TNPO2 variants associate with human developmental delays, neurologic deficits, and dysmorphic features and alter TNPO2 activity in Drosophila.

Transportin-2 (TNPO2) mediates multiple pathways including non-classical nucleocytoplasmic shuttling of >60 cargoes, such as developmental and neuronal proteins. We identified 15 individuals carrying de novo coding variants in TNPO2 who presented with global developmental delay (GDD), dysmorphic features, ophthalmologic abnormalities, and neurological features. To assess the nature of these variants, functional studies were performed in Drosophila. We found that fly dTnpo (orthologous to TNPO2) is expressed in a subset of neurons. dTnpo is critical for neuronal maintenance and function as downregulating dTnpo in mature neurons using RNAi disrupts neuronal activity and survival. Altering the activity and expression of dTnpo using mutant alleles or RNAi causes developmental defects, including eye and wing deformities and lethality. These effects are dosage dependent as more severe phenotypes are associated with stronger dTnpo loss. Interestingly, similar phenotypes are observed with dTnpo upregulation and ectopic expression of TNPO2, showing that loss and gain of Transportin activity causes developmental defects. Further, proband-associated variants can cause more or less severe developmental abnormalities compared to wild-type TNPO2 when ectopically expressed. The impact of the variants tested seems to correlate with their position within the protein. Specifically, those that fall within the RAN binding domain cause more severe toxicity and those in the acidic loop are less toxic. Variants within the cargo binding domain show tissue-dependent effects. In summary, dTnpo is an essential gene in flies during development and in neurons. Further, proband-associated de novo variants within TNPO2 disrupt the function of the encoded protein. Hence, TNPO2 variants are causative for neurodevelopmental abnormalities.

Awareness of dental complications with oral piercings.

OBJECTIVES: To investigate knowledge and awareness of dental complications with oral piercings among adults with oral piercings, a control population with no piercings and body-piercing professionals in South-East Queensland, Australia. METHODS: Participants recruited from public and private dental clinics and body-piercing professionals from piercing centres in the same region completed questionnaires to assess knowledge and attitudes. RESULTS: Respondents comprised 54 pierced individuals, 54 controls and 35 body-piercing professionals. Only 13% of the pierced group were informed about oral piercing-related complications, and 14% of body-piercing professionals did not warn their clients about long-term complications. The latter included chipped teeth (26%) and gingival recession (15%). About 76% play with their piercing, developing teeth grinding (15%), jaw clenching (15%) and sleep disturbances (4%). Interestingly, 41% of those with oral piercings advised others against getting piercings. CONCLUSIONS: There is a lack of awareness about dental-related complications among pierced individuals and body-piercing professionals. A significant proportion of pierced individuals advise others against getting piercings. Body piercers do not consistently advise clients about risks associated with oral piercings. Thus, it is recommended that programmes be developed to improve awareness of oral and dental complications associated with oral piercings.

Optical penetration models for practical prediction of femtosecond laser ablation of dental hard tissue.

OBJECTIVES: To develop and practically test high-precision femtosecond laser ablation models for dental hard tissue that are useful for detailed planning of automated laser dental restorative treatment. METHODS: Analytical models are proposed, derived, and demonstrated for practical calculation of ablation rates, ablation efficiency and ablated morphology of human dental enamel and dentin using femtosecond lasers. The models assume an effective optical attenuation coefficient for the irradiated material. To achieve ablation, it is necessary for the local energy density of the attenuated pulse in the hard tissue to surpass a predefined threshold that signifies the minimum energy density required for material ionization. A 1029 nm, 40 W carbide 275 fs laser was used to ablate sliced adult human teeth and generate the data necessary for testing the models. The volume of material removed, and the shape of the ablated channel were measured using optical profilometry. RESULTS: The models fit with the measured ablation efficiency curve against laser fluence for both enamel and dentin, correctly capturing the fluence for optimum ablation and the volume of ablated material per pulse. The detailed shapes of a 400-micrometer wide channel and a single-pulse width channel are accurately predicted using the superposition of the analytical result for a single pulse. CONCLUSIONS: The findings have value for planning automated dental restorative treatment using femtosecond lasers. The measurements and analysis give estimates of the optical properties of enamel and dentin irradiated with an infrared femtosecond laser at above-threshold fluence and the proposed models give insight into the physics of femtosecond laser processing of dental hard tissue.

