Preservation Of Structure Research Articles

Anesthesia for an infant with congenital mediastinal mass: a case report

BackgroundGiant anterior mediastinal masses in infants are one of the most challenging cases faced in pediatric anesthesia practice. They can pose unique challenges for resection such as cardiovascular collapse on induction of anesthesia and injury to surrounding structures that maybe compressed or displaced. Principles that must be followed and kept in mind during removal of giant mediastinal mass include appropriate diagnostic imaging to define mass extent, airway control during induction, a multidisciplinary team approach including cardiothoracic for sternotomy, cannulation to institute cardiopulmonary bypass, otolaryngology for rigid bronchoscopy, preservation of neurovascular structure, and complete resection whenever possible. Our patient had a mass that weighed twice his whole body weight.Case presentationHere we present a 3-month-old Middle Eastern infant weighing 3.2 kg with a large congenital teratoma who presented to the emergency room with cyanosis and respiratory distress. During his hospital course, he underwent three procedures, two of them under light-to-moderate sedation: a diagnostic computer tomography scan followed by mass content drainage by interventional radiology (Figs. 1, 2). On the third day, he had a thoracotomy with complete tumor resection under general anesthesia with the help of an epidural for pain control (Fig. 3). The resected tumor weighed 2.5 kg, which was equal to twice the patient’s total body weight (Fig. 4). After the surgery, he was extubated in the operating room and discharged home 3 days later.Fig. 1Anterior–posterior chest x-ray showing the anterior mediastinal mass (AMM)Fig. 2Cross-section computed tomography (CT) showing large anterior mediastinal mass (AMM)Fig. 3Gross anatomy of the tumor before resectionFig. 4Gross anatomy of the removed tumorConclusionAnterior mediastinal mass patients can be challenging for the anesthesiologist. They need meticulous thorough perioperative assessment to determine the extent of compression on major intramediastinal structures and to predict the complications. Planning by multidisciplinary team and discussion with the family is important. These types of cases should be preferably operated on by an experienced team in a well-equipped operation room in tertiary care institutes.

Main clinical approaches to the use of silane in restoration repairs with resin: a systematic review

Introduction: The repair in the composite clinical procedure is handy and allows the removal of the damaged portion of the restoration without requiring complete replacement of the same, allowing the preservation of sound tooth structure. Objective: The objective of this study was to review the literature evaluating the use of silane in repairs of composite resin restorations and the procedures to be followed. Methods: The PRISMA Platform systematic review rules were followed. The search was carried out from January to March 2024 in the Scopus, PubMed, Science Direct, Scielo, and Google Scholar databases. The quality of the studies was based on the GRADE instrument and the risk of bias was analyzed according to the Cochrane instrument. Results and Conclusion: A total of 117 articles were found, 43 articles were evaluated in full and 36 were included and developed in the present systematic review study. Considering the Cochrane tool for risk of bias, the overall assessment resulted in 07 studies with a high risk of bias and 21 studies that did not meet GRADE and AMSTAR-2. Most studies did not show homogeneity in their results, with X2=74.5%>50%. In the literature review was noted that the preference for maintaining part of the restoration and repair of the clinical protocol should be thoroughly evaluated and implemented to increase the predictability of the procedure. The restorative material should unite, not only to tooth structure but also already present in the resin preparation. Thus, silanes and resins without charge are traditionally used as coupling agents in repairs. The advantages of this procedure compared with total replacement, and the possibility of obtaining a good integrity and longevity of the restoration involved were discussed. It was concluded that the adhesion between the existing and the new resin increment of resin can be enhanced by employing a silane, associated or not with an adhesive system.

