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The interactions between proteins in membranes govern many cellular functions. Our ability to probe for such interactions has greatly evolved in recent years due to the introduction of new fluorescence techniques. As a result, we currently have a choice of methods that can be used to assess the spatial distribution of a membrane protein, its association state, and the thermodynamic stability of the oligomers in the native milieu. These biophysical measurements have revealed new insights into important biological processes in cellular membranes.

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The differential signaling of multiple FGF ligands through a single fibroblast growth factor (FGF) receptor (FGFR) plays an important role in embryonic development. Here, we use quantitative biophysical tools to uncover the mechanism behind differences in FGFR1c signaling in response to FGF4, FGF8, and FGF9, a process which is relevant for limb bud outgrowth. We find that FGF8 preferentially induces FRS2 phosphorylation and extracellular matrix loss, while FGF4 and FGF9 preferentially induce FGFR1c phosphorylation and cell growth arrest. Thus, we demonstrate that FGF8 is a biased FGFR1c ligand, as compared to FGF4 and FGF9. Förster resonance energy transfer experiments reveal a correlation between biased signaling and the conformation of the FGFR1c transmembrane domain dimer. Our findings expand the mechanistic understanding of FGF signaling during development and bring the poorly understood concept of receptor tyrosine kinase ligand bias into the spotlight.

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The present study endeavors to decode the details of the transcriptional autoregulation effected by the MazE9 antitoxin of the Mycobacterium tuberculosis MazEF9 toxin-antitoxin system. Regulation of this bicistronic operon at the level of transcription is a critical biochemical process that is key for the organism's stress adaptation and virulence. Here, we have reported the solution structure of the DNA binding domain of MazE9 and scrutinized the thermodynamic and kinetic parameters operational in its interaction with the promoter/operator region, specific to the mazEF9 operon. A HADDOCK model of MazE9 bound to its operator DNA has been calculated based on the information on interacting residues obtained from these studies. The thermodynamics and kinetics of the interaction of MazE9 with the functionally related mazEF6 operon indicate that the potential for intracellular cross-regulation is unlikely. An interesting feature of MazE9 is the cis ⇌ trans conformational isomerization of proline residues in the intrinsically disordered C-terminal domain of this antitoxin.

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The aim of this work is to investigate the effects produced by polymicrobial biofilm (Porphyromonas gingivalis, Streptococcus mutans, Streptococcus sanguinis, and Streptococcus salivarius) on the corrosion behavior of titanium dental implants. Pure titanium disks were polished and coated with titanium nitride (TiN) and silicon carbide (SiC) along with their quarternized versions. Next, the disks were cultivated in culture medium (BHI) with P. gingivalis, S. mutans, S. sanguinis, and S. salivarius and incubated anaerobically at 37 °C for 30 days. Titanium corrosion was evaluated through surface observation using Scanning Electron Microscope (SEM) and Atomic Force Microscopy (AFM). Furthermore, the Ti release in the medium was evaluated by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). SEM images showed that coated Ti disks exhibited lower corrosion compared to non-coated disks, except for the quartenized TiN. This was confirmed by AFM, where the roughness was higher in non-coated Ti disks. ICP showed that Ti levels were low in all coating disks. These results indicate that these SiC and TiN-based coatings could be a useful tool to reduce surface corrosion on titanium implant surfaces.

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Hydrogels have gained interest for use in tissue regeneration and wound healing because of their absorbing and swelling properties as well as their ability to mimic the natural extracellular matrix. Their use in wound healing specifically may be in the form of a patch or wound dressing or they may be administered within the wound bed as a filler, gel in situ, to promote healing. Thiolated hyaluronic acid-polyethylene diacrylate (tHA-PEGDA) hydrogels are ideal for this purpose due to their short gelation times at physiological temperature and pH. But these hydrogels alone are not enough and require added components to gain bioactivity. In this work, RGD adhesion peptides and an antivascular endothelial growth factor receptor-2 (VEGF-R2) DNA aptamer are incorporated into a tHA-PEGDA hydrogel to make a bifunctional hyaluronic acid hydrogel. RGD peptides promote attachment and growth of cells while the anti-VEGF-R2 DNA aptamer seems to improve cell viability, induce cell migration, and spur the onset of angiogenesis by tube formation by endothelial cells. This bifunctional hydrogel supports cell culture and has improved biological properties. The data suggest that these hydrogels can be used for advanced tissue regeneration applications such as in wound healing.

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Titanium nitride (TiN) and silicon carbide (SiC) adhesion properties to biofilm and the proliferation of human osteoblasts were studied. Quaternized titanium nitride (QTiN) was produced by converting the surface nitrogen on TiN to a positive charge through a quaternization process to further improve the antibacterial efficiency. The SiC required a nitridation within the plasma chamber of the surface layer before quaternization could be carried out to produce quaternized SiC (QSiC). The antimicrobial activity was evaluated on the reference strains of Porphyromonas gingivalis for 4 h by fluorescence microscopy using a live/dead viability kit. All the coatings exhibited a lower biofilm coverage compared to the uncoated samples (Ti-85.2%; TiN-24.22%; QTiN-11.4%; SiC-9.1%; QSiC-9.74%). Scanning Electron Microscope (SEM) images confirmed the reduction in P. gingivalis bacteria on the SiC and TiN-coated groups. After 24 h of osteoblast cultivation on the samples, the cell adhesion was observed on all the coated and uncoated groups. Fluorescence images demonstrated that the osteoblast cells adhered and proliferated on the surfaces. TiN and SiC coatings can inhibit the attachment of Porphyromonas gingivalis and promote osteoblast adhesion on the titanium used for implants. These coatings may possess the ability to prevent the development of peri-implantitis and stimulate osteointegration.