Exploring approaches to pulp vitality assessment: A scoping review of nontraditional methods.

INTRODUCTION: Diagnostic procedures for pulp vitality assessment are a crucial aspect of routine dental practice. This review aims to provide a comprehensive overview of nontraditional techniques and methodologies for assessing pulp vitality, specifically exploring promising approaches that are currently not used in dental practice. METHODS: The study protocol was registered a priori (https://osf.io/3m97z/). An extensive electronic search was conducted across multiple databases, including MEDLINE via PubMed, Scopus, Web of Science, and Embase. Inclusion criteria were guided by the research question based on the PCC model as follows: "What are the potential nontraditional techniques (Concept) for assessing pulp vitality (Population) in the field of endodontics or clinical practice (Context)?" Studies were included that explored possible approaches to pulp vitality assessment, utilizing a range of techniques, whilst any studies using traditional pulp tests (cold, heat, and electric stimulation) or well-known methods (pulse oximetry and laser Doppler flowmetry) were excluded. Reviewers independently screened articles and extracted data. A patent search was also performed. RESULTS: Of 3062 studies, 65 were included that described nontraditional approaches for assessing pulp vitality. These included a range of optical diagnostic methods, ultrasound Doppler flowmetry (UDF), magnetic resonance imaging (MRI), terahertz imaging, tooth temperature measurements, as well as invasive methodologies, including 133xenon washout, radioisotope-labelled tracers, hydrogen gas desaturation, intravital microscopy and fluorescent microspheres isotope clearance. The patent search included artificial intelligence and biomarkers methods. CONCLUSIONS: This review provides details for potential innovative tests that may directly describe pulp vitality. Importantly, these methods range from clinically impractical through to promising methods that may transform clinical practice. Several nontraditional techniques have the potential to enhance diagnostic accuracy and could provide valuable insights into the assessment of pulp vitality in challenging clinical scenarios.

Assessment of the onset of analgesia and length of analgesia following the use of PBM with different wavelengths: a clinical study.

This clinical study assessed photobiomodulation (PBM) induced analgesic effects of diode lasers and an LED light source on the dental pulp. Baseline responses to electric pulp testing (EPT) were recorded in 93 healthy unrestored premolar teeth in 26 adults (age range 22-63 years) attending a private dental practice. The teeth were irradiated on buccal and lingual aspects of the crown, by placing the tips on the middle third of the crown of the teeth, on separate days for each of 4 different light sources (660, 808, or 904 nm diode lasers, or a novel multi-wavelength LED light source (700-1100 nm)) using comparable parameters (100 mW, 30 s, 6 J). EPT scores were measured after a further 1-, 2-, 5- and 20-min. Discomfort caused by PBM therapy was recorded using the Wong-Baker scale. EPT changes were tracked over time using repeated measures analysis of variance. Baseline EPT scores were very consistent between different days (linear regression r2 0.9422-0.9648). All PBM devices caused a significant elevation in EPT at 5 min, with an earlier onset at 2 mins for 904 nm and LED. The LED was the only light source that elevated scores at 20 min. Across 2-20 min, when ranked by effectiveness, the greatest EPT elevations were seen for LED, followed by 904 nm, then 660 nm and finally 808 nm. Discomfort during PBM was most common with 904 nm, followed by 808 nm. No discomfort occurred from the LED. Among the light sources utilized, the LED multi-wavelength system demonstrated the largest increase in EPT readings, suggesting its potential as a non-pharmacological alternative for achieving dental analgesia compared to diode lasers.

Laser fluorescence assessment of dental caries arrest with two silver fluoride agents in patients with special needs- a preliminary report.

