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Scanning Electrochemical Microscopy (SECM) has been used as a non-invasive electrochemical technique for studying cellular processes. SECM enables the quantification of cellular metabolites in real-time providing a deeper understanding of cellular responses to external stimuli. SECM imaging of living cells requires maintaining an ideal physiological environment to ensure reliable data collection on cellular reactivity. The cellular response can be directly influenced by physicochemical parameters including cell media composition, temperature and light exposure. This research demonstrates the effect of media composition on the electrochemical current signal of adenocarcinoma cervical cancer (HeLa) cells during SECM measurements using ferrocenemethanol as a redox mediator. Investigated media that are commonly used as electrolyte, are phosphate buffered saline (PBS), and Dulbecco's modified Eagle's medium (DMEM) in the absence and presence of fetal bovine serum (FBS). In addition, this research demonstrates that fluctuating light illumination impacts the stability of the cellular electrochemical current response. Our findings reveal that media composition and illumination are important parameters that must be carefully considered and monitored during SECM live cell imaging. [ABSTRACT FROM AUTHOR]

Fluorescence labeling via fluorescent proteins (FPs) or immunofluorescence has been routinely applied for microscopic imaging of specific proteins. However, due to these over-weight and oversized labels (e.g. GFP, 238 aa, 27 kDa, ∼4 nm in size), the potential physiological malfunctions of the target proteins are largely underestimated in living cells. Herein, for living cells, we report a small and minimally-invasive Raman reporter (about 2 aa and <1 kDa), which can be site-specifically introduced into proteins by genetic codon expansion. After a single unnatural amino acid (UAA) is precisely incorporated into the target protein, the strained alkyne can rapidly undergo copper-free Diels–Alder cycloaddition reactions with the tetrazine-functionalized Raman reporter, which features a fine vibrational spectrum in contrast to fluorescence. In our experimental results, the UAA-based Raman tag was successfully incorporated into vimentin, histone 3.3 and huntingtin (Htt74Q) proteins in living HeLa cells and further utilized for stimulated Raman imaging. The site-specific bioorthogonal fusion of small Raman tags with intracellular proteins will pave the way for minimally-invasive protein labeling and multi-color imaging in living cells. [ABSTRACT FROM AUTHOR]

Fluorophores are essential tools for optical imaging and biomedical research. Their synthetic modification to incorporate new functions, however, remains a challenging task. Conventional strategies rely on linear synthesis in which a parent framework is gradually extended. We here designed and synthesized a versatile library of multi‐functional fluorophores via a scaffold‐based Ugi four‐component reaction (U‐4CR). The adaptability of the scaffold is achieved through modification of starting materials. This allows to use a small range of starting materials for the creation of fluorogenic probes that can detect reactive‐oxygen species and where the localization into subcellular organelles or membranes can be controlled. We present reaction yields ranging from 60 % to 90 % and discovered that some compounds can even function as imaging and therapeutic agents via Fenton chemistry inducing pyroptosis in living cancer cells. Our study underlines the potential of scaffold‐based synthesis for versatile creation of functional fluorophores and their applications. [ABSTRACT FROM AUTHOR]

The formation of a robust, self‐healing hydrogel of a novel pyrene‐appended dipeptide, Py‐E‐A (L‐Glutamic acid short as E; L‐Alanine short as A) is demonstrated. Detailed studies suggest that nanoscopic fibers with a length of several micrometers have formed by chiral self‐organization of Py‐E‐A gelators. Additionally, live human PBMCs imaging is shown using the Py‐E‐A fluorophore. Interestingly, electron‐rich Py‐E‐A couples with electron‐deficient NDI‐β‐A (β‐Alanine short as β‐A) by charge transfer (CT) complexation and forms stable deep violet‐colored CT super‐hydrogel. X‐ray diffraction, DFT, and 2D ROESY NMR studies suggest lamellar packing of both Py‐E‐A and the alternating CT stack in its hydrogel matrixes. Supramolecular chirality of the Py‐E‐A donor can be altered by adding an achiral acceptor NDI‐β‐A. Notably, the fibers of the CT hydrogel are found to be even thinner than the Py‐E‐A fibers, which, in turn, makes the CT hydrogel more tolerant to the applied strain. Further, the self‐healing and injectable properties of the hydrogels are shown. Finally, the magneto‐responsive behavior of the Py‐E‐A and CT hydrogels loaded with spin‐canted Cu‐ferrite (Cu0.6Zn0.4Fe2O4) nanoparticles (NPs) is demonstrated. The presence of magnetic NPs within the hydrogels has changed the fibrous morphology to rod‐like nanoclusters. [ABSTRACT FROM AUTHOR]

The brain's ability to integrate sensory and motor information allows us to maintain a sense of orientation in space, a process in which head‐direction cells play a key role. While these neurons have been studied extensively in mammals, their presence and function in non‐mammalian species remain less understood. Here, I summarize the research work for my PhD thesis, where we explore the interpeduncular nucleus (IPN) in zebrafish, a lesser known brain region, using whole‐brain electron microscopy and calcium imaging techniques. We identified a novel population of unipolar neurons, with their activity exhibiting a dynamic, rotational pattern during head movements, even in the absence of sensory cues. This population mirrors the functionality of head‐direction cells observed in mammals, suggesting a conserved mechanism for spatial orientation across vertebrates. Our findings reveal the potential of the zebrafish IPN as a vertebrate model for studying ring attractor networks, a theoretical framework previously used to explain head‐direction cell activity. These results pave the way for future research on how motor and sensory signals converge in the vertebrate brain to maintain spatial orientation. [ABSTRACT FROM AUTHOR]

Image-based cell profiling is a powerful tool that compares perturbed cell populations by measuring thousands of single-cell features and summarizing them into profiles. Typically a sample is represented by averaging across cells, but this fails to capture the heterogeneity within cell populations. We introduce CytoSummaryNet: a Deep Sets-based approach that improves mechanism of action prediction by 30–68% in mean average precision compared to average profiling on a public dataset. CytoSummaryNet uses self-supervised contrastive learning in a multiple-instance learning framework, providing an easier-to-apply method for aggregating single-cell feature data than previously published strategies. Interpretability analysis suggests that the model achieves this improvement by downweighting small mitotic cells or those with debris and prioritizing large uncrowded cells. The approach requires only perturbation labels for training, which are readily available in all cell profiling datasets. CytoSummaryNet offers a straightforward post-processing step for single-cell profiles that can significantly boost retrieval performance on image-based profiling datasets. Author summary: Image-based cell profiling experiments measure thousands of features in millions of cells in microscopy images. Measuring cell response to various treatments in this way has proven powerful for many applications in biological research and drug discovery. However, each cell population responding to a treatment is usually population-averaged, losing information about the diversity of cells within each sample. We developed CytoSummaryNet, a machine learning method that substantially improves the ability to predict a treatment's mechanism of action from cell imaging profiles. Rather than averaging all cells equally, it learns which cells are most informative; it appears to focus on large, uncluttered cells while downweighting dividing cells and debris. Training CytoSummaryNet only requires information about which samples were exposed to which treatments, making it simpler to implement than previous techniques. By learning biologically relevant ways to summarize single-cell data into treatment profiles, CytoSummaryNet could help scientists leverage cell heterogeneity to extract richer insights from cell imaging screens and ultimately accelerate the development of new therapies. [ABSTRACT FROM AUTHOR]

Manipulating and optimizing the room temperature phosphorescence (RTP) behavior of organic luminophores is highly desirable for exploring their extensive potential applications. However, achieving tailored and precise control over RTP emission colors remains a challenging task. Herein, three regioisomers, namely o‐Br, m‐Br, and p‐Br, are developed by directly bonding the nitrogen atom of commercial carbazole and the sulfur atom of the sulfonylbenzene group. This design not only features the bromine substitution at different positions but also leads to the formation of N─S═O resonance linkages. With the variation in position of bromine from ortho to para, the RTP color changes from yellow to red. Experimental investigations and theoretical calculations demonstrate that this color variation arises from the modulation of contributions from different canonical forms of N─S═O, which can be attributed to the regioisomerization of bromine atoms for fine‐tuning intra‐ and intermolecular interactions. Additionally, the in vitro cell imaging is successfully realized by employing red RTP nanoparticles prepared from the p‐Br luminophore. This study reveals a simple and effective approach for customizing RTP emission colors through the integration of isomerization and resonance variation control. [ABSTRACT FROM AUTHOR]

