Sequencing vs. amplification for the estimation of allele dosages in sugarcane (Saccharum spp.).

Premise: Detecting single-nucleotide polymorphisms (SNPs) in a cost-effective way is fundamental in any plant breeding pipeline. Here, we compare three genotyping techniques for their ability to reproduce the allele dosage of SNPs of interest in sugarcane (Saccharum spp.). Methods: To identify a reproducible technique to estimate allele dosage for the validation of SNP markers, the correlation between Flex-Seq, kompetitive allele-specific PCR (KASP), and genotyping-by-sequencing and restriction site-associated DNA sequencing (GBS+RADseq) was determined for a set of 76 SNPs. To find alternative methodologies for allele dosage estimation, the KASP and Flex-Seq techniques were compared for the same set of SNPs. For the three techniques, a population of 53 genotypes from the diverse sugarcane panel of the Centro de Investigación de la Caña de Azúcar (Cenicaña), Colombia, was selected. Results: The average Pearson correlation coefficients between GBS+RADseq and Flex-Seq, GBS+RADseq and KASP, and Flex-Seq and KASP were 0.62 ± 0.27, 0.38 ± 0.27, and 0.38 ± 0.30, respectively. Discussion: Flex-Seq reproduced the allele dosages determined using GBS+RADseq with good levels of precision because of its depth of sequencing and ability to target specific positions in the genome. Additionally, Flex-Seq outperformed KASP by allowing the conversion of a higher number of SNPs and a more accurate estimation of the allele dosage. Flex-Seq has therefore become the genotyping methodology of choice for marker validation at Cenicaña.

Premisa: Detectar polimorfismos de un único nucleótido (SNP) de forma costo­efectiva es fundamental en cualquier programa de mejoramiento genético. En este artículo nosotros comparamos tres técnicas de genotipado para medir su habilidad en reproducir las dosis alélicas de SNPs de interés en caña de azúcar (Saccharum spp.). Métodos: Para identificar una técnica reproducible para la estimación de dosis alélicas durante los pasos de validación de marcadores, la correlación entre Flex­Seq, kompetitive allele­specific PCR (KASP), y genotyping­by­sequencing and restriction site­associated DNA sequencing (GBS+RADseq) fue determinada para un set de 76 SNPs. Para identificar metodologías alternativas en la estimación de las dosis alélicas, las tecnologías KASP y Flex­Seq fueron comparadas para el mismo grupo de SNPs. Para las tres técnicas, una población de 53 genotipos fue seleccionados de la población diversa de caña de azúcar del Centro de Investigación de la Caña de Azúcar (Cenicaña), Colombia. Resultados: El promedio del coeficiente de correlación de Pearson entre GBS+RADseq y Flex­Seq, GBS+RADseq y KASP, y Flex­Seq y KASP fue de 0.62 ± 0.27, 0.38 ± 0.27, y 0.38 ± 0.30, respectivamente. Discusión: Flex­Seq reprodujo las dosis alélicas determinadas usando GBS+RADseq con buenos niveles de precisión debido a su profundidad de secuenciación y habilidad de secuenciar posiciones especificas en el genoma. Adicionalmente, Flex­Seq superó a KASP al permitir la conversión de un número mayor de SNPs y al estimar las dosis alélicas de forma más precisa. Flex­Seq por tanto se convierte en la metodología de genotipado de elección para la validación de marcadores en Cenicaña.

To Compare and Evaluate Rotary and Manual Techniques in Biomechanical Preparation of Primary Molars to Know Their Effects in Terms of Cleaning and Shaping Efficacy.

Background: The introduction of the rotary file system for children was a revolution in the field of pediatric endodontics. These files are cost-effective and help in consistent obturations with shorter instrumentation time. Aim: The present randomized controlled trial (RCT) was planned for a comparative evaluation of rotary and manual techniques in biomechanical preparation of primary molars to determine their effect in terms of cleaning and shaping efficacy, working time, quality of obturation, and postoperative pain. Materials and methods: A randomized clinical trial study was conducted in 75 children aged 5-9 years requiring pulpectomy. Each tooth was randomly assigned to one of the three treatment groups: Kedo-S files, rotary K-Flex files, and hand instruments group. Results: It was observed that Kedo-S files and rotary K-Flex files were more effective in cleaning and shaping of root canals compared to hand H/K files. The postbacterial count for hand files was higher compared to rotary files. Shorter working time was seen with rotary files (3.88-5.04 minutes) compared to hand files (15.68 minutes). Rotary files showed a reduced number of voids, with Kedo-S files in 92% of cases and rotary K-Flex files in 80% of cases. Apical seal and extent of fill were maximum with rotary files, having a grade C rating in 92% of cases. Kedo-S files and rotary K-Flex files showed a significant reduction in postoperative pain compared to hand files. Conclusion: The present study showed a significant reduction in bacterial count, working time, quality of obturation, and postoperative pain with rotary files. How to cite this article: Moudgalya MS, Tyagi P, Tiwari S, et al. To Compare and Evaluate Rotary and Manual Techniques in Biomechanical Preparation of Primary Molars to Know Their Effects in Terms of Cleaning and Shaping Efficacy. Int J Clin Pediatr Dent 2024;17(8):864-870.