Boosting toluene destruction by engineering surface acidity–alkalinity in defective cobalt oxide catalyst

The surface acidity and alkalinity of catalysts played a decisive role in the destruction of Volatile organic compounds (VOCs). In this study, a series of Co3O4 catalysts were synthesized under different temperatures by calcining metal organic frameworks (MOFs). By controlling the temperature, the organic linkers were effectively eliminated, resulting in catalysts with a high specific surface area due to the preservation of the parent structure of the MOFs. Among the catalysts tested, Co3O4−350 (calcined at 350 °C) demonstrated the highest catalytic activity for toluene oxidation (T90 =231 ℃) and reliable water resistance (3 vol.% water vapor). The efficacy was attributed to the high ratio of reactive oxygen species and high valence Co species, weaken Co−O bond strength as well as high concentration of oxygen vacancies. In situ diffuse reflectance infrared Fourier transform spectroscopy (in situ DRIFTS) demonstrate that acidity–alkalinity is crucial in the adsorption and activation, while oxygen vacancies were found to be essential for deep oxidation.

Infrared and visible image fusion based on semi-global weighted least squares and guided edge-aware filters

Most of the existing image fusion methods are dedicated to extracting the respective private features in the source image to reconstruct to get a fused image that contains rich information. However, its neglect of the enhancement of texture details in the source image as well as the preservation of background structures, which makes the fusion result look less impressive. To this end, an infrared and visible image fusion method based on a semi-global weighted least squares (SGWLS) method and guided edge-aware filter (GEAF) is proposed. First, a SGWLS method is applied to decompose the source image into base and detail layers containing different scale information. Then, the detail layer are texture-enhanced by using the scale coefficients. Due to the lack of edge features in the original base layer, for this reason, a guided edge-aware filter is designed, in which the enhanced detail layer was used as the guidance image as a way to perform a quadratic feature enhancement fusion for the base layer in order to retain richer gradient information. Finally, reconstruction of the fused detail and base layers by using inverse transformation to obtain the fusion result. Compared with nine state-of-the-art fusion algorithms, a large number of experimental results demonstrate the superior performance of the proposed method in highlighting texture details and background features, as well as the obvious advantages in SF and AG metrics.

Effect of freeze-thaw and PEF pretreatments on the kinetics and microstructure of convective and ultrasound-assisted drying of orange peel

The main waste generated by juice industry comprises orange peels, which have a great upcycling potential once stabilized. Drying is the most used method for this purpose, but the high energy consumption prompts interest in its intensification. This study assessed the influence of freeze-thaw and pulsed electric field (PEF) pretreatments in conventional and airborne ultrasound-assisted drying (50 °C) of orange peels. None of these pretreatments alone got to reduce processing times significantly, but combined with ultrasound-assisted drying produced a significant shortening of the process. This was particularly important in the lower intensity PEF pretreatment tested (0.33 kJ/kg), indicating the existence of optimum conditions to carry out the pretreatments. Microstructure analysis revealed that the application of ultrasound during drying led to better preservation of the sample structure. Thus, the integration of pretreatment techniques to ultrasound-assisted drying may not only shorten the process but also help to preserve the original structure.

Science and Islamic ethics: Navigating artificial womb technology through Quranic principles

This paper explores the complex interplay between scientific innovation and religious ethics, with a specific focus on the ethical implications of Artificial Womb Technology (AWT) as interpreted through the lens of Quranic teachings on the essence of life. The objective is to meld the burgeoning field of reproductive technologies with the foundational principles of Islamic theology through an examination of Islamic jurisprudential rulings, contemporary bioethical discourse and innovations in reproductive technology. In addition to attention given to the compatibility of AWT with Islamic teachings concerning the sanctity of life, there is also a focus on the concept of motherhood and the preservation of family structure. This study undertakes an extensive exploration of both historical and contemporary interpretations of Islamic precepts, culminating in the establishment of an ethical framework. This framework is designed to harmonise religious doctrines with the exigencies of reproductive science by proposing normative guidelines for the ethical implementation of AWT and similar technologies. This paper makes a substantial contribution to academic discourse on science and religion by integrating advancements in reproductive health technology with moral principles intrinsic to the Islamic faith.