The Chemical Communications journal, published by The Royal Society of Chemistry, features a variety of research articles on topics such as luminescent lanthanide metallopeptides, CO2 electrolysis, and supramolecular palladium complexes for cancer therapy. The journal covers a range of scientific discoveries in the field of chemistry, including studies on high-entropy alloy nanomaterials, asymmetric radical reactions, and novel nitrogen-doped porous carbon nanospheres for supercapacitors. The articles provide valuable insights into cutting-edge research in the chemical sciences, showcasing innovative approaches and applications in the field. [Extracted from the article]

Lanthanide ions display unique luminescent properties that make them particularly attractive for the development of bioprobes, including long-lived excited states that allow the implementation of time-gated experiments and the elimination of background fluorescence associated with biological media, as well as narrow emission bands in comparison with typical organic fluorophores, which allow ratiometric and multiplex assays. These luminescent complexes can be combined with peptide ligands to endow them with additional targeting, responsiveness, and selectivity, thus multiplying the opportunities for creative probe design. In this feature article we will present some of the main strategies that researchers have used to develop lanthanide metallopeptide probes for the detection of proteins and nucleic acids, as well as for monitoring enzymatic activity and cellular imaging. [ABSTRACT FROM AUTHOR]

Visual observing muscle tissue regeneration is used to measure experimental effect size in biological research to discover the mechanism of muscle strength decline due to illness or aging. Quantitative computer imaging analysis for support evaluating the recovery phase has not been established because of the localized nature of recovery and the difficulty in selecting image features for cells in regeneration. We constructed MyoRegenTrack for segmenting cells and classifying their regeneration phase in hematoxylin–eosin (HE) stained images. A straightforward approach to classification is supervised learning. However, obtaining detailed annotations for each fiber in a whole slide image is impractical in terms of cost and accuracy. Thus, we propose to learn individual recovery phase classification utilizing the proportions of cell class depending on the days after muscle injection to induce regeneration. We extract implicit multidimensional features from the HE-stained tissue images and train a classifier using weakly supervised learning, guided by their class proportion for elapsed time on recovery. We confirmed the effectiveness of MyoRegenTrack by comparing its results with expert annotations. A comparative study of the recovery relation between two different muscle injections shows that the analysis result using MyoRegenTrack is consistent with findings from previous studies. [ABSTRACT FROM AUTHOR]

Uncovering fine‐grained phenotypes of live cell dynamics is pivotal for a comprehensive understanding of the heterogeneity in healthy and diseased biological processes. However, this endeavor poses significant technical challenges for unsupervised machine learning, requiring the extraction of features that not only faithfully preserve this heterogeneity but also effectively discriminate between established biological states, all while remaining interpretable. To tackle these challenges, a self‐training deep learning framework designed for fine‐grained and interpretable phenotyping is presented. This framework incorporates an unsupervised teacher model with interpretable features to facilitate feature learning in a student deep neural network (DNN). Significantly, an autoencoder‐based regularizer is designed to encourage the student DNN to maximize the heterogeneity associated with molecular perturbations. This method enables the acquisition of features with enhanced discriminatory power, while simultaneously preserving the heterogeneity associated with molecular perturbations. This study successfully delineated fine‐grained phenotypes within the heterogeneous protrusion dynamics of migrating epithelial cells, revealing specific responses to pharmacological perturbations. Remarkably, this framework adeptly captured a concise set of highly interpretable features uniquely linked to these fine‐grained phenotypes, each corresponding to specific temporal intervals crucial for their manifestation. This unique capability establishes it as a valuable tool for investigating diverse cellular dynamics and their heterogeneity. [ABSTRACT FROM AUTHOR]

G protein-coupled receptors (GPCRs), crucial for diverse physiological responses, have traditionally been investigated in their monomeric form. However, some GPCRs can form heteromers, revealing complexity in their functional characteristics such as ligand binding properties, downstream signaling pathways, and trafficking. Understanding GPCR heteromers is crucial in both physiological contexts and drug development. Here, we review the methodologies for investigating physical interactions in GPCR heteromers, including co-immunoprecipitation, proximity ligation assays, interfering peptide approaches, and live cell imaging techniques based on resonance energy transfer and bimolecular fluorescence complementation. In addition, we discuss recent advances in live cell imaging techniques for exploring functional features of GPCR heteromers, for example, circularly permuted fluorescent protein-based GPCR biosensors, TRUPATH, and nanobody-based GPCR biosensors. These advanced biosensors and live cell imaging technologies promise a deeper understanding of GPCR heteromers, urging a reassessment of their physiological importance and pharmacological relevance. Highlights: The discovery of G protein-coupled receptor (GPCR) heteromers has revealed a new level of complexity in their functional characteristics. This review explores a variety of methodologies and live cell imaging technologies for the investigation of physical interactions in GPCR heteromers and their functional alterations. Live cell imaging technologies such as fluorescent protein-based biosensors promise a deeper understanding of GPCR heteromers, urging a reassessment of their physiological importance and pharmacological relevance. [ABSTRACT FROM AUTHOR]

Self-labeling protein (SLP) tags, such as HaloTag, have gained considerable interest as advanced tools for live cell labeling. However, the chloroalkane-based substrates that can be directly used for protein labeling are limited. Here, we report two bioorthogonal small molecule linkers, chloroalkane-tetrazine (CA-Tz) and chloroalkane-azide (CA-N3), which can penetrate cell membranes and facilitate click chemistry-based labeling in live cells. We compare their labeling capability using two clickable silicon rhodamine dyes (SiR-PEG3-TCO and SiR-PEG4-DBCO). Confocal imaging results demonstrate that using CA-Tz and SiR-PEG3-TCO dye exhibits superior intracellular labeling with low nonspecific signals. We subsequently compared the photostability of SiR dyes with that of green fluorescent proteins (mEmerald). Total internal reflection fluorescence (TIRF) imaging indicates that SiR dyes exhibit superior photostability under identical excitation conditions, making them suitable for long-term cell imaging. Furthermore, SiR dyes labeling also shows high structure retention for the fourth-order super-resolution optical fluctuation imaging (SOFI) compared to fluorescent proteins. This study presents clickable HaloTag linkers as effective tools for live cell labeling and imaging, highlighting the high-quality labeling of chloroalkane linkers and clickable dyes for live cell imaging. [ABSTRACT FROM AUTHOR]

This study reports the development of new probes RR1 ((1E)-1-(1-(6-bromo-2-oxo-2H-chromen-3-yl)ethylidene)ethyl thiosemicarbazone) and RR2 ((1E)-1-(1-(6-bromo-2-oxo-2H-chromen-3-yl)ethylidene)phenyl thiosemicarbazone), which selectively showed fluorescence turn 'OFF' response towards Cu2+ and Al3+. Further, complexes of RR1-Cu2+ and RR2-Al3+ acted as chemosensors for the detection of L -amino acids. RR1-Cu2+ selectively detected L -arginine (fluorescence turn 'ON'), and RR2-Al3+ selectively detected L -glutamic acid (fluorescence turn 'ON'). The existence of the fluorophore–quencher complexes RR1-Cu2+ and RR2-Al3+ was confirmed by theoretical studies. Further, the chemosensors RR1-Cu2+ and RR2-Al3+ have three possible structural isomers (RR1-Cu2+- L -arginine – A, B and C) and (R2-Al3+- L -glutamic acid – D, E and F), as confirmed by theoretical studies. In vitro bio-imaging of the probes (RR1 and RR2), complexes (RR1-Cu2+ and RR2-Al3+) and complexes associated with L -arginine (RR1-Cu2+- L -arginine) and L -glutamic acid (R2-Al3+- L -glutamic acid) was performed in the MDA-MB-231 cell line using their IC50 concentrations. In addition, in vivo live cell imaging studies were conducted using C. elegans as the model organism. [ABSTRACT FROM AUTHOR]

