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Purpose: Gene‐expression reporter systems, such as green fluorescent protein, have been instrumental to understanding biological processes in living organisms at organ system, tissue, cell, and molecular scales. More than 30 years of work on developing MRI‐visible gene‐expression reporter systems has resulted in a variety of clever application‐specific methods. However, these techniques have not yet been widely adopted, so a general‐purpose expression reporter is still required. Here, we demonstrate that the manganese ion transporter Zip14 is an in vivo MRI‐visible, flexible, and robust gene‐expression reporter to meet this need. Methods: Plasmid constructs consisting of a cell type–specific promoter, gene coding for human Zip14, and a histology‐visible tag were packaged into adeno‐associated viruses. These viruses were intracranially injected into the mouse brain. Serial in vivo MRI was performed using a vendor‐supplied 3D‐MPRAGE sequence. No additional contrast agents were administered. Animals were sacrificed after the last imaging timepoint for immunohistological validation. Results: Neuron‐specific overexpression of Zip14 produced substantial and long‐lasting changes in MRI contrast. Using appropriate viruses enabled both anterograde and retrograde neural tracing. Expression of Zip14 in astrocytes also enabled MRI of glia populations in the living mammalian brain. Conclusions: The flexibility of this system as an MRI‐visible gene‐expression reporter will enable many applications of serial, high‐resolution imaging of gene expression for basic science and therapy development. [ABSTRACT FROM AUTHOR]

Background: Alzheimer's disease (AD) is an irreversible primary brain disease with insidious onset. The rise of imaging genetics research has led numerous researchers to examine the complex association between genes and brain phenotypes from the perspective of computational biology. Methods: Given that most previous studies have assumed that imaging data and genetic data are linearly related and are therefore unable to explore their nonlinear relationship, our study applied a joint depth semi-supervised nonnegative matrix decomposition (JDSNMF) algorithm to solve this problem. The JDSNMF algorithm jointly decomposed multimodal imaging genetics data into both a standard basis matrix and multiple feature matrices. During the decomposition process, the coefficient matrix A multilayer nonlinear transformation was performed using a neural network to capture nonlinear features. Results: The results using a real dataset demonstrated that the algorithm can fully exploit the association between strongly correlated image genetics data and effectively detect biomarkers of AD. Our results might provide a reference for identifying biologically significant imaging genetic correlations, and help to elucidate disease-related mechanisms. Conclusions: The diagnostic model constructed by the top features of the three modality data sets mined by the algorithm has high accuracy, and these features are expected to become new therapeutic targets for AD. [ABSTRACT FROM AUTHOR]

Although mesenchyme is essential for inducing the epithelium of ectodermal organs, its precise role in organ-specific epithelial fate determination remains poorly understood. To elucidate the roles of tissue interactions in cellular differentiation, we performed single-cell RNA sequencing and imaging analyses on recombined tissues, where mesenchyme and epithelium were switched ex vivo between two types of embryonic mouse salivary glands: the parotid gland (a serous gland) and the submandibular gland (a predominantly mucous gland). We found partial induction of molecules that define gland-specific acinar and myoepithelial cells in recombined salivary epithelium. The parotid epithelium recombined with submandibular mesenchyme began to express mucous acinar genes not intrinsic to the parotid gland. While myoepithelial cells do not normally line parotid acini, newly induced myoepithelial cells densely populated recombined parotid acini. However, mucous acinar and myoepithelial markers continued to be expressed in submandibular epithelial cells recombined with parotid mesenchyme. Consequently, some epithelial cells appeared to be plastic, such that their fate could still be modified in response to mesenchymal signaling, whereas other epithelial cells appeared to be already committed to a specific fate. We also discovered evidence for bidirectional induction: transcriptional changes were observed not only in the epithelium but also in the mesenchyme after heterotypic tissue recombination. For example, parotid epithelium induced the expression of muscle-related genes in submandibular fibroblasts that began to mimic parotid fibroblast gene expression. These studies provide the first comprehensive unbiased molecular characterization of tissue recombination approaches exploring the regulation of cell fate. [ABSTRACT FROM AUTHOR]

Background: Although rare, uveal melanoma (UM) is a life-threatening malignancy. Understanding its biology is necessary to improve disease outcome. Three-dimensional (3D) in vitro culture methods have emerged as tools that incorporate physical and spatial cues that better mimic tumor biology and in turn deliver more predictive preclinical data. Herein, we comprehensively characterize UM cells under different 3D culture settings as a suitable model to study tumor cell behavior and therapeutic intervention. Methods: Six UM cell lines were tested in two-dimensional (2D) and 3D-culture conditions. For 3D cultures, we used anchorage-dependent (AD) methods where cells were embedded or seeded on top of basement membrane extracts and anchorage-free (AF) methods where cells were seeded on agarose pre-coated plates, ultra-low attachment plates, and on hanging drops, with or without methylcellulose. Cultures were analyzed for multicellular tumor structures (MCTs) development by phase contrast and confocal imaging, and cell wellbeing was assessed based on viability, membrane integrity, vitality, apoptotic features, and DNA synthesis. Vascular endothelial growth factor (VEGF) production was evaluated under hypoxic conditions for cell function analysis. Results: UM cells cultured following anchorage-free methods developed MCTs shaped as spheres. Regardless of their sizes and degree of compaction, these spheres displayed an outer ring of viable and proliferating cells, and a core with less proliferating and apoptotic cells. In contrast, UM cells maintained under anchorage-dependent conditions established several morphological adaptations. Some remained isolated and rounded, formed multi-size irregular aggregates, or adopted a 2D-like flat appearance. These cells invariably conserved their metabolic activity and conserved melanocytic markers (i.e., expression of Melan A/Mart-1 and HMB45). Notably, under hypoxia, cells maintained under 3D conditions secrete more VEGF compared to cells cultured under 2D conditions. Conclusions: Under an anchorage-free environment, UM cells form sphere-like MCTs that acquire attributes reminiscent of abnormal vascularized solid tumors. UM cells behavior in anchorage-dependent manner exposed diverse cells populations in response to cues from an enriched extracellular matrix proteins (ECM) environment, highlighting the plasticity of UM cells. This study provides a 3D cell culture platform that is more predictive of the biology of UM. The integration of such platforms to explore mechanisms of ECM-mediated tumor resistance, metastatic abilities, and to test novel therapeutics (i.e., anti-angiogenics and immunomodulators) would benefit UM care. [ABSTRACT FROM AUTHOR]

