Modular DNA Construct Design for High-Throughput Golden Gate Assembly.

Golden Gate cloning enables the modular assembly of DNA parts into desired synthetic genetic constructs. The "one-pot" nature of Golden Gate reactions makes them particularly amenable to high-throughput automation, facilitating the generation of thousands of constructs in a massively parallel manner. One potential bottleneck in this process is the design of these constructs. There are multiple parameters that must be considered during the design of an assembly process, and the final design should also be checked and verified before implementation. Doing this by hand for large numbers of constructs is neither practical nor feasible and increases the likelihood of introducing potentially costly errors. In this chapter we describe a design workflow that utilizes bespoke computational tools to automate the key phases of the construct design process and perform sequence editing in batches.

Impact of Artificial Intelligence in Endodontics: Precision, Predictions, and Prospects.

Artificial intelligence (AI) has become increasingly prevalent and significant across many industries, including the dental field. AI has shown accuracy and precision in detecting, evaluating, and predicting diseases. It can imitate human intelligence to carry out sophisticated predictions and decision-making in the health-care industry, especially in endodontics. AI models have demonstrated a wide range of applications in the field of endodontics. These include examining the anatomy of the root canal system, predicting the survival of dental pulp stem cells, gauging working lengths, identifying per apical lesions and root fractures, and predicting the outcome of retreatment treatments. Future uses of this technology were discussed in terms of robotic endodontic surgery, drug-drug interactions, patient care, scheduling, and prognostic diagnosis.

Remote Orthotic Fabrication Method Using Small Three-Dimensional Printers and Computed Tomography Data:A Technical Report.

INTRODUCTION: Computer-aided design and manufacturing (CAD/CAM) methods have gained prominence in early orthotic provision.　This study introduces an innovative approach using compact three-dimensional (3D) printers and computed tomography data to generate segmented body models for traditional Damen corset orthoses.　The goals included evaluating the comfort and fit of orthoses and assessing the effectiveness of our approach for prosthetic companies with limited financial resources. MATERIALS AND METHODS: Lumbar and thoracolumbar orthoses were crafted via CAD/CAM.　Four healthy patients wore the orthoses, with whom immediate and 1-week comfort and fit assessments were conducted.　A prosthetist assessed fit, and the Japanese edition of OPUS-CSD, the Orthotics Prosthetics Users' Survey-Client Satisfaction with Device, was used for subjective assessment of comfort and fit. RESULTS: The fit evaluations were satisfactory for all patients with positive predefined criteria.　Questionnaire responses confirmed high satisfaction and comfort, confirming a successful orthosis fit. CONCLUSION: We established an orthotic fabrication method using CAD/CAM methods with 3D printers.　Additionally, we confirmed the comfort and fit of the corsets so produced.　The initial cost of 3D printers is lower than that of traditional carving machines, enabling even small-scale orthotic fabrication facilities to utilize CAD/CAM methods.　Further research and refinements of this manufacturing approach are expected to expand its applicability.

Personalized subperiosteal implant­supported obturator for the rehabilitation of rhino­orbit­cerebral mucormycosis sequela: A case report.

Severe atrophy of the maxilla occasionally renders it impossible to place standard endosseous implants to replace absent teeth. For such cases, personalized subperiosteal implants (PSI) are presented as a treatment alternative. Due to novel design and manufacturing technologies, PSIs are fitted closely to the bone structure of the patient, after defining the anchorage areas where the bone is of higher quality and allowing a passive dental prosthesis to be attached to restore function and aesthetics to the patient. The present case report documents a patient with severe bone defects as a sequela of rhino-orbit-cerebral mucormycosis. After a failed microvascular fibula flap reconstruction, the patient was treated with a removable implant-supported prosthesis attached to a PSI, which provided occlusion with the mandible of the patient and closed the oronasal-antral communication defect. At 18 months after treatment, the patient felt well, with no biological complications and the prosthesis was well adjusted and with good function. Consequently, we consider that in some cases such as this, a customized solution of this type can avoid complex reconstruction treatments.