Neuroblastoma (NB) is the most common solid extracranial neoplasm found in children and is derived from primitive sympathoadrenal neural precursor. The disease accounts for 15% of all cancer deaths in children. The mortality rate is high in patients presenting with a metastatic tumour even with extensive treatments. This signifies the need for further research towards the development of new additional therapies that can combat not only tumour growth but metastasis, especially amongst the high-risk groups. During metastasis, primary tumour cells become migratory and travel towards a capillary within the tumour. They then degrade the matrix surrounding the pericytes and endothelial cells traversing the endothelial barrier twice to establish a secondary. This led to the hypothesis that modulation of the endothelial cell junctional stability could have an influence on tumour metastasis. To test this hypothesis, agents that modulate endothelial permeability on NB cell line migration and invasion were assessed in vitro in a tissue culture model. The cAMP agonist and its antagonists were found to have no obvious effect on both SK-N-BE2C and SK-N-AS migration, invasion and proliferation. Next, NB cells were cocultured with HDMEC cells and live cell imaging was used to assess the effect of an Epac agonist on trans-endothelial cell migration of NB cells. Epac1 agonist remarkably reduced the trans-endothelial migration of both SK-N-BE2C and SK-N-AS cells. These results demonstrate that an Epac1 agonist may perhaps serve as an adjuvant to currently existing therapies for the high-risk NB patients. [ABSTRACT FROM AUTHOR]

Nonconventional luminescent materials (NLMs) are a type of organic luminescent materials that does not contain aromatic units. Due to the simplicity of the synthesis process, mild reaction conditions, good hydrophilicity and biological compatibility, NLMs have attracted much attention. Nevertheless, numerous reports indicate that NLMs can only effectively luminesce at high concentrations and in solid state, which limits their applicability in the field of cell imaging. This study addresses this limitation by designing and synthesizing oligomers P1, P2 and P3 using ethylene glycol diglycidyl ether and amine compounds containing ethylene groups. These oligomers exhibit remarkable luminescence efficiency reaching as high as 9.2% in dilute solutions (0.1 mg/mL), making them among the best NLMs in this category. Furthermore, the synthesized oligomers exhibit excitation wavelength-dependent and concentration-dependent luminescence intensity, fluorescence response to temperature and pH changes, as well as the ability to identify Fe3+, Cu2+ and Mo5+ in dilute solutions. These characteristics render them potentially useful in the for cell imaging. [ABSTRACT FROM AUTHOR]

A multifunctional fluorescent probe P based on a naphthalimide derivative for the detection of Ag+ and Hg2+ through a dual-signal was designed and characterized. P exhibited a large Stokes shift (107 nm), high selectivity, good sensitivity, and fast response time. By adjusting the testing medium and the order of reagent addition, multifunctional detection with P was achieved. The addition of Ag+ or Hg2+ to P solution in either ethanol or an ethanol–water mixture resulted in a significant quenching of fluorescence emission at 537 nm and caused a decrease in the absorbance at 440 nm accompanied by the appearance of a new absorption peak at around 340 nm, and there was an obvious color change from yellow to colorless. In contrast, the addition of other common metal ions and anions did not produce substantial spectral or color changes. The detection limit of probe P for Ag+ and Hg2+ was calculated to be 0.33 μM. The sensing mechanism was proposed and validated through MS and 1H NMR spectrometry methods. Additionally, P demonstrated the capability to recognize Ag+ and Hg2+ in living cells with satisfactory results. [ABSTRACT FROM AUTHOR]

In this paper, LD-TCF, a targeting probe for lipid droplets (LDs) with a near-infrared emission wavelength and large Stokes shift, was fabricated for polarity detection by assembling a donor–π–acceptor (D–π–A) molecule with typical twisted intramolecular charge transfer (TICT) characteristics. Surprisingly, the fluorescence emission wavelength of the newly constructed probe LD-TCF was stretched to 703 nm, and the Stokes shift was amplified to 126 nm. Furthermore, LD-TCF could specifically answer the change in polarity efficiently and did not experience interference from other biologically active materials. Importantly, LD-TCF exhibited the ability to target lipid droplets, providing valuable insights for the early diagnosis and tracking of pathophysiological processes underlying LD polarity. [ABSTRACT FROM AUTHOR]

This study presents an advanced integration of Multi-modal Raman Light Sheet Microscopy with zero-shot learning-based computational methods to significantly enhance the resolution and analysis of complex three-dimensional biological structures, such as 3D cell cultures and spheroids. The Multi-modal Raman Light Sheet Microscopy system incorporates Rayleigh scattering, Raman scattering, and fluorescence detection, enabling comprehensive, marker-free imaging of cellular architecture. These diverse modalities offer detailed spatial and molecular insights into cellular organization and interactions, critical for applications in biomedical research, drug discovery, and histological studies. To improve image quality without altering or introducing new biological information, we apply Zero-Shot Deconvolution Networks (ZS-DeconvNet), a deep-learning-based method that enhances resolution in an unsupervised manner. ZS-DeconvNet significantly refines image clarity and sharpness across multiple microscopy modalities without requiring large, labeled datasets, or introducing artifacts. By combining the strengths of multi-modal light sheet microscopy and ZS-DeconvNet, we achieve improved visualization of subcellular structures, offering clearer and more detailed representations of existing data. This approach holds significant potential for advancing high-resolution imaging in biomedical research and other related fields. [ABSTRACT FROM AUTHOR]

Phosphate is widely used in industry and agriculture fields. However, excess accumulation of PO43− causes several adverse effects on the human body and ecological environment. Consequently, it is important to develop a simple method for the detection of PO43− concentration in the ecological environment and in vivo. Herein, two caffeic acid derivative-based fluorescence probes (BAM-HM and BAM-HH) were developed for the detection of phosphate. The BAM-HM probe could detect phosphate via fluorescence enhancement at 500 nm, with the detection limit being 0.612 µM. Meanwhile, the BAM-HH probe showed a significant turn-on signal at 450 nm after the addition of phosphate, and the detection limit was calculated to be 0.318 µM. The sensing mechanism was determined by 1H NMR and MS. Furthermore, the two probes (BAM-HM and BAM-HH) were applied for PO43−detection in living cells and water samples. [ABSTRACT FROM AUTHOR]

Sialodacryoadenitis virus (SDAV) has been identified as the etiological agent responsible for the respiratory system and salivary gland infections in rats. The existing literature on SDAV infections is insufficient to address the topic adequately, particularly in relation to the central nervous system. In order to ascertain how SDAV gains access to neuronal cells and subsequently exits, our attention was focused on the small molecule valosin-containing protein (VCP), which is an ATPase. VCP is acknowledged for its function in the ubiquitin-mediated proteasomal degradation of proteins, including those of viral origin. To ascertain the potential influence of VCP on SDAV replication and egress, high-content screening was employed to determine the viral titer and protein content. Western blot analysis was employed to ascertain the relative expression of VCP. Real-time imaging of SDAV-infected cells and confocal imaging for qualitative morphological analysis were conducted. The Eeyarestatin I (EerI) inhibitor was employed to disrupt VCP involvement in the endoplasmic reticulum-associated protein degradation pathway (ERAD) in both pre- and post-incubation systems, with concentrations of 5 μM/mL and 25 μM/mL, respectively. We demonstrated for the first time that SDAV productively replicates in cultured primary neurons. VCP expression is markedly elevated during SDAV infection. The application of 5 μM/mL EerI in the post-treatment system yielded a statistically significant inhibition of the SDAV yield. It is likely that this modulates the efficacy of virion assembly by arresting viral proteins in the submembrane area. [ABSTRACT FROM AUTHOR]

