Dynamic Volumetric Imaging of Mouse Cerebral Blood Vessels In Vivo with an Ultralong Anti-Diffracting Beam.

Volumetric imaging of a mouse brain in vivo with one-photon and two-photon ultralong anti-diffracting (UAD) beam illumination was performed. The three-dimensional (3D) structure of blood vessels in the mouse brain were mapped to a two-dimensional (2D) image. The speed of volumetric imaging was significantly improved due to the long focal length of the UAD beam. Comparing one-photon and two-photon UAD beam volumetric imaging, we found that the imaging depth of two-photon volumetric imaging (80 µm) is better than that of one-photon volumetric imaging (60 µm), and the signal-to-background ratio (SBR) of two-photon volumetric imaging is two times that of one-photon volumetric imaging. Therefore, we used two-photon UAD volumetric imaging to perform dynamic volumetric imaging of mouse brain blood vessels in vivo, and obtained the blood flow velocity.

Two loss-of-function alleles of the glutathione S-transferase (GST) gene cause anthocyanin deficiency in flower and fruit skin of peach (Prunus persica).

Flower and fruit colors are important agronomic traits. To date, there is no forward genetic evidence that the glutathione S-transferase (GST) gene is responsible for the white flower color in peach (Prunus persica). In this study, genetic analysis indicated that the white-flower trait is monogenetic, is recessive to the non-white allele, and shows pleiotropic effects with non-white-flowered types. The genetic locus underpinning this trait was mapped onto chromosome 3 between 0.421951 and 3.227115 Mb by using bulked segregant analysis in conjunction with whole-genome sequencing, and was further mapped between 0 and 1.178149 Mb by using the backcross 1 (BC1 ) population. Finally, the locus was fine-mapped within 535.974- and 552.027-kb intervals by using 151 F2 individuals and 75 individuals from a BC1 self-pollinated (BC1 S1 ) population, respectively. Pp3G013600, encoding a GST that is known to transport anthocyanin, was identified within the mapping interval. The analysis of genome sequence data showed Pp3G013600 in white flowers has a 2-bp insertion or a 5-bp deletion in the third exon. These variants likely render the GST non-functional because of early stop codons that reduce the protein length from 215 amino acids to 167 and 175 amino acids, respectively. Genetic markers based on these variants validated a complete correlation between the GST loss-of-function alleles and white flower in 128 peach accessions. This correlation was further confirmed by silencing of Pp3G013600 using virus-induced gene silencing technology, which reduced anthocyanin accumulation in peach fruit. The new knowledge from this study is useful for designing peach breeding programs to generate cultivars with white flower and fruit skin.

Fine mapping of the gene controlling the weeping trait of Prunus persica and its uses for MAS in progenies.

BACKGROUND: Fruit tree yield and fruit quality are affected by the tree's growth type, and branching angle is an important agronomic trait of fruit trees, which largely determines the crown structure. The weeping type of peach tree shows good ventilation and light transmission; therefore, it is commonly cultivated. However, there is no molecular marker closely linked with peach weeping traits for target gene screening and assisted breeding. RESULTS: First, we confirmed that the peach weeping trait is a recessive trait controlled by a single gene by constructing segregating populations. Based on BSA-seq, we mapped the gene controlling this trait within 159 kb of physical distance on chromosome 3. We found a 35 bp deletion in the candidate area in standard type, which was not lacking in weeping type. For histological assessments, different types of branches were sliced and examined, showing fiber bundles in the secondary xylem of ordinary branches but not in weeping branches. CONCLUSIONS: This study established a molecular marker that is firmly linked to weeping trait. This marker can be used for the selection of parents in the breeding process and the early screening of hybrid offspring to shorten the breeding cycle. Moreover, we preliminary explored histological differences between growth types. These results lay the groundwork for a better understanding of the weeping growth habit of peach trees.

Ultra-long anti-diffracting beam volume imaging using a single-photon excitation microscope.