Enormous success has been achieved in antitumor therapy with PD-1/PD-L1 inhibitors; however, less than 3 out of 10 patients show clinical benefits. An interesting phenomenon of patients who experienced durable clinical responses to PD-L1 blockade usually being the patients with high PD-L1 expression was noticed. Accordingly, the evaluation of PD-L1 is of great benefit to predict the immunotherapy response against PD-1/PD-L1. A small-molecule single photon emission computed tomography (SPECT) tracer [99mTc]Tc-SG2C-CBM containing a biphenyl moiety was designed and synthesized for the measurement of PD-L1 expression. The radiolabeling yield (RLY) and radiochemical purity (RCP) of [99mTc]Tc-SG2C-CBM were detected to be over 93% under the optimized labeling conditions. [99mTc]Tc-SG2C-CBM was discovered to be equipped with not only good stability but also suitable lipophilicity. In the cellular assays, [99mTc]Tc-SG2C-CBM showed moderate affinity to PD-L1 and 6.87-fold higher uptake in A375-hPD-L1 cells than in A375 cells. Then, the in vivo biodistribution results of the A375-hPD-L1 tumor-bearing mice showed that the radioactivity uptake in tumors was about 2.96 ± 0.76 and 2.31 ± 0.26 fold that in muscles and bones at 2 h post-injection, respectively. SPECT imaging was consistent with the biological distribution of [99mTc]Tc SG2C-CBM, with the uptake being mainly in the liver and less pronounced in tumors. The results of these studies indicate that [99mTc]Tc-SG2C-CBM may be a probe for imaging PD-L1 expression, but further research is needed. [ABSTRACT FROM AUTHOR]

Small molecule drugs play a pivotal role in the arsenal of anticancer pharmacological agents. Nonetheless, their small size poses a challenge when directly visualizing their localization, distribution, mechanism of action (MOA), and target engagement at the subcellular level in real time. We propose a strategy for developing triple-functioning drug beacons that seamlessly integrate therapeutically relevant bioactivity, precise subcellular localization, and direct visualization capabilities within a single molecular entity. As a proof of concept, we have meticulously designed and constructed a boronic acid fluorescence drug beacon using coumarin–hemicyanine (CHB). Our CHB design includes three pivotal features: a boronic acid moiety that binds both adenosine triphosphate (ATP) and adenosine diphosphate (ADP), thus depleting their levels and disrupting the energy supply within mitochondria; a positively charged component that targets the drug beacon to mitochondria; and a sizeable conjugated luminophore that emits fluorescence, facilitating the application of structured illumination microscopy (SIM). Our study indicates the exceptional responsiveness of our proof-of-concept drug beacon to ADP and ATP, its efficacy in inhibiting tumor growth, and its ability to facilitate the tracking of ADP and ATP distribution around the mitochondrial cristae. Furthermore, our investigation reveals that the micro-dynamics of CHB induce mitochondrial dysfunction by causing damage to the mitochondrial cristae and mitochondrial DNA. Altogether, our findings highlight the potential of SIM in conjunction with visual drug design as a potent tool for monitoring the in situ MOA of small molecule anticancer compounds. This approach represents a crucial advancement in addressing a current challenge within the field of small molecule drug discovery and validation. Drug beacon CHB inhibits energy production by binding to ATP/ADP in mitochondria, disrupting the structure and function of mitochondria, exerting anti-tumor effects. [Display omitted] [ABSTRACT FROM AUTHOR]

Animal behavior can provide useful information about animal welfare, but methods and tools used to gather behavioral data and data treatment can influence the results. Therefore, this study was carried out on dairy cow (Bos taurus) behavior and interaction with calves early post-partum aiming at comparing two sampling rules, namely continuous and instantaneous sampling at scan intervals of 30 s, 1, 2, 3, 4, 5, and 10 min, and of two methods to deal with out of sight animals. The study was based on three assumptions: (1) continuous sampling provides the most complete and accurate data, allowing the observation of seldom behaviors and short events; (2) instantaneous sampling can provide accurate measurements of frequency and duration, especially at short scan intervals; (3) differences in behavioral results may occur depending on whether a correction for time out of sight is applied or not. Thus, 10 dams were observed from videos in the 2 h post-parturition. Ruminating, stereotypies, calf-biting and calf-butting were not recorded during the observation period. Other behaviors were observed only with continuous sampling or with continuous and instantaneous at 30-s scan intervals. The recoding of several behaviors was less accurate when applying longer scan intervals. Data from continuous and instantaneous sampling at 30-s scan intervals were compared with Wilcoxon test. Results showed no significant differences for posture, position in the pen and all behaviors (p > 0.05) except vocalizing (p = 0.003). The same test did not highlight significant differences due to method of dealing with out of sight for both sampling rules (p > 0.05). Correlation between continuous and instantaneous sampling were prevalently high at 30-s intervals and they decreased as the length of scan intervals increased for most behaviors. Results confirmed the first two assumptions suggesting that continuous sampling is more accurate, in particular for short and rare behaviors, and caution against the suitability of dam behavioral data collected using instantaneous sampling at scan intervals of minutes. The third assumption was not proven by this study. Results should be considered in light of the development of new technologies that relies on data acquired by sensors and imaging to monitor cow-calf welfare and behavior post-parturition. [ABSTRACT FROM AUTHOR]