Fluorometric determination of different biologically, industrially, and environmentally important analytes is a powerful technique because this technique has excellent selectivity, high sensitivity, rapid photoluminescence response, low cost, applicability to bioimaging, and low detection limit. Fluorescence imaging is a powerful technique for screening different analytes in the living system. Heterocyclic organic compounds have been extensively used as a fluorescence chemosensor for the determination of different biologically important cations like Co2+, Zn2+, Cu2+, Hg2+, Ag+, Ni2+, Cr3+, Al3+, Pd2+, Fe3+ Pt2+, Mn2+, Sn2+, Pd2+, Au3+, Pd2+, Cd2+, Pb2+ and other ions in biological and environmental systems. These compounds also showed significant biological applications such as anti-cancer, anti-ulcerogenic, antifungal, anti-inflammatory, anti neuropathic, antihistaminic, antihypertensive, analgesic, antitubercular, antioxidant, antimalarial, antiparasitic, antiglycation, antiviral anti-obesity, and antibacterial potency. In this review, we summarize the heterocyclic organic compounds based on fluorescent chemosensors and their applications in bioimaging studies for the recognition of different biologically important metal ions. [ABSTRACT FROM AUTHOR]

[Display omitted] • Morphological Biomarkers of adipogenesis stages was established with holotomography. • Quantified lipid droplets, cellular shape, and nuclear features distinguish stages. • Human UC-MSCs were employed to create markers, offering a non-invasive approach. Adipogenesis involves complex changes in gene expression, morphology, and cytoskeletal organization. However, the quantitative analysis of live cell images to identify their stages through morphological markers is limited. Distinct adipogenesis markers on human umbilical cord-derived mesenchymal stem cells (UC-MSCs) were identified through holotomography, a label-free live cell imaging technique. In the MSC-to-preadipocyte transition, the nucleus-to-cytoplasm ratio (0.080 vs. 0.052) and lipid droplet (LD) refractive index variation decreased (0.149 % vs. 0.061 %), whereas the LD number (20 vs. 65) increased. This event was also accompanied by the downregulation and upregulation of THY1 and Preadipocyte Factor-1 (PREF-1), respectively. In the preadipocyte to immature adipocyte shift, cell sphericity (0.20 vs. 0.43) and LD number (65 vs. 200) surged, large LDs (>10 μm3) appeared, and the major axis of the cell was reduced (143.7 μm vs. 83.12 μm). These findings indicate features of preadipocyte and immature adipocyte stages, alongside the downregulation of PREF-1 and upregulation of Peroxisome Proliferator-Activated Receptor gamma (PPARγ). In adipocyte maturation, along with PPARγ and Fatty Acid-Binding Protein 4 upregulation, cell compactness (0.15 vs. 0.29) and sphericity (0.43 vs. 0.59) increased, and larger LDs (>30 μm3) formed, marking immature and mature adipocyte stages. The study highlights the distinct adipogenic morphological biomarkers of adipogenesis stages in UC-MSCs, providing potential applications in biomedical and clinical settings, such as fostering innovative medical strategies for treating metabolic disease. [ABSTRACT FROM AUTHOR]

[Display omitted] • Do Plants have intelligence system to sense different sources of sound vibrations? • SV treatment triggers short-term molecular reprogramming, long-term changes in plant architecture, and developmental and adaptive responses. • Plants use SVs for their mode of communication and alarming signal. • Role of different molecular players such as cell wall and plasma membrane and their signaling molecules involved in SV perception and signal transduction are highlighted. • High-throughput tools to decipher the molecular complexity of plant SV signaling. • How plants balance trade-offs mechanism between SV and other stimuli are addressed. How plants emit, perceive, and respond to sound vibrations (SVs) is a long-standing question in the field of plant sensory biology. In recent years, there have been numerous studies on how SVs affect plant morphological, physiological, and biochemical traits related to growth and adaptive responses. For instance, under drought SVs navigate plant roots towards water, activate their defence responses against stressors, and increase nectar sugar in response to pollinator SVs. Also, plants emit SVs during stresses which are informative in terms of ecological and adaptive perspective. However, the molecular mechanisms underlying the SV perception and emission in plants remain largely unknown. Therefore, deciphering the complexity of plant-SV interactions and identifying bonafide receptors and signaling players will be game changers overcoming the roadblocks in phytoacoustics. The aim of this review is to provide an overview of recent developments in phytoacoustics. We primarily focuss on SV signal perception and transduction with current challenges and future perspectives. Timeline breakthroughs in phytoacoustics have constantly shaped our understanding and belief that plants may emit and respond to SVs like other species. However, unlike other plant mechanostimuli, little is known about SV perception and signal transduction. Here, we provide an update on phytoacoustics and its ecological importance. Next, we discuss the role of cell wall receptor-like kinases, mechanosensitive channels, intracellular organelle signaling, and other key players involved in plant-SV receptive pathways that connect them. We also highlight the role of calcium (Ca2+), reactive oxygen species (ROS), hormones, and other emerging signaling molecules in SV signal transduction. Further, we discuss the importance of molecular, biophysical, computational, and live cell imaging tools for decoding the molecular complexity of acoustic signaling in plants. Finally, we summarised the role of SV priming in plants and discuss how SVs could modulate plant defense and growth trade-offs during other stresses. [ABSTRACT FROM AUTHOR]

Lysosomes are involved in a myriad of cellular functions, such as degradation of macromolecules, endocytosis and exocytosis, modulation of several signaling pathways, and regulation of cell metabolism. To fulfill these diverse functions, lysosomes can undergo several dynamic changes in their content, size, pH, and location within cells. Here, we studied some of these parameters during embryonic chick skeletal muscle cells. We used an anti‐lysosome‐associated membrane protein 2 (LAMP2) antibody to specifically determine the intracellular localization of lysosomes in these cells. Our data shows that lysosomes are highly enriched in the perinuclear region of chick embryonic muscle cells. We also showed that the wingless signaling pathway (Wnt)/β‐catenin signaling pathway can modulate the location of LAMP2 in chick myogenic cells. Our results highlight the role of lysosomes during muscle differentiation and particularly the presence of a subcellular population of lysosomes that are concentrated in the perinuclear region of muscle cells. [ABSTRACT FROM AUTHOR]

[Display omitted] Fluorescent microspheres are of significant interests due to their wide applications in biotechnology fields. However, their preparation presents several challenges, such as the need for dye labeling, the complexity of materials and often sophisticated preparation conditions. Here a simple process for hydrophilic and crosslinked polyurethane (CPU) microspheres, with carboxyl groups on the surface via one-step precipitation polymerization in 40 min, is presented. The microsphere size is easily adjusted by varying experimental conditions. CPU microspheres exhibit high thermal and pH stability with good redispersibility in water, and emit fluorescence without any modification or dye labeling. The emission mechanism is discussed. CPU microspheres are used as fluorescent probe to detect 4-nitrophenol (4-NP) based on their emission in UV light region, with excellent selectivity and sensitivity. In addition, they are reusable with detection limit unchanged after 7 cycles of reuses, a significant feature of this work. The mechanism of fluorescence detection is thoroughly explored and ascribed to the internal filtration effect. Based on the emission in visible light region, CPU microspheres are used as a model of PU microplastics (MPs) to visualize their biodistribution in HeLa and macrophage cells, as well as in zebrafish larvae, providing a reliable tracer for the visualization and tracking of PU MPs in organisms. [ABSTRACT FROM AUTHOR]