Is bracket bonding with guided bonding devices accurate enough for crowded dentition?

BACKGROUND: This research aimed to study whether bracket bonding using guided bonding devices (GBDs) is accurate enough for crowded dentitions in vitro. METHODS: Fifteen three-dimensionally (3D) printed resin model sets were included and divided into three groups: mild, moderate, and severe crowding. The resin models were scanned and virtually bonded with brackets. Corresponding GBDs were generated and 3D printed. Subsequently, the brackets were bonded to the resin models on a dental mannequin using the GBDs. The models with bonded brackets were scanned, and comparisons were made between the positions of the actually bonded and the planned ones to evaluate possible deviations. RESULTS: There was no immediate bonding failure in any group. The bonding duration tended to increase with crowding severity (P > 0.05). Almost all linear and angular deviations in all groups were below 0.5 mm and 2°, respectively, and no statistically significant difference was found among the different crowding degrees (P > 0.05). In all groups, the brackets tended to deviate lingually and had buccal crown torque. Brackets in the groups with mild and severe crowding showed a tendency for mesiobuccal rotation. CONCLUSION: GBDs provide high bracket bonding accuracy for dentitions with different crowding degrees and, thus, could hopefully be applied to uncrowded and crowded dentitions alike.

Energy Aware Technology Mapping of Genetic Logic Circuits.

Energy and its dissipation are fundamental to all living systems, including cells. Insufficient abundance of energy carriers─as caused by the additional burden of artificial genetic circuits─shifts a cell's priority to survival, also impairing the functionality of the genetic circuit. Moreover, recent works have shown the importance of energy expenditure in information transmission. Despite living organisms being non-equilibrium systems, non-equilibrium models capable of accounting for energy dissipation and non-equilibrium response curves are not yet employed in genetic design automation (GDA) software. To this end, we introduce Energy Aware Technology Mapping, the automated design of genetic logic circuits with respect to energy efficiency and functionality. The basis for this is an energy aware non-equilibrium steady state model of gene expression, capturing characteristics like energy dissipation─which we link to the entropy production rate─and transcriptional bursting, relevant to eukaryotes as well as prokaryotes. Our evaluation shows that a genetic logic circuit's functional performance and energy efficiency are disjoint optimization goals. For our benchmark, energy efficiency improves by 37.2% on average when comparing to functionally optimized variants. We discover a linear increase in energy expenditure and overall protein expression with the circuit size, where Energy Aware Technology Mapping allows for designing genetic logic circuits with the energetic costs of circuits that are one to two gates smaller. Structural variants improve this further, while results show the Pareto dominance among structures of a single Boolean function. By incorporating energy demand into the design, Energy Aware Technology Mapping enables energy efficiency by design. This extends current GDA tools and complements approaches coping with burden in vivo.

Fracture resistance of CAD/CAM tooth-colored versus cast metal post-and-core restorations in root filled teeth: An in vitro study.

Purpose: This study investigated the fracture resistance and failure modes of custom-fabricated post- and core dental restorations using various CAD/CAM materials. Materials and Methods: Seventy-five mandibular second premolars were allocated to five groups (n = 15) and prepared for standardized post and core restorations. The groups included a control group comprising cast metal and four CAD/CAM materials: Vita Enamic, Shofu HC, Trilor, and PEKK. Fracture resistance was assessed using a compressive force at a crosshead speed of 1 mm/min until failure occurred. Data were analyzed using one-way analysis of variance (ANOVA) and chi-square tests. Results: The metal group had the highest fracture resistance (244.41 ± 75.20 N), with a significant variance compared to that in the CAD/CAM groups (p < 0.001). No significant differences were observed among the non-metallic groups. Conclusions: While several CAD/CAM materials displayed satisfactory flexural properties, cast metal posts showed superior fracture resistance in endodontically treated teeth but were mostly associated with catastrophic failure. The clinical application of CAD/CAM materials for post-core restorations presents a viable alternative to traditional metal posts, potentially reducing the risk of unfavorable fractures.

