Splatter generated by oral surgery irrigation and its implication for infection control.

OBJECTIVES: This study aimed to evaluate the splatter contamination generated by rotary instrumentation and irrigation during simulations of surgical extractions. Specifically, comparisons of the splatters generated were made between traditional assistant-based irrigation and self-irrigating drills and between saline and hydrogen peroxide irrigant. MATERIALS AND METHODS: A fluorescein solution was infiltrated into the irrigation system of high-speed drills, and the surgical extraction procedures were performed on manikins with the typodont teeth. Filter papers were placed at the predetermined locations around the operatory to absorb the fluorescein splatters; these samples underwent photographic image analysis. RESULTS: The patient chest showed the largest area of splatters, followed by the assistant's face shield. Procedures using the hydrogen peroxide irrigant generated a larger area of splatter than those using the saline irrigant. There was no difference between the splatters produced by assistant irrigation and self-irrigating drill procedures. CONCLUSIONS: Clinicians should observe and disinfect the locations contaminated by splatters to prevent the spread of infection, since using alternative irrigant or irrigation methods did not reduce the formation of splatters. CLINICAL RELEVANCE: Oral surgery drills with irrigation generate aerosols and splatters, which have potential to spread airborne pathogens. It is important to understand the patterns of splatters to mitigate contamination.

Local application of Usag-1 siRNA can promote tooth regeneration in Runx2-deficient mice.

Runt-related transcription factor 2 (Runx2)-deficient mice can be used to model congenital tooth agenesis in humans. Conversely, uterine sensitization-associated gene-1 (Usag-1)-deficient mice exhibit supernumerary tooth formation. Arrested tooth formation can be restored by crossing both knockout-mouse strains; however, it remains unclear whether topical inhibition of Usag-1 expression can enable the recovery of tooth formation in Runx2-deficient mice. Here, we tested whether inhibiting the topical expression of Usag-1 can reverse arrested tooth formation after Runx2 abrogation. The results showed that local application of Usag-1 Stealth small interfering RNA (siRNA) promoted tooth development following Runx2 siRNA-induced agenesis. Additionally, renal capsule transplantation of siRNA-loaded cationized, gelatin-treated mouse mandibles confirmed that cationized gelatin can serve as an effective drug-delivery system. We then performed renal capsule transplantation of wild-type and Runx2-knockout (KO) mouse mandibles, treated with Usag-1 siRNA, revealing that hindered tooth formation was rescued by Usag-1 knockdown. Furthermore, topically applied Usag-1 siRNA partially rescued arrested tooth development in Runx2-KO mice, demonstrating its potential for regenerating teeth in Runx2-deficient mice. Our findings have implications for developing topical treatments for congenital tooth agenesis.

Characterisation of anterior open bite in primary school-aged children: A preliminary study with artificial neural network analysis.

BACKGROUND: Non-nutritive sucking habits, bottle feeding, and facial hyperdivergency have been suggested as the influencing factors of anterior open bite (AOB). There was inconsistent reporting of prevalence and a gap of knowledge in the literature. AIM: The aim of this study was to investigate the prevalence of AOB in 7- to 12-year-olds, with a special interest to characterise the determinants of this malocclusion. DESIGN: A sample of 203 children aged 7 to 12 years were selected from a primary school in regional Australia. The legal guardian of each child completed a self-administered questionnaire. Data were assessed using methods of univariate statistics and neural analysis. RESULTS: The prevalence of AOB and thumb sucking was 24.1% and 23.2%, respectively. AOB was associated with the habit (correlation = 0.754) and duration (correlation = 0.574) of thumb sucking. Age, gender, birth order, feeding modality, sleep patterns, history of orthodontic treatment and tonsil, adenoid or grommet surgery, and parents' level of education were not related to AOB (correlation absolute value ≤ 0.474). CONCLUSIONS: Thumb sucking, specifically for prolonged duration, increases the risk of development of AOB. Cessation of thumb sucking habits should be encouraged at an early age to avoid the development of AOB.