Background: The importance of CD11b/CD18 expression in neutrophil effector functions is well known. Beyond KINDLIN3 and TALIN1, which are involved in the induction of the high-affinity binding CD11b/CD18 conformation, the signaling pathways that orchestrate this response remain incompletely understood. Method: We performed an unbiased screening method for protein selection by biotin identification (BioID) and investigated the KINDLIN3 interactome. We used liquid chromatography with tandem mass spectrometry as a powerful analytical tool. Generation of NB4 CD18, KINDLIN3, or SKAP2 knockout neutrophils was achieved using CRISPR-Cas9 technology, and the cells were examined for their effector function using flow cytometry, live cell imaging, microscopy, adhesion, or antibody-dependent cellular cytotoxicity (ADCC). Results: Among the 325 proteins significantly enriched, we identified Src kinase-associated phosphoprotein 2 (SKAP2), a protein involved in actin polymerization and integrin-mediated outside-in signaling. CD18 immunoprecipitation in primary or NB4 neutrophils demonstrated the presence of SKAP2 in the CD11b/CD18 complex at a steady state. Under this condition, adhesion to plastic, ICAM-1, or fibronectin was observed in the absence of SKAP2, which could be abrogated by blocking the actin rearrangements with latrunculin B. Upon stimulation of NB4 SKAP2-deficient neutrophils, adhesion to fibronectin was enhanced whereas CD18 clustering was strongly reduced. This response corresponded with significantly impaired CD11b/CD18-dependent NADPH oxidase activity, phagocytosis, and cytotoxicity against tumor cells. Conclusion: Our results suggest that SKAP2 has a dual role. It may restrict CD11b/ CD18-mediated adhesion only under resting conditions, but its major contribution lies in the regulation of dynamic CD11b/CD18-mediated actin rearrangements and clustering as required for cellular effector functions of human neutrophils. [ABSTRACT FROM AUTHOR]

Single‐fiber imaging technology offers unique insights for research and inspection in difficult‐to‐reach and narrow spaces. In particular, ultra‐compact multimode fiber (MMF) imaging, has received increasing interest over the past decade. However, MMF imaging will be seriously distorted when subjected to dynamic perturbations due to time‐varying mode coupling, and the imaging of space objects via Gaussian beam will be relatively degraded at the edge due to insufficient contrast. Here, a robust super‐resolution image transmission method based on a ring core fiber (RCF) with orbital angular momentum (OAM) is proposed and experimentally demonstrated. The OAM modes propagating in the RCF form a series of weakly coupled mode groups, making the imaging system robust to external perturbations. In addition, a spiral phase plate is used as a vortex filter to produce OAM for edge enhancement, thus improving the image resolution. Furthermore, a few‐shot U‐Transformer neural network is proposed to enhance the resilience of the developed RCF‐OAM imaging system against environmental perturbations. Finally, the developed RCF‐OAM imaging system achieves biological image transmission, demonstrating the practicality of the scheme. This pioneering RCF OAM image transmission system may have broad applications, potentially revolutionizing fields such as biological imaging and industrial non‐destructive testing. [ABSTRACT FROM AUTHOR]

Four-dimensional ultrasound imaging of complex biological systems such as the brain is technically challenging because of the spatiotemporal sampling requirements. We present computational ultrasound imaging (cUSi), an imaging method that uses complex ultrasound fields that can be generated with simple hardware and a physical wave prediction model to alleviate the sampling constraints. cUSi allows for high-resolution four-dimensional imaging of brain hemodynamics in awake and anesthetized mice. [ABSTRACT FROM AUTHOR]

In recent years, the development of carbon dot‐based fluorescent nanoparticles for bioimaging applications has attracted the attention of the scientific community. However, many of these systems absorb and emit in the blue–green region of the electromagnetic spectrum, limiting their application in bioimaging. Herein, the facile design and development of highly efficient two‐photon excitable red‐emissive carbon dots are reported and their high performance in bioimaging applications is demonstrated. The importance of aromatic precursors in developing red‐emissive carbon dots is demonstrated. The optimized carbon dots are highly biocompatible and nontoxic, with remarkable photostability in cells under two‐photon near‐infrared excitation. The present study points to the great potential of two‐photon excitable red‐emissive carbon dot as an efficient bioimaging agent for cellular biolabeling, long‐term and real‐time cellular imaging, and high‐resolution deep‐tissue imaging in complex biological systems. [ABSTRACT FROM AUTHOR]