In Silico Screening, Molecular Dynamics Simulation and Binding Free Energy Identify Single-Point Mutations That Destabilize p53 and Reduce Binding to DNA.

Considering p53's pivotal role as a tumor suppressor protein, proactive identification and characterization of potentially harmful p53 mutations are crucial before they appear in the population. To address this, four computational prediction tools-SIFT, Polyphen-2, PhD-SNP, and MutPred2-utilizing sequence-based and machine-learning algorithms, were employed to identify potentially deleterious p53 nsSNPs (nonsynonymous single nucleotide polymorphisms) that may impact p53 structure, dynamics, and binding with DNA. These computational methods identified three variants, namely, C141Y, C238S, and L265P, as detrimental to p53 stability. Furthermore, molecular dynamics (MD) simulations revealed that all three variants exhibited heightened structural flexibility compared to the native protein, especially the C141Y and L265P mutations. Consequently, due to the altered structure of mutant p53, the DNA-binding affinity of all three variants decreased by approximately 1.8 to 9.7 times compared to wild-type p53 binding with DNA (14 µM). Notably, the L265P mutation exhibited an approximately ten-fold greater reduction in binding affinity. Consequently, the presence of the L265P mutation in p53 could pose a substantial risk to humans. Given that p53 regulates abnormal tumor growth, this research carries significant implications for surveillance efforts and the development of anticancer therapies.

Comparison of the Amount of Debris Extruded From the Apex During Retreatment Procedures Using Several File Systems and to Gauge the Effect of Same and Different Taper on the Debris Extrusion: An In Vitro Study.

Background: To determine the amount of apical extrusion between various file systems during the retreatment procedure and to determine the effect of taper on the same. Materials and Method: Seventy mandibular extracted premolars were taken, and preparation was performed using a Woodpecker file (Woodpecker Medical Instrument Co., Ltd., Guilin, China) (25/0.06). The gutta-percha (GP) was removed from each specimen with H file (#40/0.02) to simulate retreatment in seven groups (n = 10) using ProTaper Next (PTN;Dentsply, Maillefer, Ballaigues, Switzerland) (PTN) (30/0.07), Reciproc (25/0.08), Hyflex CM (30/0.06), Mtwo (30/0.05), Neoendo flex (30/0.04), Neoendo hybrid (30/0.04), and Flexicon file (30/0.06). The specimens were held with the help of a rubber stopper and put in the Eppendorf tube of volume 4 mL (pre-weighed before retreatment). The debris that was on the root surface of the specimen was washed with distilled water with 1 mL volume, kept in the oven, and then again weighed after 7 days. Results: Apical extrusion was found in the following order: HYFLEX CM (LEAST) < FLEXICON < PROTAPER NEXT < RECIPROC < NEOENDO FLEX < NEOENDO HYBRID < MTWO (HIGHEST). Conclusion: W Hyflex CM and Flexicon files showed lesser extrusion of debris than others.

5' Transgenes drive leaky expression of 3' transgenes in Cre-inducible bi-cistronic vectors.