Impact of Channel Thickness and Doping Concentration for Normally-Off Operation in Sn-Doped ß-Ga2O3 Phototransistors.

We demonstrate a Sn-doped monoclinic gallium oxide (ß-Ga2O3)-based deep ultraviolet (DUV) phototransistor with high area coverage and manufacturing efficiency. The threshold voltage (VT) switches between negative and positive depending on the ß-Ga2O3 channel thickness and doping concentration. Channel depletion and Ga diffusion during manufacturing significantly influence device characteristics, as validated through computer-aided design (TCAD) simulations, which agree with the experimental results. We achieved enhancement-mode (e-mode) operation in <10 nm-thick channels, enabling a zero VG to achieve a low dark current (1.84 pA) in a fully depleted equilibrium. Quantum confinement in thin ß-Ga2O3 layers enhances UV detection (down to 210 nm) by widening the band gap. Compared with bulk materials, dimensionally constrained optical absorption reduces electron-phonon interactions and phonon scattering, leading to faster optical responses. Decreasing ß-Ga2O3 channel thickness reduces VT and VG, enhancing power efficiency, dark current, and the photo-to-dark current ratio under dark and illuminated conditions. These results can guide the fabrication of tailored Ga2O3-based DUV phototransistors.

Reinforcement of 3D-printed resins for denture base by adding aramid fibers: Effect on mechanical, surface, and optical properties.

PURPOSE: The present study evaluated the mechanical, surface, and optical properties of 3D-printed resins for removable prostheses reinforced by the addition of aramid fibers. MATERIALS AND METHODS: According to ISO 20795-1:2013 standards, specimens were printed using a digital light processing 3D printer and divided into two groups (n = 06/group): 3D-printed resin for denture base as the control group, and a group with the same 3D-printed resin in addition of 5% aramid fibers as the experimental group. Red aramid fibers were chosen for aesthetic characterization. The specimens were evaluated for their mechanical properties, such as elastic modulus (GPa), flexural strength (MPa), and superficial properties by their surface microhardness (KHN), surface roughness (µm), and surface free energy (mJ/m2). Optical properties were evaluated by the color difference (∆E00) between groups. The statistical test chosen after the exploratory analysis of the data was One-way ANOVA followed by Tukey's HSD (α = 0.05). RESULTS: The results showed statistical differences in elastic modulus (p < 0.0001), flexural strength (p < 0.0001), surface free energy polar variable (p = 0.0322), total surface free energy (p = 0.0344), with higher values for the experimental. Surface hardness and surface roughness showed no statistical difference (p ≥ 0.05). The color difference (∆E00) obtained through the CIEDE2000 calculus was below the perceptibility threshold (≤1.1). CONCLUSION: Adding aramid fibers to 3D-printed resin for denture bases resulted in better mechanical properties, without major alterations in surface properties. In addition, it is an easy-to-apply choice for mechanical reinforcement and aesthetic characterization, with the expression of small blood vessels in the 3D-printed resin for removable denture bases.

Digital Simulation and Designing of a Novel Osteotomy Guide for Autotransplantation in the Anterior Region.

Autotransplantation of teeth (ATT) is a viable treatment option for replacing teeth lost due to various reasons. Periodontal ligament (PDL) on the donor tooth is one of the most crucial factors determining the success of ATT. To preserve the PDL during surgery and improve the success rate of ATT, digital dentistry has been applied to ATT. In this article, a digital workflow including surgical simulation, a three-dimensional (3D) replica fabrication, and a novel osteotomy guide design is introduced. Digital simulation of ATT on a patient's integrated model enables visualization of anatomical structures and the 3D position of the donor tooth prior to the actual surgery. The 3D-printed osteotomy guide allows the transfer of the direction and depth of the planned osteotomy into the intraoral environment. The 3D replica helps prepare the recipient site before the actual transplantation, which minimizes the extra-alveolar time of the donor tooth and decreases trauma during the try-in process. The proposed virtual planning and the use of the guide and the 3D replica can facilitate the surgical procedures while minimizing complications.

