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Field-Programmable Gate Arrays (FPGAs) play a significant and evolving role in various industries and applications in the current technological landscape. They are widely known for their flexibility, rapid prototyping, reconfigurability, and design development features. FPGA designs are often constructed as compositions of interconnected modules that implement the various features/functionalities required in an application. This work develops a novel tool FEINT, which facilitates this module composition process and automates the design-level modifications required when introducing new modules into an existing design. The proposed methodology is architected as a “template” insertion tool that operates based on a user-provided configuration script to introduce dynamic design features as plugins at different stages of the FPGA design process to facilitate rapid prototyping, composition-based design evolution, and system customization. FEINT can be useful in applications where designers need to tailor system behavior without requiring expert FPGA programming skills or significant manual effort. For example, FEINT can help insert defensive monitoring, adversarial Trojan, and plugin-based functionality enhancement features. FEINT is scalable, future-proof, and cross-platform without a dependence on vendor-specific file formats, thus ensuring compatibility with FPGA families and tool versions and being integrable with commercial tools. To assess FEINT’s effectiveness, our tests covered the injection of various types of templates/modules into FPGA designs. For example, in the Trojan insertion context, our tests consider diverse Trojan behaviors and triggers, including key leakage and denial of service Trojans. We evaluated FEINT’s applicability to complex designs by creating an FPGA design that features a MicroBlaze soft-core processor connected to an AES-accelerator via an AXI-bus interface. FEINT can successfully and efficiently insert various templates into this design at different FPGA design stages.

Study design: Clinical trial for cervical screw insertion by using individualized 3-dimensional (3D) printing screw insertion templates device. Objective: The objective of this study is to evaluate the safety and accuracy of the individualized 3D printing screw insertion template in the cervical spine. Materials and methods: Ten patients who underwent posterior cervical fusion surgery with cervical pedicle screws, laminar screws or lateral mass screws between December 2014 and December 2015 were involved in this study. The patients were examined by CT scan before operation. The individualized 3D printing templates were made with photosensitive resin by a 3D printing system to ensure the screw shafts entered the vertebral body without breaking the pedicle or lamina cortex. The templates were sterilized by a plasma sterilizer and used during the operation. The accuracy and the safety of the templates were evaluated by CT scans at the screw insertion levels after operation. Results: The accuracy of this patient-specific template technique was demonstrated. Only one screw axis greatly deviated from the planned track and breached the cortex of the pedicle because the template was split by rough handling and then we inserted the screws under the fluoroscopy. The remaining screws were inserted in the track as preoperative design and the screw axis deviated by less than 2 mm. Vascular or neurologic complications or injuries did not happen. And no infection, broken nails, fracture of bone structure, or screw pullout occurred. Conclusion: This study verified the safety and the accuracy of the individualized 3D printing screw insertion templates in the cervical spine as a kind of intraoperative screw navigation. This individualized 3D printing screw insertion template was user-friendly, moderate cost, and enabled a radiation-free cervical screw insertion.

Insertion guides are becoming popular for orthodontic mini-implant positioning. The aim of this study was to evaluate and compare the accuracy of two different mini-implant insertion guides, with or without pre-drilling, in a human cadaveric model. Maxillary casts of six fresh frozen specimens were digitized to create insertion guides. Sixty mini-implants were randomly inserted with full-arch or skeletonized guides, either with or without predrilling. Pre- and post-treatment CBCTs were superimposed using rigid registration. Transformation matrices of the planned and real positions were obtained, and distances at the mini-implant neck and apex, as well as the angular deviation, were calculated. The Kruskal–Wallis test was performed, followed by a post hoc test when indicated. Out of 60 inserted mini-implants, 46 could be evaluated. Of these, 10 initially assigned to no pre-drilling required this procedure due to very high bone density. Therefore, 32 implants were inserted with pre-drilling (n = 15 full-arch; n = 17 skeletonized) and 14 without (n = 7 full-arch; n = 7 skeletonized). The lowest mean deviation at the neck was 1.22 ± 0.6 mm, registered in the full-arch/pre-drilling group. The skeletonized/no pre-drilling group presented the lowest mean values at the apex, i.e., 1.72 ± 1.22 mm, as well as the lowest mean angular deviation, i.e., 8.23 ± 4.24°. Significant differences among groups were observed only at the neck, with higher mean deviation in the skeletonized/pre-drilling group than in the full-arch/pre-drilling one (p = 0.014). In conclusion, within the limitations of the study, rather high deviations between planned and real mini-implant positions were found. Further studies are needed on how to improve the accuracy within in vivo settings.