Awareness and knowledge of oral cancer amongst adult dental patients attending regional university clinics in New South Wales, Australia: a questionnaire-based study.

BACKGROUND: Greater awareness and knowledge of oral cancer has been shown to increase patient presentation for opportunistic screening. This study aimed to identify the level of awareness and knowledge of oral cancer amongst adult patients in regional New South Wales. METHODS: A total of 444 adult dental patients participated in a self-administered questionnaire at one of five regional university dental clinics between 23rd May and 25th July. Data analyses were performed using the chi-square test and binary logistic regression to compare sociodemographic characteristics and the self-declared awareness and knowledge of oral cancer. RESULTS: The study revealed that 73.8% of patients were aware of oral cancer; however, only 28.8% knew that they had been previously screened for oral cancer. Being female (P < 0.001, OR = 2.57), having an excellent level of oral health (P = 0.042, OR = 3.34) and previous attendance at a dental clinic of the regional university (P = 0.014, OR = 2.89) significantly enhanced awareness of oral cancer. CONCLUSION: The findings of this study have broad implications for both clinicians and public health professionals, providing a platform for discussion regarding the sociodemographic factors contributing to reduced knowledge and awareness of oral cancer amongst patients. This study also identified an essential avenue to allow a more targeted approach in future campaigns to increase education.

Molecular Modeling of ALK L1198F and/or G1202R Mutations to Determine Differential Crizotinib Sensitivity.

Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase that has been recognized as a therapeutic target for EML4-ALK fusion-positive nonsmall cell lung cancer (NSCLC) treatment using type I kinase inhibitors such as crizotinib to take over the ATP binding site. According to Shaw's measurements, ALK carrying G1202R mutation shows reduced response to crizotinib (IC50 = 382 nM vs. IC50 = 20 nM for wild-type), whereas L1198F mutant is more responsive (IC50 = 0.4 nM). Interestingly, the double mutant L1198F/G1202R maintains a similar response (IC50 = 31 nM) to the wild-type. Herein we conducted molecular modeling simulations to elucidate the varied crizotinib sensitivities in three mutants carrying L1198F and/or G1202R. Both L1198 and G1202 are near the ATP pocket. Mutation G1202R causes steric hindrance that blocks crizotinib accessibility, which greatly reduces efficacy, whereas mutation L1198F enlarges the binding pocket entrance and hydrophobically interacts with crizotinib to enhance sensitivity. With respect to the double mutant L1198F/G1202R, F1198 indirectly pulls R1202 away from the binding entrance and consequently alleviates the steric obstacle introduced by R1202. These results demonstrated how the mutated residues tune the crizotinib response and may assist kinase inhibitor development especially for ALK G1202R, analogous to the ROS1 G2302R and MET G1163R mutations that are also resistant to crizotinib treatment in NSCLC.

Molecular Modeling for Structural Insights Concerning the Activation Mechanisms of F1174L and R1275Q Mutations on Anaplastic Lymphoma Kinase.

Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase involved in various cancers. In its basal state, the structure of ALK is in an autoinhibitory form stabilized by its A-loop, which runs from the N-lobe to the C-lobe of the kinase. Specifically, the A-loop adopts an inhibitory pose with its proximal A-loop helix (αAL-helix) to anchor the αC-helix orientation in an inactive form in the N-lobe; the distal portion of the A-loop is packed against the C-lobe to block the peptide substrate from binding. Upon phosphorylation of the first A-loop tyrosine (Y1278), the αAL-helix unfolds; the distal A-loop detaches from the C-lobe and reveals the P+1 pocket that accommodates the residues immediately after their phosphorylation, and ALK is activated accordingly. Recently, two neuroblastoma mutants, F1174L and R1275Q, have been determined to cause ALK activation without phosphorylation on Y1278. Notably, F1174 is located on the C-terminus of the αC-helix and away from the A-loop, whereas R1275 sits on the αAL-helix. In this molecular modeling study, we investigated the structural impacts of F1174L and R1275Q that lead to the gain-of-function event. Wild-type ALK and ALK with phosphorylated Y1278 were also modeled for comparison. Our modeling suggests that the replacement of F1174 with a smaller residue, namely leucine, moves the αC-helix and αAL-helix into closer contact and further distorts the distal portion of the A-loop. In wild-type ALK, R1275 assumes the dual role of maintaining the αAL-helix⁻αC-helix interaction in an inactive form and securing αAL-helix conformation through the D1276⁻R1275 interaction. Accordingly, mutating R1275 to a glutamine reorients the αC-helix to an active form and deforms the entire A-loop. In both F1174L and R1275Q mutants, the A-loop rearranges itself to expose the P+1 pocket, and kinase activity resumes.

