RNA editing of 10 Didymium iridis mitochondrial genes and comparison with the homologous genes in Physarum polycephalum

Regions of the Didymium iridis mitochondrial genome were identified with similarity to typical mitochondrial genes; however, these regions contained numerous stop codons. We used RT-PCR and DNA sequencing to determine whether, through RNA editing, these regions were transcribed into mRNAs that could encode functional proteins. Ten putative gene regions were examined: atp1, atp6, atp8, atp9, cox1, cox2, cytb, nad4L, nad6, and nad7. The cDNA sequences of each gene could encode a functional mitochondrial protein that was highly conserved compared with homologous genes. The type of editing events and editing sequence features were very similar to those observed in the homologous genes of Physarum polycephalum, though the actual editing locations showed a variable degree of conservation. Edited sites were compared with encoded sites in D. iridis and P. polycephalum for all 10 genes. Edited sequence for a portion of the cox1 gene was available for six myxomycetes, which, when compared, showed a high degree of conservation at the protein level. Different types of editing events showed varying degrees of site conservation with C-to-U base changes being the least conserved. Several aspects of single C insertion editing events led to the preferential creation of hydrophobic amino acid codons that may help to minimize adverse effects on the resulting protein structure.     

Oral ontogeny of the Ayu, Plecoglossus altivelis and comparisons with the jaws of other salmoniform fishes

The Japanese Ayu, Plecoglossus altivelis Temminck & Schlegel is the sole representative of the salmoniform family Plecoglossidae. The Ayu is remarkable for its dentition which in adults comprises groups of diagonally arranged comb-like teeth in the outer tissue of the jaws. In juveniles (below 63 mm SL) the teeth are attached normally to the jaws. The transition of tooth form is correlated with a switch from zooplanktivory to algal or aufwuchs grazing. The present study follows the development of the teeth, jaws, oral cavity ethmoid and suspensorial elements in specimens ranging in size from 41–70 mm SL. The possible mode of function of the adult dentition is discussed. Comparisons are made with the jaws of other salmoniform fishes and a suite of supposed apomorphic characters are identified which are also shared with certain genera of the family Osmeridae, thus supporting the ideas of others that the Osmeridae is a paraphyletic assemblage.     

Issues related to mineralized tissue biology in human evolutionary research

This comunication has two primary aims concerned with mineralized tissue biology (e.g. hard tissue biology of bone and tooth) research in human evolutionary studies: First, to introduce the literature and the methods (at the time of this symposium) so that one has an idea of the nature of this research and where one can go for details of the methodologies, etc; Second — and of primary concern here — to discuss issues that have come to light as a result of these studies mainly because of its recent beginnings as a subfield within paleoanthropology. Issues related to skeletal studies include; 1) whether different cortical surface pattens and bone tissue types influence the appearance and interpretation of bone growth activity states; 2) if SEM analyses of cortical surfaces in fossil hominids allow one to construct meaningful representations of remodeling patterns; 3) whether these representations can be used in phylogenetic arguments; and 4) how intraspecific variability would affect these issues. Issues related to dental studies include: 1) the relationship between the rate and pattern of eraly hominid dental development; 2) experimental support for the calibration of eraly hominid dental developmental rates; and 3) whether replica techniques are suitable for microanatomical studies of these sorts.     

Hormone-responsive cells derived from human dental papilla: Characterization in vitro and in vivo in diffusion chambers

Summary Cells of the dental papilla are capable of odontoblastic, fibroblastic, and endothelial differentiation and formation of dentin and the dental pulp. In the present study dental papilla cells, obtained from human tooth buds (HDP cells), were cultured in vitro through 3 to 7 passages. After exposure to prostaglandin E₂ there was a marked decrease in intracellular cyclic AMP (cAMP) levels as compared to hormone-free controls. Parathyroid hormone and calcitonin had stimulatory effects with 1 and 2 log increases in cAMP, respectively. The HDP cells showed moderate activity of alkaline phosphatase, 1 log higher than that of hamster kidney fibroblasts (BHK 13) and 1 log lower than that of osteoblastic osteosarcoma cells (ROS 17/2). When cultured for 4 or 8 wk in diffusion chambers (DC) implanted in athymic mice, many of the HDP cells underwent odontoblastic morphodifferentiation with very long, single processes extending into the matrix. This matrix contained banded and unbanded collagen fibers. Neither light nor electron microscopy of the DC content revealed mineral deposits. These results suggest that HDP cells have an intrinsic potential for partial odontoblastic differentiation; inductive signals like those originating from odontogenic epithelium are probably essential for the completion of hard tissue formation.     

Study of physiopathological phenomena in dental enamel by neutron activation analysis

An epithermal neutron activation method is used to determine the concentration of mineral elements in human dental enamel. A large number (252) of samples from ancient and modern origins are analyzed. The analytical results are mathematically processed using a statistical multivariant method. This allows to differentiate deciduous from permanent teeth and decayed from sound enamel. It is also possible to distinguish the teeth coming from two different necropoles. The origin and the localization of determined elements in the mineralized part, or in the aqueous-organic part, of enamel is suggested. Their role, as witnessed in the physiopathological phenomena of dental enamel, is discussed.     

