ABSTRACT
In the field of nucleic acid amplification assays, developing enzyme-free, easy-to-use, and highly sensitive amplification approaches remains a challenge. In this work, we synthesized a heterogeneous Cu2O nanocatalyst (hnCu2O) with different particle sizes and shapes, which was used for developing enzyme- and label-free nucleic acid amplification methods based on the nucleic acid-templated azide-alkyne cycloaddition (AAC) reaction catalyzed by hnCu2O. The hnCu2O exhibited size- and shape-dependent catalytic activity, with smaller sizes and spherical-like shapes exhibiting superior activity. Spherical-like hnCu2O (61 ± 8 nm) not only achieved a ligation yield of up to 84.2 ± 3.9 % in 3 min but also exhibited faster kinetics in the nucleic acid-templated hnCu2O-catalyzed AAC reaction, with a high reaction rate of 0.65 min-1 and a half-life of 1.07 ± 0.09 min. Based on this result, we developed nucleic acid-templated click ligation linear amplification reaction (NA-CLLAR) and nucleic acid-templated click ligation exponential amplification reaction (NA-CLEAR) approach. By combining the recognition (complementary to the target sequence) and signal output (split G-quadruplex sequence) elements into a DNA probe, the NA-CLLAR and NA-CLEAR fluorescence assays achieved highly specific detection of target nucleic acids, with a detection limit of 2.8 aM based on G-quadruplex-enhanced fluorescence. This work is a valuable reference and will inspire researchers to design enzyme-free nucleic acid signal amplification strategies by developing different types of Cu(I) catalysts with improved catalytic activity.
ABSTRACT
Nucleic acids play a pivotal role in the diagnosis of diseases. However, rapid, cost-efficient, and ultrasensitive identification of nucleic acid targets still represents a significant challenge. Herein, we describe an enzyme-free DNA amplification method capable of achieving accurate and ultrasensitive nucleic acid detection via DNA-templated click ligation chain reaction (DT-CLCR) catalyzed by a heterogeneous nanocatalyst made of Cu2O (hnCu2O). This hnCu2O-DT-CLCR method is built on two cross-amplifying hnCu2O-catalyzed DNA-templated azide-alkyne cycloaddition-driven DNA ligation reactions that boast a fast reaction rate and a high DNA ligation yield in minutes, enabling rapid exponential amplification of specific DNA targets. This newly developed hnCu2O-DT-CLCR-enabled DNA amplification strategy is further integrated with two signal reporting mechanisms to achieve low-cost and easy-to-use biosensors: an electrochemical sensor through the conjugation of a methylene blue redox reporter to a DNA probe used in hnCu2O-DT-CLCR and a colorimetric sensor through the incorporation of the split-to-intact G-quadruplex DNAzyme encoded into hnCu2O-DT-CLCR. Both sensors are able to achieve specific detection of the intended DNA target with a limit of detection at aM ranges, even when challenged in complex biological matrices. The combined hnCu2O-DT-CLCR and sensing strategies offer attractive universal platforms for enzyme-free and yet efficient detection of specific nucleic acid targets.
Subject(s)
Click Chemistry , Copper , DNA , Nucleic Acid Amplification Techniques , Copper/chemistry , DNA/chemistry , Catalysis , Humans , Biosensing Techniques/methods , Limit of Detection , DNA, Catalytic/chemistry , DNA, Catalytic/metabolism , Azides/chemistry , Colorimetry/methods , Electrochemical Techniques/methods , Cycloaddition ReactionABSTRACT
Over the past decade, advancements in technology and methodology within the field of cryogenic electron microscopy (cryo-EM) single-particle analysis (SPA) have substantially improved our capacity for high-resolution structural examination of biological macromolecules. This advancement has ushered in a new era of molecular insights, replacing X-ray crystallography as the dominant method and providing answers to longstanding questions in biology. Since cryo-EM does not depend on crystallization, which is a significant limitation of X-ray crystallography, it captures particles of varying quality. Consequently, the selection of particles is crucial, as the quality of the selected particles directly influences the resolution of the reconstructed density map. An innovative iterative approach for particle selection, termed CryoSieve, significantly improves the quality of reconstructed density maps by effectively reducing the number of particles in the final stack. Experimental evidence shows that this method can eliminate the majority of particles in final stacks, resulting in a notable enhancement in the quality of density maps. This article outlines the detailed workflow of this approach and showcases its application on a real-world dataset.