PURPOSE: While silver diamine fluoride has been used extensively for caries arrest and desensitising, silver fluoride (AgF) at neutral pH may also have value as a minimally invasive dental caries treatment. This study explored the effectiveness of two AgF products (AgF/KI and AgF/SnF2) when used in adult patients with special needs (SN) who had high caries risk and salivary gland hypofunction. METHODS: This split-mouth clinical study, over two appointments 3-months apart, compared the impact of a single application of AgF/KI (Riva Star Aqua, SDI) and AgF/SnF2 (Creighton Dental CSDS, Whiteley) on matched carious lesions in the same arch, by clinical visual-tactile (cVT) assessment of caries status and laser fluorescence (LF, DIAGNOdent) evaluation of bacterial load in the lesions, using repeated measures analysis. RESULTS: Twelve participants were recruited in the study. A total of 56 teeth (28 pairs) were included. Both AgF products gave a significant decrease in caries activity as measured by cVT (P < 0.0001) and LF (P = 0.0027). There were no statistically significant differences between the two AgF treatments, with response rates for improvements in active lesions of 92% in the AgF/KI arm, and 96% in the AgF/SnF2 arm. There was no effect of tooth type, lesion type, arch type, plaque metabolism and plaque area at the site level on outcomes, nor was there a clustering effect of sites in a patient level analysis. Overall, LF was superior to cVT for detecting lesions that still progressed despite treatment (P = 0.0027). CONCLUSION: A single application of AgF/KI or AgF/SnF2 has high predictability (over 90%) for achieving arrest in active caries lesions in adult patients with SN and high caries risk. Clinical assessment should use visual-tactile examination combined with LF readings to detect lesions that are still progressing and that require additional treatments. Future studies should compare these AgF modalities with SDF and explore factors such as time between applications and the need for repeated applications. TRIAL REGISTRATION: The study was registered with the Australian Clinical Trials Registry (ACTRN12621001139864p) on 23/08/2021.

Singapore dentists' attitudes toward dental care provision for older adults with disabilities.

OBJECTIVES: This study aimed to assess dentists' attitudes toward providing dental care for older adults with disabilities in Singapore and the factors influencing their willingness to provide care. BACKGROUND: Dentists face a rapidly ageing patient demographic. Practitioner unwillingness to circumvent the attending challenges of care provision prevents older adults from accessing the dental care necessary for better oral and systemic health. Previous studies have reported on dentists' attitudes toward caring for older adults with disabilities; however, these are mostly limited to dependent older adults and exclude frail, community-dwelling older persons. METHODS: A cross-sectional study was conducted in 2020 using a self-administered questionnaire. Quantitative data analysis was presented in the form of descriptive statistics, followed by bivariate analyses. RESULTS: There were 193 respondents, a response rate of under 9%. Dentists were less willing, confident and involved in the care of older adults with disabilities as the individual's severity of impairments increased. Less than half of respondents were willing to treat individuals with severe disabilities in cooperation (23.3%), swallowing (30.6%), mobility (33.7%) and communication (45.1%). Dentists with training in geriatric dentistry were more willing to provide care. However, younger dentists, general dental practitioners, private practitioners and public sector dentists had specific restraining and driving forces, which further modulated their willingness to provide care. CONCLUSIONS: The current areas of training needs among Singapore dentists are in severe impairments of swallowing, communication, cooperation and mobility. Further geriatric dentistry training may effectively increase dentists' willingness to provide care; however, additional specific targeted interventions are also needed.

Loss-of-Function Variants in PPP1R12A: From Isolated Sex Reversal to Holoprosencephaly Spectrum and Urogenital Malformations.

In two independent ongoing next-generation sequencing projects for individuals with holoprosencephaly and individuals with disorders of sex development, and through international research collaboration, we identified twelve individuals with de novo loss-of-function (LoF) variants in protein phosphatase 1, regulatory subunit 12a (PPP1R12A), an important developmental gene involved in cell migration, adhesion, and morphogenesis. This gene has not been previously reported in association with human disease, and it has intolerance to LoF as illustrated by a very low observed-to-expected ratio of LoF variants in gnomAD. Of the twelve individuals, midline brain malformations were found in five, urogenital anomalies in nine, and a combination of both phenotypes in two. Other congenital anomalies identified included omphalocele, jejunal, and ileal atresia with aberrant mesenteric blood supply, and syndactyly. Six individuals had stop gain variants, five had a deletion or duplication resulting in a frameshift, and one had a canonical splice acceptor site loss. Murine and human in situ hybridization and immunostaining revealed PPP1R12A expression in the prosencephalic neural folds and protein localization in the lower urinary tract at critical periods for forebrain division and urogenital development. Based on these clinical and molecular findings, we propose the association of PPP1R12A pathogenic variants with a congenital malformations syndrome affecting the embryogenesis of the brain and genitourinary systems and including disorders of sex development.