Loss of Stemness, EMT, and Supernumerary Tooth Formation in Cebpb-/-Runx2+/- Murine Incisors.

Adult Cebpb KO mice incisors present amelogenin-positive epithelium pearls, enamel and dentin allopathic hyperplasia, fewer Sox2-positive cells in labial cervical loop epitheliums, and reduced Sox2 expression in enamel epithelial stem cells. Thus, Cebpb acts upstream of Sox2 to regulate stemness. In this study, Cebpb KO mice demonstrated cementum-like hard tissue in dental pulp, loss of polarity by ameloblasts, enamel matrix in ameloblastic layer, and increased expression of epithelial-mesenchymal transition (EMT) markers in a Cebpb knockdown mouse enamel epithelial stem cell line. Runx2 knockdown in the cell line presented a similar expression pattern. Therefore, the EMT enabled disengaged odontogenic epithelial stem cells to develop supernumerary teeth. Cebpb and Runx2 knockdown in the cell line revealed higher Biglycan and Decorin expression, and Decorin-positive staining in the periapical region, indicating their involvement in supernumerary tooth formation. Cebpb and Runx2 acted synergistically and played an important role in the formation of supernumerary teeth in adult incisors.

Molecular modeling on HIF-2α-ARNT dimer destabilization caused by R171A and/or V192D mutations in HIF-2α.

Oxygen homeostasis in normal and tumor cells is mediated by hypoxia-inducible factors (HIFs), which are active as heterodimer complexes, such as HIF-2α-aryl hydrocarbon receptor nuclear translocator (ARNT) and HIF-1α-ARNT. A series of mutations on the interfaces between HIF-2α and ARNT and on the domain-domain interface within HIF-2α has been reported to exert varying effects on HIF-2α-ARNT dimerization. In the present study, molecular dynamic simulations were conducted to evaluate HIF-2α mutations, namely R171A, V192D, and R171A/V192D, which are not involved in the interaction with ARNT but impede HIF-2α-ARNT dimerization. Our results indicate that these mutations induct local conformation leading to a shortened (by V192D) or widened (by R171A and R171A/V192D) Y91-E346 separation distance, where E346 and Y91 are located on the HIF-2α and interact with ARNT according to electrostatic and geometrical shape complementarity, respectively.

Antagonistic Functions of USAG-1 and RUNX2 during Tooth Development.

Supernumerary teeth and tooth agenesis are common morphological anomalies in humans. We previously obtained evidence that supernumerary maxillary incisors form as a result of the successive development of the rudimentary maxillary incisor tooth germ in Usag-1 null mice. The development of tooth germs is arrested in Runx2 null mice, and such mice also exhibit lingual epithelial buds associated with the upper molars and incisors. The aim of this study is to investigate the potential crosstalk between Usag-1 and Runx2 during tooth development. In the present study, three interesting phenomena were observed in double null Usag-1-/-/Runx2-/- mice: the prevalence of supernumerary teeth was lower than in Usag-1 null mice; tooth development progressed further compared than in Runx2 null mice; and the frequency of molar lingual buds was lower than in Runx2 null mice. Therefore, we suggest that RUNX2 and USAG-1 act in an antagonistic manner. The lingual bud was completely filled with odontogenic epithelial Sox2-positive cells in the Usag-1+/+/Runx2-/- mice, whereas almost no odontogenic epithelial Sox2-positive cells contributed to supernumerary tooth formation in the rudimentary maxillary incisors of the Usag-1-/-/Runx2+/+ mice. Our findings suggest that RUNX2 directly or indirectly prevents the differentiation and/or proliferation of odontogenic epithelial Sox2-positive cells. We hypothesize that RUNX2 inhibits the bone morphogenetic protein (BMP) and/or Wnt signaling pathways regulated by USAG-1, whereas RUNX2 expression is induced by BMP signaling independently of USAG-1.