Acute Lymphoblastic Leukemia (ALL) is one of the most common cancers among children under the age of 15. An increasing awareness to detect early leukemia can be avoided preventable deaths from leukemia. Accurate leukemia cells image classification plays a crucial role in reducing health risks by enabling early diagnosis, which helps in mitigating the severity of the disease through timely and targeted treatment The detection of Acute Lymphoblastic Leukemia incurs significant costs in terms of time. We have developed an alternative method, A Modified EfficientNetV2-S to detect Acute Lymphoblastic Leukemia quickly, accurately, and affordably. We proposed a new architecture called Hybrid Kernel on EfficientNetV2-S. Our architecture integrates three convolutions with different kernels: regular convolution, dilated convolution, and depth-wise convolution. Three convolutions work together in one layer to provide detailed information about the object. The Network Encapsulation method uses convolution results to find object features from an image based on different locations and directions and provide combined convoluted information. We update the weight to customize the Networks Encapsulation feature map results iteratively. We continuously repeat this operation until the updated weight difference falls below the specified threshold. The approach we suggested is unique because it uses several kernels for convolutions to help capture various feature variations. We have evaluated our new architecture using the C-NMC-2019 dataset at three learning and epoch pairs: {0.001;5}, {0.0001;10}, and {0.00001;15}. We used 12,528 images to assess the reliability of our proposed new architecture. The results showed that the best accuracy, precision, recall, and F1-Score were 97.44%, 97.35%, 98.88%, and 97.48%, respectively. Our proposed method performance outperformed VGG19, Xception, RestNet50, and EfficientNetB0. [ABSTRACT FROM AUTHOR]

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 labeling 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 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 Arabidopsis centromere-specific guide RNA, we demonstrate target-specific labeling with a fluorescence-labeled NbALFA nanobody. The dCas9 protein possessing multiple copies of the ALFA-tag, in combination with a minibody and fluorescence-labeled 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 Nicotiana 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 method with CRISPR–FISH, demonstrating effective labeling of Arabidopsis centromeres. [ABSTRACT FROM AUTHOR]

Individual cells grown in culture exhibit remarkable differences in their growth, with some cells capable of forming large clusters, while others are limited or fail to grow at all. While these differences have been observed across cell lines and human samples, the growth dynamics and associated cell states remain poorly understood. In this study, we performed clonal tracing through imaging and cellular barcoding of an in vitro model of esophageal epithelial cells (EPC2-hTERT). We found that about 10% of clones grow exponentially, while the remaining have cells that become non-proliferative leading to a halt in the growth rate. Using mathematical models, we demonstrate two distinct growth behaviors: exponential and logistic. Further, we discovered that the propensity to grow exponentially is largely heritable through four doublings and that the less proliferative clones can become highly proliferative through increasing plating density. Combining barcoding with single-cell RNA-sequencing (scRNA-seq), we identified the cellular states associated with the highly proliferative clones, which include genes in the WNT and PI3K pathways. Finally, we identified an enrichment of cells resembling the highly proliferative cell state in the proliferating healthy human esophageal epithelium. Author summary: Single cells derived from tissues and cell lines can have variable abilities to grow in culture. Understanding the molecular differences that lead to this variability has the potential to reveal regulators of growth and to improve in vitro culture for tissue regeneration. In this study, we investigated differences in growth between esophageal epithelial cells and found that these differences are non-genetic in origin. Through quantitative analysis of clonal growth at different time points, we show that the heterogeneity in growth can be modeled as two separate populations, one that is highly proliferative and grows exponentially, and another that is less proliferative and is able to transition into a non-proliferative state. Tracking clones through cell barcoding with scRNA-seq, we define the molecular differences between these populations and their associated signaling pathways. Together, this work highlights the contributions of non-genetic heterogeneity in proliferation, which has applications for tissue regeneration and stem cell biology. [ABSTRACT FROM AUTHOR]

Supramolecular assemblies of stimuli-responsive amphiphilic molecules have been of utmost interest in targeted drug delivery applications, owing to their capability of sequestering drug molecules in one set of conditions and releasing them in another. To minimize undesired disassembly and stabilize noncovalently encapsulated drug molecules, the strategy of core or shell cross-linking has become a fascinating approach to constructing cross-linked polymeric or small molecule-based nanoassemblies. In this article, we discuss the design and synthetic strategies for cross-linked nanoassemblies from small molecule-based amphiphiles, with robust stability and enhanced drug encapsulation capability. We highlight their potential biomedical applications, particularly in drug or gene delivery, and cell imaging. This feature article offers a comprehensive overview of the recent developments in the application of small molecule-based covalently cross-linked nanocarriers for materials and biomedical applications, which may inspire the use of these materials as a potential drug delivery system for future chemotherapeutic applications. [ABSTRACT FROM AUTHOR]

The correction notice in the Scientific Reports journal addresses errors in the original article regarding the assembly of figures and images. Specifically, Figure 4B and supplementary Figures S4A, S4C, S4B, and S4D contain overlapping images due to errors during preparation and assembly. The corrected figures show the effect of temperature on exosome uptake by cancer cells and the impact of different uptake inhibitors on labeled exosomes. The authors of the article are Sherif E. Emam, Hidenori Ando, Amr S. Abu Lila, Taro Shimizu, Keiichiro Okuhira, Yu Ishima, Mahmoud A. Mahdy, Fakhr-eldin S. Ghazy, Ikuko Sagawa, and Tatsuhiro Ishida. [Extracted from the article]

Most fluorogens with aggregation-induced emission (AIE) characteristics are hydrophobic and most common sugars are hydrophilic and naturally nontoxic. The combination of AIEgens and sugars can construct glycosyl AIEgens with the advantages of good water-solubility, low fluorescent background and satisfactory biocompatibility. Based on the specific reaction or binding with analytes to change the conjugate system or restrict intramolecular motions, glycosyl AIEgens can be used as powerful tools for detecting bioactive molecules or imaging living cells. In this feature article, we summarize recent advances in sugar-based AIE luminogens and their applications in biosensing and imaging. The sugar units could significantly increase the solubility, biocompatibility, target activity, and chemical modifying capacity and often decrease the background fluorescence of the AIE probes. Corresponding studies not only expand the application fields of AIEgens but also provide effective tools for broad carbohydrate research. [ABSTRACT FROM AUTHOR]

ABSTRACT Clusters of similar or dissimilar objects are encountered in many fields. Frequently used approaches treat each cluster's central object as latent. Yet, often objects of one or more types cluster around objects of another type. Such arrangements are common in biomedical images of cells, in which nearby cell types likely interact. Quantifying spatial relationships may elucidate biological mechanisms. Parent‐offspring statistical frameworks can be usefully applied even when central objects (“parents”) differ from peripheral ones (“offspring”). We propose the novel multivariate cluster point process (MCPP) to quantify multi‐object (e.g., multi‐cellular) arrangements. Unlike commonly used approaches, the MCPP exploits locations of the central parent object in clusters. It accounts for possibly multilayered, multivariate clustering. The model formulation requires specification of which object types function as cluster centers and which reside peripherally. If such information is unknown, the relative roles of object types may be explored by comparing fit of different models via the deviance information criterion (DIC). In simulated data, we compared a series of models' DIC; the MCPP correctly identified simulated relationships. It also produced more accurate and precise parameter estimates than the classical univariate Neyman–Scott process model. We also used the MCPP to quantify proposed configurations and explore new ones in human dental plaque biofilm image data. MCPP models quantified simultaneous clustering of Streptococcus and Porphyromonas around Corynebacterium and of Pasteurellaceae around Streptococcus and successfully captured hypothesized structures for all taxa. Further exploration suggested the presence of clustering between Fusobacterium and Leptotrichia, a previously unreported relationship. [ABSTRACT FROM AUTHOR]