Background Corynebacterium glutamicum is an important industrial workhorse and advanced genetic engineering tools are urgently demanded. Recently, the clustered regularly interspaced short palindromic repeats (CRISPR) and their CRISPR-associated proteins (Cas) have revolutionized the field of genome engineering. The CRISPR/Cas9 system that utilizes NGG as protospacer adjacent motif (PAM) and has good targeting specificity can be developed into a powerful tool for efficient and precise genome editing of C. glutamicum . Results Herein, we developed a versatile CRISPR/Cas9 genome editing toolbox for C. glutamicum . Cas9 and gRNA expression cassettes were reconstituted to combat Cas9 toxicity and facilitate effective termination of gRNA transcription. Co-transformation of Cas9 and gRNA expression plasmids was exploited to overcome high-frequency mutation of cas9 , allowing not only highly efficient gene deletion and insertion with plasmid-borne editing templates (efficiencies up to 60.0 and 62.5%, respectively) but also simple and time-saving operation. Furthermore, CRISPR/Cas9-mediated ssDNA recombineering was developed to precisely introduce small modifications and single-nucleotide changes into the genome of C. glutamicum with efficiencies over 80.0%. Notably, double-locus editing was also achieved in C. glutamicum . This toolbox works well in several C. glutamicum strains including the widely-used strains ATCC 13032 and ATCC 13869. Conclusions In this study, we developed a CRISPR/Cas9 toolbox that could facilitate markerless gene deletion, gene insertion, precise base editing, and double-locus editing in C. glutamicum . The CRISPR/Cas9 toolbox holds promise for accelerating the engineering of C. glutamicum and advancing its application in the production of biochemicals and biofuels.

Background Recent advances in CRISPR technology have enabled us to perform gene knock-in in various species and cell lines. CRISPR-mediated knock-in requires donor DNA which serves as a template for homology-directed repair (HDR). For knock-in of short sequences or base substitutions, ssDNA donors are frequently used among various other forms of HDR donors, such as linear dsDNA. However, partly due to the complexity of long ssDNA preparation, it remains unclear whether ssDNA is the optimal type of HDR donors for insertion of long transgenes such as fluorescent reporters in human cells. Results In this study, we established a nuclease-based simple method for the preparation of long ssDNA with high yield and purity, and comprehensively compared the performance of ssDNA and dsDNA donors with 90 bases of homology arms for endogenous gene tagging with long transgenes in human diploid RPE1 and HCT116 cells. Quantification using flow cytometry revealed lower efficiency of endogenous fluorescent tagging with ssDNA donors than with dsDNA. By analyzing knock-in outcomes using long-read amplicon sequencing and a classification framework, a variety of mis-integration events were detected regardless of the donor type. Importantly, the ratio of precise insertion was lower with ssDNA donors than with dsDNA. Moreover, in off-target integration analyses using donors without homology arms, ssDNA and dsDNA were comparably prone to non-homologous integration. Conclusions These results indicate that ssDNA is not superior to dsDNA as long HDR donors with relatively short homology arms for gene knock-in in human RPE1 and HCT116 cells.

Purpose Implant dentistry is an established therapy option with sufficient long-term success for the replacement of missing teeth. Education in implant dentistry should not only focus on theory but also on practical courses. The purpose of the current examination was to assess the accuracy of fully guided and pilot-drill guided implant insertion applying plotted static guides in a cohort of undergraduate dental students. Methods Matching a three-dimensional set of radiographic data and surface scans of 51 artificial mandibular models, 51 surgical templates were produced by plotting. Metal sleeves allowing either a pilot-drill or fully guided implant insertion were inserted alternatively in region 36 and 46. A total of 102 implants were inserted by 51 undergraduates. Subsequently, the positions of the implants were analyzed radiographically considering the accuracy. Additionally, the time required for implant insertion was recorded and a questionnaire was completed. Statistical analysis followed. Results In general, the accuracy of fully guided implant insertion was higher compared to pilot-drill guided. Mean three-dimensional deviation was 2.24 ± 0.38 degrees for fully guided vs. 4.51 ± 2.20 degrees for pilot-drill guided implant insertion. Time required for fully guided implant insertion was statistically significant higher compared to pilot-drill guided (15:22 ± 5:22 vs. 9:35 ± 3:58 min, p  < 0.01). The returned questionnaires reported a high interest but a self-assessed minor previous knowledge in implant dentistry. Conclusion The examination could show that inexperienced undergraduates benefited from fully guided implant insertion in a laboratory set-up. Based on the questionnaires there is a distinct demand for an extended education in implant dentistry among undergraduate students. Graphical abstract

In order to meet the clinical requirements of spine surgery, this paper proposes the fabrication of the customized template for spine surgery through computer-aided design. A 3D metal printing-selective laser melting (SLM) technique was employed to directly fabricate the 316L stainless steel template, and the metal template with tiny locating holes was used as an auxiliary tool to insert spinal screws inside the patient’s body. To guarantee accurate fabrication of the template for cervical vertebra operation, the contact face was placed upwards to improve the joint quality between the template and the cervical vertebra. The joint surface of the printed template had a roughness of Ra = 13 ± 2 μm. After abrasive blasting, the surface roughness was Ra = 7 ± 0.5 μm. The surgical metal template was bound with the 3D-printed Acrylonitrile Butadiene Styrene (ABS) plastic model. The micro-hardness values determined at the cross-sections of SLM-processed samples varied from HV0.3 250 to HV0.3 280, and the measured tensile strength was in the range of 450 MPa to 560 MPa, which showed that the template had requisite strength. Finally, the metal template was clinically used in the patient’s surgical operation, and the screws were inserted precisely as the result of using the auxiliary template. The geometrical parameters of the template hole (e.g., diameter and wall thickness) were optimized, and measures were taken to optimize the key geometrical units (e.g., hole units) in metal 3D printing. Compared to the traditional technology of screw insertion, the use of the surgical metal template enabled the screws to be inserted more easily and accurately during spinal surgery. However, the design of the high-quality template should fully take into account the clinical demands of surgeons, as well as the advice of the designing engineers and operating technicians.