Environmental and perceived stress in Australian dental undergraduates: Preliminary outcomes.

Background. Dental students have reported a high prevalence of psychological stress and the causes are associated with the challenging dental environmental and demographic factors. This study aimed to conduct a preliminary investigation on dental students' stress status, using a sample of first-to-third-year Bachelor of Dental Surgery students in an Australian university. Special interests included causes of dental environmental stress and access to help services. Methods. A sample of 145 students was surveyed with a modified Dental Environmental Survey and Depression Anxiety Stress Scale in 2014. The participants' demographic information was also collected. Results. The response rate was 95.4%. Second-year (P = 0.042), third-year (P < 0.001) and employed students (P = 0.027) were more likely to report stress resulting from transition to clinical learning. Third-year students were more often stressed about communicating and approaching staff (P = 0.023) as well as different opinions between staff (P < 0.001) and reduced holidays (P < 0.001). Students that were younger than 21 years of age (P = 0.001), that were first years (P < 0.001), and that were not in a relationship (P = 0.010) more often found difficulty of course work stressful. Students who were not in a relationship more often considered learning manual dexterity a source of stress (P = 0.034). Students previously seeking professional help were more likely to be stressed (P = 0.010). Conclusion. Causes of dental environment stress varied among years of study and demographic backgrounds. Professional support to stressed students should be enhanced. Further investigation is indicated.

ANKH Polymorphisms and Clicking of the Temporomandibular Joint in Dental Residents.

AIM: This study aimed to carry out a case-control research study to assess occurrence of clicking of the temporomandibular joint (TMJ) in order to establish the relationship between TMJ clicking and the genotype of "ANKH inorganic pyrophosphate transport regulator" (ANKH) polymorphisms. MATERIALS AND METHOD: A sample of 41 first-year dental residents was selected. Each was examined using standard clinical procedures and genotyping techniques. RESULTS: The participation rate was 91.8 %. The prevalence of TMJ clicking was 51.2 % (95 % CI: 35.7-66.7 %). Occurrence of TMJ clicking was not related to age, gender and genotypes of ANKH-OR as well as ANKH-TR polymorphisms (p ≥ 0.165). CONCLUSION: A similar distribution of ANKH genotypes in TMJ clicking and asymptomatic individuals has been demonstrated by this study. A high percentage of TMJ clicking has been confirmed. Future investigations are indicated.

Prospective signs of cleidocranial dysplasia in Cebpb deficiency.