Oral parafunction and bruxism in Rett syndrome and associated factors: An observational study.

OBJECTIVES: To explore patterns of parafunction, and bruxism, and its relationships with genotype and snoring in individuals with Rett syndrome (RTT). METHODS: Retrospective observational data of those with confirmed MECP2 mutations in the InterRett database (n = 216) were used to investigate experience of parafunctional habits, and bruxism and their relationships with genotype and snoring using multivariable linear regression. RESULTS: The prevalence of parafunction was 98.2%. Bruxism was reported (66.2%) with the patterns mostly both diurnal and nocturnal (44.1%) and exclusively diurnal (42.7%). Compared to individuals with C-terminal deletion, individuals with p.Arg106Trp mutations were less likely to have bruxism reported (aOR = 0.15; 95% CI 0.02-0.98, p = 0.05) and those with p.Arg168* mutation were more likely to have frequent bruxism than none or occasional bruxism reported (aROR 3.4; 95% CI 1.1-10.7 p = 0.04). The relative odds of having nocturnal bruxism constantly, compared to none/occasionally, were higher among those 'always' snoring (aROR 6.24; 95% CI 2.1-18.2, p = 0.001) than those with no snoring. CONCLUSIONS: There appeared to be genotypic association with bruxism in p.Arg168* and p.Arg106Trp mutations and association between nocturnal bruxism and frequent snoring in an international sample of individuals with RTT. Clinical significance of the high prevalence of bruxism should be highlighted in relation to difficulty communicating pain and increased dental treatment need in RTT.

The application of lasers in vital pulp therapy: a review of histological effects.

Vital pulp therapy (VPT) is primarily intended to preserve the vitality of pulp tissues, which have been exposed for any reason. Various materials and techniques have been proposed to improve treatment outcomes, including the use of lasers. This study aimed to review the histological results of different dental lasers including low-level lasers, carbon dioxide (CO2), erbium-doped yttrium aluminum garnet laser (Er:YAG), neodymium-doped yttrium aluminum garnet (Nd:YAG), erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) lasers, and diode lasers for VPT. This focused review included a comprehensive electronic search of Scopus, MEDLINE, Web of Science, and Google Scholar databases from 2000 to 2022 by two independent investigators. Different combinations of keywords were used, and reference mining of related papers was done. The review included studies related to histologic evaluation of laser-assisted vital pulp therapy that stated the laser parameters that were used. Articles with radiographic or clinical assessments or articles lacking necessary data were excluded. Non-English articles were excluded unless their abstract was in English and encompassed the necessary data. Most studies indicated the efficacy of lasers for reduction of inflammation, acceleration of healing, and increasing the thickness of dentinal bridge. According to the evidence, lasers used in combination with pulp capping agents are beneficial to enhance the success rate of VPT.

Positive oral health outcomes: A partnership model improves care in a rural Indigenous community.

OBJECTIVE: The purpose of this study was to explore the benefits of a partnership between a university dental school and a Community Controlled Health Service, specifically in relation to improving the oral health of an underserved rural Indigenous community. We sought community opinions on health and social outcomes arising from the service provided by the dental student clinical outplacement. SETTING: In Dalby, Queensland. PARTICIPANTS: In total, 38 participants in five focus groups were representative of local Indigenous community Elders, community health support group members and management and staff. DESIGN: A descriptive qualitative study employing semi-structured audio-recorded focus group discussions conducted with purposefully selected Indigenous community groups to explore participant views and experience of the partnership model. Qualitative data were analysed using thematic content analysis. RESULTS: The Indigenous community representatives expressed positive benefit in both their general and oral health awareness, in improved access to dental care provided in their own safe space, while they were pleased to assist with students' learning. They viewed the partnership as mutually beneficial. Suggestions for enhancement of the oral health service were also offered. CONCLUSION: This partnership between a university and a Community Controlled Health Service provides sustainable positive social and health benefits for the targeted Indigenous community and for the wider local population, while simultaneously providing enhanced educational benefits for students on clinical outplacement. Translation and uptake of this successful model of care would benefit both underserved communities and dental and other health care professional educators worldwide.