We studied a novel volumetric single-photon excitation microscope with an ultralong anti-diffracting (UAD) beam as illumination. Volumetric fluorescence image direct mapping showed that the axial imaging range of the UAD beam was approximately 14 times and 2 times that of conventional Gaussian and Airy beams, respectively, while maintaining a narrow lateral width. We compared the imaging capabilities of the Gaussian, Airy, and UAD modes through a strongly scattering environment mixed with fluorescent microspheres and agarose gel. Thick samples were scanned layer by layer in the Gaussian, Airy, and UAD modes, and then the three-dimensional structural information was projected onto a two-dimensional image. Benefiting from the longer focal length of the UAD beam, a deeper axial projection was provided, and the volume imaging speed was vastly increased. To demonstrate the performances of the UAD microscope, we performed dynamic volumetric imaging on the cardiovascular system of zebrafish labeled with green fluorescent proteins in the three modes and dynamically monitored substance transport in zebrafish blood vessels. In addition, the symmetrical curve trajectory of the UAD beam and the axial depth of the lateral position can be used for localization of micro-objects.

Implementation of a fluorescence spatiotemporal modulation super-resolution microscope.

Super-resolution microscopy (SRM) unveils details of subcellular organelles and provides a technical foundation for cellular biology research. Long-term, non-invasive live-cell super-resolution imaging requires low-intensity illumination and high image quality. Here, we present a new, to the best of our knowledge, method based on time-resolved detection termed fluorescence spatiotemporal modulation, in which highly spatially resolved photons in the beam center are extracted by taking the difference of the photons in the beam periphery with a weighted coefficient. The experimental results show a sub-100 nm resolution at tens of microwatts of laser power. Our proposed method requires only one laser, laying a foundation for a lower-cost multi-color super-resolution imaging system.

Molecular and morphological analyses clarify species delimitation in section Costatae and reveal Betula buggsii sp. nov. (sect. Costatae, Betulaceae) in China.

BACKGROUND AND AIMS: Delineating closely related and morphologically similar species is difficult. Here, we integrate morphology, genetics, ploidy and geography to resolve species and subspecies boundaries in four trees of section Costatae (genus Betula): Betula ashburneri, B. costata, B. ermanii and B. utilis, as well as multiple subspecies and polyploid races. METHODS: We genotyped 371 individuals (20-133 per species) from 51 populations at 15 microsatellite markers, as well as a subset of individuals, using restriction-site associated DNA sequencing and nuclear internal transcribed spacers. We determined the ploidy level of eight individuals using flow cytometry and characterized leaf variation for a subset of 109 individuals by morphometric analysis. KEY RESULTS: Integration of multiple lines of evidence suggested a series of revisions to the taxonomy of section Costatae. Betula costata and B. ermanii were found to be valid. Molecular and leaf morphology analyses revealed little differentiation between diploid B. albosinensis and some samples of B. utilis ssp. utilis. By contrast, other B. utilis ssp. utilis samples and ssp. albosinensis formed a morphological continuum but differed based on genetics. Specifically, B. utilis ssp. albosinensis was divided into two groups with group I genetically similar to B. utilis ssp. utilis and group II, a distinct cluster, proposed as the new diploid species Betula buggsii sp. nov. Phylogenomic analysis based on 2285 620 single nucleotide polymorphisms identified a well-supported monophyletic clade of B. buggsii. Morphologically, B. buggsii is characterized by elongated lenticels and a distinct pattern of bark peeling and may be geographically restricted to the Qinling-Daba Mountains. CONCLUSIONS: Our integrated approach identifies six taxa within section Costatae: B. ashburneri, B. buggsii, B. costata, B. utilis ssp. utilis, B. utilis ssp. albosinensis and B. ermanii. Our research demonstrates the value of an integrative approach using morphological, geographical, genetic and ploidy-level data for species delineation.

Aberration Correction to Optimize the Performance of Two-Photon Fluorescence Microscopy Using the Genetic Algorithm.

Due to less light scattering and a better signal-to-noise ratio in deep imaging, two-photon fluorescence microscopy (TPFM) has been widely used in biomedical photonics since its advent. However, optical aberrations degrade the performance of TPFM in terms of the signal intensity and the imaging depth and therefore restrict its application. Here, we introduce adaptive optics based on the genetic algorithm to detect the distorted wavefront of the excitation laser beam and then perform aberration correction to optimize the performance of TPFM. By using a spatial light modulator as the wavefront controller, the correction phase is obtained through a signal feedback loop and a process of natural selection. The experimental results show that the signal intensity and imaging depth of TPFM are improved after aberration correction. Finally, the method was applied to two-photon fluorescence lifetime imaging, which helps to improve the signal-to-noise ratio and the accuracy of lifetime analysis. Furthermore, the method can also be implemented in other experiments, such as three-photon microscopy, light-sheet microscopy, and super-resolution microscopy.

