Antimicrobial activity and biologic potential of silver-substituted calcium phosphate constructs produced with self-propagating high-temperature synthesis.

There is significant demand for synthetic bone substitute materials that can decrease the incidence of implant-based bacterial infections. The intent of this research was to evaluate the antimicrobial activity and biologic potential of calcium phosphate (CaP) constructs substituted with silver (Ag) that were produced via self-propagating high-temperature synthesis (SHS). SHS is a combustion synthesis technique that has successfully generated porous CaP bioceramics intended for use in bone repair. SHS reactions are highly versatile; dopants can be added to the reactant powders to alter product chemistry and morphology. In this research, Ag powder was added to the reactants generating porous CaP constructs containing 0.5, 1, or 2 wt% Ag. Antibacterial performance of the constructs was assessed against Escherichia coli, a representative model for Gram-negative bacteria. Liquid solutions (1 µg/mL) of CaP-Ag particles to phosphate buffered saline were incubated with 10(5) cells/mL. After 24 h, 10 µL of solution were spread on an LB agar plate and cultured for 24 h at 37 °C. Samples cultured with CaP-Ag showed complete bacterial inhibition while the controls (E. coli only and CaP without Ag) exhibited significant colony formation. The effects of Ag concentration on cytotoxicity and biocompatibility were tested in vitro. At 7 days, osteoblasts uniformly enveloped the CaP-Ag particles and displayed a healthy flattened morphology suggesting the concentrations of Ag incorporated into constructs were not cytotoxic. CaP-Ag constructs produced via SHS represent a source of synthetic bone substitute materials that could potentially inhibit, or reduce the incidence of post-operative bacterial infections.

Developing genomic resources for the common bottlenose dolphin (Tursiops truncatus): isolation and characterization of 153 single nucleotide polymorphisms and 53 genotyping assays.

Although single nucleotide polymorphisms (SNPs) are commonly used in human genetics, they have only recently been incorporated into genetic studies of non-model organisms, including cetaceans. SNPs have several advantages over other molecular markers for studies of population genetics: they are quicker and more straightforward to score, cross-laboratory comparisons of data are less complicated, and they can be used successfully with low-quality DNA. We screened portions of the genome of one of the most abundant cetaceans in U.S. waters, the common bottlenose dolphin (Tursiops truncatus), and identified 153 SNPs resulting in an overall average of one SNP every 463 base pairs. Custom TaqMan(®) Assays were designed for 53 of these SNPs, and their performance was tested by genotyping a set of bottlenose dolphin samples, including some with low-quality DNA. We found that in 19% of the loci examined, the minor allele frequency (MAF) estimated during initial SNP ascertainment using a DNA pool of 10 individuals differed significantly from the final MAF after genotyping over 100 individuals, suggesting caution when making inferences about MAF values based on small data sets. For two assays, we also characterized the basis for unusual clustering patterns to determine whether their data could still be utilized for further genetic studies. Overall results support the use of these SNPs for accurate analysis of both poor and good-quality DNA. We report the first SNP markers and genotyping assays for use in population and conservation genetic studies of bottlenose dolphins.