Comprehensive Guide to Transoral Endoscopic Thyroidectomy Vestibular Approach: Step-by-Step Technique

AbstractTransoral endoscopic thyroidectomy vestibular approach (TOETVA) is a minimally invasive surgical technique that offers safety comparable to conventional open thyroidectomy, with the added benefit of superior cosmetic outcomes due to the absence of visible scars. This innovative approach is gaining popularity worldwide as an alternative for patients seeking thyroid surgery without postoperative scarring. TOETVA involves positioning the patient supine with slight neck extension under general anesthesia. The procedure uses three trocars placed via midline and lateral incisions, with a 10-mm 30-degree laparoscope for visualization. Preoperative care includes cleaning the surgical area and administering intravenous antibiotics to prevent infection. The technique mirrors conventional thyroidectomy but utilizes standard laparoscopic instruments. It emphasizes the precise identification and preservation of critical anatomical structures, such as the cricoid cartilage, thyroid isthmus, trachea, carotid artery, superior laryngeal nerve, and recurrent laryngeal nerve. The use of an energy device and careful dissection help minimize bleeding and reduce nerve injury risks. Superior pole management and meticulous handling of the thyroid lobe ensures optimal outcomes. The growing popularity of TOETVA indicates a shift toward minimally invasive thyroid surgery, with TOETVA emerging as a leading method. This paper provides a comprehensive overview of the procedure, highlighting its benefits and critical aspects of its successful implementation.

Oral nicotinamide provides robust, dose-dependent structural and metabolic neuroprotection of retinal ganglion cells in experimental glaucoma

A compromised capacity to maintain NAD pools is recognized as a key underlying pathophysiological feature of neurodegenerative diseases. NAD acts as a substrate in major cell functions including mitochondrial homeostasis, cell signalling, axonal transport, axon/Wallerian degeneration, and neuronal energy supply. Dendritic degeneration is an early marker of neuronal stress and precedes cell loss. However, little is known about dendritic structural preservation in pathologic environments and remodelling in mature neurons. Retinal ganglion cell dendritic atrophy is an early pathological feature in animal models of the disease and has been demonstrated in port-mortem human glaucoma samples. Here we report that a nicotinamide (a precursor to NAD through the NAD salvage pathway) enriched diet provides robust retinal ganglion cell dendritic protection and preserves dendritic structure in a rat model of experimental glaucoma. Metabolomic analysis of optic nerve samples from the same animals demonstrates that nicotinamide provides robust metabolic neuroprotection in glaucoma. Advances in our understanding of retinal ganglion cell metabolic profiles shed light on the energetic shift that triggers early neuronal changes in neurodegenerative diseases. As nicotinamide can improve visual function short term in existing glaucoma patients, we hypothesize that a portion of this visual recovery may be due to dendritic preservation in stressed, but not yet fully degenerated, retinal ganglion cells.

RSTC: Residual Swin Transformer Cascade to approximate Taylor expansion for image denoising

Traditional denoising methods establish mathematical models by employing different priors, which can achieve preferable results but they are usually time-consuming and their outputs are not adaptive on regularization parameters. While the success of end-to-end deep learning denoising strategies depends on a large amount of data and lacks a theoretical interpretability. In order to address the above problems, this paper proposes a novel image denoising method, namely Residual Swin Transformer Cascade (RSTC), based on Taylor expansion. The key procedures of our RSTC are specified as follows: Firstly, we discuss the relationship between image denoising model and Taylor expansion, as well as its adjacent derivative parts. Secondly, we use a lightweight deformable convolutional neural network to estimate the basic layer of Taylor expansion and a residual network where swin transformer block is selected as a backbone for pursuing the solution of the derivative layer. Finally, the results of the two networks contribute to the approximation solution of Taylor expansion. In the experiments, we firstly test and discuss the selection of network parameters to verify its effectiveness. Then, we compare it with existing advanced methods in terms of visualization and quantification, and the results show that our method has a powerful generalization ability and performs better than state-of-the-art denoising methods on performance improvement and structure preservation.