Background: Botrytis cinerea is a broad-host-range pathogen causing gray mold disease and significant yield losses of numerous crops. However, the mechanisms underlying its rapid invasion and efficient killing of plant cells remain unclear. Results: In this study, we elucidated the dynamics of B. cinerea infection in Arabidopsis thaliana by live cell imaging and dual RNA sequencing. We found extensive transcriptional reprogramming events in both the pathogen and the host, which involved metabolic pathways, signaling cascades, and transcriptional regulation. For the pathogen, we identified 591 candidate effector proteins (CEPs) and comprehensively analyzed their co-expression, sequence similarity, and structural conservation. The results revealed temporal co-regulation patterns of these CEPs, indicating coordinated deployment of effectors during B. cinerea infection. Through functional screening of 48 selected CEPs in Nicotiana benthamiana, we identified 11 cell death-inducing proteins (CDIPs) in B. cinerea. Conclusions: The findings provide important insights into the transcriptional dynamics and effector biology driving B. cinerea pathogenesis. The rapid infection of this pathogen involves the temporal co-regulation of CEPs and the prominent role of CDIPs in host cell death. This work highlights significant changes in gene expression associated with gray mold disease, underscoring the importance of a diverse repertoire of effectors crucial for successful infection. [ABSTRACT FROM AUTHOR]

ABSTRACT A highly selective fluorescent sensor Por‐RBO for determination of Hg2+ was designed and synthesized with Rhodamine B and Tetraphenylporphyrin. The sensor can determine the Hg2+ of a solution within the pH 5.0–8.0, free from interference of other metal ions. When detected in buffer solution, the fluorescence was the strongest when five times the concentration of Hg2+ ion was added. Calculated by fluorescence titration method, the detection limit of sensor Por‐RBO for Hg2+ ion was as low as 0.12 μmol/L. The geometries of Por‐RBO and Por‐RBO‐Hg2+ were optimized at the B3LYP/6‐31G** level by density functional theory. The charge distribution, orbital interactions, and bonding characteristics were analyzed and compared in detail to discuss the recognition mechanism and structure–fluorescence property relationships. The results of cell fluorescence imaging and CCK‐8 experiments indicate that the sensor Por‐RBO exhibits lower cytotoxicity. Por‐RBO can be used for fluorescence distribution imaging of Hg2+ in living cells. [ABSTRACT FROM AUTHOR]

We explore the potential of fluorine‐containing small Mn2+ chelates as alternatives to perfluorinated nanoparticles, widely used as 19F MRI probes. In MnL1, the cyclohexanediamine skeleton and two piperidine rings, involving each a metal‐coordinating amide group and an appended CF3 moiety, provide high rigidity to the complex. This allows for good control of the Mn−F distance (rMnF=8.2±0.2 Å determined from 19F relaxation data), as well as for high kinetic inertness (a dissociation half‐life of 1285 h is estimated for physiological conditions). The paramagnetic Mn2+ leads to a ~150‐fold acceleration of the longitudinal 19F relaxation, with moderate line‐broadening effect, resulting in T2/T1 ratios of 0.8 (9.4 T). Owing to its inner sphere water molecule, MnL1 is a good 1H relaxation agent as well (r1=5.36 mM−1 s−1 at 298 K, 20 MHz). MnL1 could be readily visualized in 19F MRI by using fast acquisition techniques, both in phantom images and living mice following intramuscular injection, with remarkable signal‐to‐noise ratios and short acquisition times. While applications in targeted imaging or cell therapy monitoring require further optimisation of the molecular structure, these results argue for the potential of such small, monohydrated and fluorinated Mn2+ complexes for combined 19F and 1H MRI detection. [ABSTRACT FROM AUTHOR]

The mesoscope has emerged as a powerful imaging tool in biomedical research, yet its high cost and low resolution have limited its broader application. Here, we introduce the Omni-Mesoscope, a high-spatial-temporal and multimodal mesoscopic imaging platform built from cost-efficient off-the-shelf components. This system uniquely merges the capabilities of label-free quantitative phase microscopy to capture live-cell morphodynamics across thousands of cells with highly multiplexed fluorescence imaging for comprehensive molecular characterization. This Omni-Mesoscope offers a mesoscale field of view of ~5 square millimeters with a high spatial resolution down to 700 nanometers, enabling the capture of detailed subcellular features. We demonstrate its capability in delineating molecular characteristics underlying rare morphodynamic cellular phenomena, including cancer cell responses to chemotherapy and the emergence of polyploidy in drug-resistant cells. We also integrate expansion technique to enhance three-dimensional volumetric super-resolution imaging of thicker tissues, opening the avenues for biological exploration at unprecedented scales and resolutions. [ABSTRACT FROM AUTHOR]

Fluorescent Carbon Quantum Dots (CQDs) are being used in medical applications, particularly in theranostics. These Carbon Quantum Dots have been gaining more attention lately due to their potential as an effective replacement for hazardous synthetic organic dyes in a variety of biomedical applications, including live cell imaging and diagnostics. In this study, highly fluorescent Carbon Quantum Dots by one pot microwave based green route with a size of less than 10 nm, was prepared from commercially available almond resin, Prunus dulcis and conjugated with honey as additional reagent for surface functionalization. They exhibit a deep blue emission on excitation at 350 nm with an elevated quantum yield at 61%. They possess atomic nature and basic features such as high photo-stability, varying fluorescence, greater biocompatibility, and better water solubility. These fluorescent labels exhibit faster cellular invagination without disturbing the cell stability. The CQDs present cell imaging capacity with multi-coloration for visualizing the fine architecture of the nucleus naming, the nuclear membrane and nucleolus, which is linked with their varied, surface structures such as amphiphilic property and higher positive charges. These characteristics with minimal invasion have made carbon quantum dots to become the spotlight in theranostics. They can be used as alternatives to synthetic dyes for fluorescence- related cell-imaging. The intriguing fact about this approach is that it opens the possibility of combining therapy and diagnostics into one unit, which can alter how some diseases are handled and, in turn, transform the field of healthcare. [ABSTRACT FROM AUTHOR]

Image-based models that use features extracted from cell microscopy images can estimate the activity of small molecules in various biological assays. Typically, models are trained on images stained by an optimized protocol (e.g. Cell Painting) after exposure to a fairly high small molecule concentration (referred to as 'image concentration') of 10 μ M or higher. Low concentration images (e.g. 0.16 μM, 0.8 μM, 4 μM) tend to yield models with worse performance. In this work, we nevertheless report a practical use for low image concentration data. We propose the combination of well-performing models trained at higher image concentrations, with lower image concentration for inference to identify more potent compounds. We show that this approach improves on the conventional method (directly training a high-potency model) in 65 % of assays investigated in terms of AUC-ROC, and 75 % of assays in terms of RIPtoP-corrected AUC-PR. [ABSTRACT FROM AUTHOR]

Darkfield microscopy is an effective technique and well suited for use involving live and unstained biological samples, where a bulky condenser should be precisely aligned in the optical path to produce a cone of illumination light. Here, a compact darkfield illuminator is invented that has a planar shape and can work at broadband wavelengths. It can be easily incorporated into a conventional microscope as a substrate to produce hollow cones of light for large‐area darkfield illumination, resulting in a high‐contrast high‐sensitivity widefield microscopy images. Taking advantages of its planar and compact structure, this darkfield illuminator is successfully integrated with a microfluidic chip; thus, the actions and reactions of biological and chemical specimens flowing inside micrometer‐sized channels, can be real‐time imaged with high contrast and sensitivity in a large field‐of‐view (FoV). For the first time, the work integrates a compact darkfield illuminator with microfluidic chips, which produces a new platform for applications in biology, chemistry, and biomedical engineering. [ABSTRACT FROM AUTHOR]

