Caerin 4 is a family of AMPs isolated from the frog called Litoria caerulea. In silico drug designing methods and using machine learning algorithms for AMPs design can reduce their usage restrictions such as production costs and the time required for investigation of their activity and toxicity. In this study, two short peptides were designed based on direct and reverse mirror repeats of GLWQKI conserved sequence from Caerin 4 family that called dCar12 and rCar12. Also, Caerin 4.1 was synthesized without primary GLWQKI sequence and named Car7‐23. Following the synthesis of peptides, their antimicrobial properties, cytotoxicity, secondary structure, and mode of action were further evaluated. Results indicated that rCar12 had a good antibacterial activity (at an MIC of 3.9–62.5 µg/ml), while Car7‐23 did not have any antimicrobial properties. Cytotoxicity of rCar12 at MICs range was <5%, which is much less than Caerin 4.1. In conclusion, rCar12 with reverse mirror repeat has different functional properties compared with dCar12. These results corroborate the fact that in two peptides with identical residues and length, the position and arrangement of amino acids are very important concerning peptide function. Moreover, GLWQKI sequence is highly crucial for the antimicrobial activity of Caerin 4 antimicrobial peptide family.
The number of 88 rats was chosen, after 21 days, they were ablactated and categorized them as following groups:
The control group 1 has a cage of 8 rats in a social condition. After 6 weeks of saline injection (1mg / kg), the yawning behavior is recorded for 60 minutes. Group 2 in social conditions of receiving ethanol for 2 months, each serving of 600 mg / kg. Group 3 social isolation without treatment with agonist and antagonist, Opioid and alcohol drugs in a separate cage, each cage has one rat. After 6 weeks, It is recorded the yawning was cured for 60 minutes. Group 4 social isolation for 6 weeks and then treatment with alcohol agonist (1200 mg / kg) after 30 minutes of yawing behavior is recorded for 60 minutes. Group 5 Social isolation after 6 weeks of treatment with opioid drugs (5mg / kg of morphine) after 30 minutes of yawing behavior is recorded for 60 minutes. Group 6 social isolation after 6 weeks of morphine + alcohol antagonist treatment after 30 minutes of yawing behavior is recorded for 60 minutes. Group 7 social isolation followed by 6 weeks of treatment with antagonist alcohol (naltrexone), 10 mg / kg, after 30 minutes of yawing behavior is recorded for 60 minutes. Group 8 Social isolation after 6 weeks of treatment with antagonist morphine (naloxone) 1 mg / kg of bad breath after 30 minutes of yawing behavior is recorded for 60 minutes. Group 9 social isolation followed by 6 weeks of treatment with antagonist alcohol + morphine after 30 minutes for 60 minutes of yawing behavior is recorded for 60 minutes. Group 10 social isolation followed by 6 weeks of treatment with antagonist alcohol + morphine antagonist after 30 minutes for 60 minutes of yawing behavior is recorded for 60 minutes. Group 11 Social isolation after 6 treatments with antagonist morphine + alcohol followed after 30 minutes for 60 minutes of yawing behavior is recorded for 60 minutes.
Injection of alcohol chronically reduces yawning, but alcohol injections in conditions of social isolation increase the number of yawns. Antagonist injection of naltrexone prior morphine in conditions of social isolation of yawning. Injection of morphine in conditions of social isolation does not change the number of yawning. Injection of morphine antagonist (naloxone) prior to antagonist of alcohol (naltrexone) in conditions of social isolation increases yawn. Also, the injection of morphine antagonist (naloxone) prior to alcohol into social isolation causes an increase in the number of yawns. Alcohol injection before morphine in conditions of social isolation increases the number of yawns.
It seems that the stress of social isolation by decreasing alcohol and morphine function increases the number of yawns.
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