Cadherin-related neuronal receptors in incisor development.

Cadherins are cell adhesion molecules that are critical for tissue development. In this report, we identified members of the cadherin family cadherin-related neuronal receptors (CNRs) 1 and 5 expressed in rat incisors by the differential display method. Quantitative RT-PCR revealed that CNR1 mRNA is expressed in the secretory stage but reduced in the early-maturation stage, while CNR5 mRNA is expressed in both these stages. In situ hybridization showed that strong expression of CNR1 is strong in the secretory stage, but reduced in the early phase and diminished in the late phase of the early-maturation stage. CNR5 mRNA is expressed almost at the same levels in the secretory and in the early phase of the early-maturation stages but is absent in the late phase of the early-maturation stage. Both CNR1 and 5 mRNA are continuously expressed in odontoblasts. Immunohistology showed that CNR proteins are expressed in the secretory and early-maturation stages of ameloblasts, but no protein expression at the late-maturation stage was observed. CNR proteins were continuously expressed in odontoblasts. We found that recombinant CNR1 binds dental epithelial and mesenchymal cells through N-terminal domain EC1 in vitro. These results suggest that CNR1 and CNR5 may play an important role in enamel and dentin formation, probably through cell-cell and/or cell-matrix interactions.

In situ acid resistance of in vivo formed white spot lesions.

Thin sections of natural enamel lesions, so-called white spots (WS), and areas of sound enamel (SEn) adjacent to the WS were exposed to an intraoral environment for 2 weeks. Thin sections of WS samples, clamped in a PMMA holder, were microradiographed before and after exposure to intraoral conditions. Acid resistance was evaluated by lesion depth and mineral changes during the cariogenic challenge. The results show that there were statistically significant differences in lesion depth, mineral loss and mineral volume percent at the surface before and after the intraoral cariogenic challenge at least at p<0.05, except for a change in mineral volume percent at the surface of WS samples. This exception indicates that no mineral change occurred in the surface layer of WS. The fact of 2.8 and 1.8 times higher ratios of SEn over WS of mean changes in lesion depth and mineral loss data, respectively, seems to indicate a quantitative difference in acid resistance level of WS lesions compared with the areas of SEn. Regarding the site of mineral changes, a distinctive feature of WS samples is that mineral loss occurs at the bottom of lesions. In contrast, areas of SEn produce a typical subsurface type of lesions. From this in situ study, it can be concluded that the surface of WS samples was apparently much more acid-resistant (at least approximately 2 times) than the areas of SEn that received a similar intraoral acid challenge.

Relationship between mineral distributions in dentine lesions and subsequent remineralization in vitro.

Though the mineral distribution of the dentine carious lesion varies largely from tooth to tooth and from patient to patient, there are two main distribution profiles that characterize natural carious lesions in dentine. These profiles include softened and subsurface lesion types. The mineral distribution relationship between the starting profile and the profile after remineralization is not known. In order to study the relational aspects, we have produced demineralized dentine samples in vitro with mineral profiles similar to those of typical natural carious lesions, and subsequently remineralized the samples in a remineralizing solution with various fluoride concentrations (0, 2 and 10 ppm F). The mineral distributions were obtained by using an improved microradiographic technique. In addition, the nature of deposited mineral was analyzed by diamond-coupled total internal reflectance spectroscopy. Definite relationship was observed between the original lesion mineral distribution and the mineral distributions following remineralization. The amount of mineral present in approximately the first 50 microm of the lesion influenced the overall mineral profile after remineralization, possibly through influencing ion transport. If the amount was high (> approximately 10 vol%), the deposited mineral was confined to the surface (0-50 microm). The original mineral at those depths acted like a nucleus of mineral regrowth when the amount of residual mineral was intermediate, and like a transport barrier when the surface layer was well mineralized. If a surface barrier was not present, mineral was deposited at deeper depths in the lesion. Fluoride effect on dentine remineralization was dependent on the original mineral content and its distribution in the lesion. Although a high concentration of fluoride was very effective in low-mineral lesions, it produced hyperremineralization on well-mineralized subsurface lesions so that it prevented effective remineralization especially in deeper lesions.

Crystallographic analysis of demineralized human enamel treated by laser-irradiation or remineralization.