Influence of eight debridement techniques on three different titanium surfaces: A laboratory study.

OBJECTIVES: Debridement methods may damage implant surfaces. This in vitro study investigated eight debridement protocols across three implant surfaces to assess both biofilm removal and surface alterations. MATERIAL AND METHODS: One hundred sixty commercially pure titanium discs were treated to simulate commercially available titanium implant surfaces-smooth, abraded and abraded and etched. Following inoculation with whole human saliva to create a mixed species biofilm, the surfaces were treated with eight debridement methods currently used for clinical peri-implantitis (n = 10). This included air abrasion using powders of glycine, sodium bicarbonate and calcium carbonate; conventional mechanical methods-piezoelectric scaler, carbon and stainless steel scalers; and a chemical protocol using 40% citric acid. Following treatment, remaining biofilm was analysed using scanning electron microscopy and crystal violet assays. For statistical analysis, ANOVA was applied (p < 0.05). RESULTS: All debridement techniques resulted in greater than 80% reduction in biofilm compared with baseline, irrespective of the surface type. Glycine powder delivered through an air polishing system eliminated the most biofilm. Mechanical instruments were the least effective at eliminating biofilm across all surfaces and caused the greatest surface alterations. Citric acid was comparable with mechanical debridement instruments in terms of biofilm removal efficacy. Titanium surfaces were least affected by air abrasion protocols and most affected by mechanical methods. CONCLUSIONS: Mechanical protocols for non-surgical debridement should be approached with caution. Glycine powder in an air polisher and 40% citric acid application both gave minimal alterations across all implant surfaces, with glycine the superior method in terms of biofilm removal.

Oral health education and promotion in special needs children: Systematic review and meta-analysis.

OBJECTIVE: To review the effectiveness of oral health education and oral health promotion interventions for children and adolescents with intellectual and developmental disabilities (IDD), in ensuring optimal gingival health, caries experience and oral health-related quality of life, compared to no interventions or alternative interventions. METHODS: A systematic review was conducted to identify published studies from four databases (Medline, PsycINFO, CINAHL and ERIC). Randomised or quasi-randomised controlled trials were included. Two independent reviewers performed risk of bias and qualitative analysis. Meta-analysis was performed as appropriate. RESULTS: Eight treatment comparisons were identified. There was low certainty evidence that fluoride interventions provided long-term reductions in caries in those with IDD; and there was some evidence that chlorhexidine albeit with low certainty provided short-term and long-term reductions in plaque and gingivitis. There was moderate certainty evidence for short-term reductions in dental plaque from the use of modified toothbrushes, but not compelling evidence for powered toothbrushes. CONCLUSIONS: Most studies provided a low quality of evidence, and so any adaptations made to oral health practices of individuals with IDD need to consider their individual needs. PROSPERO registration: CRD42019145784.

A critical analysis of research methods and experimental models to study tooth discolouration from endodontic materials.

As a range of materials used in endodontics may cause tooth discolouration, exploration of factors responsible for the darkening of the tooth crown or root is warranted. This narrative review paper discusses the range of technical factors that apply in laboratory studies that assess endodontic discolouration. As an example of how these factors operate, particular examples relating to discolouration caused by endodontic medicaments containing tetracycline antibiotics are used. Following the PRILE 2021 guideline approach, a summary of key variables to be addressed in the methodology for laboratory studies is presented, to inform future work.

The influence of delivery power losses and full operating parametry on the effectiveness of diode visible-near infra-red (445-1064 nm) laser therapy in dentistry-a multi-centre investigation.