Background/Objectives: Imaging studies have transformed the diagnosis of large vessel vasculitis (LVV) involvement in giant cell arteritis (GCA). A positron emission tomography/computed tomography (PET/CT) scan with 18-fluorodeoxyglucose (18F-FDG) has emerged as a valuable tool for assessing LVV. We aimed to determine the utility of an 18F-FDG-PET/CT scan in detecting LVV in GCA in the ARTESER registry. Methods: The ARTESER study is a large multicenter, retrospective, longitudinal, and observational study, promoted by the Spanish Society of Rheumatology. It included patients newly diagnosed with GCA across 26 tertiary hospitals from 1 June 2013 to 29 March 2019. Patients with a diagnosis of incidental GCA were included if they fulfilled specific criteria, including the ACR 1990 criteria, positive imaging examinations, or the expert clinical opinion of investigators. Differences between patients with positive and negative 18F-FDG-PET/CT scan results were analyzed using a bivariate model. A regression model assessed associations in patients with a positive scan, and the predictive capacity of the cumulative dose of glucocorticoids (GC) on PET scan outcomes was evaluated using ROC curve analysis. Results: Out of 1675 GCA patients included in the registry, 377 met the inclusion criteria of having an 18F-FDG-PET/CT scan. The majority were diagnosed with a cranial GCA phenotype, and 65% had LVV. The thoracic aorta was the most frequently affected. Cardiovascular disease, diabetes, and older age had a negative association with a positive scan outcome. The OR for having a positive 18F-FDG-PET/CTC scan was lower as the number of days increased. Depending on the cumulative dosage of the GC, the 18F-FDG-PET/CT scan showed an AUC of 0.74, with a Youden index > 60 mg/day. Conclusions: Younger patients showed a higher probability of presenting LVV as detected by the 18F-FDG-PET/CT scan. The timing of the examination and the cumulative dosage of the GC influenced the likelihood of a positive result, with earlier tests being more likely to detect inflammation. [ABSTRACT FROM AUTHOR]

The current study aimed to investigate the physicochemical properties of the natural eggshell membrane (NEM) and its protective effects against H2O2-induced oxidative stress in human chondrocytes (SW-1353). Bioactive components from NEM related to cartilage were profiled, consisting of 1.1 ± 0.07% hyaluronic acid, 1.2 ± 0.25% total sulfated glycosaminoglycans as chondroitin sulfate, 3.1 ± 0.33% collagen, and 54.4 ± 2.40% total protein. Protein was hydrolyzed up to 43.72 ± 0.76% using in vitro gastro–intestinal digestive enzymes. Peptides eluted at 9.58, 12.46, and 14.58 min using nano-LC-ESI-MS were identified as TEW, SWVE, and VYL peptides with an M/Z value of 435.1874, 520.2402, and 394.2336, respectively. Radical scavenging activity of NEM at 10 mg/mL using the ABTS assay was revealed to be 2.1 times higher than that of the positive control. NEM treatment significantly enhanced cellular SOD expression (p < 0.05). Pre-treatment with NEM (0.1, 1, and 10 mg/mL) dose-dependently reduced H2O2-induced ROS levels in SW-1353. Cell live imaging confirmed that NEM pre-treatment led to a significant reduction in apoptosis expression compared to control. Results from the present study suggest that NEM rich in cartilage protective components including hyaluronic acid, collagen, and chondroitin antioxidative peptides could be a potential therapeutic agent for osteoarthritis (OA) by scavenging oxidative stress. [ABSTRACT FROM AUTHOR]

Microscopy is essential for detecting, identifying, analyzing, and measuring small objects. Access to modern microscopy equipment is crucial for scientific research, especially in the biomedical and analytical sciences. However, the high cost of equipment, limited availability of parts, and challenges associated with transporting equipment often limit the accessibility and operational capabilities of these tools, particularly in field sites and other remote or resource-limited settings. Thus, there is a need for affordable and accessible alternatives to traditional microscopy systems. We address this challenge by investigating the feasibility of using a simple microcontroller board not only as a portable and field-ready digital microscope, but furthermore as a versatile platform which can easily be adapted to a variety of imaging applications. By adding a few external components, we demonstrate that a low-cost ESP32 camera board can be used to build an autonomous in situ platform for digital time-lapse imaging of cells. Our prototype of this approach, which we call ESPressoscope, can be adapted to applications ranging from monitoring incubator cell cultures in the lab to observing ecological phenomena in the sea, and it can be adapted for other techniques such as microfluidics or spectrophotometry. Our prototype of the ESPressoscope concept achieves a low power consumption and small size, which makes it ideal for field research in environments and applications where microscopy was previously infeasible. Its Wi-Fi connectivity enables integration with external image processing and storage systems, including on cloud platforms when internet access is available. Finally, we present several web browser-based tools to help users operate and manage our prototype's software. Our findings demonstrate the potential for low-cost, portable microscopy solutions to enable new and more accessible experiments for biological and analytical applications. [ABSTRACT FROM AUTHOR]

Background: Estimation of beta cell mass is currently restricted to evaluating pancreatic tissue samples, which provides limited information. A non-invasive imaging technique that reliably quantifies beta cell mass enables monitoring of changes of beta cell mass during the progression of diabetes mellitus and may contribute to monitoring of therapy effectiveness. We assessed the specificity of radiolabelled exendin for beta cell mass quantification in humans. Fourteen adults with pancreas tumours were injected with 111In-labeled exendin-4 prior to pancreatic resection. In resected pancreas tissue, endocrine-exocrine ratios of tracer uptake were determined by digital autoradiography and accumulation of 111In-labeled exendin-4 was compared to insulin and GLP-1 receptor staining. Of four participants, abdominal single photon emission computed tomography/computed tomography (SPECT/CT) images were acquired to quantify pancreatic uptake in vivo Results: Tracer uptake was predominantly present in the endocrine pancreas (endocrine-exocrine ratio: 3.6 [2.8–10.8]. Tracer accumulation showed overlap with insulin-positive regions, which overlapped with GLP-1 receptor positive areas. SPECT imaging showed pancreatic uptake of radiolabelled exendin in three participants. Conclusion: Radiolabelled exendin specifically accumulates in the islets of Langerhans in human pancreas tissue. The clear overlap between regions positive for insulin and the GLP-1 receptor substantiate the beta cell specificity of the tracer. Radiolabelled exendin is therefore a valuable imaging agent for human beta cell mass quantification and has the potential to be used for a range of applications, including improvement of diabetes treatment by assessment of the effects of current and novel diabetes therapies on the beta cell mass. Trial registration: ClinicalTrials.gov NCT03889496, registered 26,032,019, URL https://clinicaltrials.gov/study/NCT03889496?term=NCT03889496. ClinicalTrials.gov NCT04733508, registered 02022021, URL https://clinicaltrials.gov/study/NCT04733508. [ABSTRACT FROM AUTHOR]

To explore the potential mechanisms which O‐linked‐N‐acetylglucosaminylation (O‐GlcNAcylation) regulates osteogenesis, a publicly RNA‐seq dataset was re‐analyzed with literature‐mining and showed the primary targets of O‐GlcNAcylation in osteoblasts are mitochondria/cytoskeleton. Although the O‐GlcNAcylation‐regulated mitochondria/cytoskeleton has been extensively studied, its specific role during osteogenesis remains unclear. To address this, we knocked out Ogt (Ogt‐KO) in MC3T3‐E1 osteoblastic cells. Then, significantly reduced osteoblast differentiation, motility, proliferation, mitochondria–endoplasmic reticulum (Mito–ER) coupling, volume of ER, nuclear tubulins, and oxygen metabolism were observed in Ogt‐KO cells. Through artificial intelligence (AI)‐predicted cellular structures, the time‐lapse live cells imaging with reactive‐oxygen‐species/hypoxia staining showed that lower cell proliferation and altered oxygen metabolism in the Ogt‐KO cells were correlated with the Mito–ER coupling. Bioinformatics analysis, combined with correlated mRNA and protein expression, suggested that Ezh2 and its downstream targets (Opa1, Gsk3a, Wnt3a, Hif1a, and Hspa9) may be involved in O‐GlcNAcylation‐regulated Mito–ER coupling, ultimately impacting osteoblast differentiation. In conclusion, our findings indicate that O‐GlcNAcylation‐regulated osteoblast differentiation is linked to morphological changes in mitochondria, cytoskeleton, and ER, with Ezh2 potentially playing a crucial role. [ABSTRACT FROM AUTHOR]