Spinal surgery demands exceptional theoretical knowledge and practical skills, with pedicle screw procedures posing significant risks due to proximity to critical anatomical structures. This study validates a virtual reality (VR) simulation platform for training in pedicle screw arthrodesis using patient-specific vertebral drilling templates. The practical simulation of a specific case study was evaluated with both neurosurgical residents and medical students. Results demonstrate that while experienced residents completed simulated procedures significantly faster than students ( p  < 0.01), these latter ones showed marked improvement across consecutive training sessions ( p  < 0.001). The most substantial performance gains occurred between the first and second trials, highlighting the rapid learning curve facilitated by the VR environment. System Usability Scale assessments revealed high satisfaction with the simulation platform, with participants emphasizing the value of risk-free repetitive practice. Qualitative feedback from experienced participants confirmed the precision and realism of the training environment as critical factors contributing to their efficiency. This validation confirms that VR simulation platforms offer an effective training environment for complex spinal procedures, providing a safe space for skill development before clinical application, particularly when incorporating innovative surgical aids such as patient-specific drilling templates.

Although customized three-dimensional (3D) templates have shown advantages in brachytherapy, widespread application is still full of challenges. The present work proposed the use of a commercial 3D standardized template-guided intracavitary/interstitial brachytherapy (IC/ISBT) that could provide simple and reproducible needles' insertion. 43 patients received external beam radiotherapy (EBRT) with 45-50.4 Gy and subsequent IC/ISBT with 28 Gy in 4 fractions. In terms of IC/ISBT, 24 patients were treated with 3D standardized templates (ST group), and 19 patients were treated using free-hand implantation (FH group). Consistency of implantation for all needles and dosimetric differences for target and organs at risk (OARs) were then compared between two groups. The mean variation of tip position between insertions for needles was 1.41 mm and 2.74 mm in ST group and FH group, respectively ( < 0.001). ST group was superior in terms of dosimetric conformity index (CI) and D for high-risk clinical target volume (HR-CTV), significantly improving to 23.21% ( < 0.001) and 3.58% ( = 0.031) compared with FH group. The D of the bladder and sigmoid in the ST group were lower than those in the FH group ( < 0.05). Meanwhile, a strong correlation between the volume of HR-CTV and its CI in the ST group ( = 0.865, < 0.001) was found with Spearman's correlation analysis. The implementation of 3D standardized template can potentially improve the precision and consistency in the needle insertion procedure that may replace some customized 3D templates, and achieve clinical satisfied dose distribution in IC/ISBT plans for patients with LACC.

Background Most 3D-printed guiding templates require dissection of soft tissues to match the corresponding surfaces of the guiding templates. This study sought to explore the accuracy and acceptability of the novel 3D printed individualized guiding templates based on cutaneous fiducial markers in minimally invasive screw placement for pelvic fractures. Methods The printed template was tested on five high-fidelity biomimetic phantom models of the bony pelvis and its surrounding soft tissues as well as on two fresh frozen cadavers. Four cutaneous fiducial markers were transfixed on each phantom model prior to performing CT scans to reconstruct their 3D models. Personalized templates for guiding screw insertion were designed based on the positions of the fiducial markers and virtually planned target screw channels after scanning, followed by 3D printing of the guide. Phase 1 consisted of five expert surgeons inserting one anterograde supra-pubic screw and one sacroiliac screw percutaneously into each phantom model using the 3D-printed guide. The deviation of screw positions between the pre-operative planned and post-operative actual ones was measured after registering their 3D modelling. A Likert scale questionnaire was completed by the expert surgeons to assess their satisfaction and acceptability with the guiding template. Phase 2 consisted of repeating the same procedures on the fresh frozen cadavers in order to demonstrate face, content and concurrent validity. Results In Phase 1, all ten screws were successfully implanted with the assistance of the guiding template. Postoperative CT scans confirmed that all screws were safely positioned within the bony pelvic channels without breaching the far cortex. The mean longitudinal deviation at the bony entry point and screw tip between the pre-operative planned and post-operative actual screw paths were 2.83 ± 0.60 mm and 3.12 ± 0.81 mm respectively, with a mean angular deviation of 1.25 ± 0.41°. Results from the Likert questionnaire indicated a high level of satisfaction for using the guiding template among surgeons. In Phase 2, results were similar to those in Phase 1. Conclusions The 3D-printed guiding template based on cutaneous fiducial markers shows potential for assisting in the accurate insertion of percutaneous screws in the pelvis.