Molecular cloning techniques enabling contemporaneous expression of two or more protein-coding sequences provide an invaluable tool for understanding the molecular regulation of cellular functions. The Cre-lox system is used for inducing the expression of recombinant proteins encoded within a bi-/poly-cistronic cassette. However, leak expression of transgenes is often observed in the absence of Cre recombinase activity, compromising the utility of this approach. To investigate the mechanism of leak expression, we generated Cre-inducible bi-cistronic vectors to monitor the expression of transgenes positioned either 5' or 3' of a 2A peptide or internal ribosomal entry site (IRES) sequence. Cells transfected with these bi-cistronic vectors exhibited Cre-independent leak expression specifically of transgenes positioned 3' of the 2A peptide or IRES sequence. Similarly, AAV-FLEX vectors encoding bi-cistronic cassettes or fusion proteins revealed the selective Cre-independent leak expression of transgenes positioned at the 3' end of the open reading frame. Our data demonstrate that 5' transgenes confer promoter-like activity that drives the expression of 3' transgenes. An additional lox-STOP-lox cassette between the 2A sequence and 3' transgene dramatically decreased Cre-independent transgene expression. Our findings highlight the need for appropriate experimental controls when using Cre-inducible bi-/poly-cistronic constructs and inform improved design of vectors for more tightly regulated inducible transgene expression.

Long term surgical results and safety profile of the novel CM T Flex scleral fixated intraocular lens.

PURPOSE: To report the long term visual outcomes and complications with use of the novel CM-T Flex scleral fixated intraocular lens (CMT-SFIOL). MATERIALS AND METHODS: 116 eyes that underwent CMT-SFIOL were reviewed and 57 eyes with CMT-SFIOL that completed a 2-year follow-up were included. Main outcome measures noted were best-corrected visual acuity (BCVA) and complications. Postoperatively, follow-ups were done at 1 week (1w), 1 month (1 m), 1 year (1y) and 2-year (2y) intervals. RESULTS: 40 (70.17%) of 57 eyes received CMT-SFIOL for surgical aphakia. Mean follow up was 39.77 ± 8.44 months. BCVA for distance & near improved from 1.26 ± 0.84 to 0.76 ± 0.77, 0.50 ± 0.72 and 0.51 ± 0.73 & 1.28 ± 0.58 to 0.98 ± 0.49, 0.92 ± 0.44 and 0.89 ± 0.40 at 1 m, 1y and 2y respectively (p < 0.001 for all). At 1w, 10 eyes (17.54%) had corneal edema (CE) and 8 eyes (14.03%) had anterior chamber (AC) reaction. Two eyes (3.50%) had IOP > 30 mm Hg and 1 eye (1.75%) had vitreous hemorrhage (VH). At 1 m, 3 eyes (5.26%) had CE and 5 eyes (8.77%) had AC reaction. Two eyes (3.50%) had IOP > 30 mm Hg and 1 eye (1.75%) had VH. Cystoid macular edema was noted in 3 eyes (5.26%). At 1y and 2y, 2 eyes (3.5%) and 1 eye (1.75%), had CE. No decentration, dislocation, haptic exposure or retinal detachment was noted. No eyes required resurgery. CONCLUSION: CM-T Flex SFIOL is an effective method to correct aphakia, with reliable and safe long-term results.

Characterization of a commercial plastic scintillator for electron FLASH dosimetry.

PURPOSE: This study investigated the potential of a commercially available plastic scintillator, the Exradin W2, as a real-time dosimeter for ultra-high-dose-rate (UHDR) electron beams. This work aimed to characterize this system's performance under UHDR conditions and addressed limitations inherent to other conventional dosimetry systems. METHODS AND MATERIALS: We assessed the W2's performance as a UHDR electron dosimeter using a 16 MeV UHDR electron beam from the FLASH research extension (FLEX) system. Additionally, the vendor provided a beta firmware upgrade to better handle the processing of the high signal generated in the UHDR environment. We evaluated the W2 regarding dose-per-pulse, pulse repetition rate, charge versus distance, and pulse linearity. Absorbed dose measurements were compared against those from a plane-parallel ionization chamber, optically stimulated luminescent dosimeters and radiochromic film. RESULTS: We observed that the 1 × 1 mm W2 scintillator with the MAX SD was more suitable for UHDR dosimetry compared to the 1 × 3 mm W2 scintillator, capable of matching film measurements within 2% accuracy for dose-per-pulse up to 3.6 Gy/pulse. The W2 accurately ascertained the inverse square relationship regarding charge versus virtual source distance with R2 of ∼1.00 for all channels. Pulse linearity was accurately measured with the W2, demonstrating a proportional response to the delivered pulse number. There was no discernible impact on the measured charge of the W2 when switching between the available repetition rates of the FLEX system (18-180 pulses/s), solidifying consistent beam output across pulse frequencies. CONCLUSIONS: This study tested a commercial plastic scintillator detector in a UHDR electron beam, paving the way for its potential use as a real-time, patient-specific dosimetry tool for future FLASH radiotherapy treatments. Further research is warranted to test and improve the signal processing of the W2 dosimetry system to accurately measure in UHDR environments using exceedingly high dose-per-pulse and pulse numbers.