The transcriptional factor Apt regulates neuroblast differentiation through activating CycE expression.

In Drosophila, the thoracic neuroblast 6-4 (NB6-4T) divides asymmetrically into a glial precursor and a neuronal precursor, while the abdominal neuroblast 6-4 (NB6-4A) divides symmetrically to produce two glial cells. The underlying mechanism by which NB6-4T and NB6-4A undergo distinct differentiation is still elusive. Here, we find that the transcription factor Apontic (Apt) exclusively expresses in NB6-4T cells and is involved in regulating NB6-4T differentiation. Loss of Apt results in neuronal precursor loss. Epistasis analysis shows that Apt controls NB6-4T differentiation through activating CycE expression. On the other hand, Gcm suppresses Apt expression in the NB6-4A cell, thus inhibiting CycE expression. Taken together, our findings reveal a Gcm-Apt-CycE axis that regulates neuroblast and glia cell differentiation.

A nanozyme with switchable enzyme-like activity for the logic gates detection of thymol and hydrogen peroxide in honey.

A new nanozyme (CuGaa) with switchable enzyme-like activity of peroxidase and polyphenol oxidase was successfully prepared based on guanidinoacetic acid and copper. The two enzyme-like activities can be easily switched by changing temperature or adding MnCl2. At 4 °C, polyphenol oxidase-like activity decreased to nearly 1%, and the material is mainly characterized by peroxidase-like activity at this point. However, at 60 °C in the presence of 20 mM MnCl2, the peroxidase-like activity decreased to nearly 10%, and the polyphenol oxidase-like activity of the materials increased to 140%. Based on the switchable enzyme-like activity of CuGaa, detection methods for thymol and hydrogen peroxide were developed. In addition, a rapid combination strategy was further established combined with logic gate technology for the facile identification of complex contamination in honey, which provided new ideas for low-cost and rapid honey identification.

Modulation of the enzyme-like activity of CuAsp nanozyme by gallic acid and the selective detection of bisphenol A in infant food packaging.

The activity modulation of nanozymes with multi-enzymatic activities has both opportunities and challenges in practical applications. In this study, we found firstly that gallic acid erosion had a significant inhibitory effect on the peroxidase-catalyzed colorimetric reaction process of copper aspartate nanozyme prepared based on aspartic acid and copper (CuAsp), and the laccase-like catalytic activity remained almost unchanged. A sensing strategy for bisphenol A was then developed based on the laccase-like activity of GA-CuAsp synthesized by gallic acid (GA) acid erosion of CuAsp, which may have less interference due to the peroxidase-like activity. The developed sensing strategy had good selectivity and interference resistant ability, with a detection limit of 0.75 µmol L-1. In addition, the method was successfully applied to detecting BPA in plastic bottled drinking water samples and infant food packaging.

Lysine Deacetylation Is a Key Function of the Lysyl Oxidase Family of Proteins in Cancer.

Mammalian members of the lysyl oxidase (LOX) family of proteins carry a copper-dependent monoamine oxidase domain exclusively within the C-terminal region, which catalyzes ε-amine oxidation of lysine residues of various proteins. However, recent studies have demonstrated that in LOX-like (LOXL) 2-4 the C-terminal canonical catalytic domain and N-terminal scavenger receptor cysteine-rich (SRCR) repeats domain exhibit lysine deacetylation and deacetylimination catalytic activities. Moreover, the N-terminal SRCR repeats domain is more catalytically active than the C-terminal oxidase domain. Thus, LOX is the third family of lysine deacetylases in addition to histone deacetylase and sirtuin families. In this review, we discuss how the LOX family targets different cellular proteins for deacetylation and deacetylimination to control the development and metastasis of cancer.

Rhodamine-based fluorescent probe for dynamic STED imaging of mitochondria.