The development of protocols for laser-assisted therapy demands strict compliance with comprehensive operating parametry. The purpose of this investigation was to examine the accuracy of correlation between laser control panel and fibre emission power values in a selection of diode dental lasers. Through retrospective analysis using successive systematic review and meta-analysis, it is clear that there is inconsistency in the details, and possible inaccuracies in laser power applied and associated computed data. Through a multi-centre investigation, 38 semi-conductor ("diode") dental laser units were chosen, with emission wavelengths ranging from 445 to 1064 nm. Each unit had been recently serviced according to manufacturer's recommendations, and delivery fibre assembly checked for patency and correct alignment with the parent laser unit. Subject to the output capacity of each laser, four average power values were chosen using the laser control panel-100 mW, 500 mW, 1.0 W, and 2.0 W. Using a calibrated power meter, the post-fibre emission power value was measured, and a percentage power loss calculated. For each emission, a series of six measurements were made and analysed to investigate sources of power losses along the delivery fibre, and to evaluate the precision of power loss determinations. Statistical analysis of a dataset comprising % deviations from power setting levels was performed using a factorial ANOVA model, and this demonstrated very highly significant differences between devices tested and emission power levels applied (p < 10-142 and < 10-52 respectively). The devices × emission power interaction effect was also markedly significant (p < 10-66), and this confirmed that differences observed in these deviations for each prior power setting parameter were dependent on the device employed for delivery. Power losses were found to be negatively related to power settings applied. Significant differences have emerged to recommend the need to standardize a minimum set of parameters that should form the basis of comparative research into laser-tissue interactions, both in vitro and in vivo.

Simulated and clinical aerosol spread in common periodontal aerosol-generating procedures.

OBJECTIVES: This study evaluated particle spread associated with various common periodontal aerosol-generating procedures (AGPs) in simulated and clinical settings. MATERIALS AND METHODS: A simulation study visualized the aerosols, droplets, and splatter spread with and without high-volume suction (HVS, 325 L/min) during common dental AGPs, namely ultrasonic scaling, air flow prophylaxis, and implant drilling after fluorescein dye was added to the water irrigant as a tracer. Each procedure was repeated 10 times. A complementary clinical study measured the spread of contaminated particles within the dental operatory and quantified airborne protein dispersion following 10 min of ultrasonic supragingival scaling of 19 participants during routine periodontal treatment. RESULTS: The simulation study data showed that air flow produced the highest amount of splatters and the ultrasonic scaler generated the most aerosol and droplet particles at 1.2 m away from the source. The use of HVS effectively reduced 37.5-96% of splatter generation for all three dental AGPs, as well as 82-93% of aerosol and droplet particles at 1.2 m for the ultrasonic scaler and air polisher. In the clinical study, higher protein levels above background levels following ultrasonic supragingival scaling were detected in fewer than 20% of patients, indicating minimal particle spread. CONCLUSIONS: While three common periodontal AGPs produce aerosols and droplet particles up to at least 1.2 m from the source, the use of HVS is of significant benefit. Routine ultrasonic supragingival scaling produced few detectable traces of salivary protein at various sites throughout the 10-min dental operatory. CLINICAL RELEVANCE: The likelihood of aerosol spread to distant sites during common periodontal AGPs is greatly reduced by high-volume suction. Clinically, limited evidence of protein contaminants was found following routine ultrasonic scaling, suggesting that the the majority of the contamination consisits of the irrigant rather than organic matter from the oral cavity.

Effects of decontamination protocols on the aesthetic, morphological, and material composition of preformed paediatric crowns.