A 48-month clinical performance of hybrid ceramic fragment restorations manufactured in CAD/CAM in non-carious cervical lesions: case report.

From the restorative perspective, various methods are available to prevent the progression of non-carious cervical lesions. Direct, semi-direct, and indirect composite resin techniques and indirect ceramic restorations are commonly recommended. In this context, semi-direct and indirect restoration approaches are increasingly favored, particularly as digital dentistry becomes more prevalent. To illustrate this, we present a case report demonstrating the efficacy of hybrid ceramic fragments fabricated using computer-aided design (CAD)/computer-aided manufacturing (CAM) technology and cemented with resin cement in treating non-carious cervical lesions over a 48-month follow-up period. A 24-year-old male patient sought treatment for aesthetic concerns and dentin hypersensitivity in the cervical region of the lower premolar teeth. Clinical examination confirmed the presence of two non-carious cervical lesions in the buccal region of teeth #44 and #45. The treatment plan involved indirect restoration using CAD/CAM-fabricated hybrid ceramic fragments as a restorative material. After 48 months, the hybrid ceramic material exhibited excellent adaptation and durability provided by the CAD/CAM system. This case underscores the effectiveness of hybrid ceramic fragments in restoring non-carious cervical lesions, highlighting their long-term stability and clinical success.

Influence of impression method and shoulder design on the marginal adaptation of CAD/CAM nanoceramic resin onlay restorations.

Objective: This in-vitro study investigates the influence of two different impression techniques and two shoulder designs on the marginal adaptation of computer-aided design/computer-aided manufacturing restorations. Methods: Forty mandibular first premolars were cast into dental arch models for this in vitro study. Fragile cusps and concavities on the mesial-buccal-occlusal surfaces were treated, with 2 mm of the occlusal surface removed. Teeth were categorised into two groups based on shoulder preparation. Digital scanning using a 3Shape 3D scanner identified them further for allocation into conventional and digital impression subgroups. The restorations were created from nanoceramic resin blocks using prescribed guidelines. Microscopic evaluation assessed the restoration's marginal adaptation, with data analysed using SPSS 27.0. The level of significance was set at p ≤ 0.05. Results: Digital intraoral scanning consistently demonstrated smaller marginal gaps than the traditional impression method, regardless of shoulder preparation, with the differences being statistically significant (p < 0.05). Furthermore, shoulder preparation significantly reduced the marginal gaps in both the digital and traditional impression groups (p < 0.05). Conclusions: The onlay preparation design with a shoulder led to restorations with improved marginal adaptation compared with the design with no shoulder. Direct digital impression techniques produced restorations within a better marginal discrepancy than traditional impressions.

Improved stability of open bite deformities: taking control of the transverse width using digital technology and robotic archwire bending.

Many factors need to be considered when selecting treatment protocol for surgical correction of skeletal open bite deformities. In order to achieve stable long-term results, it is essential to explore the origin of the open bite, including dysfunction of the temporomandibular joint, tongue and compromised nasal breathing, in addition to the skeletal deformity. Recurrence of skeletal open bite is associated with relapse of the expanded transverse width. Three-dimensional virtual planning allows different treatment options to be explored and final decisions to be made together with the orthodontist. This study presents a treatment protocol for predictable and stable widening of the maxillary transverse width over the long term, involving premolar extraction and rounding and shortening of the upper dental arch by advancing the molar segments. The stability of inter-canine, inter-premolar, and inter-molar distances, as well as overjet and overbite, were measured in 16 patients treated with this technique; measurements were obtained pre- and post-surgery, and the mean follow-up was 43 months. Orthodontic treatment was designed digitally and finished with robotically bent wires (SureSmile), which allowed exact planning of the overall treatment, thus making orthognathic surgery more predictable for the patient. The changes in transverse width were significant and stable over time.

Rapid transformation of nanobodies affinity based on AlphaFold2's high-accuracy predictions and interaction analysis for enrofloxacin detection in coastal fish.

