VicRK (WalRK or YycFG) is a conserved 2-component regulatory system (TCS) that regulates cell division, cell wall biosynthesis, and homeostasis in low-GC Gram-positive bacteria. VicRK is also associated with biofilm formation of Streptococcus mutans on the tooth surface as it directly regulates the extracellular polysaccharide (EPS) synthesis. Of the 2 components, VicK possesses both autokinase and phosphatase activities, which regulate the phosphorylation and dephosphorylation of the regulator VicR in response to environmental cues. However, the dual mechanism of VicK as the autokinase/phosphatase in regulating S. mutans’ responses is not well elucidated. Previously, it has been shown that the phosphatase activity depends on the PAS domain and residues in the DHp domain of VicK in S. mutans. Specifically, mutating proline at 222 in the PAS domain inhibits VicK phosphatase activity. We generated a VicKP222A mutant to determine the level of VicR-P in the cytoplasm by Phos-tag sodium dodecyl sulfate polyacrylamide gel electrophoresis. We show that in VicKP222A phosphatase, attenuation increased phosphorylated VicR (VicR-P) that downregulated glucosyltransferases, gtfBC, thereby reducing the synthesis of water-insoluble polysaccharides (WIS-EPS) in the biofilm. In addition, VicKP222A presented as long-rod cells, reduced growth, and displayed asymmetrical division. A major adhesin of S. mutans, SpaP was downregulated in VicKP222A, making it unable to agglutinate in saliva. In summary, we have confirmed that VicK phosphatase activity is critical to maintain optimal phosphorylation status of VicR in S. mutans, which is important for cell growth, cell division, EPS synthesis, and bacterial agglutination in saliva. Hence, VicK phosphatase activity may represent a promising target to modulate S. mutans’ pathogenicity.

The nerve growth factor (NGF) and calcitonin gene-related peptide (CGRP) play a crucial role in the regulation of orofacial pain. It has been demonstrated that CGRP increases orofacial pain induced by NGF. V-type proton ATPase subunit an isoform 1 (Atp6v0a1) is involved in the exocytosis pathway, especially in vesicular transport in neurons. The objective was to examine the role of Atp6v0a1 in NGF-induced upregulation of CGRP in orofacial pain induced by experimental tooth movement. Orofacial pain was elicited by ligating closed-coil springs between incisors and molars in Sprague–Dawley rats. Gene and protein expression levels were determined through real-time polymerase chain reaction, immunostaining, and fluorescence in situ hybridization. Lentivirus vectors carrying Atp6v0a1 shRNA were used to knockdown the expression of Atp6v0a1 in TG and SH-SY5Y neurons. The release of vesicles in SH-SY5Y neurons was observed by using fluorescence dye FM1-43, and the release of CGRP was detected by Enzyme-Linked Immunosorbent Assy. Orofacial pain was evaluated through the rat grimace scale. Our results revealed that intraganglionic administration of NGF and Atp6v0a1 shRNA upregulated and downregulated CGRP in trigeminal ganglia (TG) and trigeminal subnucleus caudalis (Vc), respectively, and the orofacial pain was also exacerbated and alleviated, respectively, following administration of NGF and Atp6v0a1 shRNA. Besides, intraganglionic administration of NGF simultaneously caused the downregulation of Atp6v0a1 in TG. Moreover, the release of vesicles and CGRP in SH-SY5Y neurons was interfered by NGF and Atp6v0a1 shRNA. In conclusion, in the orofacial pain induced by experimental tooth movement, NGF induced the upregulation of CGRP in TG and Vc, and this process is dependent on Atp6v0a1 and vesicle release, suggesting that they are involved in the transmission of nociceptive information in orofacial pain.

No abstract

Objectives To develop an artificial intelligence (AI) system for automatic palate segmentation through CBCT, and to determine the personalized available sites for palatal mini implants by measuring palatal bone and soft tissue thickness according to the AI‐predicted results. Materials and Methods Eight thousand four hundred target slices (from 70 CBCT scans) from orthodontic patients were collected, labelled by well‐trained orthodontists and randomly divided into two groups: a training set and a test set. After the deep learning process, we evaluated the performance of our deep learning model with the mean Dice similarity coefficient (DSC), average symmetric surface distance (ASSD), sensitivity (SEN), positive predictive value (PPV) and mean thickness percentage error (MTPE). The pixel traversal method was proposed to measure the thickness of palatal bone and soft tissue, and to predict available sites for palatal orthodontic mini implants. Then, an example of available sites for palatal mini implants from the test set was mapped. Results The average DSC, ASSD, SEN, PPV and MTPE for the segmented palatal bone tissue were 0.831%, 1.122%, 0.876%, 0.815% and 6.70%, while that for the palatal soft tissue were 0.741%, 1.091%, 0.861%, 0.695% and 12.2%, respectively. Besides, an example of available sites for palatal mini implants was mapped according to predefined criteria. Conclusions Our AI system showed high accuracy for palatal segmentation and thickness measurement, which is helpful for the determination of available sites and the design of a surgical guide for palatal orthodontic mini implants.

In the process of orthodontic treatment, root parallelism is related to the relapse of extraction spaces, black triangles, and periodontal health. However, there are few studies on root parallelism in extraction cases with different types of appliances. The objective was to compare the root parallelism in extraction cases treated with clear aligners vs. those treated with fixed appliances by measuring the differences per extraction site and to access the root-movement control capacity of both orthodontic appliances. A retrospective study was conducted on cases in which the first premolars were extracted, treated by clear aligner appliance (“Invisalign” system, 28 patients) or fixed appliance (30 patients). The angulations of the tooth axis (canines, the second premolars, the first molars, and the second molars) were measured to analyze and compare the difference in root parallelism between the two orthodontic appliances. The percentage of root parallelism between the canine and second premolar in both groups had no significant difference. However, judging from the average angle difference value, the fixed appliance might have a greater range in root angulation adjustment than the aligner. In the angulation categories, the aligner mainly showed root apical divergence, while the fixed appliance mainly showed root apical convergence. The dental crown of the molars was mainly non-tipping in both groups, but it could be observed that the dental crown of the maxillary molars had a tendency toward mesial inclination. In conclusion, the same root parallelism and root inclination were obtained in tooth extraction cases whether treated by clear aligners or fixed appliances. This study provides clinicians with more information on the performance of clear aligners and fixed appliances.