C-terminus of PIEZO1 governs Ca2+ influx and intracellular ERK1/2 signaling pathway in mechanotransduction.

Cells sense and respond to extracellular mechanical stress through mechanotransduction receptors and ion channels, which regulate cellular behaviors such as cell proliferation and differentiation. Among them, PIEZO1, piezo-type mechanosensitive ion channel component 1, has recently been highlighted as a mechanosensitive ion channel in various cell types including mesenchymal stem cells. We previously reported that PIEZO1 is essential for ERK1/2 phosphorylation and osteoblast differentiation in bone marrow-derived mesenchymal stem cells (BMSCs), induced by hydrostatic pressure loading and treatment with the PIEZO1-specific activator Yoda1. However, the molecular mechanism underlying how PIEZO1 induces mechanotransduction remains unclear. In this study, we investigated that the role of the C-terminus in regulating extracellular Ca2+ influx and activating the ERK1/2 signaling pathway. We observed the activation of Fluo-4 AM in the Yoda1-stimulated human BMSC line UE7T-13, but not in a calcium-depleted cell culture medium. Similarly, Western blotting analysis revealed that Yoda1 treatment induced ERK1/2 phosphorylation, but this induction was not observed in calcium-depleted cell culture medium. To investigate the functional role of the C-terminus of PIEZO1, we generated HEK293 cells stably expressing the full-length mouse PIEZO1 (PIEZO1-FL) and a deletion-type PIEZO1 lacking the C-terminal intracellular region containing the R-Ras-binding domain (PIEZO1-ΔR-Ras). We found that Yoda1 treatment predominantly activated Flou-4 AM and ERK1/2 in PIEZO1-FL-trasfected cells but neither in PIEZO1-ΔR-Ras-transfected cells nor control cells. Our results indicate that the C-terminus of PIEZO1, which contains the R-Ras binding domain, plays an essential role in Ca2+ influx and activation of the ERK1/2 signaling pathway, suggesting that this domain is crucial for the mechanotransduction of osteoblastic differentiation in BMSCs.

Assessment of psychological changes in young children during dental treatment: Analysis of the autonomic nervous activity and electroencephalogram.

BACKGROUND: Understanding the emotional changes in children during dental treatment is necessary for providing stress-free care. AIM: To objectively assess the stress associated with dental treatment in children, based on the autonomic nervous activity and the electroencephalogram (EEG). DESIGN: Twenty-two children aged 4-9 years were recruited from outpatients of a paediatric clinic. Electrocardiogram and EEG were recorded throughout the treatment to analyze the autonomic nervous activities and the powers of brain waves, respectively. Changes in these measurements during each treatment process were evaluated in two age groups: 4-6 years and 6-9 years. RESULTS: Elevations in sympathetic activities accompanied by decreased parasympathetic activities induced by stress were observed during rubber dam setting (RD) in the age group of 4-6 years and during infiltration anesthesia (IA), RD, and cavity preparation with a dental turbine (CP-T) in the age group of 6-9 years. Stress-related beta wave increments in EEG were observed during IA and CP-T in the age group of 6-9 years but not in the age group of 4-6 years. CONCLUSION: Monitoring the autonomic nervous activities during treatments is useful in assessing stresses in a wide age of young children, whereas EEG monitoring is applicable only to children older than 6 years.

Lantibiotics Produced by Oral Inhabitants as a Trigger for Dysbiosis of Human Intestinal Microbiota.

Lantibiotics are a type of bacteriocin produced by Gram-positive bacteria and have a wide spectrum of Gram-positive antimicrobial activity. In this study, we determined that Mutacin I/III and Smb (a dipeptide lantibiotic), which are mainly produced by the widespread cariogenic bacterium Streptococcus mutans, have strong antimicrobial activities against many of the Gram-positive bacteria which constitute the intestinal microbiota. These lantibiotics also demonstrate resistance to acid and temperature. Based on these features, we predicted that lantibiotics may be able to persist into the intestinal tract maintaining a strong antimicrobial activity, affecting the intestinal microbiota. Saliva and fecal samples from 69 subjects were collected to test this hypothesis and the presence of lantibiotics and the composition of the intestinal microbiota were examined. We demonstrate that subjects possessing lantibiotic-producing bacteria in their oral cavity exhibited a tendency of decreased species richness and have significantly reduced abundance of the phylum Firmicutes in their intestinal microbiota. Similar results were obtained in the fecal microbiota of mice fed with S. mutans culture supernatant containing the lantibiotic bacteriocin Mutacin I. These results showed that lantibiotic bacteriocins produced in the oral cavity perturb the intestinal microbiota and suggest that oral bacteria may be one of the causative factors of intestinal microbiota dysbiosis.

Eruption disturbance caused by an odontoma that was partially attached to the crown of the adjacent maxillary first molar, and step lesion excavation for the tooth preservation: A case report.

Despite the partial attachment of a complex odontoma to an impacted tooth, it is possible to guide the tooth to erupt normally and preserve it by step lesion excavation and the prevention of infection.

Intentional Replantation of an Immature Incisor with a Transverse Root Fracture and Endo-Perio Condition: 4 Year Follow-Up.

This case report describes the importance of continual examination for dental trauma and the efficacy of intentional replantation with retrograde root canal filling for a transverse root fracture in an immature incisor accompanied by subsequent periodontal-endodontic disease. In the treatment of traumatically fractured roots in immature incisors, continual examination is indispensable for the final diagnosis because roots in immature teeth are less calcified, resulting in less detailed radiological examinations. Thus, common dental trauma complications such as pulp necrosis may appear months after the initial examination. Endodontic treatment for transverse root fracture is mainly determined according to radiographic examination findings; for fractured immature roots, apexification with calcium hydroxide of the coronal fragment is generally applied. However, this method requires removal of considerable amounts of enamel and dentin to allow access to the cavity for preparation, which may increase the risk of future fractures. In contrast, intentional replantation with retrograde root canal filling does not require the removal or long-term application of calcium hydroxide. However, it requires careful extraction of the tooth, maintenance of root wetness during the extraoral procedure, rigid splinting, and oral hygiene control. Management of tooth mobility is also important in the post-replantation course.