Validation study of the prognostic value of cyclin-dependent kinase (CDK)-based risk in Caucasian breast cancer patients.

In a Japanese study, cyclin-dependent kinase (CDK) based risk determined by CDK 1 and 2 activities was associated with risk of distance recurrence in early breast cancer patients. The aim of our study was to validate this risk categorization in European early breast cancer patients. We retrospectively analyzed frozen breast cancer specimens of 352 Dutch patients with histologically confirmed primary invasive early breast cancer. CDK-based risk was determined in tumour tissues by calculating a risk score (RS) according to kinases activity and protein mass concentration assay without the knowledge of outcome. Determination of CDK-based risk was feasible in 184 out of 352 (52%) tumours. Median follow-up of these patients was 15 years. In patients not receiving systemic treatment, the proportions of risk categories were 44% low, 16% intermediate, and 40% high CDK-based risk. These groups remained significant after univariate and multivariate Cox-regression analysis. Factors associated with a shorter distant recurrence-free period were positive lymph nodes, mastectomy with radiotherapy, and high CDK-based risk. There was no significant correlation with overall survival (OS). CDK-based risk is a prognostic marker of distance recurrence of patients with early breast cancer. More validation would be warranted to use of CDK-based risk into clinical practice.

Streptococcus mutans lipoteichoic acid-induced apoptosis in cultured dental pulp cells from human deciduous teeth.

Herein, we suggest that Streptococcus mutans lipoteichoic acid-induced death of dental pulp cells on human deciduous teeth is caused by apoptosis. We provide evidence for the causal role of apoptosis in this process by demonstrating an increase in the proportion of fragmented DNA in such dental pulp cells, which results in a ladder pattern of DNA fragmentation. Additionally, Streptococcus mutans lipoteichoic acid-induced apoptotic cell death is suppressed by caspase inhibitor. Collectively, these findings suggest that Streptococcus mutans lipoteichoic acids may cause apoptosis in human dental pulp cells, and serve as an important factor in pulpitis.

IL-10 inhibits Porphyromonas gingivalis LPS-stimulated human gingival fibroblasts production of IL-6.

Lipopolysaccharides (LPS) of Porphyromonas gingivalis have been implicated in the initiation and development of periodontal diseases. In a previous study, we investigated the signal transduction pathway of P. gingivalis and demonstrated that LPS stimulates the production of interleukin (IL)-6 in human gingival fibroblasts (HGFs), which in turn activates osteoclasts in vitro. The cytokine, IL-10, was initially described as cytokine synthesis inhibitory factor. In this study, we examined that effect of IL-10 on P. gingivalis LPS-induced human gingival fibroblast production of IL-6. LPS-induced IL-6 production was inhibited by IL-10 in a dose-dependent manner. Flow cytometric analysis showed that HGFs bind to fluorescein-isothiocyanate (FITC) labeled IL-10. Western blotting analysis demonstrated the expression of IL-10 receptor on the cell surface of these cells. Engagement of LPS initiated the protein tyrosine phosphorylation of several intracellular proteins including extracellular signal-regulated kinase 2 (ERK2), and these events were suppressed by IL-10. These results suggest that IL-10 inhibits the inflammatory response via the IL-10 receptor in P. gingivalis LPS-initiated periodontal diseases.

Purification and characterization of a trypsin-like protease from the culture supernatant of Actinobacillus actinomycetemcomitans Y4.

We purified and characterized a protease from Actinobacillus actinomycetemcomitans. The protease was isolated from the culture supernatant by sonication in phosphate-buffered 3-[(3 cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). The protease was purified by acetone precipitation, followed by column chromatography with Arginine Sepharose 4B, DEAE Sepharose CL-6B, Sephacryl S-200HR and HiTrap Q. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified protease showed a clear band at approximately 50 kDa. The protease showed trypsin-like activity with hydrolytic activity for the synthetic substrates N alpha-benzoyl-DL-arginine p-nitroanilide (BApNA) and N alpha benzoyl-DL-lysine p-nitroanilide (BLpNA). The activity of the protease was stable at pH 7.0 to approximately 8.0. The activity of the protease was inhibited by leupeptin, phenylmethylsulfonyl fluoride (PMSF), and EDTA, but was not affected by dithiothreitol (DTT), cysteine, 2-mercaptoethanol, pepstatin or soybean trypsin inhibitor. These data suggest that this protease is a serine protease or metallo protease. This enzyme extensively degraded collagen type I and fibronectin.

Induction of apoptosis in human gingival fibroblasts by a Porphyromonas gingivalis protease preparation.

