Search

Cell size and density are tightly controlled in mammalian cells. They impact a wide range of physiological functions, including osmoregulation, tissue homeostasis, and growth regulation. Compared to size, density variation for a given cell type is typically much smaller, implying that cell-type-specific density plays an important role in cell function. However, little is known about how cell density affects cell function or how it is regulated. Current tools for intracellular cell density measurements are limited to either suspended cells or cells grown on 2D substrates, neither of which recapitulate the physiology of single cells in intact tissue. While optical measurements have the potential to noninvasively measure cell density

Wild loquats (Eriobotrya japonica Lindl.) provide remarkable genetic resources for studying domestication and breeding improved varieties. Herein, we generate the first high-quality chromosome-level genome assembly of wild loquat, with 45 791 predicted protein-coding genes. Analysis of comparative genomics indicated that loquat shares a common ancestor with apple and pear, and a recent whole-genome duplication event occurred in loquat prior to its divergence. Genome resequencing showed that the loquat germplasms can be distinctly classified into wild and cultivated groups, and the commercial cultivars have experienced allelic admixture. Compared with cultivated loquats, the wild loquat genome showed very few selected genomic regions and had higher levels of genetic diversity. However, whole-genome scans of selective sweeps were mainly related to fruit quality, size, and flesh color during the domestication process. Large-scale transcriptome and metabolome analyses were further performed to identify differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) in wild and cultivated loquats at various fruit development stages. Unlike those in wild loquat, the key DEGs and DAMs involved in carbohydrate metabolism, plant hormone signal transduction, flavonoid biosynthesis, and carotenoid biosynthesis were significantly regulated in cultivated loquats during fruit development. These high-quality reference genome, resequencing, and large-scale transcriptome/metabolome data provide valuable resources for elucidating fruit domestication and molecular breeding in loquat.

In the Chinese society, border agents in channel transactions will choose different opportunistic behavior response strategies to the tolerance of other members based on the relationship between the two parties. Based on 206 valid questionnaires collected, structural equation model and regression analysis were used to investigate the influence of opportunistic behavior tolerance on response strategy selection. The results show that the channel boundary personnel's tolerance to opportunistic behavior (based on work or personal) negatively influences their choice of a positive response strategy and positively influences their choice of a negative response strategy. Among the mediating effects of contract formulation, transaction terms have a positive effect on the choice of negative response strategies based on the work and individual opportunistic behavior tolerance and have no mediating effect on the choice of positive response strategies; the contingency clause has no mediating effect on the choice of positive response strategies based on individual opportunistic behavior tolerance.

Label-free multiphoton microscopy is a powerful platform for biomedical imaging. Recent advancements have demonstrated the capabilities of transient absorption microscopy (TAM) for label-free quantification of hemoglobin and stimulated Raman scattering (SRS) microscopy for pathological assessment of label-free virtual histochemical staining. We propose the combination of TAM and SRS with two-photon excited fluorescence (TPEF) to characterize, quantify, and compare hemodynamics, vessel structure, cell density, and cell identity in vivo between age groups. In this study, we construct a simultaneous nonlinear absorption, Raman, and fluorescence (SNARF) microscope with the highest reported in vivo imaging depth for SRS and TAM at 250-280 μm to enable these multimodal measurements. Using machine learning, we predict capillary-lining cell identities with 90% accuracy based on nuclear morphology and capillary relationship. The microscope and methodology outlined herein provides an exciting route to study several research topics, including neurovascular coupling, blood-brain barrier, and neurodegenerative diseases.

Hyperspectral imaging is a technique that provides rich chemical or compositional information not regularly available to traditional imaging modalities such as intensity imaging or colour imaging based on the reflection, transmission or emission of light. Analysis of hyperspectral imaging often relies on machine learning methods to extract information. Here we present a new flexible architecture—the U-within-U-Net—that can perform classification, segmentation and prediction of orthogonal imaging modalities on a variety of hyperspectral imaging techniques. Specifically, we demonstrate feature segmentation and classification on the Indian Pines hyperspectral dataset and simultaneous location prediction of multiple drugs in mass spectrometry imaging of rat liver tissue. We further demonstrate label-free fluorescence image prediction from hyperspectral stimulated Raman scattering microscopy images. The applicability of the U-within-U-Net architecture on diverse datasets with widely varying input and output dimensions and data sources suggest that it has great potential in advancing the use of hyperspectral imaging across many different application areas ranging from remote sensing, to medical imaging, to microscopy. Imaging over a wide spectral profile allows for detailed chemical characterization, but processing this data is not trivial. A new neural network architecture can process hyperspectral information from a variety of imaging techniques effectively by using the popular U-net motive around another U-net processing spatial patterns.