Lipid droplets (LDs) represent highly dynamic organelles found in almost all organisms and play a pivotal role in regulating the balance of intracellular lipid homeostasis. Viscosity directly affects the metabolism of lipid droplets, and the abnormal change of viscosity will lead to a series of related diseases. Therefore, developing a tool that can monitor changes in intracellular lipid droplet viscosity in real-time is of great significance for disease prevention, diagnosis, and treatment. In this study, a viscosity-sensitive fluorescent probe CA-LD based on an intramolecular twisted charge transfer (TICT) mechanism was developed. The CA-LD probe exhibited strong fluorescence emission at approximately 640 nm and showed a 161-fold increase in fluorescence intensity in 99% glycerol compared to water. Additionally, the probe CA-LD demonstrated the advantages of good anti-interference, excellent biocompatibility and good photostability. Significantly, because of the probe's sensitivity to viscosity, the probe CA-LD successfully distinguished normal cells from cancer cells and has been able to visualize changes in lipid droplets' viscosity induced by oleic acid. In addition, the probe CA-LD was effectively employed to monitor changes in the viscosity of cells and zebrafish induced by different drugs. Therefore, the probe CA-LD can provide an effective means for the detection of lipid droplet viscosity changes and is of great value for physiological diagnosis, pathological analysis and medical research on related diseases. [ABSTRACT FROM AUTHOR]

Fourier ptychographic microscopy (FPM) breaks through the resolution limitations of conventional optical systems, which offer a full‐field view and high resolution without additional mechanical scanning. However, conventional image‐domain optimizations require trade‐offs between correction efficacy, data redundancy, and reconstruction accuracy. Furthermore, the existing linear time‐invariant model for actual nonlinear, time‐varying optical systems leads to forward model mismatch, complicating the corrections of the vignetting effect. To overcome these challenges and achieve stitching‐free FPM, a family of forward wavelet‐transform models (WL‐FPM) is proposed. WL‐FPM employs the reversibility of the wavelet transform for high‐fidelity reconstruction in the multiscale feature domain. The wavelet loss function is updated in each iteration, and non‐convex optimization is solved by complex back diffraction. WL‐FPM offers stitching‐free, high‐resolution, and robust reconstruction under various challenging conditions, including vignetting effects, LED position mismatch, intensity fluctuations, and high‐level noise environments, which outperform conventional FPM methods. Under a 4X objective with NA 0.1, WL‐FPM achieves a 435‐nm resolution and stitching‐free full‐field reconstruction of a 3.328 × 3.328 mm2 pathological section with distinct subcellular organelles. In live cell imaging, it provides a full‐field observation with distinct lipids in a single cell. A large number of simulation and experimental results demonstrate its potential for biomedical applications. [ABSTRACT FROM AUTHOR]

Rationale: Chronic inflammation plays an important role in alveolar tissue damage in emphysema, but the underlying immune alterations and cellular interactions are incompletely understood. Objectives: To explore disease-specific pulmonary immune cell alterations and cellular interactions in emphysema. Methods: We used single-cell mass cytometry (CyTOF) to compare the immune compartment in alveolar tissue from 15 patients with severe emphysema and 5 control subjects. Imaging mass cytometry (IMC) was applied to identify altered cell–cell interactions in alveolar tissue from patients with emphysema (n = 12) compared with control subjects (n = 8). Measurements and Main Results: We observed higher percentages of central memory CD4 T cells in combination with lower proportions of effector memory CD4 T cells in emphysema. In addition, proportions of cytotoxic central memory CD8 T cells and CD127+CD27+CD69− T cells were higher in emphysema, the latter potentially reflecting an influx of circulating lymphocytes into the lungs. Central memory CD8 T cells, isolated from alveolar tissue from patients with emphysema, exhibited an IFN-γ response upon anti-CD3 and anti-CD28 activation. Proportions of CD1c+ dendritic cells, expressing migratory and costimulatory markers, were higher in emphysema. Importantly, IMC enabled us to visualize increased spatial colocalization of CD1c+ dendritic cells and CD8 T cells in emphysema in situ. Conclusions: Using CyTOF, we characterized the alterations of the immune cell signature in alveolar tissue from patients with chronic obstructive pulmonary disease stage III or IV emphysema versus control lung tissue. These data contribute to a better understanding of the pathogenesis of emphysema and highlight the feasibility of interrogating the immune cell signature using CyTOF and IMC in human lung tissue. Clinical trial registered with (NCT04918706). [ABSTRACT FROM AUTHOR]

Activity-dependent modulations of N-methyl-D-aspartate glutamate receptor (NMDAR) trapping at synapses regulate excitatory neurotransmission and shape cognitive functions. Although NMDAR synaptic destabilization has been associated with severe neurological and psychiatric conditions, tuning NMDAR synaptic trapping to assess its clinical relevance for the treatment of brain conditions remains a challenge. Here, we report that ketamine (KET) and other clinically relevant NMDAR open channel blockers (OCBs) promote interactions between NMDAR and PDZ-domain-containing scaffolding proteins and enhance NMDAR trapping at synapses. We further show that KET-elicited trapping enhancement compensates for depletion in synaptic receptors triggered by autoantibodies from patients with anti-NMDAR encephalitis. Preventing synaptic depletion mitigates impairments in NMDAR-mediated CaMKII signaling and alleviates anxiety- and sensorimotor-gating-related behavioral deficits provoked by autoantibodies. Altogether, these findings reveal an unexpected dimension of OCB action and stress the potential of targeting receptor anchoring in NMDAR-related synaptopathies. [Display omitted] • Open channel blockers (OCBs) such as ketamine promote NMDAR synaptic trapping • OCB binding enhances interactions between NMDARs and scaffolding proteins • Ketamine prevents NMDAR depletion at synapses exposed to anti-NMDAR autoantibodies • Ketamine alleviates behavioral deficits provoked by anti-NMDAR autoantibodies Villéga, Fernandes, Jézéquel et al. report that NMDAR open channel blockers, such as ketamine, favor receptor trapping at excitatory synapses through enhanced interactions with scaffolding proteins. They further show that ketamine-elicited trapping enhancement alleviates NMDAR synaptic impairments and behavioral deficits caused by autoantibodies from patients with anti-NMDAR encephalitis. [ABSTRACT FROM AUTHOR]

γ-Aminobutyric acid (GABA) is one of the primary inhibitory neurotransmitters and encompasses 10–30% of the neurons in the CNS. Fundamentally, GABA is responsible for decreasing the nervous system activity by inhibiting the action potential and hence excitotoxicity of neurons. Owing to these neuroregulatory features, GABA has been known to be involved in numerous diseases such as schizophrenia, epilepsy, and other psychiatric conditions. Therefore, from a neuroanatomical as well as a pathological perspective, the visual localization and relative abundance research of neurotransmitters is of great importance. The intricacies of GABA release from neurons are still largely unknown; hence, it will be necessary to analyze endogenous GABA concentrations. Here, we synthesized nitrogen-doped carbon quantum dots (N-CQDs), which employ a turn-off mechanism to detect GABA in vivo. High-resolution transmission electron microscopy (HRTEM), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), and energy-dispersive X-ray spectroscopy (EDX) have been used to verify the structural characteristics and size of the N-CQDs. Using the Stern–Volmer equation, UV–vis and fluorescence spectroscopies, and time-correlated single-photon counting (TCSPC), the mechanism of fluorescence quenching was thoroughly explained. Moreover, 0.16 μM was determined to be the N-CQD detection limit. Furthermore, exogenous GABA in human cell lines and endogenous GABA in the zebrafish forebrain (telencephalon) and midbrain (optic tectum) were both successfully detected by our sensor. Thus, this probe may be used as a competent nanosensor to detect comparative GABA levels among healthy and diseased animal models with GABA neurotransmitter imbalance and neurodegeneration. [ABSTRACT FROM AUTHOR]