BACKGROUND: Although runt-related transcription factor 2 (RUNX2) has been considered a determinant of cleidocranial dysplasia (CCD), some CCD patients were free of RUNX2 mutations. CCAAT/enhancer-binding protein beta (Cebpb) is a key factor of Runx2 expression and our previous study has reported two CCD signs including hyperdontia and elongated coronoid process of the mandible in Cebpb deficient mice. Following that, this work aimed to conduct a case-control study of thoracic, zygomatic and masticatory muscular morphology to propose an association between musculoskeletal phenotypes and deficiency of Cebpb, using a sample of Cebpb-/-, Cebpb+/- and Cebpb+/+ adult mice. Somatic skeletons and skulls of mice were inspected with soft x-rays and micro-computed tomography (µCT), respectively. Zygomatic inclination was assessed using methods of coordinate geometry and trigonometric function on anatomic landmarks identified with µCT. Masseter and temporal muscles were collected and weighed. Expression of Cebpb was examined with a reverse transcriptase polymerase chain reaction (RT-PCR) technique. RESULTS: Cebpb-/- mice displayed hypoplastic clavicles, a narrow thoracic cage, and a downward tilted zygomatic arch (p < 0.001). Although Cebpb+/- mice did not show the phenotypes above (p = 0.357), a larger mass percentage of temporal muscles over masseter muscles was seen in Cebpb+/- littermates (p = 0.012). The mRNA expression of Cebpb was detected in the clavicle, the zygoma, the temporal muscle and the masseter muscle, respectively. CONCLUSIONS: Prospective signs of CCD were identified in mice with Cebpb deficiency. These could provide an additional aetiological factor of CCD. Succeeding investigation into interactions among Cebpb, Runx2 and musculoskeletal development is indicated.

Interactions between BMP-7 and USAG-1 (uterine sensitization-associated gene-1) regulate supernumerary organ formations.

Bone morphogenetic proteins (BMPs) are highly conserved signaling molecules that are part of the transforming growth factor (TGF)-beta superfamily, and function in the patterning and morphogenesis of many organs including development of the dentition. The functions of the BMPs are controlled by certain classes of molecules that are recognized as BMP antagonists that inhibit BMP binding to their cognate receptors. In this study we tested the hypothesis that USAG-1 (uterine sensitization-associated gene-1) suppresses deciduous incisors by inhibition of BMP-7 function. We learned that USAG-1 and BMP-7 were expressed within odontogenic epithelium as well as mesenchyme during the late bud and early cap stages of tooth development. USAG-1 is a BMP antagonist, and also modulates Wnt signaling. USAG-1 abrogation rescued apoptotic elimination of odontogenic mesenchymal cells. BMP signaling in the rudimentary maxillary incisor, assessed by expressions of Msx1 and Dlx2 and the phosphorylation of Smad protein, was significantly enhanced. Using explant culture and subsequent subrenal capsule transplantation of E15 USAG-1 mutant maxillary incisor tooth primordia supplemented with BMP-7 demonstrated in USAG-1+/- as well as USAG-1-/- rescue and supernumerary tooth development. Based upon these results, we conclude that USAG-1 functions as an antagonist of BMP-7 in this model system. These results further suggest that the phenotypes of USAG-1 and BMP-7 mutant mice reported provide opportunities for regenerative medicine and dentistry.

Social nicotine dependence in Australian dental undergraduate students.

OBJECTIVES: This study aimed to investigate dental undergraduate students' tobacco usage and social nicotine dependence in Australia. A special interest was to identify the role of factors such as age, gender, year of dental education and cohabitants' smoking status for social nicotine dependence. MATERIALS AND METHODS: A sample of 252, first-to-fifth year undergraduate students in an Australian dental school was used. Each completed a self-administered questionnaire. RESULTS: The smoking rate was 4.8%. Current smokers displayed higher social nicotine dependence than those that had never smoked (t=3.1, df=244, P=0.002). Dental undergraduate students that showed higher social nicotine dependence (P=0.001, OR=1.3, 95% CI: 1.1-1.6), or that had smoking cohabitants (P=0.016, OR=4.8, 95% CI: 1.3-17.0), were more likely to smoke. Students' social nicotine dependence increased with year of dental study (P=0.043, ß=0.4, t=2.0). Social nicotine dependence enhanced tobacco usage among Year-1-to-4 students (P=0.005, OR=1.4, 95% CI: 1.1-1.7) but not Year-5 undergraduates (P=0.432). CONCLUSIONS: Social nicotine dependence has become a developing issue in dental education. Tobacco control should be highlighted in the dental curriculum. Future investigations into the effects of dental education on social nocotine dependence and tobacco usage are indicated.