Crystallographic information on dental hard tissue is helpful in evaluating whether incipient caries that has received preventative treatments is resistant to subsequent attack. The aim here was to analyse crystallographically by means of high-resolution electron microscopy (HREM) demineralized human enamel that had been laser-irradiated or remineralized. Electron-microscopic observation identified a distinct layer at a depth of 100 nm in the demineralized and laser-irradiated enamel. The crystallinity in the shallower area was inferior to that in the deeper area. Comparison of the HREM findings from the deeper area with the data provided by the Joint Committee on Powdered Diffraction Standards revealed that the mineral in the demineralized and laser-irradiated enamel was either alpha- or beta-tricalcium phosphate, while that in the remineralized enamel was thought to be tetracalcium diphosphate monoxide.

The remineralization of EDTA-treated human dentine.

The remineralization of whole human dentine treated with a neutral EDTA solution was investigated: the treatment periods were 15, 30 and 120 min. From the literature it is known that EDTA removes noncollagenous proteins (NCPs) from dentine powders. In order to extract more phosphoproteins from dentine lesions, in part of this work samples were also treated with 1 M NaCl or 4 M guanidine chloride solutions. All the dentine samples after the treatments mentioned were immersed subsequently in a remineralizing solution without fluoride for 2 weeks and microradiographed. To investigate the effects of fluoride, samples treated with EDTA for 120 min were also remineralized with 2 ppm fluoride in solution. The results presented show that: (1) Measurable remineralization did not occur when fluoride was free in the remineralizing solution. Because remineralization occurred neither at the lesion front nor in the nonmineral part of the surface-softened tissue, presumably the NCPs (inhibitors) of underlying dentine diffused into the tissue during the remineralization period. (2) 2 ppm fluoride caused remineralization at the lesion front. In this case we assume fluoride either acted as nucleating agent or overcame the NCP effects at the lesion front.

In vitro remineralization of in vivo and in vitro formed enamel lesions.

Thin sections of natural white spot enamel lesions (WS) and of artificial in vitro lesions (VL) were remineralized simultaneously in vitro. The sections, clamped in a PMMA holder, were microradiographed at baseline and after remineralization in a calcium- and phosphate-containing solution (pH = 7.0; 1 ppm F) after 2 and 4 weeks. All data were analyzed with respect to baseline. The results show that the lesion depth values did not change significantly during 2 and 4 weeks of remineralization. The mineral accumulation (change in DeltaZ), however, was substantial and significant in WS and VL. In WS the change in mineral accumulation was roughly proportional to the amount of mineral at baseline. The WS accumulated more than two times the amount of mineral than VL in the same periods of remineralization. After 4 weeks of remineralization the maximum mineral value Vmax in the surface layer of the WS was nearly up to the sound enamel level thick approximate87 vol%. This study shows that the technique and calculation procedure described make this single section method attractive for longitudinal demineralization-remineralization studies in vitro or in situ. Both WS and VL samples obviously remineralized in vitro similarly with respect to the baseline. Furthermore, this in vitro work indicates that remineralization inhibitors present in saliva, and previously penetrated into the enamel tissue, do not influence remineralization later on.

Influence of nail varnish on the remineralization of enamel single sections assessed by microradiography and confocal laser scanning microscopy.

Single-section techniques are attractive in enamel de- and remineralization investigations because they allow longitudinal studies in which mineral changes can be assessed by microradiography (TMR). Nail varnish (NV) is in general applied to coat the cut thin-section sides. The aims of this study were to investigate: (1) NV penetration depth in cut surfaces of demineralized enamel, (2) the influence of NV on cut surfaces of demineralized enamel on TMR, (3) the influence of NV penetration on a following remineralization. Cut surfaces of thin sections of demineralized enamel were NV coated; the NV was peeled off and the penetration depth assessed by confocal laser scanning microscopy. The NV penetration was 18+/-5 micrometer (mean+/-SD) in demineralized enamel. To evaluate the possible influence of NV on TMR, cut surfaces of thin sections of demineralized enamel were coated (twice) and microradiographed before and after nail varnishing. The NV effect (total effect of penetrated and surface NV) on the main parameters of TMR, Ld and DeltaZ, was less than 5% of the mean values. In the remineralization experiment (remineralization with 1.5 mM Ca2+, 0.9 mM phosphate, pH 7, 1 ppm F for 1 and 2 weeks), lesions in bulk samples, lesions in thin sections with NV-coated cut surfaces and lesions in thin sections in a PMMA (polymethylmethacrylate) holder were compared. (1) The remineralization of bulk samples and of NV-coated thin sections is different in one aspect. The amounts of mineral deposited in the lesions expressed as DeltaZ are comparable after 1 week. But because the NV penetrates part of the lesion outside, there was an Ld difference. The lesion depth difference between bulk lesions and NV-coated lesions in thin sections was statistically significant and was about 19% less in NV-coated lesions after 1 week; after 2 weeks of remineralization there was no difference in Ld between bulk- and NV-coated lesions any more. (2) There was no difference in remineralization efficacy between lesions in bulk samples and lesions in thin sections in the PMMA holder.

