SARS-CoV-2 Nucleocapsid Protein Is Not Responsible for Over-Activation of Complement Lectin Pathway.

The nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a viral structural protein that is abundant in the circulation of infected individuals. Previous published studies reported controversial data about the role of the N protein in the activation of the complement system. It was suggested that the N protein directly interacts with mannose-binding lectin-associated serine protease-2 (MASP-2) and stimulates lectin pathway overactivation/activity. In order to check these data and to reveal the mechanism of activation, we examined the effect of the N protein on lectin pathway activation. We found that the N protein does not bind to MASP-2 and MASP-1 and it does not stimulate lectin pathway activity in normal human serum. Furthermore, the N protein does not facilitate the activation of zymogen MASP-2, which is MASP-1 dependent. Moreover, the N protein does not boost the enzymatic activity of MASP-2 either on synthetic or on protein substrates. In some of our experiments, we observed that MASP-2 digests the N protein. However, it is questionable, whether this activity is biologically relevant. Although surface-bound N protein did not activate the lectin pathway, it did trigger the alternative pathway in 10% human serum. Additionally, we detected some classical pathway activation by the N protein. Nevertheless, we demonstrated that this activation was induced by the bound nucleic acid, rather than by the N protein itself.

Individual recognition and long-term memory of inanimate interactive agents and humans in dogs.

Investigation of individual recognition (IR) is difficult due to the lack of proper control of cues and previous experiences of subjects. Utilization of artificial agents (Unidentified Moving Objects: UMOs) may offer a better approach than using conspecifics or humans as partners. In Experiment 1, we investigated whether dogs are able to develop IR of UMOs (that is stable for at least 24 h) or that they only retain a more generalised memory about them. The UMO helped dogs to obtain an unreachable ball and played with them. One day, one week or one month later, we tested whether dogs display specific behaviour toward the familiar UMO over unfamiliar ones (four-way choice test). Dogs were also re-tested in the same helping context and playing interaction. Subjects did not approach the familiar UMO sooner than the others; however, they gazed at the familiar UMO earlier during re-testing of the problem solving task, irrespectively of the delay. In Experiment 2, we repeated the same procedure with human partners, applying a two-way choice test after a week delay, to study whether lack of IR was specific to the UMO. Dogs did not approach the familiar human sooner than the unfamiliar, but they gazed at the familiar partner earlier during re-testing. Thus, dogs do not seem to recognise an individual UMO or human after a short experience, but they remember the interaction with the novel partner in general, even after a long delay. We suggest that dogs need more experience with a specific social partner for the development of long-term memory.