Causation of Oxidative Stress and Defense Response of a Yeast Cell Model after Treatment with Orthodontic Alloys Consisting of Metal Ions

Misaligned teeth have a tremendous impact on oral and dental health, and the most efficient method of correcting the problem is orthodontic treatment with orthodontic appliances. The study was conducted to investigate the metal composition of selected orthodontic alloys, the release of metal ions, and the oxidative consequences that the metal ions may cause in the cell. Different sets of archwires, stainless steel brackets, and molar bands were incubated in artificial saliva for 90 days. The composition of each orthodontic material and quantification of the concentration of metal ions released were evaluated. Metal ion mixtures were prepared to determine the occurrence of oxidative stress, antioxidant enzyme defense system, and oxidative damage to proteins. The beta titanium alloy released the fewest metal ions and did not cause oxidative stress or protein damage. The metal ions from stainless steel and the cobalt-chromium alloy can cause oxidative stress and protein damage only at high concentrations. All metal ions from orthodontic alloys alter the activity of antioxidant enzymes in some way. The determined amounts of metal ions released from orthodontic appliances in a simulated oral environment are still below the maximum tolerated dose, and the concentrations of released metal ions are not capable of inducing oxidative stress, although some changes in antioxidant enzyme activity were observed at these concentrations.

Staphylococcal ClpXP protease targets the cellular antioxidant system to eliminate fitness-compromised cells in stationary phase

The transition from growth to stationary phase is a natural response of bacteria to starvation and stress. When stress is alleviated and more favorable growth conditions return, bacteria resume proliferation without a significant loss in fitness. Although specific adaptations that enhance the persistence and survival of bacteria in stationary phase have been identified, mechanisms that help maintain the competitive fitness potential of nondividing bacterial populations have remained obscure. Here, we demonstrate that staphylococci that enter stationary phase following growth in media supplemented with excess glucose, undergo regulated cell death to maintain the competitive fitness potential of the population. Upon a decrease in extracellular pH, the acetate generated as a byproduct of glucose metabolism induces cytoplasmic acidification and extensive protein damage in nondividing cells. Although cell death ensues, it does not occur as a passive consequence of protein damage. Instead, we demonstrate that the expression and activity of the ClpXP protease is induced, resulting in the degeneration of cellular antioxidant capacity and, ultimately, cell death. Under these conditions, inactivation of either clpX or clpP resulted in the extended survival of unfit cells in stationary phase, but at the cost of maintaining population fitness. Finally, we show that cell death from antibiotics that interfere with bacterial protein synthesis can also be partly ascribed to the corresponding increase in clpP expression and activity. The functional conservation of ClpP in eukaryotes and bacteria suggests that ClpP-dependent cell death and fitness maintenance may be a widespread phenomenon in these domains of life.

Protocol for subcellular targeted microthermal protein damage in cells cultivated on plasmonic nanosilver-modified surfaces

Despite proteotoxic stress and heat shock are implicated in diverse pathologies, currently no methodology to inflict defined, subcellular thermal damage exists. Here, we present a protocol for such a single-cell method compatible with laser-scanning microscopes, adopting the plasmon resonance principle. The method is based on modified microscopic cell culture plates, pre-coated by a layer of anisotropic silver NPs allowing excitation through targeted irradiation by conventional lasers used in the laser scanning microscopes and allowing controllable heating. Dose-defined heat causes protein damage in subcellular compartments, rapid heat-shock chaperones recruitment and stress signalling, thereby allowing unprecedented spatiotemporal analysis of thermal damage with broad applicability in biomedicine.

Influence of spatial structure on protein damage susceptibility: a bioinformatics approach

Aging research is a very popular field of research in which the deterioration or decline of various physiological features is studied. Here we consider the molecular level, which can also have effects on the macroscopic level. The proteinogenic amino acids differ in their susceptibilities to non-enzymatic modification. Some of these modifications can lead to protein damage and thus can affect the form and function of proteins. For this, it is important to know the distribution of amino acids between the protein shell/surface and the core. This was investigated in this study for all known structures of peptides and proteins available in the PDB. As a result, it is shown that the shell contains less susceptible amino acids than the core with the exception of thermophilic organisms. Furthermore, proteins could be classified according to their susceptibility. This can then be used in applications such as phylogeny, aging research, molecular medicine, and synthetic biology.