Differentiation of clinical isolates of Actinobacillus actinomycetemcomitans using an insertion sequence, ISAa1.

We previously identified an IS200-like sequence (ISAa1) in the genome of Actinobacillus actinomycetemcomitans FDC Y4. One or more hybridizing bands to the ISAa1 probe were detected in each of several reference strains, representing three of the serotypes (a through c) of A. actinomycetemcomitans. In this study, we examined whether a restriction fragment-length polymorphism (RFLP) with ISAa1 as a probe could differentiate clinical isolates. One or more hybridizing bands were detected in each of the 27 strains examined, which could be divided into seven groups according to restriction fragment-length polymorphism pattern. Several strains were observed with identical restriction fragment-length polymorphism types but with different serotypes. Conversely, strains were also observed with differing restriction fragment-length polymorphism types and identical serotypes.

Ageing behaviour in a dental low-gold alloy with high copper content.

Phase transformation behaviour in a dental low-gold alloy with high copper content during continuous heating was investigated by hardness tests, electrical resistivity measurements, X-ray diffraction, scanning and transmission electron microscopies. Two kinds of solution treatment conditions (at 873 K and 1073 K) followed by iced-brine quench, represented different ageing behaviours. Although subsequent anisothermal annealing produced same phase separation of face-centred cubic disordered and ordered (Cu3Au) phases in both specimens, the specimen quenched from 1073 K had already been hardened by a spinodal decomposition.

Cloning and sequencing of part of the S10 operon from Actinobacillus actinomycetemcomitans FDC Y4.

We have cloned and sequenced the 5.2 kb EcoRI fragment that contained part of the S10 operon from Actinobacillus actinomycetemcomitans FDC Y4. The order of the ribosomal protein genes was identical to that of the S10 operon of Haemophilus influenzae and Escherichia coli. The deduced amino acid sequences of ribosomal proteins in this operon displayed significant homologies (65.3%-100%) to those of H. influenzae, E. coli, Yersinia enterocolitica and Yersinia pseudotuberculosis. Phylogenetic trees obtained for these ribosomal proteins were similar to that obtained for 16S rRNA.

A computer-assisted videodensitometric method to visualize mineral distributions in in vitro and in vivo formed root caries lesions.

A computer-assisted videodensitometry (CAV) method is presented for the microradiographical mineral determination and mineral visualization of in vitro and in vivo formed dentin lesions. The method employs as essential steps (1) image digitizing by a CCD camera, (2) conversion of gray values to equivalent aluminium thicknesses and (3) conversion of aluminium thickness to vol% of mineral. Procedure and accuracy are described. The CAV method is illustrated for dentin lesions formed in vitro as well as for clinical root caries lesions. The CAV method is especially advantageous for clinical lesions because it can be employed for very complex mineral distributions and irregular lesion outlines. In given areas of interest, the mineral distribution (in vol%) can be plotted in 3D form in pseudo-colors. Conventional microradiographical profiles can be obtained directly from the CAV images. For in vitro lesions, the mineral distribution parameters, lesion depth, mineral loss, and mineral acquisition values can be generated from CAV images with a reproducibility of better than 7%. The CAV method is a valuable tool in quantitative mineral studies of clinical lesions in extracted teeth as well as in in situ lesion investigations.

Molecular analysis of a new insertion sequence from Actinobacillus (Haemophilus) actinomycetemcomitans FDC Y4.

We have found a new insertion sequence (IS), designated ISAa1, downstream of the S10 operon in Actinobacillus (Haemophilus) actinomycetemcomitans FDC Y4. ISAa1, the first IS element characterized in this organism, is 705 bp long and lacks terminal inverted repeats. This element displayed significant homology with IS200. Hybridization patterns of genomic DNA of seven A. actinomycetemcomitans strains with an internal ISAa1 probe varied depending on the serotypes, suggesting that ISAa1 might be a useful tool for epidemiological studies.