Coherent anti-Stokes Raman scattering (CARS) implemented as a vibrational micro-spectroscopy modality eradicates the need for potentially perturbative fluorescent labeling while still providing high-resolution, chemically specific images of biological samples. Isotopic substitution of hydrogen atoms with deuterium introduces minimal change to molecular structures and can be coupled with CARS microscopy to increase chemical contrast. Here, we investigate HeLa cells incubated with non-deuterated or deuterium-labeled fatty acids, using an in-house-developed hyperspectral CARS microscope coupled with an unsupervised quantitative data analysis algorithm, to retrieve Raman susceptibility spectra and concentration maps of chemical components in physically meaningful units. We demonstrate that our unsupervised analysis retrieves the susceptibility spectra of the specific fatty acids, both deuterated and non-deuterated, in good agreement with reference Raman spectra measured in pure lipids. Our analysis, using the cell-silent spectral region, achieved excellent chemical specificity despite having no prior knowledge and considering the complex intracellular environment inside cells. The quantitative capabilities of the analysis allowed us to measure the concentration of deuterated and non-deuterated fatty acids stored within cytosolic lipid droplets over a 24 h period. Finally, we explored the potential use of deuterium-labeled lipid droplets for non-invasive cell tracking, demonstrating an effective application of the technique for distinguishing between cells in a mixed population over a 16 h period. These results further demonstrate the chemically specific capabilities of hyperspectral CARS microscopy to characterize and distinguish specific lipid types inside cells using an unbiased quantitative data analysis methodology. [ABSTRACT FROM AUTHOR]

Intense sun exposure is a major risk factor for the development of melanoma, an abnormal proliferation of skin cells. Yet, this more prevalent type of skin cancer can also develop in less-exposed areas, such as those that are shaded. Melanoma is the sixth most common type of skin cancer. In recent years, computer-based methods for imaging and analyzing biological systems have made considerable strides. This work investigates the use of advanced machine learning methods, specifically ensemble models with Auto Correlogram Methods, Binary Pyramid Pattern Filter, and Color Layout Filter, to enhance the detection accuracy of Melanoma skin cancer. These results suggest that the Color Layout Filter model of the Attribute Selection Classifier provides the best overall performance. Statistics for ROC, PRC, Kappa, F-Measure, and Matthews Correlation Coefficient were as follows: 90.96% accuracy, 0.91 precision, 0.91 recall, 0.95 ROC, 0.87 PRC, 0.87 Kappa, 0.91 F-Measure, and 0.82 Matthews Correlation Coefficient. In addition, its margins of error are the smallest. The research found that the Attribute Selection Classifier performed well when used in conjunction with the Color Layout Filter to improve image quality. [ABSTRACT FROM AUTHOR]

Geographic atrophy (GA) is characterized by atrophy of the retina, retinal pigment epithelium and choriocapillaris, causing a gradual loss of vision over time. Treatment options to prevent initiation or progression of GA are limited; two recently FDA‐approved inhibitors of the complement system (pegcetacoplan, avacincaptad pegol) showed a modest decrease in GA lesion growth in phase 3 clinical trials. Exploration of genetic and molecular biomarkers in GA plays a critical role in our battle against this blinding disease to improve early disease detection, to find more effective therapies, and to provide personalized care to patients. In this review, we provide a comprehensive overview of the current literature investigating genetic and molecular biomarkers for GA. Genetic studies identified multiple genes and variants that play a role in progression to GA and GA lesion growth, involving pathways such as complement activation, extracellular matrix interaction and lipid metabolism. The number of published studies assessing molecular biomarkers for GA initiation and progression in ocular matrices is limited. Several studies evaluated molecular biomarkers in the systemic circulation, showing higher levels of complement activation and a causal role of lipid subfractions in GA. Larger, well‐powered studies are needed to identify novel and validate existing biomarkers, and to investigate the potential of combining genetic and molecular markers with imaging techniques for more accurate diagnosis and monitoring of GA. The development of personalized medicine approaches based on individual genetic and molecular profiles could hold promise for more effective and targeted treatments for this devastating disease. [ABSTRACT FROM AUTHOR]

• The review explores the world of functional chromophores synthesized via MCRs. • MCR-synthesized functional chromophores are reshaping imaging research. • Bioimaging probes synthesized by MCRs with tunable properties. This review aims to provide a comprehensive overview of recent advancements and applications of fluorescence imaging probes synthesized via MCRs (multicomponent reactions). These probes, also known as functional chromophores, belong to a currently investigated class of fluorophores that are presently being successfully applied in bioimaging experiments, especially in various living cell lineages. We describe some of the MCRs that have been employed in the synthesis of these probes and explore their applications in biological imaging, with an emphasis on cellular imaging. The review also discusses the challenges and future perspectives in the field, particularly considering the potential impact of MCR-based fluorescence imaging probes on advancing this field of research in the coming years. Considering that this area of research is relatively new and nearly a decade has passed since the first publication, this review also provides a historical perspective on this class of fluorophores, highlighting the pioneering works published between 2011 and 2016. [ABSTRACT FROM AUTHOR]

High-altitude climates are characterized by intense solar and UV (ultraviolet) stress, long periods of sunlight, low temperatures, and a significant temperature differential between day and night. These traits affect high-altitude plants' physical structure and physiological and metabolic functions. Abiotic stress responses are critical for understanding how plants adapt to high altitudes. Thus, the adverse environment of mountainous areas makes organisms an ideal natural laboratory for examining speciation and adaptive evolution. Alpine plants employ morphological modifications to adapt to their high-altitude habitat, but the molecular and physiological mechanisms that underpin these adaptations remain unclear. According to recent plant climate adaptation findings, climate change may drive fresh selection and interfere with local adaptation. Genome/transcriptome sequencing has shown that energy consumption and positive selection response to various stresses depend on certain genes. RNA sequencing may also aid in the development of molecular markers such as co-dominant microsatellite markers and single-copy nuclear genes. Phytoconstituents are predominantly alkaloids, glycosides, polyphenols, and terpenes. Secondary metabolites from medicinal plants are plentiful and widely employed in traditional medicine. In drug development, huge datasets are analyzed using whole-genome sequencing and cellular protein expression patterns. Along with genomics, proteomics, and metabolomics, OMICs are used to find and characterize targeted drugs. Morphological statistics and high-throughput multi-omics have been widely employed to explore how plants adapt to high-altitude settings. These technologies can now capture images of a biological system's underlying biology at a resolution never before possible. This review delves into the assessment of the transcriptome and metabolome in plant adaptation along altitude. The insights gained from this exploration can be of significant assistance to researchers engaged in the fields of medicinal plant biotechnology, chemical genetics, and adaption biology. [ABSTRACT FROM AUTHOR]