High-affinity antibodies are crucial in biosensors, disease diagnostics, therapeutic drug development, and immunological analysis, making the enhancement of antibody affinity a key research focus within the field. Computer-aided design is recognized as a time-saving and labor-efficient method for nanobodies in vitro affinity maturation. Compared to experimental mutagenesis techniques, it is advantageous due to the elimination of the need for laborious library construction and screening processes. However, these approaches are constrained by structural prediction since inaccuracy in structure could readily result in maturation failures. Herein, a novel nanobodies modification method for in vitro affinity maturation, utilizing the high accuracy prediction of AlphaFold2, was employed to rapidly transform a low affinity nanobody against enrofloxacin (ENR) into one with high affinity. The molecular docking results revealed a 1.5- to 2.5-fold increase in the number of noncovalent interactions of modified nanobodies, accompanied by a reduction in binding free energy ranging from 14.1 to 62.6%. The evaluation results from ELISA and BLI indicated that the affinity of the modified nanobodies had been enhanced by 6.2-91.6 times compared to the template nanobody. Furthermore, the modified nanobodies were employed for the detection of ENR-spiked coastal fish samples. In summary, this research proposed a nanobodies modification method from a new perspective, endowing its great application potential in biosensors, food safety, and environmental monitoring.

PSI: Planner-specific, physician-specific, or patient-specific implant for orbital reconstruction?

This study aimed to identify and quantify the variations in PSI designs intended for an identical patient. Records from 10 patients with an orbital fracture involving two walls, for which a primary orbital reconstruction was indicated, were retrospectively included. Clinical engineers from two centers independently generated proposal designs for all patients. Following web meeting(s) with the surgeon from the same institute, the PSI designs were finalized by the engineer. A cross-over of the engineer with the surgeon of the other center created two new design teams. In total, 20 proposal and 40 final PSI designs were produced. A three-dimensional comparison between different PSI designs for the same patient was performed by computing a difference score. Initially, the design proposals of the two engineers showed a median difference score of 37%, which was significantly reduced to a median difference score of 26% for the final designs with different engineers. The median difference score of 22% between surgeons demonstrated that both parties introduced notable user variations to the final designs. Evidence supporting the advantages of an experienced design team was found, with significantly fewer modifications, fewer meetings, and less time required to complete the design (up to 40% time reduction). The findings of the study underline the dependency of PSI design on the surgeon and engineer, and support the need for a more evidence-based protocol for PSI design.

Digital technology implementation in prosthodontics postgraduate programs in Saudi Arabia: a multi-institutional survey of program directors.

OBJECTIVE: To assess the current status of digital technology (DT) implementation in prosthodontics post-graduate (PG) programs in Saudi Arabian dental institutions. METHODOLOGY: A 19-item survey was created using Google forms and the link was emailed to the 29 prosthodontics PG program directors (advanced clinical training and joint program) in Saudi Arabia. The questionnaire contained five Sect. 1) didactic training and DT usage in PG preclinical, clinical and laboratory training, 2) incorporation of DT in PG program, 3) utilization of DT and the types of cases treated by PG students during their course, 4) information on the faculties involved in prosthodontics PG education, and 5) potential challenges encountered and program directors' satisfaction of the PG program. Descriptive statistics was used to present the frequencies and proportions, and Chi square inferential test was used to compare the participants' response based on the type of PG programs (α = 0.05). RESULTS: Only 28 program directors responded to the survey, yielding a 95.5% response rate. Among the program directors, 24 (85.7%) and 4 (14.3%) directors supervised the advanced clinical training and joint programs, respectively. Among the DT, CAD/CAM was the most used technology (50-80%), followed by intra-oral scanning (28-96%). Digital technology implementation was largely limited by lack of resources or equipment (67%), cost (53%) and scarcity of PG teaching staff (46%). A significant difference was observed between the programs regarding the mandatory use of DT for their cases (p = 0.03). CONCLUSIONS: This multi-institutional survey of program directors revealed that DT is continuously implemented in the last few years. Saudi dental institutions must prioritize implementing and utilizing DT in PG training to graduate competent prosthodontists in this fast-paced digital era. While DT is pivotal in dental education, its implementation is limited in many institutions due to resources or equipment, cost, and lack of trained faculty.