Stimulated emission depletion (STED) microscopy holds tremendous potential and practical implications in the field of biomedicine. However, the weak anti-bleaching performance remains a major challenge limiting the application of STED fluorescent probes. Meanwhile, the main excitation wavelengths of most reported STED fluorescent probes were below 500 nm or above 600 nm, and few of them were between 500-600 nm. Herein, we developed a new tetraphenyl ethylene-functionalized rhodamine dye (TPERh) for mitochondrial dynamic cristae imaging that was rhodamine-based with an excitation wavelength of 560 nm. The TPERh probe exhibits excellent anti-bleaching properties and low saturating stimulated radiation power in mitochondrial STED super-resolution imaging. Given these outstanding properties, the TPERh probe was used to measure mitochondrial deformation, which has positive implications for the study of mitochondria-related diseases.

ScrollTimes: Tracing the Provenance of Paintings as a Window Into History.

The study of cultural artifact provenance, tracing ownership and preservation, holds significant importance in archaeology and art history. Modern technology has advanced this field, yet challenges persist, including recognizing evidence from diverse sources, integrating sociocultural context, and enhancing interactive automation for comprehensive provenance analysis. In collaboration with art historians, we examined the handscroll, a traditional Chinese painting form that provides a rich source of historical data and a unique opportunity to explore history through cultural artifacts. We present a three-tiered methodology encompassing artifact, contextual, and provenance levels, designed to create a "Biography" for handscroll. Our approach incorporates the application of image processing techniques and language models to extract, validate, and augment elements within handscroll using various cultural heritage databases. To facilitate efficient analysis of non-contiguous extracted elements, we have developed a distinctive layout. Additionally, we introduce ScrollTimes, a visual analysis system tailored to support the three-tiered analysis of handscroll, allowing art historians to interactively create biographies tailored to their interests. Validated through case studies and expert interviews, our approach offers a window into history, fostering a holistic understanding of handscroll provenance and historical significance.

Coexistence of a novel chromosomal integrative and mobilizable element Tn7548 with two blaKPC-2-carrying plasmids in a multidrug-resistant Aeromonas hydrophila strain K522 from China.

OBJECTIVES: Herein, we detected one multidrug-resistant Aeromonas hydrophila strain K522 co-carrying two blaKPC-2 genes together with a novel chromosomal integrative and mobilizable element (IME) Tn7548 from China. To reveal the genetic characteristics of the novel reservoir of blaKPC-2 and IME in Aeromonas, a detailed genomic characterization of K522 was performed, and a phylogenetic analysis of Tn7412-related IMEs was carried out. METHODS: Carbapenemases were detected by using the immunocolloidal gold technique and antimicrobial susceptibility was tested by using VITEK 2. The whole-genome sequences of K522 were analysed using phylogenetics, detailed dissection, and comparison. RESULTS: Strain K522 carried a Tn7412-related chromosomal IME Tn7548 and three resistance plasmids pK522-A-KPC, pK522-B-KPC, and pK522-MOX. A phylogenetic tree of 82 Tn7412-related IMEs was constructed, and five families of IMEs were divided. These IMEs shared four key backbone genes: int, repC, and hipAB, and carried various profiles of antimicrobial resistance genes (ARGs). pK522-A-KPC and pK522-B-KPC carried blaKPC-2 and belonged to IncG and unclassified type plasmid, respectively. The blaKPC-2 regions of these two plasmids were the truncated version derived from Tn6296, resulting in the carbapenem resistance of K522. CONCLUSION: We first reported A. hydrophila harbouring a novel Tn7412-related IME Tn7548 together with two blaKPC-2 carrying plasmids and a MDR plasmid. Three of these four mobile genetic elements (MGEs) discovered in A. hydrophila K522 were novel. The emergence of novel MGEs carrying ARGs indicated the rapid evolution of the resistance gene vectors in A. hydrophila under selection pressure and would contribute to the further dissemination of various ARGs in Aeromonas.

MiR-383-5p inhibits the proliferation and migration of lung adenocarcinoma cells by targeting SHMT2.