Microscale thermometry of aqueous solutions is essential to understand the dynamics of local heat generation and dissipation in chemical and biological systems. A wide variety of fluorescent probes have been developed to map temperature changes with submicrometer resolution, but they often suffer from the uncertainty associated with microenvironment-dependent fluorescent properties. In this work, we develop a label-free ratiometric stimulated Raman scattering (SRS) microscopy technique to quantify microscale temperature by monitoring the O-H Raman stretching modes of water. By tracking the ratio changes of the hydrogen-bonding O-H band and the isosbestic band, we can directly quantify the temperature of water-based environments in real time without exogenous contrast agents. We demonstrate real-time measurement of localized intracellular and extracellular temperature changes due to laser absorption. This high-speed nonlinear optical imaging technique has the potential for

Flower development is a vital developmental process in the life cycle of woody perennials, especially fruit trees. Herein, we used transcriptomic, proteomic, and hormone analyses to investigate the key candidate genes/proteins in loquat (Eriobotrya japonica) at the stages of flower bud differentiation (FBD), floral bud elongation (FBE), and floral anthesis (FA). Comparative transcriptome analysis showed that differentially expressed genes (DEGs) were mainly enriched in metabolic pathways of hormone signal transduction and starch and sucrose metabolism. Importantly, the DEGs of hormone signal transduction were significantly involved in the signaling pathways of auxin, gibberellins (GAs), cytokinin, ethylene, abscisic acid (ABA), jasmonic acid, and salicylic acid. Meanwhile, key floral integrator genes FLOWERING LOCUS T (FT) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1) and floral meristem identity genes SQUAMOSA PROMOTER BINDING LIKE (SPL), LEAFY (LFY), APETALA1 (AP1), and AP2 were significantly upregulated at the FBD stage. However, key floral organ identity genes AGAMOUS (AG), AP3, and PISTILLATA (PI) were significantly upregulated at the stages of FBE and FA. Furthermore, transcription factors (TFs) such as bHLH (basic helix-loop-helix), NAC (no apical meristem (NAM), Arabidopsis transcription activation factor (ATAF1/2) and cup-shaped cotyledon (CUC2)), MYB_related (myeloblastosis_related), ERF (ethylene response factor), and C2H2 (cysteine-2/histidine-2) were also significantly differentially expressed. Accordingly, comparative proteomic analysis of differentially accumulated proteins (DAPs) and combined enrichment of DEGs and DAPs showed that starch and sucrose metabolism was also significantly enriched. Concentrations of GA3 and zeatin were high before the FA stage, but ABA concentration remained high at the FA stage. Our results provide abundant sequence resources for clarifying the underlying mechanisms of the flower development in loquat.

Double-flower Eriobotrya japonica, of which one phenotype is homeotic transformation of sepals into petals, is a new germplasm for revealing the molecular mechanisms underlying the floral organ transformation. Herein, we analyzed the sequence, expression pattern and functional characterization of EjPI, which encoded a B-class floral homeotic protein referred to as PISTILLATA ortholog, from genetically cognate single-flower and double-flower E. japonica. Phylogenetic analysis suggested that the EjPI gene was assigned to the rosids PI/GLO lineage. Analysis of protein sequence alignments showed that EjPI has typical domains of M, I, K, and C, and includes a distinctive PI motif at the C-terminal region. Compared with asterids PI/GLO lineage, the K1 and K3 subdomains of EjPI both contain a single amino acid difference. Subcellular localization of EjPI was determined to be in the nucleus. Expression pattern analysis revealed that EjPI expressed not only in petals, filament, and anther in single-flower E. japonica, but also in petaloid sepals in double-flower E. japonica. Meanwhile, there were high correlation between EjPI transcript level and petaloid area within a sepal. Furthermore, 35S::EjPI transgenic wild-type Arabidopsis caused the homeotic transformation of the first whorl sepals into petaloid sepals. Ectopic expression of EjPI in transgenic pi-1 mutant Arabidopsis rescued normal petals and stamens. These results suggest expression pattern of EjPI is associated with the formation of petaloid sepal. Our study provides the potential application of EjPI for biotechnical engineering to create petaloid sepals or regulate floral organ identity in angiosperms.