This article, published in the journal Neuropediatrics, discusses the role of autophagy in cellular clearance and the degradation of specific substrates. The authors focus on the RNaseL pathway and its activation by 2′,5′-oligoadenylate synthetase proteins (OAS) in response to viral RNA. They present clinical, molecular, and imaging data from four patients with biallelic variants in OAS2, highlighting the association of these variants with skeletal dysplasia, neurodevelopmental disorders, and immunodeficiency. The study also includes experimental findings from cellular assays and proteomics, as well as single cell RNA sequencing. The authors conclude that biallelic OAS2 variants may be a novel candidate gene for developmental disorders and suggest a close relationship between deficient innate immune response and autophagy in monogenic neuroimmunological disorders. [Extracted from the article]

Gliomas are histologically and genetically heterogeneous tumors. However, classical histopathological typing often ignores the high heterogeneity of tumors and thus cannot meet the requirements of precise pathological diagnosis. Here, proximity‐anchored in situ spectral coding amplification (ProxISCA) is proposed for multiplexed imaging of RNA mutations, enabling visual typing of brain gliomas with different pathological grades at the single‐cell and tissue levels. The ligation‐based padlock probe can discriminate one‐nucleotide variations, and the design of proximity primers enables the anchoring of amplicons on target RNA, thus improving localization accuracy. The DNA module‐based spectral coding strategy can dramatically improve the multiplexing capacity for imaging RNA mutations through one‐time labelling, with low cost and simple operation. One‐target‐one‐amplicon amplification confers ProxISCA the ability to quantify RNA mutation copy number with single‐molecule resolution. Based on this approach, it is found that gliomas with higher malignant grades express more genes with high correlation at the cellular and tissue levels and show greater cellular heterogeneity. ProxISCA provides a tool for glioma research and precise diagnosis, which can reveal the relationship between cellular heterogeneity and glioma occurrence or development and assist in pathological prognosis. [ABSTRACT FROM AUTHOR]

Introduction: Complete surgical tumor resection is paramount in the management of soft tissue sarcoma (STS) in humans, dogs, and cats alike. Near-infrared targeted tracers for fluorescence-guided surgery (FGS) could facilitate intraoperative visualization of the tumor and improve resection accuracy. Target identification is complicated in STS due to the rarity and heterogeneity of the disease. This study aims to validate the expression of fibroblast activation protein alpha (FAP) in selected human, canine, and feline STS subtypes to assess the value of FAP as a target for FGS and to validate companion animals as a translational model. Methods: Formalin-fixed and paraffin-embedded tissue samples from 53 canine STSs (perivascularwall tumor (PWT), canine fibrosarcoma (cFS), and STS not further specified (NOS)), 24 feline fibrosarcomas, and 39 human STSs (myxofibrosarcoma, undifferentiated pleomorphic sarcoma, dermatofibrosarcoma protuberans, and malignant peripheral nerve sheath tumor) as well as six canine and seven feline healthy controls and 10 inflamed tissue samples were immunohistochemically stained for their FAP expression. FAP labeling in tumor, peritumoral, healthy skin, and inflamed tissue samples was quantified using a visually assessed semiquantitative expression score and digital image analysis. Target selection criteria (TASC) scoring was subsequently performed as previously described. Results: Eighty-five percent (85%) of human (33/39), 76% of canine (40/53), and 92% of feline (22/24) STSs showed FAP positivity in over 10% of the tumor cells. A high expression was determined in 53% canine (28/53), 67% feline (16/24), and 44% human STSs (17/39). The average FAP-labeled area of canine, feline, and human STSs was 31%, 33%, and 42%, respectively (p > 0.8990). The FAP-positive tumor area was larger in STS compared to healthy and peritumoral tissue samples (p < 0.0001). TASC scores were above 18 for all feline and human STS subtypes and canine PWTs but not for canine STS NOS and cFS. Conclusion: This study represents the first cross-species target evaluation of FAP for STS. Our results demonstrate that FAP expression is increased in various STS subtypes compared to non-cancerous tissues across species, thereby validating dogs and cats as suitable animal models. Based on a TASC score, FAP could be considered a target for FGS. [ABSTRACT FROM AUTHOR]