Direct versus fully digital indirect bracket bonding: a split-mouth randomized clinical trial on accuracy.

OBJECTIVES: The primary aim is to assess differences in accuracy of orthodontic bracket positioning between fully digital indirect bracket bonding (IDB) and conventional direct bracket bonding (DBB). The secondary aims are to assess differences in bracket bonding failures, bracket repositioning need, clinician experience and patient satisfaction. MATERIALS AND METHODS: This prospective study was designed as a split-mouth randomized clinical trial. In total, 35 patients were analyzed with a six month follow-up period. Translational and orientational deviations from the planned bracket position were determined. Clinician experience and patient satisfaction were evaluated by means of a survey. RESULTS: The difference in translation was 0.34 mm (95% CI: 0.238-0.352, p = 0.017), the difference in orientation was 4.80˚ (95% CI: 3.858-5.727, p < 0.001), both in favour of IDB. IDB showed significantly more immediate (IDB: 3.9%, DBB: 0%) and late (IDB: 5.4%, DBB: 2.5%, p = 0.008) bonding failures. Clinicians and patients experienced a shorter clinical chair time with indirect bonding over direct bonding. CONCLUSIONS: IDB bracket positioning leads to significant smaller translation and orientation deviations from digital IDB planning, than DBB bracket positioning. However, IDB leads to more immediate bonding failures than DBB. The majority of patients preferred IDB over DBB, due to a shorter clinical chair time. CLINICAL RELEVANCE: This study adds to the knowledge of IDB in orthodontics and contributes to evidence on this technique. This evidence is applicable in everyday orthodontics, with respect to patient satisfaction and technical limits of IDB. The trial was registered in the Dutch Trial Register and the International Clinical Trials Registry Platform (ICTRP) of the World Health Organization (WHO), number NL9411.

Establishing complexity targets to enhance artificial reef designs.

Artificial reefs (AR), which are integral tools for fish management, ecological reconciliation and restoration efforts, require non-polluting materials and intricate designs that mimic natural habitats. Despite their three-dimensional complexity, current designs nowadays rely on empirical methods that lack standardised pre-immersion assessment. To improve ecosystem integration, we propose to evaluate 3-dimensional Computer-aided Design (3D CAD) models using a method inspired by functional ecology principles. Based on existing metrics, we assess geometric (C-convexity, P-packing, D-fractal dimension) and informational complexity (R-specific richness, H- diversity, J-evenness). Applying these metrics to different reefs constructed for habitat protection, biomass production and bio-mimicry purposes, we identify potential complexity target points (CTPs). This method provides a framework for improving the effectiveness of artificial reef design by allowing for the adjustment of structural properties. These CTPs represent the first step in enhancing AR designs. We can refine them by evaluating complexity metrics derived from 3D reconstructions of natural habitats to advance bio-mimicry efforts. In situ, post-immersion studies can help make the CTPs more specific for certain species of interest by exploring complexity-diversity or complexity-species distribution relationships at the artificial reef scale.

Successful use of a 3D-printed surgical guide to facilitate transiliosacral fixation in a cat with bilateral sacroiliac luxation.

Case summary: A 7-year-old male castrated domestic shorthair cat was presented for treatment of a bilateral sacroiliac luxation (SIL). CT was performed and the data were extracted in a stereolithography (STL) file, after which a 3D-printed drill guide (3DPDG) was devised, using computer-aided design (CAD) software, and printed. Using an open surgical approach, the guide was used as an aid for drilling the sacrum. The ilial wings were drilled free-hand later and a transiliosacral pin (TP) was inserted to realign and stabilise the SIL. The cat exhibited an early return to normal limb function and a CT scan performed at the postoperative follow-up showed early signs of bone remodelling at the sacroiliac joint. Relevance and novel information: To the authors' knowledge, this is the first report using a 3DPDG for implant placement in the feline sacrum without intraoperative imaging.