Proteases produced by Porphyromonas gingivalis are believed to contribute to the pathogenesis of periodontal diseases. Here the cytotoxic effects of a purified preparation of a P. gingivalis protease with trypsin-like specificity were tested on human gingival fibroblasts in vitro. The active protease induced apoptotic cell death in the fibroblasts, as indicated by DNA fragmentation and the expression of 7A6 antigen. Thus, the production of proteases by periodontopathic bacteria could be an important factor in the induction of apoptosis of host cells in the aetiology of periodontal diseases.

Salivary amylase activity of rats fed a low calcium diet.

Wistar strain rats fed low calcium diets (1, 2, 3, 4, 6 and 8 weeks) exhibited changes in secretory function of whole saliva. In particular, there were changes in salivary flow rate, total salivary protein, amylase enzyme activity, salivary amylase content and the level of cyclic AMP in the parotid gland acinar cell. Although there were no changes for the first 3 weeks, the levels increased at week 4 and decreased at week 6. The wet weight of the parotid gland started to decrease at week 4. These results suggest that when fed low calcium diets for long periods of time, rats develop defective salivary secretion.

Effects of various collagen crosslinking techniques on mechanical properties of collagen film.

We examined the relationship between collagen crosslinking techniques and dentinum reinforcement. In the present study, the tensile tests, imbibition test and antienzyme test were performed on collagen films mainly composed of type I collagen after ultraviolet irradiation and immersion in tannic acid and glutaraldehyde. The elastic modulus increased with the reaction time, but reduction of elongation at breaking point and increased stiffness were observed. This tendency was enhanced by ultraviolet irradiation. Tannic acid treatment resulted in slower crosslinking than the other two techniques. Antienzyme activity was markedly increased by treatment with tannic acid and glutaraldehyde.

Obturation of dentinal tubules with tannin-fluoride preparation (HY agent) incorporated into glass ionomer cement.

The degree of obturation of dentinal tubules as well as the dentinal uptake of F, Zn and Sr was investigated after placement of glass ionomer cement (GIC) containing variable proportions of the tannin-fluoride preparation, HY agent, into freshly prepared cavities. It was found that when HY agent was incorporated into the cement powder, there was both increase in electrical resistance in the dentinal floor of the GIC-restored cavity in addition to inhibition of dye penetration. Further, these changes were directly proportional to the concentration of HY agent. Electron probe microanalysis of the principal constituents of HY agent (F, Zn and Sr) revealed that the penetration depth for F and Zn was also directly proportional to the relative amount of incorporated HY agent, whereas Sr could not be detected regardless of HY concentration. Results clearly demonstrated that the higher the concentration of HY agent incorporated into GIC, the greater the degree of dentinal tubular obturation.

Clinical observations on the development of third molars.

In order to investigate the developmental conditions of third molars in Japanese, we studied the panoramic radiographs taken during dental treatment at the Pediatric Outpatient Section of Osaka Dental University Hospital of 9,111 children (4,646 boys and 4,465 girls) between the ages of 7 years 0 months and 16 years 11 months, in addition to 2,769 panoramic radiographs of students of this university (2,312 men 457 women) kept by the Department of Oral Radiology. The following results were obtained. 1. Calcification of the third molars in both boys and girls began as early as 7 years 6 months in the maxilla and 7 years 0 months in the mandible. The average age for initiation of calcification in the maxilla was 9 years 4 months for boys, and 9 years 2 months for girls, while the ages in the mandible were 9 years 1 month and 8 years 9 months, respectively. 2. The average age for completion of the third molar crowns in the maxilla was 11 years 8 months for boys and 11 years 5 months for girls, while in the mandible it was 12 years 4 months and 12 years 3 months, respectively. 3. At greater than 13 years of age, the tooth germ could be found in boys about 70% of the time in the maxilla and 75% of the time in the mandible, while these figures for girls were 65 and 80%, respectively. 4. All four third molars were present in 52.3% of the males and 45.5% of the females, while 9.5% of the males and 12.0% of the females had no third molars at all. 5. The direction of eruption was classified as either vertical, mesial, horizontal, distal, or buccal/lingual type. The rate for the vertical type in males was 70% in the maxilla and 45% in the mandible, while these figures for females were 50 and 40%, respectively. 6. Microdontia of the third molars appeared only in the maxilla. 7. Congenital absence of the third molars was more common in females than males, and occurred more frequently in the maxilla than in the mandible. The results of this study allow a better understanding of the third molars of Japanese and provide an aid to planning in such areas of clinical pediatric practice as guiding occlusal development.

Radiographic observations on root resorption in the primary dentition.