Effect of sodium hypochlorite treatment on remineralization of human root dentine in vitro.

Dentine consists simplified of mineral and of several organic components. Sodium hypochlorite (NaOCl) is a well-known nonspecific proteolytic agent capable to remove organic material. The aim of this study was to investigate the influence of organic material removal from artificial dentine lesions by means of NaOCl pretreatment on subsequent remineralization with and without fluoride. Human root dentine samples were demineralized in an acidic gel (pH = 5) at 37 degrees C for 2 weeks. After 2 min of pretreatment with a 0.4, 2 or 10% NaOCl solution, the samples were remineralized in a 20 mM HEPES buffer (pH = 7) containing 1.5 mM Ca(2) and 0.9 mM phosphate with or without addition of 10 ppm F(-) as NaF at 37 degrees C for 8 days. Mineral profiles were assessed by means of transversal microradiography after diol treatment to avoid shrinkage caused by drying. In a separate experiments the dentine contraction caused by 10% NaOCl was assessed. The contraction (negligible for sound dentine) was found to be about 12% for the lesions. The remineralization results showed that pretreatment with a 10% NaOCl solution for 2 min, increased lesion remineralization. After NaOCl treatment, the amount of accumulated mineral increased by about 27% without F in the remineralization solution, and by about 4% with 10 ppm in solution. The in vitro results suggest that removal of organic materials from dentine lesions is an interesting approach to enhance remineralization.

The effects of a sodium hypochlorite treatment on demineralized root dentin.

The effects of a 10% NaOCl treatment for 2 min on demineralized human root dentin were investigated by means of: microradiography (MR), scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM) and secondary ion mass spectroscopy (SIMS). MR measurements revealed that NaOCl caused a tissue contraction not related to water loss but to removal of organic substance(s), resulting in reductions of the lesion depth and mineral loss values by 15% and 42%, respectively. CLSM observations on wet dentin showed that the dentinal tubules underneath the surface are clearly observable and not deformed substantially by the NaOCl, except near the outermost surface. This indicates the importance of wet as well as of dried (high vacuum) observations. SEM micrographs (high vacuum) showed definite changes in the outer dentin surface structure; 85% of the originally open dentinal tubules were closed after NaOCl treatment. No marked changes were observed in the dentin ultrastructure inside lesions, as shown by SEM on fractured surfaces. SIMS data, pertaining to samples in high vacuum, showed a remarkable increase of chlorine (Cl) content in the entire lesion due to the NaOCl, indicating deep penetration of the original OC1 ions. The results suggest that the 2-min treatment of demineralized dentin by NaOCl solutions removes and/or changes part of the dentin matrix in nearly the whole lesion. As a consequence the mineral is somewhat redistributed, the outermost surface of a few mu m is changed, but the main dentin structure and element composition are still intact. These findings indicate that NaOCl treatments are of interest in remineralization and hyper-remineralization studies of dentin.

The influence of air-drying on hyper-remineralization of demineralized dentine: a study on bulk as well as on thin wet section of bovine dentine.

The influence of air-drying on the remineralization of demineralized bovine dentine was examined in wet bulk samples, in dried bulk samples as well as in wet thin sections. Bulk samples of bovine dentine were first demineralized in an acidic gel (pH = 5) at 37 degrees C for 3 weeks. After 24-hour pre-treatment of either air-drying or immersion in water, the bulk samples were remineralized in a solution containing 1.5 mM Ca, 0.9 mM phosphate and 10 ppm F (pH = 7) at 37 degrees C for 2, 4 or 8 days. Separately thin sections prepared from demineralized bulk dentine were immersed in water for 24 h and were also exposed to the remineralization solution for 2, 4 or 8 days. The results show that air-drying of the bulk samples increased remineralization of dentine considerably; the microradiographic parameters (ld, delta Z and la) show that the degree of remineralization ranks: thin wet sections > bulk dried > bulk wet. Especially, the remineralization inside lesions was greatly enhanced in thin sections and dried samples. It is presumed that the increased remineralization in dried samples is caused by a 'sponge effect', in which the remineralization fluid is sucked up in a dried shrunken lesion, resulting in fast and deep penetration of remineralization solution and/or presumably increased nucleation.

[A case of interstitial pneumonitis associated with polymyositis complicated by renal cell carcinoma].