Investigation of four nickel titanium endodontic instruments, with cyclic fatigue resistance, scanning electron microscopy, and energy dispersive x-ray spectroscopy.

The aim of this study was to compare of four different nickel-titanium (Ni-Ti) endodontic files and evaluate in terms of cyclic fatigue resistance and metallurgical properties. Four different type Ni-Ti root canal files Protaper Next X2 (PTN) (Dentsply Maillefer, Ballaigues, Switzerland), One Curve (OC) #25.06 (Micro Mega, Besancon, France), EndoPlus Flex Plus Gold X2 (EPG) (Turkuaz Dental, Denizli, Turkey), and EndoPlus Flex Plus Blue #25.06 (EPB) (Turkuaz Dental, Denizli, Turkey) files were tested for cyclic fatigue resistance (n = 20). During experiments artificial zirconia block canal was used. The artificial canal designed with curvature 60° and 5-mm radius. The number of cyclic to fracture (NCF) was noted. Fractured length (FL) parts of Ni-Ti files were recorded to assessment of fracture volumetric point. All fractured surfaces of Ni-Ti files were assessed by scanning electron microscope (SEM) to confirm the type of fractures. Descriptive evaluation become accomplished for the fundamental composition of units with the aid of using energy-dispersive x-ray spectroscopy (EDX). NCF data were evaluated via Bonferroni test with post hoc multiple comparison method. OC showed the highest NCF values (p < .05). The standardization of the study was confirmed as the FL of files was statistically similar in length (p > .05). SEM analysis confirmed that all scanned samples were fractured due to cyclic fatigue. EDX analysis confirmed that EPB established the poorest Ni content file. RESEARCH HIGHLIGHTS: The cyclic fatigue-related failure of One Curve was significantly more resistant than Protaper Next and EndoPlus files. Scanning electron microscopy images showed that One Curve and Protaper Next have round tips Energy dispersive x-ray spectroscopy showed that all four endodontic instruments mainly have Nickel and Titanium elements.

Dosimetric Evaluation of Semiflex Three-dimensional Chamber under Unflatten Beam in Comparison among Different Detectors.

Purpose: The goal of this study is to investigate the dosimetric properties of a Semiflex three-dimensional (3D) chamber in an unflatten beam and compare its data from a small to a large field flattening filter-free (FFF) beam with different radiation detectors. Methods: The sensitivity, linearity, reproducibility, dose rate dependency, and energy dependence of a Semiflex 3D detector in flattening filter and filter-free beam were fully investigated. The minimum radiation observed field widths for all detectors were calculated using lateral electronic charged particle equilibrium to investigate dosimetric characteristics such as percentage depth doses (PDDs), profiles, and output factors (OPFs) for Semiflex 3D detector under 6FFF Beam. The Semiflex 3D measured data were compared to that of other detectors employed in this study. Results: The ion chamber has a dosage linearity deviation of +1.2% for <10 MU, a dose-rate dependency deviation of +0.5%, and significantly poorer sensitivity due to its small volume. There is a difference in field sizes between manufacturer specs and derived field sizes. The measured PDD, profiles, and OPFs of the Semiflex 3D chamber were within 1% of each other for all square field sizes set under linac for the 6FFF beam. Conclusion: It was discovered to be an appropriate detector for relative dose measurements for 6 FFF beams with higher dose rates for field sizes more than or equal to 3 cm × 3 cm.

A Versatile Approach for Adaptive Grid Mapping and Grid Flex-Graph Exploration with a Field-Programmable Gate Array-Based Robot Using Hardware Schemes.

