The teeth and faces of twins: providing insights into dentofacial development and oral health for practising oral health professionals.

The continuing studies of the teeth and faces of Australian twins and their families in the Craniofacial Biology Research Group in the School of Dentistry at the University of Adelaide began 30 years ago. Three main cohorts of twins have been recruited, enabling various objectives and specific hypotheses to be addressed about the roles of genetic, epigenetic and environmental influences on human dentofacial growth and development, as well as oral health. This paper highlights some key findings arising from these studies, emphasizing those of direct relevance to practising oral health professionals. We also draw on published literature to review the significant developments in relation to the use of precision 2D and 3D imaging equipment, the application of modern molecular techniques, and the development of sophisticated computer software for analysing genetic relationships and comparing complex shapes. Such developments are valuable for current and future work. Apart from the classical or traditional twin model, there are several other twin models that can be used in research to clarify the relative contributions of genetic, epigenetic and environmental contributions to phenotypic variation. The monozygotic (MZ) co-twin model is one particularly valuable method, given that examination of only one pair of MZ twins can provide considerable insights into underlying causes of observed variation. This model can be used in a dental practice environment, with oral health professionals having the opportunity to explore differences in orofacial structures between MZ co-twins who are attending as patients. As researchers have become more aware of the complexities of the interactions between the genome, the epigenome and the environment during development, there is the need to collect more phenotypic data and define new phenotypes that will better characterize variations in growth processes and health status. When coupled with powerful new genetic approaches, including genome-wide association studies and linkage analyses, exciting opportunities are opening up to unravel the causes of problems in craniofacial growth and common oral diseases in human populations.

Primary tooth emergence in Australian children: timing, sequence and patterns of asymmetry.

BACKGROUND: Information on the timing and sequence of human tooth emergence is valuable when analysing human growth and development, predicting the age of individuals, and for understanding the effects of genetic and environmental influences on growth processes. This paper provides updated data on the timing and sequence of primary tooth emergence in Australian children for both clinicians and researchers. METHODS: Twins were recruited from around Australia with data collected through parental recording of twins' primary tooth emergence. One twin from each pair was then randomly selected to enable the calculation of descriptive statistics for timing, sequence and asymmetry in tooth emergence. RESULTS: The first and last primary teeth emerged, on average, at 8.6 months and 27.9 months, respectively, with teeth emerging in the order: central incisor, lateral incisor, first molar, canine, second molar. Left-side antimeric teeth were more likely to emerge before their right-side counterparts but this was not statistically significant. At least 35% of all antimeric pairs had emerged within two weeks of each other, serving as a useful guideline for assessing symmetrical versus asymmetrical development. CONCLUSIONS: Primary tooth emergence in Australian twins is occurring later than reported previously for Australian singletons but is consistent with findings for singletons in other ethnic groups. The most common sequence of primary tooth emergence appears to be consistent in twins and singletons and has not changed over time.

Patterns of asymmetry in primary tooth emergence of Australian twins.

AIMS: This study is part of a larger investigation of genetic and environmental influences on primary tooth emergence in Australian twins. Our aims were to describe patterns of emergence asymmetry, including directional and fluctuating components (DA, FA), and to test for a genetic basis to observed asymmetry. METHODS: The study sample consisted of 131 twin pairs. Using one randomly-selected twin from each pair, dental asymmetry was examined by analysing the number of days between emergence of antimeres (Delta), with dates of emergence provided through parental recording. Scatterplots were used for assessment of DA and FA, followed by paired t-tests to detect significant differences in mean Delta from zero (evidence of DA). FA was assessed by calculating means and variances of the absolute value of Delta. A range of intervals (0, 7, 14, 21, 28 days) was used to define symmetrical emergence of antimeres. RESULTS: Although a trend in left-side advancement for tooth emergence was detected, this was not statistically significant. Relatively low levels of FA were noted through -out the primary dentition, with maxillary and mandibular lateral inisors displaying the highest values, but no evidence of a genetic influence on FA was noted. Around 50% of all antimeric pairs of primary teeth were found to emerge within 14 days of each other, although time differences of more than 50 days were noted in some cases. CONCLUSION: Studies of dental asymmetry provide insights into the biological basis of lateralisation in humans and the results can also assist clinicians to discriminate between normal and abnormal developmental patterns.

Strong genetic control of emergence of human primary incisors.

Our understanding of tooth eruption in humans remains incomplete. We hypothesized that genetic factors contribute significantly to phenotypic variation in the emergence of primary incisors. We applied model-fitting to data from Australian twins to quantify contributions of genetic and environmental factors to variation in timing of the emergence of human primary incisors. There were no significant differences in incisor emergence times between zygosity groups or sexes. Emergence times of maxillary central incisors and mandibular lateral incisors were less variable than those of maxillary lateral incisors and mandibular central incisors. Maxillary lateral incisors displayed significant directional asymmetry, the left side emerging earlier than the right. Variation in timing of the emergence of the primary incisors was under strong genetic control, with a small but significant contribution from the external environment. Estimates of narrow-sense heritability ranged from 82 to 94% in males and 71 to 96% in females.