Purpose: To explore the effects of miR-383-5p and serine hydroxymethyltransferase 2 (SHMT2) on the proliferation and migration of lung adenocarcinoma cells. Methods: SHMT2 expression in lung adenocarcinoma and normal tissues was investigated using The Cancer Genome Atlas database. Immunohistochemical analysis was performed to confirm SHMT2 expression in lung adenocarcinoma and adjacent normal lung tissues. Bioinformatics analysis and luciferase reporter assays were used to analyze the relationship between miR-383-5p and SHMT2 expression. The protein expression levels of SHMT2, vimentin, N-cadherin, E-cadherin, Bcl-2, and cyclinD1 were analyzed using western blotting. The reverse transcription-quantitative polymerase chain reaction was used to detect SHMT2 knockdown efficiency, miR-383-5p overexpression, and inhibition efficiency. The proliferative ability of cells was detected using the Cell Counting Kit-8 assay. The Transwell assay was used to detect the migration ability of cells. Results: SHMT2 expression was significantly increased in patients with lung adenocarcinoma compared to that in control patients; the higher the SHMT2 expression the worse the outcomes were in patients with lung adenocarcinoma. SHMT2 knockdown inhibited the proliferation, migration, and epithelial-mesenchymal transition of lung adenocarcinoma A549 and H1299 cells. MiR-383-5p directly targeted and downregulated SHMT2 in A549 and H1299 cells. The effects of miRNA-383-5p on the proliferation and migration of these cells differed from those of SHMT2. Exogenous overexpression of SHMT2 reversed the miR-383-5p-induced proliferation and migration inhibition in A549 and H1299 cells. Conclusion: MiR-383-5p inhibits the proliferation and migration of lung adenocarcinoma cells by targeting and downregulating SHMT2.

All-organic transparent plant e-skin for noninvasive phenotyping.

Real-time in situ monitoring of plant physiology is essential for establishing a phenotyping platform for precision agriculture. A key enabler for this monitoring is a device that can be noninvasively attached to plants and transduce their physiological status into digital data. Here, we report an all-organic transparent plant e-skin by micropatterning poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) on polydimethylsiloxane (PDMS) substrate. This plant e-skin is optically and mechanically invisible to plants with no observable adverse effects to plant health. We demonstrate the capabilities of our plant e-skins as strain and temperature sensors, with the application to Brassica rapa leaves for collecting corresponding parameters under normal and abiotic stress conditions. Strains imposed on the leaf surface during growth as well as diurnal fluctuation of surface temperature were captured. We further present a digital-twin interface to visualize real-time plant surface environment, providing an intuitive and vivid platform for plant phenotyping.

Horizontal CO2 Compensation in the Yangtze River Delta Based on CO2 Footprints and CO2 Emissions Efficiency.

PURPOSE: In this study, we attempted to reduce the negative economic externalities related to Carbon Dioxide (CO2) emissions in the Yangtze River Delta region (YRD) and designed a cross-municipality responsibility-sharing mechanism. METHODS: We estimated the municipal CO2 footprints in the YRD from 2000 to 2019 based on nighttime light data and measured CO2 emissions efficiency using a super slack-based measurement (super-SBM) model. Based on this, we designed a scenario of horizontal CO2 compensation among the YRD's municipalities from the perspectives of both CO2 footprints and CO2 trading (CO2 unit prices in trading were determined based on CO2 emissions efficiency). RESULTS: The results showed the following: (1) The CO2 footprints evolution of the YRD municipalities could be divided into four categories, among which, eleven municipalities showed a decreasing trend. Thirteen municipalities stabilized their CO2 footprints. Thirteen municipalities exhibited strong growth in their CO2 footprints, whereas four municipalities maintained a low level of slow growth. (2) Spatially, CO2 emissions efficiency evolved from a broad distribution of low values to a mosaic distribution of multi-type zones. (3) After 2011, the ratio of CO2 footprint compensation amounts to local Gross Domestic Product (GDP) in most municipalities was less than 0.01%, with its center of gravity shifting cyclically. It was appropriate to start charging the CO2 footprint compensation amounts after 2011, with a dynamic adjustment of 3 years. (4) After 2007, the supply-demand relationship of CO2 trading continued to deteriorate, and it eased in 2016. However, its operational mechanism was still very fragile and highly dependent on a few pioneering municipalities. INNOVATIONS: In this study, we designed a horizontal CO2 compensation mechanism from the binary perspective of CO2 footprints and CO2 trading. In this mechanism, the former determines the CO2 footprint compensation amounts paid by each municipality based on whether the CO2 footprint exceeds its CO2 allowance. The latter determines the CO2 trading compensation amounts paid by the purchasing municipalities based on their CO2 emissions efficiency. This system balances equity and efficiency and provides new ideas for horizontal CO2 compensation.