Subject(s)
Cryoelectron Microscopy , Cryoelectron Microscopy/methods , Image Processing, Computer-Assisted/methodsABSTRACT
Diffusion, a fundamental internal mechanism emerging in many physical processes, describes the interaction among different objects. In many learning tasks with limited training samples, the diffusion connects the labeled and unlabeled data points and is a critical component for achieving high classification accuracy. Many existing deep learning approaches directly impose the fusion loss when training neural networks. In this work, inspired by the convection-diffusion ordinary differential equations (ODEs), we propose a novel diffusion residual network (Diff-ResNet), internally introduces diffusion into the architectures of neural networks. Under the structured data assumption, it is proved that the proposed diffusion block can increase the distance-diameter ratio that improves the separability of inter-class points and reduces the distance among local intra-class points. Moreover, this property can be easily adopted by the residual networks for constructing the separable hyperplanes. Extensive experiments of synthetic binary classification, semi-supervised graph node classification and few-shot image classification in various datasets validate the effectiveness of the proposed method.
ABSTRACT
Cryogenic electron microscopy (cryo-EM) is widely used to determine near-atomic resolution structures of biological macromolecules. Due to the low signal-to-noise ratio, cryo-EM relies on averaging many images. However, a crucial question in the field of cryo-EM remains unanswered: how close can we get to the minimum number of particles required to reach a specific resolution in practice? The absence of an answer to this question has impeded progress in understanding sample behavior and the performance of sample preparation methods. To address this issue, we develop an iterative particle sorting and/or sieving method called CryoSieve. Extensive experiments demonstrate that CryoSieve outperforms other cryo-EM particle sorting algorithms, revealing that most particles are unnecessary in final stacks. The minority of particles remaining in the final stacks yield superior high-resolution amplitude in reconstructed density maps. For some datasets, the size of the finest subset approaches the theoretical limit.
Subject(s)
Algorithms , Single Molecule Imaging , Cryoelectron Microscopy/methods , Macromolecular Substances/chemistryABSTRACT
Collecting paired training data is difficult in practice, but the unpaired samples broadly exist. Current approaches aim at generating synthesized training data from unpaired samples by exploring the relationship between the corrupted and clean data. This work proposes LUD-VAE, a deep generative method to learn the joint probability density function from data sampled from marginal distributions. Our approach is based on a carefully designed probabilistic graphical model in which the clean and corrupted data domains are conditionally independent. Using variational inference, we maximize the evidence lower bound (ELBO) to estimate the joint probability density function. Furthermore, we show that the ELBO is computable without paired samples under the inference invariant assumption. This property provides the mathematical rationale of our approach in the unpaired setting. Finally, we apply our method to real-world image denoising, super-resolution, and low-light image enhancement tasks and train the models using the synthetic data generated by the LUD-VAE. Experimental results validate the advantages of our method over other approaches.
ABSTRACT
Remarkable achievements have been obtained by deep neural networks in the last several years. However, the breakthrough in neural networks accuracy is always accompanied by explosive growth of computation and parameters, which leads to a severe limitation of model deployment. In this paper, we propose a novel knowledge distillation technique named self-distillation to address this problem. Self-distillation attaches several attention modules and shallow classifiers at different depths of neural networks and distills knowledge from the deepest classifier to the shallower classifiers. Different from the conventional knowledge distillation methods where the knowledge of the teacher model is transferred to another student model, self-distillation can be considered as knowledge transfer in the same model - from the deeper layers to the shallow layers. Moreover, the additional classifiers in self-distillation allow the neural network to work in a dynamic manner, which leads to a much higher acceleration. Experiments demonstrate that self-distillation has consistent and significant effectiveness on various neural networks and datasets. On average, 3.49 and 2.32 percent accuracy boost are observed on CIFAR100 and ImageNet. Besides, experiments show that self-distillation can be combined with other model compression methods, including knowledge distillation, pruning and lightweight model design.
