30 research outputs found
Application of MLST and Pilus Gene Sequence Comparisons to Investigate the Population Structures of Actinomyces naeslundii and Actinomyces oris
Actinomyces naeslundii and Actinomyces oris are members of the oral biofilm. Their identification using 16S rRNA sequencing is problematic and better achieved by comparison of metG partial sequences. A. oris is more abundant and more frequently isolated than A. naeslundii. We used a multi-locus sequence typing approach to investigate the genotypic diversity of these species and assigned A. naeslundii (n = 37) and A. oris (n = 68) isolates to 32 and 68 sequence types (ST), respectively. Neighbor-joining and ClonalFrame dendrograms derived from the concatenated partial sequences of 7 house-keeping genes identified at least 4 significant subclusters within A. oris and 3 within A. naeslundii. The strain collection we had investigated was an under-representation of the total population since at least 3 STs composed of single strains may represent discrete clusters of strains not well represented in the collection. The integrity of these sub-clusters was supported by the sequence analysis of fimP and fimA, genes coding for the type 1 and 2 fimbriae, respectively. An A. naeslundii subcluster was identified with both fimA and fimP genes and these strains were able to bind to MUC7 and statherin while all other A. naeslundii strains possessed only fimA and did not bind to statherin. An A. oris subcluster harboured a fimA gene similar to that of Actinomyces odontolyticus but no detectable fimP failed to bind significantly to either MUC7 or statherin. These data are evidence of extensive genotypic and phenotypic diversity within the species A. oris and A. naeslundii but the status of the subclusters identified here will require genome comparisons before their phylogenic position can be unequivocally established
Characterization of Granulations of Calcium and Apatite in Serum as Pleomorphic Mineralo-Protein Complexes and as Precursors of Putative Nanobacteria
Calcium and apatite granulations are demonstrated here to form in both human and
fetal bovine serum in response to the simple addition of either calcium or
phosphate, or a combination of both. These granulations are shown to represent
precipitating complexes of protein and hydroxyapatite (HAP) that display marked
pleomorphism, appearing as round, laminated particles, spindles, and films.
These same complexes can be found in normal untreated serum, albeit at much
lower amounts, and appear to result from the progressive binding of serum
proteins with apatite until reaching saturation, upon which the mineralo-protein
complexes precipitate. Chemically and morphologically, these complexes are
virtually identical to the so-called nanobacteria (NB) implicated in numerous
diseases and considered unusual for their small size, pleomorphism, and the
presence of HAP. Like NB, serum granulations can seed particles upon transfer to
serum-free medium, and their main protein constituents include albumin,
complement components 3 and 4A, fetuin-A, and apolipoproteins A1 and B100, as
well as other calcium and apatite binding proteins found in the serum. However,
these serum mineralo-protein complexes are formed from the direct chemical
binding of inorganic and organic phases, bypassing the need for any biological
processes, including the long cultivation in cell culture conditions deemed
necessary for the demonstration of NB. Thus, these serum granulations may result
from physiologically inherent processes that become amplified with calcium
phosphate loading or when subjected to culturing in medium. They may be viewed
as simple mineralo-protein complexes formed from the deployment of
calcification-inhibitory pathways used by the body to cope with excess calcium
phosphate so as to prevent unwarranted calcification. Rather than representing
novel pathophysiological mechanisms or exotic lifeforms, these results indicate
that the entities described earlier as NB most likely originate from calcium and
apatite binding factors in the serum, presumably calcification inhibitors, that
upon saturation, form seeds for HAP deposition and growth. These calcium
granulations are similar to those found in organisms throughout nature and may
represent the products of more general calcium regulation pathways involved in
the control of calcium storage, retrieval, tissue deposition, and disposal