Background: The liver plays an important role in maintaining copper homeostasis. Copper ion accumulation was elevated in HCC tissue samples. Copper homeostasis is implicated in cancer cell proliferation and angiogenesis. The potential of copper homeostasis as a new theranostic biomarker for molecular imaging and the targeted therapy of HCC has been demonstrated. Recent studies have reported a novel copper-dependent nonapoptotic form of cell death called cuproptosis, strikingly different from other known forms of cell death. The correlation between cuproptosis and hepatocellular carcinoma (HCC) is not fully understood. Materials and Methods: The transcriptomic data of patients with HCC were retrieved from the Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA-LIHC) and were used as a discovery cohort to construct the prognosis model. The gene expression data of patients with HCC retrieved from the International Cancer Genome Consortium (ICGC) and Gene Expression Omnibus (GEO) databases were used as the validation cohort. The Least Absolute Shrinkage and Selection Operator (LASSO) regression analysis was used to construct the prognosis model. A principal component analysis (PCA) was used to evaluate the overall characteristics of cuproptosis regulator genes and obtain the PC1 and PC2 scores. Unsupervised clustering was performed using the ConsensusClusterPlus R package to identify the molecular subtypes of HCC. Cox regression analysis was performed to identify cuproptosis regulator genes that could predict the prognosis of patients with HCC. The receiver operating characteristics curve and Kaplan–Meier survival analysis were used to understand the role of hub genes in predicting the diagnosis and prognosis of patients, as well as the prognosis risk model. A weighted gene co-expression network analysis (WGCNA) was used for screening the cuproptosis subtype-related hub genes. The functional enrichment analysis was performed using Metascape. The 'glmnet' R package was used to perform the LASSO regression analysis, and the randomForest algorithm was performed using the 'randomForest' R package. The 'pRRophetic' R package was used to estimate the anticancer drug sensitivity based on the data retrieved from the Genomics of Drug Sensitivity in Cancer database. The nomogram was constructed using the 'rms' R package. Pearson's correlation analysis was used to analyze the correlations. Results: We constructed a six-gene signature prognosis model and a nomogram to predict the prognosis of patients with HCC. The Kaplan–Meier survival analysis revealed that patients with a high-risk score, which was predicted by the six-gene signature model, had poor prognoses (log-rank test p < 0.001; HR = 1.83). The patients with HCC were grouped into three distinct cuproptosis subtypes (Cu-clusters A, B, and C) based on the expression pattern of cuproptosis regulator genes. The patients in Cu-cluster B had poor prognosis (log-rank test p < 0.001), high genomic instability, and were not sensitive to conventional chemotherapeutic treatment compared to the patients in the other subtypes. Cancer cells in Cu-cluster B exhibited a higher degree of the senescence-associated secretory phenotype (SASP), a marker of cellular senescence. Three representative genes, CDCA8, MCM6, and NCAPG2, were identified in patients in Cu-cluster B using WGCNA and the "randomForest" algorithm. A nomogram was constructed to screen patients in the Cu-cluster B subtype based on three genes: CDCA8, MCM6, and NCAPG2. Conclusion: Publicly available databases and various bioinformatics tools were used to study the heterogeneity of cuproptosis in patients with HCC. Three HCC subtypes were identified, with differences in the survival outcomes, genomic instability, senescence environment, and response to anticancer drugs. Further, three cuproptosis-related genes were identified, which could be used to design personalized therapeutic strategies for HCC. [ABSTRACT FROM AUTHOR]

Identifying biomarkers for Alzheimer's disease with a goal of early detection is a fundamental problem in clinical research. Both medical imaging and genetics have contributed informative biomarkers in literature. To further improve the performance, recently, there is an increasing interest in developing analytic approaches that combine data across modalities such as imaging and genetics. However, there are limited methods in literature that are able to systematically combine high-dimensional voxel-level imaging and genetic data for accurate prediction of clinical outcomes of interest. Existing prediction models that integrate imaging and genetic features often use region level imaging summaries, and they typically do not consider the spatial configurations of the voxels in the image or incorporate the dependence between genes that may compromise prediction ability. We propose a novel integrative Bayesian scalar-on-image regression model for predicting cognitive outcomes based on high-dimensional spatially distributed voxel-level imaging data, along with correlated transcriptomic features. We account for the spatial dependencies in the imaging voxels via a tensor approach that also enables massive dimension reduction to address the curse of dimensionality, and models the dependencies between the transcriptomic features via a Graph-Laplacian prior. We implement this approach via an efficient Markov chain Monte Carlo (MCMC) computation strategy. We apply the proposed method to the analysis of longitudinal ADNI data for predicting cognitive scores at different visits by integrating voxel-level cortical thickness measurements derived from T1w-MRI scans and transcriptomics data. We illustrate that the proposed imaging transcriptomics approach has significant improvements in prediction compared to prediction using a subset of features from only one modality (imaging or genetics), as well as when using imaging and transcriptomics features but ignoring the inherent dependencies between the features. Our analysis is one of the first to conclusively demonstrate the advantages of prediction based on combining voxel-level cortical thickness measurements along with transcriptomics features, while accounting for inherent structural information. [ABSTRACT FROM AUTHOR]

A novel fluorescent probe based on azamonardine (Aza) fluorophore was designed and synthesized for the highly selective detection of cysteine (Cys) in vivo and in vitro. After reacting with acryloyl chloride, the fluorescence of Aza is effectively quenched, resulting in the formation of the Aza-acryl probe. Upon the addition of Cys, the ester bond of Aza-acryl is cleaved, releasing a new compound (Compound 1) with strong fluorescence, thereby achieving fluorescence turn-on detection of Cys. The structure of Aza-acryl was characterized using X-ray crystallography and NMR spectroscopy. Additionally, density functional theory was employed to elucidate the quenching mechanism of the acyl group on the Aza. Aza-acryl exhibits high selectivity towards Cys and distinguishes it from other biothiols such as homocysteine (Hcy) and glutathione (GSH). The mechanism of Aza-acryl for detecting Cys was investigated through HPLC, NMR spectroscopy, high-resolution mass spectrometry, and reaction kinetics experiments. Aza-acryl demonstrates excellent imaging capabilities for Cys in cells and zebrafish, providing a reliable and selectable tool for the detection and imaging of Cys in biological systems. [ABSTRACT FROM AUTHOR]