Subject(s)
Algorithms , Neural Networks, Computer , HumansABSTRACT
The structural variations of multidomain proteins with flexible parts mediate many biological processes, and a structure ensemble can be determined by selecting a weighted combination of representative structures from a simulated structure pool, producing the best fit to experimental constraints such as interatomic distance. In this study, a hybrid structure-based and physics-based atomistic force field with an efficient sampling strategy is adopted to simulate a model di-domain protein against experimental paramagnetic relaxation enhancement (PRE) data that correspond to distance constraints. The molecular dynamics simulations produce a wide range of conformations depicted on a protein energy landscape. Subsequently, a conformational ensemble recovered with low-energy structures and the minimum-size restraint is identified in good agreement with experimental PRE rates, and the result is also supported by chemical shift perturbations and small-angle X-ray scattering data. It is illustrated that the regularizations of energy and ensemble-size prevent an arbitrary interpretation of protein conformations. Moreover, energy is found to serve as a critical control to refine the structure pool and prevent data overfitting, because the absence of energy regularization exposes ensemble construction to the noise from high-energy structures and causes a more ambiguous representation of protein conformations. Finally, we perform structure-ensemble optimizations with a topology-based structure pool, to enhance the understanding on the ensemble results from different sources of pool candidates.
Subject(s)
Molecular Dynamics Simulation , Poly(A)-Binding Proteins/chemistry , Saccharomyces cerevisiae Proteins/chemistry , Amino Acids/chemistry , Binding Sites , Electron Spin Resonance Spectroscopy , Protein Binding , Protein Domains , Protein Structure, Secondary , Saccharomyces cerevisiae , Structure-Activity Relationship , ThermodynamicsABSTRACT
A sensor pixel integrates optical intensity across its extent, and we explore the role that this integration plays in phase space tomography. The literature is inconsistent in its treatment of this integration-some approaches model this integration explicitly, some approaches are ambiguous about whether this integration is taken into account, and still some approaches assume pixel values to be point samples of the optical intensity. We show that making a point-sample assumption results in apodization of and thus systematic error in the recovered ambiguity function, leading to underestimating the overall degree of coherence. We explore the severity of this effect using a Gaussian Schell-model source and discuss when this effect, as opposed to noise, is the dominant source of error in the retrieved state of coherence.
ABSTRACT
The human cerebellum has recently been discovered to contribute to cognition and emotion beyond the planning and execution of movement, suggesting its functional heterogeneity. We aimed to identify the functional parcellation of the cerebellum using information from resting-state functional magnetic resonance imaging (rs-fMRI). For this, we introduced a new data-driven decomposition-based functional parcellation algorithm, called Sparse Dictionary Learning Clustering (SDLC). SDLC integrates dictionary learning, sparse representation of rs-fMRI, and k-means clustering into one optimization problem. The dictionary is comprised of an over-complete set of time course signals, with which a sparse representation of rs-fMRI signals can be constructed. Cerebellar functional regions were then identified using k-means clustering based on the sparse representation of rs-fMRI signals. We solved SDLC using a multi-block hybrid proximal alternating method that guarantees strong convergence. We evaluated the reliability of SDLC and benchmarked its classification accuracy against other clustering techniques using simulated data. We then demonstrated that SDLC can identify biologically reasonable functional regions of the cerebellum as estimated by their cerebello-cortical functional connectivity. We further provided new insights into the cerebello-cortical functional organization in children.
ABSTRACT
In recent years, sparse coding has been widely used in many applications ranging from image processing to pattern recognition. Most existing sparse coding based applications require solving a class of challenging non-smooth and non-convex optimization problems. Despite the fact that many numerical methods have been developed for solving these problems, it remains an open problem to find a numerical method which is not only empirically fast, but also has mathematically guaranteed strong convergence. In this paper, we propose an alternating iteration scheme for solving such problems. A rigorous convergence analysis shows that the proposed method satisfies the global convergence property: the whole sequence of iterates is convergent and converges to a critical point. Besides the theoretical soundness, the practical benefit of the proposed method is validated in applications including image restoration and recognition. Experiments show that the proposed method achieves similar results with less computation when compared to widely used methods such as K-SVD.