Basement membrane component laminin-5 is a target of the tumor suppressor Smad4.

The tumor suppressor Smad4 is involved in carcinogenesis mainly of the pancreas and colon. Functional inactivation of Smad4 is a genetically late event that occurs upon transition from premalignant stages to invasive and metastatic growth. Smad4 encodes an intracellular messenger common to all signalling cascades induced by members of the transforming growth factor-beta (TGF-beta) superfamily of cytokines. Despite extensive knowledge about the mechanisms of TGF-beta/Smad signal transduction, little is known about Smad4 targets involved in the transition to malignancy. The hallmark of invasive growth is a breakdown of the basement membrane (BM), a specialized sheet of extracellular matrix produced through cooperation of epithelial and stromal cells. Laminin-5, a heterotrimeric epithelial-derived BM component, is commonly lost in carcinomas but not in premalignant tumors. Herein, we report that in human colon and pancreatic tumor cells, Smad4 functions as a positive transcriptional regulator of all three genes encoding laminin-5. Coordinate re-expression of the three laminin-5 chains induced by reconstitution of Smad4 leads to secretion and deposition of the heterotrimeric molecule in BM-like structures. These data define the expression control of an essential BM component as a novel function for the tumor suppressor Smad4.

Analysis of adenovirus gene transfer into adult neural stem cells.

Adult neural stem cells (aNSCs) represent an attractive source for the production of specific types of neurons in degenerative CNS diseases and for the development of new regenerative gene therapies. However, the use of adult NSCs for transplantation and gene replacement strategies requires efficient gene expression in the cells. Due to the low pathogenicity of adenovirus (Ad) for humans, its large delivery capacity, and long-term transgene expression, Ad vectors are widely used. Here, we tested the potential of the Ad vector system to transduce adult NSCs. Analysis of Ad receptor expression in primary aNSCs revealed a complete lack of the coxsackie-adenovirus receptor and no or low expression of alphanu- and beta5-integrins, respectively, on mRNA and protein level. Consistently, transduction at different multiplicities of infection using an Ad vector expressing the enhanced green fluorescent protein (GFP) showed that adult NSCs are particularly resistant to Ad infection even at highest MOI (1000) in contrast to differentiated types of neural cells.

Buoyancy-driven instability of an autocatalytic reaction front in a Hele-Shaw cell.

An autocatalytic reaction-diffusion front between two reacting species may propagate as a solitary wave, namely, at constant velocity and with a stationary concentration profile. Recent experiments on such reactions have been reported to be buoyancy unstable, under certain conditions. We calculate the linear dispersion relation of the resulting instability, by applying our recent analysis of the Rayleigh-Taylor instability of two miscible fluids in a Hele-Shaw cell. The computed dispersion relation as well as our three-dimensional lattice Bhatnagar-Gross-Krook (BGK) simulations fit reasonably well experimental growth rates reported previously.

Growth rates of the buoyancy-driven instability of an autocatalytic reaction front in a narrow cell

Experimental studies were performed on the buoyancy-driven instability of an autocatalytic reaction front in a quasi-2D cell. The unstable density stratification at an ascending front leads to convection that results in a fingerlike front deformation. The growth rates of the spatial modes of the instability are determined at the initial stage. A stabilization is found at higher wave numbers, while the system is unstable against low wave number perturbations. Whereas comparison with a reported model governed by Hele-Shaw flow fails, a two-dimensional Navier-Stokes model yields more satisfactory results. Still, present deviations suggest the presence of an additional mechanism that suppresses the growth.

Morphological evidence for a close interaction of chromaffin cells with cortical cells within the adrenal gland.

The adrenal medulla appears to exert a regulatory influence on adrenocortical steroidogenesis. We have therefore studied the morphology of rat, porcine and bovine adrenals in order to characterize the contact zones of adrenomedullary and adrenocortical tissues. The distribution of chromaffin cells located within the adrenal cortex and of cortical cells located within the adrenal medulla was investigated. Chromaffin cells were characterized by immunostaining for synaptophysin and chromogranin A, both being considered specific for neuroendocrine cells. Cortical cells were characterized by immunostaining for 17 alpha-hydroxylase, an enzyme of the steroid pathway. Cellular contacts of chromaffin cells and cortical cells were examined at the electron microscopical level. In rat and porcine adrenals, rays of chromaffin cells, small cell clusters and single chromaffin cells or small invaginations from the medulla could be detected in all three zones of the cortex. Chromaffin cells often spread in the subcapsular space of the zona glomerulosa. In porcine and bovine adrenals, 17 alpha-hydroxylase immunoreactive cells were localized within the medulla. Single cortical cells and small accumulations of cells were spread throughout this region. At the ultrastructural level, the chromaffin cells located within the cortex in pig and rat adrenals formed close cellular contacts with cortical cells in all three zones. Our morphological data provide evidence for a possible paracrine role of chromaffin cells; this may be important for the neuroregulation of the adrenal cortex.