RESUMO
Gasdermins execute programmatory cell death, known as pyroptosis, by forming medium-sized membrane pores. Recently, the molecular structure of those pores as well as the diversity in their shape and size have been revealed by cryoTEM and atomic force microscopy, respectively. Even though a growth of smaller to larger oligomers and reshaping from slits to rings could be documented, the initiation of the gasdermin pore formation remains a mystery. In one hypothesis, gasdermin monomers insert into membranes before associating into oligomeric pores. In the other hypothesis, gasdermin oligomers preassemble on the membrane surface prior to membrane insertion. Here, by studying the behavior of monomeric membrane-inserted gasdermin-A3 (GSDMA3), we unveil that a monomeric gasdermin prefers the membrane-adsorbed over the membrane-inserted state. Our results thus support the hypothesis of oligomers preassembling on the membrane surface before membrane penetration. At the same time, our simulations of small membrane-inserted arcs of GSDMA3 suggest that the inserting oligomer can be small and does not have to comprise a full ring of approximately 26-30 subunits. Moreover, our simulations have revealed an astonishingly large impact of salt-bridge formation and protein surroundings on the transmembrane passage of charged residues, reducing the energetic cost by up to 53% as compared to their free forms. The here observed free energy barrier of mere 5.6 kcal/mol for the membrane insertion of monomeric GSDMA3 explains the surprising ability of gasdermins to spontaneously self-insert into cellular membranes.
RESUMO
Aim of this randomized clinical trial was to assess the development of root caries lesions with and without (adjuvant) professional prevention treatment over 24 months. 20 participants with two or three non-cavitated root carious lesions were included (n = 52), whereby lesions were randomly assigned to one out of three groups depending on varnish application (CF: Cervitec F [n = 20], P: placebo [n = 20], DP: Duraphate [n = 12]). All lesions were assessed by quantitative light-induced fluorescence (QLF; QRayCam); following outcome parameters were analyzed: fluorescence loss (ΔF %), lesion volume (ΔQ %µm2) and bacterial activity (ΔR %). Professional tooth cleaning and adjuvant varnish application were performed at baseline, after 3, 6, and 9 months. A follow-up examination was performed 1 year after preventive care with varnish application 24 months after baseline. ∆F showed a significant time effect in CF (p = 0.03), which was not confirmed in post hoc analysis (p > 0.05). For P and DP, no time effect was detected (p > 0.05). ∆Q was significantly higher 12 months after baseline in CF (p = 0.02). In P, a significant time effect occurred (p = 0.01), without significant results in post hoc testing. ∆R showed higher values at baseline vs. 12 months in CF (p = 0.03) and 24 months compared to 12 months in DP (p = 0.02). Professional preventive treatment inhibited the progression of root caries lesions beyond their termination for 12 months, irrespective of an adjunctive varnish application. Preventive measures have a long-term effect on root carious lesions, even 1 year after their termination.