Robotic exploration in dynamic and complex environments requires advanced adaptive mapping strategies to ensure accurate representation of the environments. This paper introduces an innovative grid flex-graph exploration (GFGE) algorithm designed for single-robot mapping. This hardware-scheme-based algorithm leverages a combination of quad-grid and graph structures to enhance the efficiency of both local and global mapping implemented on a field-programmable gate array (FPGA). This novel research work involved using sensor fusion to analyze a robot's behavior and flexibility in the presence of static and dynamic objects. A behavior-based grid construction algorithm was proposed for the construction of a quad-grid that represents the occupancy of frontier cells. The selection of the next exploration target in a graph-like structure was proposed using partial reconfiguration-based frontier-graph exploration approaches. The complete exploration method handles the data when updating the local map to optimize the redundant exploration of previously explored nodes. Together, the exploration handles the quadtree-like structure efficiently under dynamic and uncertain conditions with a parallel processing architecture. Integrating several algorithms into indoor robotics was a complex process, and a Xilinx-based partial reconfiguration approach was used to prevent computing difficulties when running many algorithms simultaneously. These algorithms were developed, simulated, and synthesized using the Verilog hardware description language on Zynq SoC. Experiments were carried out utilizing a robot based on a field-programmable gate array (FPGA), and the resource utilization and power consumption of the device were analyzed.

Retrieval of Embolized WATCHMAN® Flex Atrial Appendage Occlusion Device.

This case report documents the management of a 66-year old man with atrial fibrillation with recent placement of a WATCHMAN® Flex atrial appendage occlusion device. The patient presented with renal failure, abdominal pain, and difficulty walking 2 months after placement. The WATCHMAN® Flex device was found to have embolized to his abdominal aorta at the level of the renal arteries with associated thrombus. Extensive workup revealed reduced left ventricular cardiac function and decreased renal function, both of which were felt to be potentially reversible with device removal. The patient then underwent retrieval of the device and all associated thrombus via an open retroperitoneal approach. This case demonstrates a potential consequence of implanting devices such as an atrial appendage occlusion device and describes a technique for removal.

Role of Flex-Dose Delivery Program in Patients Affected by HCC: Advantages in Management of Tare in Our Experience.

Background: Introduced in the latest BCLC 2022, endovascular trans-arterial radioembolization (TARE) has an important role in the treatment of unresectable hepatocellular carcinoma (HCC) as a "bridge" or "downstaging" of disease. The evolution of TARE technology allows a more flexible and personalized target treatment, based on the anatomy and vascular characteristics of each HCC. The flex-dose delivery program is part of this perspective, which allows us to adjust the dose and its radio-embolizing power in relation to the size and type of cancer and to split the therapeutic dose of Y90 in different injections (split-bolus). Methods: From January 2020 to January 2022, we enrolled 19 patients affected by unresectable HCC and candidates for TARE treatment. Thirteen patients completed the treatment following the flex-dose delivery program. Response to treatment was assessed using the mRECIST criteria with CT performed 6 and 9 months after treatment. Two patients did not complete the radiological follow-up and were not included in this retrospective study. The final cohort of this study counts eleven patients. Results: According to mRECIST criteria, six months of follow-up were reported: five cases of complete response (CR, 45.4% of cases), four cases of partial response (PR, 36.4%), and two cases of progression disease (PD, 18.2%). Nine months follow-up reported five cases of complete response (CR, 45.4%), two cases of partial response (PR, 18.2%), and four cases of progression disease (PD, 36.4%). No intra and post-operative complications were described. The average absorbed doses to the hepatic lesion and to the healthy liver tissue were 319 Gy (range 133-447 Gy) and 9.5 Gy (range 2-19 Gy), respectively. Conclusions: The flex-dose delivery program represents a therapeutic protocol capable of "saving" portions of healthy liver parenchyma by designing a "custom-made" treatment for the patient.

The development and testing of a smart sensorized guide wire for catheterization in a "blood" vessel phantom to support aortic valve implementation.

PURPOSE: Heart valve disease is commonly treated by minimally invasive procedures with guide wires and catheterization. The main purpose of this study is to find out whether an extension of the guide wire with a sensor can support the surgeon within the blood vessel to reduce X-ray necessity. METHODS: A smart guide wire is developed by an extension with a flex-bending sensor to evaluate the sensor signal with and without "blood" flow at a constant compression force. Various surgically relevant investigations are performed. For assessment, the mean temporal average of the moving averaged filtered ADC signal and a subsequent FFT are carried out. RESULTS: Results show that there is a smaller sensor signal when the applied force or bending at the sensor is higher. In all investigations, there was a different sensor signal. The flex-bending sensor can detect the effect of pulsatile flow. The smallest temporal averaged signal difference between reference and clamp in the front wire's tip is 1.09%. For example, the mean temporal average of the filtered ADC signal for different clinically relevant scenarios is between 2550 and 2900. CONCLUSIONS: The results show that the sensorized guide wire developed for catheterization can support aortic valve implementation. The sensor sensitivity is sufficient to detect even very small variations within the blood vessel and therefore is promising to support catheterization heart valve surgeries in future.

