recombinant HN proteins were detected by Western blot analysis using a polyclonal antibody against GST-HN fusion proteins. The highest level of expression of HN in leaves of transgenic plants was approximately 0.069% of the total soluble protein. ELISA assay showed that the recombinant protein extracted from transformants has normal immunoactivity. Transgenic tobacco expressing HN of NDV with sterilized PBS was fed to 6-week-old chickens. Immunized chickens developed slightly high titers of anti-HN serum IgG compared with those of the wild type plant. These results suggest that oral immunization with HN-transgenic tobacco provides a potential means of protecting chickens from NDV. Further modified animal experiments would be needed to increase the immunity of HN by co-administration of classical adjuvants or other trials.
The purpose of this study is to determine the difference in sleep-related factors and metabolites between normal sleep (NS) and sleep deficiency (SD) and to analyze the variations in metabolites according to the intensity of aerobic exercise under SD conditions. This study was conducted on 32 healthy male university students. Participants experienced both NS (8 h of sleep per night for 3 consecutive days) and SD (4 h of sleep per night for 3 consecutive days). After the SD period, the participants underwent treatment for 30 min by the assigned group [sleep supplement after SD (SSD), low-intensity aerobic exercise after SD (LES), moderate-intensity aerobic exercise after SD (MES), high-intensity aerobic exercise after SD (HES)]. For analysis, sleep-related factors were measured, and metabolites were analyzed by untargeted metabolite analysis using gas chromatography-time-of-flight mass spectrometry. As a result, SD showed that total sleep time (TST), duration of rapid eye movement (REM), duration of light sleep, and duration of deep sleep were significantly decreased compared to NS, whereas the Pittsburgh sleep quality index (PSQI), Epworth sleepiness scale (ESS), and visual analogue scale (VAS) were significantly increased compared to NS. The difference in metabolites between NS and SD showed that there were significant changes in the seven metabolites. There were 18 metabolites that changed according to the treatment groups in SD conditions. In summary, SD can exacerbate sleep quality, induce daytime sleepiness, increase fatigue, and increase metabolites that cause insulin resistance. Aerobic exercise under SD conditions can reduce metabolites that induce insulin resistance and increase the metabolites that help relieve depression caused by SD. However, HES has a negative effect, which increases fatigue, whereas LES has no negative effect. Thus, this study suggests that LES is the most appropriate exercise method under SD conditions.
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