Iron and Manganese concentrations in leaf tissues of Rhizophora mangle (Rhizophoraceae): implications for energetic metabolism

Introduction: Iron (Fe) and manganese (Mn) are bioessential micronutrients for plants but can impair the energetic metabolism when present at high levels. Objective: To assess the photosynthetic performance and oxidative damages in Rhizophora mangle L. leaf tissues at low and high concentrations of Fe (74 and 195 mg kg-1; Feleaf) and Mn (65 and 414 mg kg-1; Mnleaf). Methods: Photosynthetic pigments, chlorophyll a fluorescence, leaf CO2 assimilation and gas exchange and DPPH• radical scavenging capacity were sampled in R. mangle growing in estuarine forests in the North region of Espírito Santo State and the extreme South of Bahia State (Brazil) showing low and high Feleaf and Mnleaf. Results: Effects of high Fe and Mn were not identified on pigment levels. The increase in Feleaf and Mnleaf at the levels observed in this assessment had a positive effect on the number of reaction centers and on the efficiency of the oxygen-evolving complex, evaluated as K-band, while no changes were found in the parameters related to the excitation trapping efficiency at the active center of photosystem II. Distinct interference patterns of Fe and Mn on the functional processes of photosynthesis were identified, especially on CO2 assimilation and reactive oxygen species metabolism, with major effects on CO2 assimilation and carboxylation efficiency of Rubisco at high Mnleaf. Conclusion: These findings demonstrate the efficiency of R. mangle in positively regulating the electron transport chain in response to high Fe and Mn, at least in terms of the preservation of structure and functionality of the plant photosynthetic apparatus. Moreover, interference of high Mnleaf in R. mangle occurs at non-stomatal and biochemical levels. There is an antagonistic interference of these trace elements with the physiology of R. mangle, which is a dominant species in Brazilian mangroves.

Cross-domain manifold structure preservation for transferable and cross-machine fault diagnosis

To address the decline or failure in the autonomous learning capability of traditional transfer learning methods when training and test samples come from different machines, resulting in low cross-machine fault diagnosis rates, we propose a cross-domain manifold structure preservation (CDMSP) method for diagnosing rolling bearing faults across machines. The CDMSP method can induce the manifold space projection matrices of the source and target domains more effectively. This method maps high-dimensional features into a low-dimensional manifold, preserving non-linear relationships and aligning distribution differences while maintaining cross-domain manifold structure consistency. Additionally, highly confidently labeled target domain samples are selected from each mapping result and added to the training dataset to enhance subspace learning in subsequent iterations. The CDMSP method is both simple and effective at capturing the underlying structures and patterns in the data. The CWRU dataset and our self-built test platform dataset were used to validate this method. Experimental results show that CDMSP, as a non-deep domain adaptation method of transfer learning, outperforms similar methods in cross-machine fault identification, achieving a maximum fault identification accuracy of 100 % with excellent convergence performance. Furthermore, simulated diagnostic experiments under noise interference indicate that CDMSP maintains high fault identification accuracy, even in noisy environments. Overall, CDMSP is an efficient and reliable new method for diagnosing cross-machine bearing faults.

Outcome of Patients with Early Glottic Laryngeal Carcinoma Treated with Radical Radiotherapy