The Heterogeneous Effects of Formal and Informal Environmental Regulation on Green Technology Innovation-An Empirical Study of 284 Cities in China.

Promoting green technology innovation (GTI) through environmental regulation is a key measure in reducing the severity of environmental problems. However, the effects of formal environmental regulation (FER) and informal environmental regulation (IER) on GTI have not been clarified. Through theoretical analysis, this paper analyzes the effects of FER and IER on GTI based on OLS and GTWR models. The results show the following: (1) In all Chinese cities, both FER and IER have had a positive impact on GTI. The impact of FER has been much stronger than that of IER. They show a linkage effect, and their interaction (TER) has had a positive impact on GTI. (2) In terms of spatial heterogeneity, the impact of FER, IER, and TER on GTI has decreased across the east-west gradient and has been supplemented by a core-periphery structure. (3) In terms of urban heterogeneity, the impact of FER, IER, and TER has decreased with the size of the city. This study has the potential to strengthen the effect of environmental regulation on GTI. It can provide a decision-making reference for cities to coordinate FER and IER strategies, and provides evidence for adopting regionally differentiated environmental regulation strategies.

Short-Term Dendritic Dynamics of Neonatal Cortical Neurons Revealed by In Vivo Imaging with Improved Spatiotemporal Resolution.

Individual neurons in sensory cortices exhibit specific receptive fields based on their dendritic patterns. These dendritic morphologies are established and refined during the neonatal period through activity-dependent plasticity. This process can be visualized using two-photon in vivo time-lapse imaging, but sufficient spatiotemporal resolution is essential. We previously examined dendritic patterning from spiny stellate (SS) neurons, the major type of layer 4 (L4) neurons, in the mouse primary somatosensory cortex (barrel cortex), where mature dendrites display a strong orientation bias toward the barrel center. Longitudinal imaging at 8 h intervals revealed the long-term dynamics by which SS neurons acquire this unique dendritic pattern. However, the spatiotemporal resolution was insufficient to detect the more rapid changes in SS neuron dendrite morphology during the critical neonatal period. In the current study, we imaged neonatal L4 neurons hourly for 8 h and improved the spatial resolution by uniform cell surface labeling. The improved spatiotemporal resolution allowed detection of precise changes in dendrite morphology and revealed aspects of short-term dendritic dynamics unique to the neonatal period. Basal dendrites of barrel cortex L4 neurons were highly dynamic. In particular, both barrel-inner and barrel-outer dendrites (trees and branches) emerged/elongated and disappeared/retracted at similarly high frequencies, suggesting that SS neurons acquire biased dendrite patterns through rapid trial-and-error emergence, elongation, elimination, and retraction of dendritic trees and branches. We also found correlations between morphology and behavior (elongation/retraction) of dendritic tips. Thus, the current study revealed short-term dynamics and related features of cortical neuron dendrites during refinement.

Low-power compact continuous-wave stimulated emission depletion microscopy.

Stimulated emission depletion (STED) microscopy can break the optical diffraction barrier and provide subdiffraction resolution. According to the STED superresolution imaging principle, the resolution of STED is positively related to the power of the depletion laser. However, high-laser power largely limits the study of living cells or living bodies. Moreover, the high complexity and high cost of conventional pulsed STED microscopy limit the application of this technique. Therefore, this paper describes a simple continuous-wave STED (CW-STED) system constructed on a 45 × 60 cm breadboard and combined with digitally enhanced (DE) technology; low-power superresolution imaging is realized, which has the advantages of reducing system complexity and cost. The low-system complexity, low cost, and low-power superresolution imaging features of CW-STED have great potential to advance the application of STED microscopy in biological research.