BACKGROUND: Preformed crowns (PCs) are exposed to potentially infectious bodily fluids during the trial fit; however, whether they can be safely re-used remains uncertain. This study determined the effects of decontamination methods on various PCs. The key criteria examined were aesthetic, morphological, and elemental characteristics. DESIGN: This is a laboratory-based experimental study. Stainless steel, pre-veneered stainless steel, and zirconia PCs underwent either: (a) ultrasonic cleaning with EmPower® followed by steam sterilisation, (b) thermal disinfection in an instrument washer with Asepti® Neutraliser/Asepti® followed by steam sterilisation, or (c) immersion in an ortho-phthalaldehyde (OPA) instrument-level disinfectant. The methods were performed for 1, 5, or 10 cycles, and scanning electron microscopy (SEM); red, green, and blue (RGB) value; and luminosity value were tracked (using a histogram of individual colour reference chips in Adobe Photoshop® ) to assess colour changes, and energy-dispersive X-ray (EDX) spectroscopy to characterise the material composition. RESULTS: Pre-veneered stainless steel crowns showed isolated zones of darkening with loss of luminosity and variations in green and blue values after ultrasonic or thermal washer disinfection combined with steam autoclaving at 5 and 10 cycles. No changes occurred with OPA. Both SEM imaging and EDX analysis identified no changes to crown materials from any of the three reprocessing methods. CONCLUSIONS: Reprocessing protocols need to reflect the differing responses of various types of PCs. Repeated steam sterilisation causes slight aesthetic alterations by way of colour instability and loss of luminosity to pre-veneered stainless steel crowns, but no issues with stainless steel or zirconia PCs.

Functional Dysregulation of CDC42 Causes Diverse Developmental Phenotypes.

Exome sequencing has markedly enhanced the discovery of genes implicated in Mendelian disorders, particularly for individuals in whom a known clinical entity could not be assigned. This has led to the recognition that phenotypic heterogeneity resulting from allelic mutations occurs more commonly than previously appreciated. Here, we report that missense variants in CDC42, a gene encoding a small GTPase functioning as an intracellular signaling node, underlie a clinically heterogeneous group of phenotypes characterized by variable growth dysregulation, facial dysmorphism, and neurodevelopmental, immunological, and hematological anomalies, including a phenotype resembling Noonan syndrome, a developmental disorder caused by dysregulated RAS signaling. In silico, in vitro, and in vivo analyses demonstrate that mutations variably perturb CDC42 function by altering the switch between the active and inactive states of the GTPase and/or affecting CDC42 interaction with effectors, and differentially disturb cellular and developmental processes. These findings reveal the remarkably variable impact that dominantly acting CDC42 mutations have on cell function and development, creating challenges in syndrome definition, and exemplify the importance of functional profiling for syndrome recognition and delineation.

Disruption of RFX family transcription factors causes autism, attention-deficit/hyperactivity disorder, intellectual disability, and dysregulated behavior.

PURPOSE: We describe a novel neurobehavioral phenotype of autism spectrum disorder (ASD), intellectual disability, and/or attention-deficit/hyperactivity disorder (ADHD) associated with de novo or inherited deleterious variants in members of the RFX family of genes. RFX genes are evolutionarily conserved transcription factors that act as master regulators of central nervous system development and ciliogenesis. METHODS: We assembled a cohort of 38 individuals (from 33 unrelated families) with de novo variants in RFX3, RFX4, and RFX7. We describe their common clinical phenotypes and present bioinformatic analyses of expression patterns and downstream targets of these genes as they relate to other neurodevelopmental risk genes. RESULTS: These individuals share neurobehavioral features including ASD, intellectual disability, and/or ADHD; other frequent features include hypersensitivity to sensory stimuli and sleep problems. RFX3, RFX4, and RFX7 are strongly expressed in developing and adult human brain, and X-box binding motifs as well as RFX ChIP-seq peaks are enriched in the cis-regulatory regions of known ASD risk genes. CONCLUSION: These results establish a likely role of deleterious variation in RFX3, RFX4, and RFX7 in cases of monogenic intellectual disability, ADHD and ASD, and position these genes as potentially critical transcriptional regulators of neurobiological pathways associated with neurodevelopmental disease pathogenesis.

Finite element analysis of rotary nickel-titanium endodontic instruments: A critical review of the methodology.

Finite element analysis has been a valuable research tool for rotary nickel-titanium endodontic files over the last two decades. This review aims to summarise and critique the methodology as used in currently available endodontic literature for finite element analysis of rotary nickel-titanium instruments. An electronic literature research in PubMed and Scopus databases was performed using the appropriate search terms, and the titles and abstracts were screened for relevance. The review revealed an inconsistent approach to the finite element method, particularly with regards to the boundary conditions in which the instruments are tested. Moreover, there is a lack of experimental data to validate in silico findings. A standardised protocol to finite element analysis of rotary endodontic instruments could be considered for future studies.