Introduction: Glottic cancer is the most common laryngeal cancer and it has the most favorable prognosis, mainly if diagnosed early. Ideally, early glottic cancer (T1–T2 N0) should be managed with a single modality either microsurgery or definitive Radiotherapy. The local control rate with definitive radiotherapy is approximately 83–95% for T1 and 70–80% for T2 lesions. This study intends to find out the treatment outcome in early glottic cancer treated with radical radiation treatment from 1st January 2014 to 31st December 2019 at our center. Methods: This was a retrospective analysis of medical records and radiation charts of all patients of early stage glottis cancer who underwent radiation treatment from 1st January 2014 to 31st December 2019 at our centre. The statistical analysis was done using SPSS version 20.0.Continuous variables such as age was summarized in terms of either mean ± Standard Deviation or median (inter quartile range) depending on the statistical distribution of data. Categorical variables were e pressed in terms of frequency and percentage. Survival analysis was done using Kaplan Meier curves. Results: A total of 91 patients underwent radical radiation for early glottic carcinoma at our centre. The median age was 63 years (42-85 years). Out of them 94% were males and 6% were females. 78% of patients had stage 1 disease and 22% had stage 11 disease. 52% of patients had co morbidities and 48% did not have any comorbidity All cases had Squamous cell carcinoma histology. 59% of patients had anterior commissure involvement. 12 out of 91 (13%) patients developed local recurrence on follow up. The five year disease free survival was 84.3%. The five year overall survival was 87.7%. There was a statistically significant difference in survival between stage I and stage II. Conclusion: For T1–T2 glottic carcinomas, definitive RT can provide a good term local control and survival with the preservation of the normal structure and function of larynx.

VEGFD signaling balances stability and activity-dependent structural plasticity of dendrites

Mature neurons have stable dendritic architecture, which is essential for the nervous system to operate correctly. The ability to undergo structural plasticity, required to support adaptive processes like memory formation, is still present in mature neurons. It is unclear what molecular and cellular processes control this delicate balance between dendritic structural plasticity and stabilization. Failures in the preservation of optimal dendrite structure due to atrophy or maladaptive plasticity result in abnormal connectivity and are associated with various neurological diseases. Vascular endothelial growth factor D (VEGFD) is critical for the maintenance of mature dendritic trees. Here, we describe how VEGFD affects the neuronal cytoskeleton and demonstrate that VEGFD exerts its effects on dendrite stabilization by influencing the actin cortex and reducing microtubule dynamics. Further, we found that during synaptic activity-induced structural plasticity VEGFD is downregulated. Our findings revealed that VEGFD, acting on its cognate receptor VEGFR3, opposes structural changes by negatively regulating dendrite growth in cultured hippocampal neurons and in vivo in the adult mouse hippocampus with consequences on memory formation. A phosphoproteomic screening identified several regulatory proteins of the cytoskeleton modulated by VEGFD. Among the actin cortex-associated proteins, we found that VEGFD induces dephosphorylation of ezrin at tyrosine 478 via activation of the striatal-enriched protein tyrosine phosphatase (STEP). Activity-triggered structural plasticity of dendrites was impaired by expression of a phospho-deficient mutant ezrin in vitro and in vivo. Thus, VEGFD governs the equilibrium between stabilization and plasticity of dendrites by acting as a molecular brake of structural remodeling.

Dynamic 4D facial capture pipeline with appearance driven progressive retopology based on optical flow

This paper presents a production-oriented 4D facial reconstruction pipeline designed to produce high-fidelity facial mesh sequences with a consistently structured topology, while preserving the wireframe structure specified by artists. We have designed and developed a compact, efficient, and fast optical capture system based on synchronized camera arrays for high-precision dynamic 3D facial imaging. Unlike prevailing methods that primarily concentrate on single-frame reconstruction, often reliant on labor-intensive manual annotation, our framework exploits the constraint of appearance consistency to autonomously establish feature correspondence and uphold temporal coherence within the mesh. Consequently, our approach eliminates mesh drifting and jitter, enabling full parallelization for dynamic facial expression capture. The proposed pipeline decouples the non-linear deformation of facial expressions from the rigid movements of the skull through a stable external device. Leveraging progressive retopology, our methodology employs artist-guided templates as priors, ensuring the preservation of wireframe structures across the result sequence. Progressive retopology is achieved by constraining different fine-grained features of 3D landmarks, scan surface shapes, and appearance textures. The results of our study showcase facial mesh sequences with production-quality topology, adept at faithfully reproducing character expressions from photographs while achieving artist-friendly stable facial movements.