Cytosolic Ca2+ signals are organized in complex spatial and temporal patterns that underlie their unique ability to regulate multiple cellular functions. Changes in intracellular Ca2+ concentration ([Ca2+]i) are finely tuned by the concerted interaction of membrane receptors and ion channels that introduce Ca2+ into the cytosol, Ca2+-dependent sensors and effectors that translate the elevation in [Ca2+]i into a biological output, and Ca2+-clearingmechanisms that return the [Ca2+]i to prestimulation levels and prevent cytotoxic Ca2+ overload. The assortment of the Ca2+ handling machinery varies among different cell types to generate intracellular Ca2+ signals that are selectively tailored to subserve specific functions. The advent of novel high-speed, 2D and 3D time-lapse imaging techniques, single-wavelength and genetic Ca2+ indicators, as well as the development of novel genetic engineering tools to manipulate single cells and whole animals, has shed novel light on the regulation of cellular activity by the Ca2+ handlingmachinery. A symposium organized within the framework of the 72nd Annual Meeting of the Italian Society of Physiology, held in Bari on 14-16th September 2022, has recently addressed many of the unexpected mechanisms whereby intracellular Ca2+ signalling regulates cellular fate in healthy and disease states. Herein, we present a report of this symposium, in which the following emerging topics were discussed: 1) Regulation of water reabsorption in the kidney by lysosomal Ca2+ release through Transient Receptor Potential Mucolipin 1 (TRPML1); 2) Endoplasmic reticulum-to-mitochondria Ca2+ transfer in Alzheimer's disease-related astroglial dysfunction; 3) The non-canonical role of TRPMelastatin 8 (TRPM8) as a Rap1A inhibitor in the definition of some cancer hallmarks; and 4) Non-genetic optical stimulation of Ca2+ signals in the cardiovascular system. [ABSTRACT FROM AUTHOR]

• Review of the role of the enteric nervous system (ENS) in autism spectrum disorders. • Key neuroimmune interactions within the ENS may influence intestinal homeostasis. • Zebrafish are an ideal model to assess cellular interactions. • State of the art methods can help uncover complex mechanisms of cell interactions. Autism spectrum disorders (ASD) affect about 1% of the population and are strongly associated with gastrointestinal diseases creating shortcomings in quality of life. Multiple factors contribute to the development of ASD and although neurodevelopmental deficits are central, the pathogenesis of the condition is complex and the high prevalence of intestinal disorders is poorly understood. In agreement with the prominent research establishing clear bidirectional interactions between the gut and the brain, several studies have made it evident that such a relation also exists in ASD. Thus, dysregulation of the gut microbiota and gut barrier integrity may play an important role in ASD. However, only limited research has investigated how the enteric nervous system (ENS) and intestinal mucosal immune factors may impact on the development of ASD-related intestinal disorders. This review focuses on the mechanistic studies that elucidate the regulation and interactions between enteric immune cells, residing gut microbiota and the ENS in models of ASD. Especially the multifaceted properties and applicability of zebrafish (Danio rerio) for the study of ASD pathogenesis are assessed in comparison to studies conducted in rodent models and humans. Advances in molecular techniques and in vivo imaging, combined with genetic manipulation and generation of germ-free animals in a controlled environment, appear to make zebrafish an underestimated model of choice for the study of ASD. Finally, we establish the research gaps that remain to be explored to further our understanding of the complexity of ASD pathogenesis and associated mechanisms that may lead to intestinal disorders. [ABSTRACT FROM AUTHOR]

Age-related hearing loss, or presbyacusis, is a common degenerative disorder affecting communication and quality of life for millions of older adults. Multiple pathophysiologic manifestations, along with many cellular and molecular alterations, have been linked to presbyacusis; however, the initial events and causal factors have not been clearly established. Comparisons of the transcriptome in the lateral wall (LW) with other cochlear regions in a mouse model (of both sexes) of “normal” age-related hearing loss revealed that early pathophysiological alterations in the stria vascularis (SV) are associated with increased macrophage activation and a molecular signature indicative of inflammaging, a common form of immune dysfunction. Structure-function correlation analyses in mice across the lifespan showed that the age-dependent increase in macrophage activation in the stria vascularis is associated with a decline in auditory sensitivity. High-resolution imaging analysis of macrophage activation in middle-aged and aged mouse and human cochleas, along with transcriptomic analysis of age-dependent changes in mouse cochlear macrophage gene expression, support the hypothesis that aberrant macrophage activity is an important contributor to age-dependent strial dysfunction, cochlear pathology, and hearing loss. Thus, this study highlights the SV as a primary site of age-related cochlear degeneration and aberrant macrophage activity and dysregulation of the immune system as early indicators of age-related cochlear pathology and hearing loss. Importantly, novel new imaging methods described here now provide a means to analyze human temporal bones in a way that had not previously been feasible and thereby represent a significant new tool for otopathological evaluation. [ABSTRACT FROM AUTHOR]