A 55-year-old woman was referred to the department of urology in our hospital with left renal tumor, discovered during examinations at another hospital for fever and dyspnea on exertion. Because surgery was difficult due to severe hypoxemia, pulmonary function impairment (restrictive) and bilateral diffuse interstitial shadows on chest X-ray film, the patient was referred to our department. Interstitial pneumonitis was found on transbronchial lung biopsy, and serum GOT, LDH and CPK values were elevated. These symptoms and abnormalities of laboratory data were improved by administration of prednisolone 60 mg/day, and left nephrectomy was performed without any complications. Pathological examination of the surgical specimen showed clear cell carcinoma (Grawitz). Steroid therapy was tapered off and her clinical course was good. Six months after surgery, the patient developed a recurrence of fever, which was not responsive to antibiotics. Polymyositis was diagnosed on the basis of elevated serum GOT, LDH and CPK, electromyogram and muscle biopsy findings and positive anti-Jo-1 antibody. Polymyositis/dermatomyositis is sometimes associated with interstitial pneumonitis or malignant neoplasms, but rarely with both simultaneously. Moreover, renal cell carcinoma is very rare among the malignant neoplasms associated with polymyositis/dermatomyositis, and we therefore report this unusual case.

Caries experience in subjects 18-22 years of age after 13 years' discontinued water fluoridation in Okinawa.

The purpose of this study is to compare the state of teeth in young adults who had consumed fluoridated water from birth to 5-8 yr of age with the subjects who had non-fluoridated water. The sample consisted of 117 18-22-yr-old female nursing school students. The data obtained in dental examinations were sorted according to their place of residence, and then divided into groups depending on whether their place of residence had been supplied with fluoridated water or not. The fluoride concentration in the water had been 0.7-1.0 ppm in the six water supply systems which were under the control of the US military base. In the subjects who had fluoridated water, the following differences were found when compared to the control subjects: missing teeth and highly progressed caries occurred less frequently; significantly lower caries prevalence was present on free smooth and approximal surfaces; the overall DMFT differences in favor of the fluoridated subjects were small and non-significant.

[A case of exogenous lipoid pneumonia showing a coin lesion with cavities].

A 26-year-old male was referred to our department because of an abnormal chest X-ray, obtained during a routine physical examination in the hospital admitted after a traffic accident. The P-A chest film revealed a coin lesion with cavities in the left S9. He complained of no symptoms except for expectoration of bloody sputum. Transbronchial lung biopsy specimens showed destruction of normal alveolar pattern and collections of lipid-laden macrophages enclosed by fibrous tissue. The lipid material of macrophages in sputum was stained with Sudan III. He had no difficulty in swallowing and no history of regular use of oily drugs. Fiberoptic bronchoscopy and chest CT showed no obstruction of bronchi. He had been working in a repair shop for motorcycles for years, therefore the lesion could be an occupational exogenous lipoid pneumonia. The diameter of the coin lesion has decreased without any therapy.

[Three cases of pneumothorax due to lung cancer].

Spontaneous pneumothorax due to primary lung cancers are increasingly reported as of lung cancer incidence increased. We experienced one lung cancer case with pneumothorax as a primary symptom and two who developed pneumothorax after chemotherapy, among 238 cases of primary lung cancer. In the literature, 76 cases of pneumothorax due to primary lung cancers are described. The mechanism of pneumothorax is discussed.

[Cognitive changes in decision making process underlying prosocial behavior].

Using a method of monitoring information acquisition, 76 subjects were instructed to simulate the information search process in which they selected a behavior from available behavioral alternatives which were expected to occur in a situation where donating behavior was needed. In order to measure the cognitive changes, they were asked to rate the importance of behavioral attributes both before and after the decision task. After the decision task, they were asked to rate the inner states. (1) Defensive cognitive changes were found which increased the importance of behavioral costs and decreased the importance of personal moral obligation feelings. This pattern of changes was consistent with the Schwartz & Howard model (1981, 1982, 1984). (2) The defensive cognitive changes were related to the information search strategies. This pattern of relationship partly confirmed the prediction derived from the Schwartz & Howard model (1981, 1982, 1984). (3) The result that the cognitive changes were not related to the inner states was inconsistent with the model of either Piliavin, Dovidio, Gaertner, & Clark (1981, 1982) or Schwartz & Howard (1981, 1982, 1984). An alternative model was proposed and discussed.