Validation of Roberts' method using root canal width patterns as a mandibular maturity marker in determining the 18-year threshold.

The South African Bill of Rights and the Children's Act define a 'child' as a person under the age of 18 years. The age of 18 years is therefore significant for legal purposes in South Africa. The third molar is an important indicator in determining the 18-year threshold. Human biological growth markers are accepted indicators of a subject attaining the age of 18 years. A recent study by Roberts et al. suggested that the relative width of the distal root canals (RCW) of the lower left permanent molars (Fédération Dentaire Internationale notation 36, 37 and 38) as visualised on dental panoramic radiographs can be used as such a growth marker. This study aimed to validate this human biological growth marker in both black and white populations living in South Africa according to Roberts' method. The findings of this validation study were in agreement with this method which showed that individuals with Demirjian stage H left molars and category RCW-C were indeed over the 18-year threshold. Important aspects regarding the rationality and application of Roberts' method are discussed. The authors conclude that this method should not be used in isolation but as an adjunct with other age estimation methods.

Hydroxycinnamate conjugates as potential monolignol replacements: in vitro lignification and cell wall studies with rosmarinic acid.

The plasticity of lignin biosynthesis should permit the inclusion of new compatible phenolic monomers, such as rosmarinic acid (RA) and analogous catechol derivatives, into cell-wall lignins that are consequently less recalcitrant to biomass processing. In vitro lignin polymerization experiments revealed that RA readily underwent peroxidase-catalyzed copolymerization with monolignols and lignin oligomers to form polymers with new benzodioxane inter-unit linkages. Incorporation of RA permitted extensive depolymerization of synthetic lignins by mild alkaline hydrolysis, presumably by cleavage of ester intra-unit linkages within RA. Copolymerization of RA with monolignols into maize cell walls by in situ peroxidases significantly enhanced alkaline lignin extractability and promoted subsequent cell wall saccharification by fungal enzymes. Incorporating RA also improved cell wall saccharification by fungal enzymes and by rumen microflora even without alkaline pretreatments, possibly by modulating lignin hydrophobicity and/or limiting cell wall cross-linking. Consequently, we anticipate that bioengineering approaches for partial monolignol substitution with RA and analogous plant hydroxycinnamates would permit more efficient utilization of plant fiber for biofuels or livestock production.

Fluorescence-tagged monolignols: synthesis, and application to studying in vitro lignification.

Fluorescence-tagged coniferyl alcohols, coniferyl alcohol γ-coupled by ethylenediamine spacers to dimethylaminocoumarin or nitrobenzofuran fluorophores, were tested as photoprobes to study the oxidase-mediated polymerization of monolignols. The fluorescent coniferyl alcohol derivatives readily underwent peroxidase-catalyzed in vitro copolymerization with coniferyl alcohol to yield fluorescent dehydrogenation polymers, the backbone polymers of which were structurally indistinguishable from polymers formed solely from coniferyl alcohol. To illustrate the use of the photoprobes, we successfully monitored in real time the complexation of coniferyl alcohol with horseradish apoperoxidase by Förster resonance energy transfer (FRET) using the protein-tryptophan near the active site and a dimethylaminocoumarin moiety as donor and acceptor fluorophores. Furthermore, mixtures of fluorescence-tagged and normal coniferyl alcohols readily diffused into isolated maize cell walls and reacted with wall-bound peroxidases to form in muro artificial lignins that could be visualized by fluorescence microscopy. Thus we anticipate that fluorescence-tagged monolignols will be useful for in vitro and in vivo studies of cell wall lignification.