Fast Prediction Method for Scattering Parameters of Rigid-Flex PCBs Based on ANN.

InGaAs detection systems have been increasingly used in the aerospace field, and due to the high signal-to-noise ratio requirements of short-wave infrared quantitative payloads, there is an urgent need for methods for the rapid and precise evaluation and the optimal design of these systems. The rigid-flex printed circuit board (PCB) is a vital component of InGaAs detectors, as its grid ground plane design parameters impact parasitic capacitance and thus affect weak infrared analog signals. To address the time-intensive and costly nature of design optimization achieved with simulations and experimental measurements, we propose an innovative method based on a neural network to predict the scattering parameters of rigid-flex boards for InGaAs detection links. This is the first study in which the effects of rigid-flex boards on weak infrared detection signals have been considered. We first obtained sufficient samples with software simulation. A backpropagation (BP) neural network prediction model was trained on existing sample sets and then verified on a rigid-flex board used in a crucial aerospace short-wave infrared quantitative mission. The model efficiently and accurately predicted high-speed interconnect scattering parameters under various rigid-flex board grid plane parameter conditions. The prediction error was less than 1% compared with a 3D field solver, indicating an overcoming of the iterative optimization inefficiency and showing improved design quality for InGaAs detection circuits.

Comparison between prefabricated ankle-foot orthoses, Dyna Ankle and UD Flex, in patients with hemiplegia.

OBJECTIVE: To compare the kinematic effects of two widely-used prefabricated ankle-foot orthoses (AFOs), the Dyna Ankle (DA) and UD Flex (UD), on the gait cycle of patients with hemiplegia due to cerebral palsy or acquired brain injury. METHODS: This was a retrospective cohort study involving 29 patients. Gait analysis results were assessed under three conditions: barefoot, with the DA, and with the UD. Friedman tests and post hoc analysis with Bonferroni correction were performed to assess differences between the three conditions. RESULTS: The DA significantly improved ankle dorsiflexion during the mid-swing phase, making it more effective in correcting foot drop compared with the UD (DA: 2.28°, UD: 0.44°). Conversely, the UD was more effective in preventing knee flexion during the loading response (DA: 28.11°, UD: 26.72°). CONCLUSIONS: The DA improved ankle dorsiflexion during the swing phase significantly more than that with the UD in patients with hemiplegia. Compared with the DA, the UD more effectively prevented increased knee flexion during the loading response. The choice to prescribe these orthoses should consider individual patient characteristics.

Comparative evaluation of Borrelia burgdorferi antibody detection between the VetScan Flex4 and SNAP 4Dx Plus.

Two studies were developed to compare Borrelia burgdorferi antibody detection between the VetScan Flex4 and SNAP 4Dx Plus tests. The objective of the first study was to evaluate the diagnostic sensitivity (Se) and specificity (Sp) of VetScan Flex4 and SNAP 4Dx Plus B. burgdorferi results using field sourced samples compared to a Western Blot reference method. The sensitivity and specificity of VetScan Flex4 were 81.9 % (95 % CI: 71.9 %-89.5 %) and 89.3 % (95 % CI: 85.2 %-92.9 %) respectively, and SNAP 4Dx Plus's sensitivity and specificity were 80.7 % (95 % CI: 70.6 %-88.6 %) and 92.8 % (95 % CI: 89.1 %-95.5 %) respectively. When comparing VetScan Flex4 and Snap 4Dx Plus, the Simple Kappa Coefficient estimate was 0.76 (95 % CI: 0.69-0.84) indicating substantial agreement between the two methods. McNemar's Test revealed concordance between the two methods was not statistically significant (P = 0.05). The objective of the second study was to evaluate whether VetScan Flex4 differentiates between B. burgdorferi antibodies derived from infection versus vaccination with commonly used canine Lyme vaccines. The sensitivity and specificity of the VetScan Flex4 in differentiating canine Lyme vaccination from infection with Borrelia burgdorferi were 100 % (Se 95 % CI: 78.2 %-100 %; Sp 95 % CI: 91.2 %-100 %). In conclusion, the VetScan Flex4 is a reliably sensitive and specific point-of-care test that is similar to Snap 4Dx Plus, can differentiate between infection and Lyme vaccination, and can be utilized by veterinarians for Lyme disease diagnosis and surveillance of B. burgdorferi exposure.

