Suitability of Biomorphic Silicon Carbide Ceramics as Drug Delivery Systems against Bacterial Biofilms.

The present work is aimed at getting a new insight into biomorphic silicon carbides (bioSiCs) as bone replacement materials. BioSiCs from a variety of precursors were produced, characterized, and loaded with a broad-spectrum antibiotic. The capacity of loaded bioSiCs for preventing and/or treating preformed S. aureus biofilms has been studied. The differences in precursor characteristics are maintained after the ceramic production process. All bioSiCs allow the loading process by capillarity, giving loaded materials with drug release profiles dependent on their microstructure. The amount of antibiotic released in liquid medium during the first six hours depends on bioSiC porosity, but it could exceed the minimum inhibitory concentration of Staphylococcus aureus, for all the materials studied, thus preventing the proliferation of bacteria. Differences in the external surface and the number and size of open external pores of bioSiCs contribute towards the variations in the effect against bacteria when experiments are carried out using solid media. The internal structure and surface properties of all the systems seem to facilitate the therapeutic activity of the antibiotic on the preformed biofilms, reducing the number of viable bacteria present in the biofilm compared to controls.

Development of a novel AMX-loaded PLGA/zein microsphere for root canal disinfection.

The aim of this study was to develop polymeric biodegradable microspheres (MSs) of poly(D-L lactide-co-glycolide) (PLGA) and zein capable of delivering amoxicillin (AMX) at significant levels for root canal disinfection. PLGA/zein MSs were prepared using a spray-drying technique. The systems were characterized in terms of particle size, morphology, drug loading and in vitro release. Drug levels were reached to be effective during the intracanal dressing in between visits during the endodontic treatment. In vitro release studies were carried out to understand the release profile of the MSs. Antimicrobial activity of AMX was performed by antibiograms. Enterococcus faecalis was the bacteria selected due to its prevalence in endodontic failure. Drug microencapsulation yielded MSs with spherical morphology and an average particle size of between 5 and 38 µm. Different drug-release patterns were obtained among the formulations. Release features related to the MSs were strongly dependent on drug nature as it was demonstrated by using a hydrophobic drug (indomethacin). Finally, AMX-loaded MSs were efficient against E faecalis as demonstrated by the antibiogram results. In conclusion, PLGA/zein MSs prepared by spray drying may be a useful drug delivery system for root canal disinfection.

Properties of the protein kinase that phosphorylates prothymosin alpha.

The prothymosin alpha kinase (ProTalphaK) is an apparently novel enzyme that is responsible for the phosphorylation of prothymosin alpha (ProTalpha), involved in the proliferation of mammalian cells. The present study investigated the properties of this enzyme. ProTalphaK is more effectively activated by Mn2+ than by other divalent cations, and its activity is unaffected by RNA. Its principal substrate in proliferating cells appears to be ProTalpha. Both in vivo and in vitro, it is unable to phosphorylate the peptides thymosin alphaI and thymosin alphaII, derived from the amino terminus of ProTalpha, despite the fact that the sites of phosphorylation of ProTalpha are contained within this part of its sequence. In trials in vivo, inhibition of gene expression abolished both phosphorylation of ProTalpha and ProTalphaK activity. ProTalphaK is located in the cytosolic fractions throughout the cell cycle. Its activity, which is dependent on cell proliferation, increases markedly during S phase and begins to decline as the cell enters G2. Studies of the effects of activators and inhibitors of protein kinases involved in signal transduction pathways suggest that ProTalphaK is activated by phosphorylation in a mitogen-initiated pathway that is dependent on PKC; however, PKC does not itself phosphorylate ProTalphaK, which is therefore presumably phosphorylated by another kinase.

A 180-kDa protein kinase seems to be responsible for the phosphorylation of prothymosin alpha observed in proliferating cells.

Prothymosin alpha (ProTalpha) is an acidic protein involved in cell proliferation. Its phosphorylation status is correlated with proliferative activity. Here we report the isolation and characterization of a ProTalpha-phosphorylating kinase (ProTalphaK) from mouse splenocytes that seems to be responsible for the in vivo phosphorylation of ProTalpha and that differs from other protein kinases reported to date. This enzyme, mainly located in the cytosol, has an molecular mass of 180 kDa and appears to be made up of two proteins of 64 and 60 kDa. Its activity was markedly enhanced by mitogenic activation of cells. The ProTalpha residues phosphorylated by the enzyme in vitro are a Thr at position 7 and another Thr at positions 12 or 13, both located within casein kinase 2 (CK-2) consensus motifs; these are the same residues as are phosphorylated in vivo. The new enzyme shows a number of clear structural and catalytic differences from CK-2. It phosphorylates histones H2B and H3, although with weaker activity than ProTalpha. An enzyme with the same characteristics was also found in other murine tissues and cell lines.

Prothymosin alpha binds histones in vitro and shows activity in nucleosome assembly assay.

Prothymosin alpha (Pro Talpha) is a polypeptide which appears to be involved in cell proliferation, though its precise function has yet to be identified. Here, we report experiments which show that calf Pro Talpha selectively binds to core histones and histone H1 in vitro. Characterization of these interactions by various procedures (including affinity chromatography on Pro T alpha-Sepharose columns, immunoblotting assay and investigation of the behaviour of mixtures of Pro T alpha and histones in solution) indicated that Pro T alpha has higher affinity for core histones (particularly H3 and H4) than for H1. Similarities between the histone-binding patterns of Pro T alpha and of poly(glutamic acid) suggest that the observed histone-binding capacity resides largely in the acidic central region of Pro T alpha. However, all five histones were also bound by T alpha 1 (a peptide corresponding to the first 28 amino acids of Pro T alpha); histone binding by the N-terminal region of Pro T alpha thus cannot be ruled out. Phosphorylation of Pro T alpha does not appear to affect these interactions. In accordance with the observed capacity for histone binding, Pro T alpha (in conjunction with ATP and some Pro T alpha-binding factor/s in a thymocyte extract) was able to induce in vitro nucleosome assembly. We discuss the possibility that Pro T alpha plays a role in chromatin remodelling.