Comparison of a Bone Criterion and a dental Criterion for estimation of age at death in the study of an Ancient Cemetery

Two methods, a visual method based on the appearance of the auricular surface of the ilium and a metric method based on two dental criteria, were used in conjunction for estimation of skeletal age at death in paleodemographic study of an ancient cemetery, and were found to be coherent. However, the paleodemographic profiles differed according to sex, indicating a sexual difference in the evolution of the sacroiliac joint.     

A tooth crown morphology framework for interpreting the diversity of primate dentitions

Variation in tooth crown morphology plays a crucial role in species diagnoses, phylogenetic inference, and the reconstruction of the evolutionary history of the primate clade. While a growing number of studies have identified developmental mechanisms linked to tooth size and cusp patterning in mammalian crown morphology, it is unclear (1) to what degree these are applicable across primates and (2) which additional developmental mechanisms should be recognized as playing important roles in odontogenesis. From detailed observations of lower molar enamel–dentine junction morphology from taxa representing the major primate clades, we outline multiple phylogenetic and developmental components responsible for crown patterning, and formulate a tooth crown morphology framework for the holistic interpretation of primate crown morphology. We suggest that adopting this framework is crucial for the characterization of tooth morphology in studies of dental development, discrete trait analysis, and systematics.     

Histological Techniques to Improve Testing of Pulpal Response of Teeth to Filling Material

Two fixation fluids, two fixation techniques and two embedding methods were investigated for their effects on the quality of sections of teeth for pulpal response to filling materials to improve evaluation of pulpal responses. Sections from 32 baboon teeth were prepared, half with experimental cavities and half without, using either 10% formaldehyde or 4% glutaraldehyde, longitudinal tooth splitting or removal of the tooth apex, and paraffin or K plast resin embedding; decalcification in a formic acid mixture was a constant throughout. Histometric analysis showed that paraffin embedding produced less shrinkage than the K Plast resin embedding although the difference was not statistically significant. Six parameters of separation at the pu1p:dentine interface were studied: embedding, fixative, presence or absence of a cavity, cutting technique and individual animal tooth type. Statistical investigation revealed that fixative, cutting technique, and fixative and cutting technique combined had significant influences on the separation artifact. Of the combinations tested the choice of embedding method depends on which of the two artifacts, shrinkage or separation, is more adverse in the opinion of the investigator. Four percent glutaraldehyde together with the longitudinal split technique of fixation. processed by either K Plast resin embedding or paraffin embedding produced satisfactory pulpal sections.     

Evaluation of Preparation, Staining and Microscopic Techniques for Counting Incremental Lines in Cementum of Human Teeth

Apposition of cementum occurs in phases resulting in two types of layers with different optical and staining properties that can be observed by light microscopy. Narrow, dark staining incremental lines are separated by wider bands of pale staining cementum. The distance from one line to the next represents a yearly increment deposit of cementum in many mammals, and counting these lines has been used routinely to estimate the age of the animals. Incremental lines in cementum have also been observed in sections of human teeth, and the object of the present investigation was to examine a number of methods for preparing and staining them for counting. Longitudinal and transverse sections, either ground or decalcified, were cut from formalin fixed human dental roots, paraffin embedded or frozen, and stained using several techniques. The cementum was investigated using conventional light, fluorescence, polarized light, confocal laser scanning, interference contrast, phase contrast, and scanning electron microscopy. Incremental lines in the cementum could be observed in ground sections and, following decalcification, in both frozen and paraffin embedded sections. Toluidine blue, cresyl violet, hematoxylin, or periodic acid Schiff (PAS) stained incremental lines allowing differentiation by conventional light microscopy. Contrast was best using fluorescence microscopy and excitation by green light since the stained cemental bands, but not the incremental lines, fluoresced after staining with cresyl violet, PAS or hematoxylin and eosin. The results with other microscopic techniques were unsatisfactory. Since incremental lines are not destroyed by acids and stain differently than the remaining cementum, it is likely that they possess an organic structure which differs from the cementum. Incremental lines in human dental cementum could be observed best using decalcified sections stained with cresyl violet excited by green light.     

Phylogenetic position of Dictyostelium inferred from multiple protein data sets

The phylogenetic position of Dictyostelium inferred from 18S rRNA data contradicts that from protein data. Protein trees always show the close affinity of Dictyostelium with animals, fungi, and plants, whereas in 18S rRNA trees the branching of Dictyostelium is placed at a position before the massive radiation of protist groups including the divergence of the three kingdoms. To settle this controversial issue and to determine the correct position of Dictyostelium, we inferred the phylogenetic relationship among Dictyostelium and the three kingdoms Animalia, Fungi, and Plantae by a maximum-likelihood method using 19 different protein data sets. It was shown at the significance level of 1 SE that the branching of Dictyostelium antedates the divergence of Animalia and Fungi, and Plantae is an outgroup of the Animalia-Fungi-Dictyostelium clade.Correspondence to: T. Miyata