Initial Experience and Clinical Assessment of the Karl Storz FLEX-XC1 Single-Use Flexible Ureteroscope.

INTRODUCTION: The Karl Storz FLEX-XC1 is a novel single-use flexible ureteroscope that uses the same videographics platform as its reusable digital counterpart. We evaluated the technical performance of the FLEX-XC1 in its initial clinical use. METHODS: We reviewed a series of consecutive ureteroscopy procedures performed by 2 endourologists using the FLEX-XC1 for indications for which we typically use a single-use device: total stone burden > 15 mm or > 10 mm in the lower pole, anticipated case duration > 60 minutes, bilateral procedure, or upper tract urothelial cancer procedures. We assessed device tip deflection, intraoperative mechanical failure, and clinical outcomes for each case. Surgeons rated visual clarity, image quality, and maneuverability on a 1 to 5 Likert scale. RESULTS: Of 29 procedures using FLEX-XC1, 27 (93%) were successfully completed. Preoperative upward deflection was < 270° in 6 (21%) cases, and downward deflection was < 270° in 9 (31%) cases. Three types of intraoperative malfunctions occurred: rotational twisting of deflectable tip (4 cases, 13%), device not advancing through distal ureter (1 case, 3%), and working channel not accommodating a 365-µm laser (1 case, 3%). Visual clarity, image quality, and maneuverability were rated as 5 "very good" or 4 "good" in 100%, 100%, and 97% of cases, respectively. No device-specific or general 30-day complications were observed. CONCLUSIONS: The FLEX-XC1 showed comparable image quality and maneuverability to reusable digital devices. We observed incomplete deflection in up to 31% of cases and mechanical failure in 2 cases. The FLEX-XC1 may be advantageous in prolonged cases where maintaining visual clarity is paramount.

Validation and Implementation of OptiView and EnVision FLEX Detection Systems for Immunocytochemical Staining Protocols of the Ten Most Commonly Used Diagnostic Markers in Routine Cytopathological Practice.

The withdrawal of the iView detection system (iV) forced many cytopathology laboratories, including ours, to substitute immunocytochemical (ICC) staining protocols for routine practice with other detection systems. Our objective was to optimize, validate, and implement ICC protocols using OptiView (OV) and EnVision FLEX (EnV) detection systems, comparing the results with those obtained using iV. Residual cytologic samples with known diagnoses were used, testing antibodies for the ten most common markers in routine cytopathology diagnostics (calretinin, Ber-EP4, MOC-31, CKAE1/AE3, CK5/6, CD68, LCA, desmin, HBME-1, and WT1). Different staining parameters were tested using OV on BenchMark ULTRA and EnV on Dako Omnis immunostainer, respectively. Optimal staining protocols were then selected and validated on 10 positive and 10 negative cases. The staining results were compared with iV protocols through evaluation of UK NEQAS and internal scores. The optimal staining protocols with OV and EnV demonstrated similar or superior results compared to the existing iV protocols, with slightly stronger intensity regarding positive cells. We have successfully established and validated optimal ICC staining protocols for commonly used markers in routine cytopathology practice. These protocols may benefit other laboratories using similar staining platforms. However, the challenge regarding standardizing ICC protocols across different cytopathology laboratories remains unresolved.

FLEX: genetically encodable enzymatic fluorescence signal amplification using engineered peroxidase.

Fluorescent tagging of biomolecules enables their sensitive detection during separation and determining their subcellular location. In this context, peroxidase-based reactions are actively utilized for signal amplification. To harness this potential, we developed a genetically encodable enzymatic fluorescence signal amplification method using APEX (FLEX). We synthesized a fluorescent probe, Jenfluor triazole (JFT1), which effectively amplifies and restricts fluorescence signals under fixed conditions, enabling fluorescence-based detection of subcellularly localized electron-rich metabolites. Moreover, JFT1 exhibited stable fluorescence signals even under osmium-treated and polymer-embedded conditions, which supported findings from correlative light and electron microscopy (CLEM) using APEX. Using various APEX-conjugated proteins of interest (POIs) targeted to different organelles, we successfully visualized their localization through FLEX imaging while effectively preserving organelle ultrastructures. FLEX provides insights into dynamic lysosome-mitochondria interactions upon exposure to chemical stressors. Overall, FLEX holds significant promise as a sensitive and versatile system for fluorescently detecting APEX2-POIs in multiscale biological samples.