In order to construct standardized charts for root resorption in the primary dentition of Japanese children, we investigated the stages of resorption (1/4, 1/2 and 3/4 of the root) based on the criterion of Fanning, et al. using 11,167 panoramic radiographs (5,759 of boys and 5,408 of girls). The following information was obtained. 1) The root resorption for a tooth on one side was the same as for its counterpart on the other side regardless of differences in sex or arch. 2) The average age for each root resorption stage was lower in girls than in boys regardless of differences in sex or arch. 3) The average age for each root resorption stage was lower in the mandible than in the maxilla for every tooth. 4) In the maxilla, the period between 1/4 to 3/4 resorption of the root was greater in the primary molars than in the primary anterior teeth. However, the opposite was true in the mandible. 5) Although no difference was observed between the maxilla and the mandible in the resorption period for the primary incisors and canines, the resorption time for the primary molars was greater in the maxilla than in the mandible. 6) When the relationship was observed between the resorption stages for each tooth and the calcification stages for the corresponding successional permanent tooth. a) With the exception of the maxillary and mandibular canines, the average age for 1/4 resorption of the primary root corresponded to the period between completion of the permanent crown and initial root formation of the successional permanent tooth. b) With the exception of the maxillary and mandibular canines, and the mandibular second premolars, the average age for 3/4 resorption of the primary root corresponded to the period between 1/4 and 1/2 formation of the root of the successional permanent tooth. From these results, standardized charts were constructed for the resorption of the roots of primary teeth in Japanese children. These standardized charts are useful in daily clinical practice in such areas as endodontic treatment of primary teeth and orthodontics.

[Case report of osteogenesis imperfecta].

Osteogenesis imperfecta (OGI) is a rare genetic disease which, as a result of a disorder in the formation of the organic stroma of the bone due to a defect in osteogenic function, induces brittle bones, whereby only weak forces bring about multiple, repeated pathological fractures. This disease is thought to entail various problems with regards to carrying out pediatric dentistry due to the ease with which bones may be fractured. We report here the findings obtained as a result of the careful examination of a 1-year-3-month-old girl encountered in our practice and who was diagnosed as having osteogenesis imperfecta. 1) Out of the three major symptoms for osteogenesis imperfecta, this case showed signs of fragile bones and blue scleras, but did not reveal signs of deafness. 2) There was retardation in system growth and development. 3) Aside from a high level of alkaline phosphatase, there were no notable abnormalities revealed in the biochemical blood tests. 4) Dentinogenesis imperfecta was observed throughout the erupted teeth. 5) There was a definite improvement in cooperation with each visit to the clinic.

Morphological characteristics of the teeth of Newar children.

The morphology of the teeth of Newar children was investigated in order to determine racial characteristics of the teeth in comparison with those of Japanese. The results were as follows. 1) The incidence of a C type labial surface pattern on the maxillary incisors was greater in Newars than Japanese for both the maxillary central and lateral incisors. 2) The total incidence of shovel-shaped and semi-shovel-shaped maxillary incisors was greater than 80% in Japanese, although at most about 25% in Newars. 3) The incidence of interstitial tubercles on the maxillary premolars in Newars was less than in Japanese. 4) The incidence of the cusp of Carabelli on maxillary first molars was greater in Newars than in Japanese. 5) Although the incidence of protostylids on the mandibular first molars was less in Newars than in Japanese, the incidence of the + type occlusal groove pattern was higher in Newars. Even though four-cusp mandibular first molars were more common in Newars than in Japanese, the incidence of a sixth cusp was lower in Newars. Also, the incidence of a sixth cusp was lower in Newars. Also, the incidence of medial of medial lingual accessory tubercles was slightly higher in Newars than in Japanese. Examination of the Newar teeth confirmed previous findings that they are racially Caucasoid, although they have some similarities with Japanese.

Calcification of the permanent first molars observed in panoramic radiographs.

We investigated the development (calcification) stages of the permanent first molars of Japanese children by classifying them into 14 stages by the criterion of Moorrees, et al., using 11,167 panoramic radiographs, 5,759 of boys and 5,408 of girls, between the ages of 2 years 0 months and 14 years 11 months. The following information was obtained from calculations of the average chronological ages for the development stages of these teeth. 1) The development of first molars was earlier in girls than boys. This was particularly true during the middle phase of development. 2) A first molar on one side developed at the same rate as its counterpart on the other side, regardless of differences in sex or arch. 3) In both boys and girls, the development of the mandibular first molars was about one month earlier than that of the maxillary first molars at each stage. 4) When these results were compared with those of Nolla, Moorrees and Haavikko, who pioneered this field of study, it was clear that the development of the teeth in Japanese is different from that in Caucasians. 5) We made charts of the development of the first molars in Japanese which can be applied in routine clinical practice.