Decalcification-free technique on the analysis of dental pulp tissue: histological and immunohistochemical analysis.

Histological techniques are essential for the microscopic study and investigation of the human dental pulp. The aim of this study was to evaluate the impact of decalcification-free technique by examining dental pulp morphology by histological staining with haematoxylin and eosin and immunohistochemistry. The sample consisted of 30 healthy third molars extracted for orthodontic indication, the pulp tissue was obtained by removing the mineralized tissues, separating the enamel and dentine and by marking with a flexible diamond disc on the coronal surface and longitudinal axis of the root. These guides made it possible to separate the fragments and obtain the pulp tissue for fixation and staining with H&E and subsequent immunohistochemistry with CD34 and S-100 antibodies. The technique showed preservation of pulp morphology with adequate preservation of microscopic structures. No alterations in tissue viability were observed. The staining allowed an accurate assessment of vascular and nervous components by means of CD34 and S-100 markers, respectively. This technique allows preservation of pulp tissue, maintaining viable tissue for histological analysis and immunohistochemistry tests, as well as reducing sample processing time.

Nonlinear dynamic transfer partial least squares for domain adaptive regression

Aiming to address soft sensing model degradation under changing working conditions, and to accommodate dynamic, nonlinear, and multimodal data characteristics, this paper proposes a nonlinear dynamic transfer soft sensor algorithm. The approach leverages time-delay data augmentation to capture dynamics and projects the augmented data into a latent space for constructing a nonlinear regression model. Two regular terms, distribution alignment regularity and first-order difference regularity, are introduced during data projection to address data distribution disparities. Laplace regularity is incorporated into the nonlinear regression model to ensure geometric structure preservation. The final optimization objective is formulated within the framework of partial least squares, and hyperparameters are determined using Bayesian optimization. The effectiveness of the proposed algorithm is demonstrated through experiments on three public datasets.

Cardiomyocyte Regeneration in Human Myocarditis.

Newly generated cardiomyocytes (NGCs) concur with the recovery of human myocarditis occurring spontaneously in around 50% of cases. However, NGCs decline with age, and their modality of myocardial homing and integration are still unclear. We retrospectively assessed NGCs in 213 consecutive patients with endomyocardial biopsy denoting acute myocarditis, with normal coronaries and valves. Tissue samples were processed for histology (H&E), immunohistochemistry for the evaluation of inflammatory infiltrates, immunostaining for alpha-sarcomeric-actin, junctional connexin-43, Ki-67, and phosphorylated STAT3 (p-STAT3), and Western blot (WB) for HMGB1. Frozen samples were analyzed using polymerase chain reaction (PCR) for cardiotropic viruses. Controls included 20 normal surgical biopsies. NGCs were defined as small myocytes (diameter < 10 µm) with nuclear positivity to Ki-67 and p-STAT3 and positive immunostaining for cytoplasmic α-sarcomeric actin and connexin-43. Their number/mm2 in relation to age and pathway of integration was evaluated. NGCs crossed the membrane and grew integrated within the empty necrotic myocytes. NGC mean diameter was 6.6 ± 3.34 vs. 22.5 ± 3.11 µm adult cells; their number, in comparison to LVEF, was 86.3 ± 10.3/mm2 in patients between 18 and 40 years, 50.4 ± 13.8/mm2 in those between 41 and 60, and 15.1 ± 5.7/mm2 in those between 61 and 80. Control NGCs' mean diameter was 0.2 ± 0.2 mm2. PCR was positive for viral genomes in 16% of cases; NGCs were not statistically different in viral and non-viral myocarditis. WB analysis revealed a higher expression of HMGB1 in myocarditis compared to myocardial controls. NGCs are constantly recognizable in acute human myocarditis. Their number declines with age. Their integration within necrotic myocytes allows for the preservation of the cardiac structure and function.

