Thyroid cancer therapy is based on surgery followed by radioiodine treatment. The incorporation of radioiodine by cancer cells is mediated by sodium iodide symporter (NIS) (codified by the SLC5A5 gene), that is functional only when targeted to the cell membrane. We aimed to evaluate if NIS expression in thyroid primary tumors would be helpful in predicting tumor behavior, response to therapy and prognosis. NIS expression was addressed by qPCR and immunohistochemistry. In order to validate our data, we also studied SLC5A5 expression on 378 primary papillary thyroid carcinomas from The Cancer Genome Atlas (TCGA) database. In our series, SLC5A5 expression was lower in carcinomas with vascular invasion and with extrathyroidal extension and in those harboring BRAFV600E mutation. Analysis of SLC5A5 expression from TCGA database confirmed our results. Furthermore, it showed that larger tumors, with locoregional recurrences and/or distant metastases or harboring RAS, BRAF and/or TERT promoter (TERTp) mutations presented significantly less SLC5A5 expression. Regarding immunohistochemistry, 12/211 of the cases demonstrated NIS in the membrane of tumor cells, those cases showed variable outcomes concerning therapy success, prognosis and all but one were wild type for BRAF, NRAS and TERTp mutations. SLC5A5 mRNA lower expression is associated with features of aggressiveness and with key genetic alterations involving BRAF, RAS and TERTp. Mutations in these genes seem to decrease protein expression and its targeting to the cell membrane. SLC5A5 mRNA expression is more informative than NIS immunohistochemical expression regarding tumor aggressiveness and prognostic features.
A novel hydrogel, based on an alginate/hyaluronate mixture and Ce(III) ions, with effective bioactive and antimicrobial ability was developed to be used as vehicle of a synthetic bone substitute producing an injectable substitute (IBS). Firstly, three different IBSs were prepared using three developed alginate-based hydrogels, the hydrogel Alg composed by alginate, the hydrogel Alg/Ch composed by an alginate/chitosan mixture and the hydrogel Alg/HA composed by an alginate/hyaluronate mixture. MG63 cells viability on the IBSs was evaluated, being observed a significantly higher cell viability on the Alg/HA_IBS at all time points, which indicates a better cell adaptation to the material, increasing their predisposition to produce extracellular matrix and thus allowing a better bone regeneration. Moreover, SEM analysis showed evident filopodia and a spreader shape of MG63 cells when seeded on Alg/HA_IBS. This way, based upon the in vitro results, the hydrogel Alg/HA was chosen to the in vivo study by subcutaneous implantation in an animal model, promoting a slight irritating tissue response and visible tissue repairing. The next step was to grant antimicrobial properties to the hydrogel that showed the best biological behavior by incorporation of Ce(III) ions into the Alg/HA, producing the hydrogel Alg/HA2. The antimicrobial activity of these hyaluronate-based hydrogels was evaluated against Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Candida albicans. Results showed that Ce(III) ions can significantly enhance the hydrogel antimicrobial ability without compromising the osteoconductivity improvement promoted by the vehicle association to the synthetic bone substitute.
Titanium samples of different roughness R(a) and morphology were prepared using a combination of mechanical (grinding with a SiC paper or blasting with aluminum oxide particles with 65 or 250 microm) and chemical (attack with a sulphuric acid based solution or a hydrofluoric acid based solution) treatments. The biological performance of the prepared surfaces was evaluated using human bone marrow osteoblastic cell cultures. Mechanically treated samples presented different R(a) values and surface morphology. The hydrofluoric acid solution was more effective than the sulphuric acid solution in smoothing titanium surface and also in eliminating aluminum contamination resulting from the blasting process. Bone marrow cells seeded on the different titanium samples showed a similar pattern of behavior during cell attachment and spreading. Cells proliferated very well on all the titanium surfaces and cell growth was observed during approximately two to three weeks. The samples treated with the hydrofluoric acid solution presented higher alkaline phosphatase activity. Only the blasted samples treated with the acid solutions allowed seeded bone marrow cells to form a mineralized extracellular matrix. The best biological performance was found in the blasted samples treated with the hydrofluoric acid solution, which could be related to the characteristic microtopography of these samples that presented a homogeneous and smooth roughness.
Objective: Porphyromonas gingivalis and Prevotella intermedia are thought to be pathogens in adult periodontitis. Antibiotherapy is usually needed in the treatment of periodontitis being often prescribed empirically. To allow prescription of a specific antibiotic treatment, identification of resistance genes should be performed. The aim of this study was the identification of the presence of TetM, TetQ, TEM, cfxA, MefA, ErmB and Nim resistance genes in previously identified P. intermedia and P. gingivalis isolated from samples collected from periodontal infections.Method: PCR was used for the identification of TetM, TetQ, TEM, cfxA, MefA, ErmB and Nim resistance genes in strains isolated from samples collected from periodontal infections.Results: It was seen that 8% of isolates had one of the tested tetracycline resistance genes. A total of 32% of β-lactamases resistance genes was observed in isolated strains. It was also observed that 2% of isolates had one of the analysed erythromycin resistance genes. None of the isolates showed the presence of the metronidazole resistance gene.
Conclusions:Most strains harboring β-lactamase resistance genes had been previously identified as P. intermedia. No tetracycline resistance gene and a very low percentage of β-lactamase resistance genes were observed in P. gingivalis strains.
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