Preservation of retinal structure and function in two mouse models of inherited retinal degeneration by ONL1204, an inhibitor of the Fas receptor

Due to the large number of genes and mutations that result in inherited retinal degenerations (IRD), there has been a paucity of therapeutic options for these patients. There is a large unmet need for therapeutic approaches targeting shared pathophysiologic pathways in a mutation-independent manner. The Fas receptor is a major activator and regulator of retinal cell death and inflammation in a variety of ocular diseases. We previously reported the activation of Fas-mediated photoreceptor (PR) cell death in two different IRD mouse models, rd10 and P23H, and demonstrated the protective effect of genetic Fas inhibition. The purpose of this study was to examine the effects of pharmacologic inhibition of Fas in these two models by intravitreal injection with a small peptide inhibitor of the Fas receptor, ONL1204. A single intravitreal injection of ONL1204 was given to one eye of rd10 mice at P14. Two intravitreal injections of ONL1204 were given to the P23H mice, once at P14 and again at 2-months of age. The fellow eyes were injected with vehicle alone. Fas activation, rate of PR cell death, retinal function, and the activation of immune cells in the retina were evaluated. In both rd10 and P23H mice, ONL1204 treatment resulted in decreased number of TUNEL (+) PRs, decreased caspase 8 activity, enhanced photoreceptor cell counts, and improved visual function compared with vehicle treated fellow eyes. Treatment with ONL1204 also reduced immune cell activation in the retinas of both rd10 and P23H mice. The protective effect of pharmacologic inhibition of Fas by ONL1204 in two distinct mouse models of retinal degeneration suggests that targeting this common pathophysiologic mechanism of cell death and inflammation represents a potential therapeutic approach to preserve the retina in patients with IRD, regardless of the genetic underpinning.

Global Structure Preservation and Self-Representation-Based Supervised Feature Selection

Feature selection aims to select a subset of features from high-dimensional data, which can overcome the curse of dimensionality for the next dealing steps. However, the feature selection itself could face the curse of dimensionality. To overcome the above problem, in this paper, a new feature selection framework is designed according to a human processing in our daily life. In our daily life, to evaluate a candidate's ability to work, the related professional knowledge and the comprehensive ability of a candidate should be both evaluated. Actually, a candidate only with good professional knowledge often hardly solves new problems in the work. Based on the above analysis, in our new designed framework, the features are selected by evaluating its ability of global structure preservation and self-representation, which are respectively similar to the professional knowledge and comprehensive ability in evaluating candidate. As a result, the selected features can accommodate larger changes in test data. The conducted experiments validate the effectiveness of our feature selection.

The potential of ALFA-tag and tyramide-based fluorescence signal amplification to expand the CRISPR-based DNA imaging toolkit.

Understanding the spatial organization of genomes within chromatin is crucial for deciphering gene regulation. A recently developed CRISPR-dCas9-based genome labeling tool, known as CRISPR-FISH, allows efficient labelling of repetitive sequences. Unlike standard fluorescence in situ hybridization (FISH), CRISPR-FISH eliminates the need for global DNA denaturation, allowing for superior preservation of chromatin structure. Here, we report on the further development of the CRISPR-FISH method, which has been enhanced for increased efficiency through the engineering of a recombinant dCas9 protein containing an ALFA-tag. Using an ALFA-tagged dCas9 protein assembled with an A. thaliana centromere-specific gRNA, we demonstrate target-specific labelling with a fluorescence-labeled NbALFA nanobody. The dCas9 protein possessing multiple copies of the ALFA-tag, in combination with a minibody and fluorescence-labelled anti-rabbit secondary antibody, resulted in enhanced target-specific signals. The dCas9-ALFA-tag system was also instrumental in live cell imaging of telomeres in N. benthamiana. This method will further expand the CRISPR imaging toolkit, facilitating a better understanding of genome organization. Furthermore, we report the successful integration of the highly sensitive Tyramide Signal Amplification (TSA) method with CRISPR-FISH, demonstrating effective labeling of A. thaliana centromeres.