Summary: Traditionally, botanists study plant anatomy by carefully sectioning samples, histological staining to highlight tissues of interests, then imaging slides under light microscopy. This approach generates significant details; however, this workflow is laborious, particularly in woody vines (lianas) with heterogeneous anatomies, and ultimately yields two‐dimensional (2D) images. Laser ablation tomography (LATscan) is a high‐throughput imaging system that yields hundreds of images per minute. This method has proven useful for studying the structure of delicate plant tissues; however, its utility in understanding the structure of woody tissues is underexplored.We report LATscan‐derived anatomical data from several stems of lianas (c. 20 mm) of seven species and compare these results with those obtained through traditional anatomical techniques.LATscan successfully allows the description of tissue composition by differentiating cell type, size, and shape, but also permits the recognition of distinct cell wall composition (e.g. lignin, suberin, cellulose) based on differential fluorescent signals on unstained samples.LATscan generate high‐quality 2D images and 3D reconstructions of woody plant samples; therefore, this new technology is useful for both qualitative and quantitative analyses. This high‐throughput imaging technology has the potential to bolster phenotyping of vegetative and reproductive anatomy, wood anatomy, and other biological systems. [ABSTRACT FROM AUTHOR]

Zebrafish (Danio rerio) has been successfully used for decades in developmental studies and disease modelling. The remarkable uptake of zebrafish as a model system is partly due to its transparency during the early weeks of its development, allowing in vivo imaging of cellular and molecular processes. However, this key advantage wears off when tissues become opaque as the animal reaches juvenile and adult stages, rendering access to tissues for live imaging and longitudinal studies difficult. Here we provide a novel approach to image and assess tissue integrity of adult zebrafish using MRI on live zebrafish suitable for longitudinal studies. We built a 3D‐printed life support chamber and designed a protocol‐directed sedation regime to recover adult zebrafish after scanning in a 9.4 T MRI scanner. Our life support chamber is cheap and easy to create using 3D printing, allowing other groups to copy our template for quick setup. Additionally, we optimized the delivery of contrast agent to enhance brain signals in order to refine current delivery, usually delivered intravenously in rodents. We show here that immersion in gadolinium was a viable alternative to intraperitoneal injection to reduce T1 relaxation times. This resulted in protocol refinement as per the 3Rs guidelines and improved image contrast in adult zebrafish disease models. In conclusion, we provide here a detailed methodology to allow longitudinal studies of brain tissue integrity of adult zebrafish, combining safe and efficient delivery of contrast agent and live MRI. This technique can be used to bridge the gap between in vivo studies and longitudinal brain analysis in adult zebrafish, and can be applied to the ever‐growing number of adult zebrafish models of ageing and neurodegenerative diseases. [ABSTRACT FROM AUTHOR]

Here, π‐Extended BTD derivatives were successfully synthesized by Heck coupling reaction, which exhibited the the advantages of simplicity and efficiency, wild substrate scope, easily available substrates and high yield. The fluorescent probe PEG‐BTDAr targeting LDs was successfully prepared by the nucleophilic substitution reaction between the Heck coupling reaction product 3 h and Amino polyethylene glycol monomethyl ether (Mn=2000). PEG‐BTDAr exhibited the advantages of high selectivity, good stability and pH resistance. The use of PEG as a substrate gave PEG‐BTDAr good biocompatibility. It was worth mentioning that PEG‐BTDAr could not only track LDs in cells under different physiological conditions, but also distinguish between living and dead cells in biological systems. [ABSTRACT FROM AUTHOR]

Introduction: The cerebellum is organized into functional regions each dedicated to process different motor or sensory inputs for controlling different locomotor behaviors. This functional regionalization is prominent in the evolutionary conserved single-cell layered Purkinje cell (PC) population. Fragmented gene expression domains suggest a genetic organization of PC layer regionalization during cerebellum development. However, the establishment of such functionally specific domains during PC differentiation remained elusive. Methods and results: We show the progressive emergence of functional regionalization of PCs from broad responses to spatially restricted regions in zebrafish by means of in vivo Ca2+-imaging during stereotypic locomotive behavior. Moreover, we reveal that formation of new dendritic spines during cerebellar development using in vivo imaging parallels the time course of functional domain development. Pharmacological as well as cell-type specific optogenetic inhibition of PC neuronal activity results in reduced PC dendritic spine density and an altered stagnant pattern of functional domain formation in the PC layer. Discussion: Hence, our study suggests that functional regionalization of the PC layer is driven by physiological activity of maturing PCs themselves. [ABSTRACT FROM AUTHOR]

Single cell technologies, lineage tracing mouse models and advanced imaging techniques unequivocally improved the resolution of the cellular landscape of atherosclerosis. Although the discovery of the heterogeneous nature of the cellular plaque architecture has undoubtedly improved our understanding of the specific cellular states in atherosclerosis progression, it also adds more complexity to current and future research and will change how we approach future drug development. In this review, we will discuss how the revolution of new single cell technologies allowed us to map the cellular networks in the plaque, but we will also address current (technological) limitations that confine us to identify the cellular drivers of the disease and to pinpoint a specific cell state, cell subset or cell surface antigen as new candidate drug target for atherosclerosis. [Display omitted] • Single cell omics unveiled the heterogeneous cellular landscape of atherosclerosis. • Intraplaque heterogeneity adds complexity to current and future (drug) research. • Studying cell activation and cell-cell interactions is the next step forward. • More disease and site-specific therapies for atherosclerosis are needed. [ABSTRACT FROM AUTHOR]

The optimization of the radiolabeling yield of carvedilol with iodine-131 was described. Dependence of the labeling yield of [131I]iodocarvedilol on the concentration of carvedilol, chloramine-T content, pH of the reaction mixture and reaction time was studied in details. Carvedilol was labeled with iodine-131 at pH 6 with a labeling yield of 92.6 ± 2.77% by using 100 µg carvedilol, 200 µg chloramin-T (CAT) and 30 min reaction time. The formed [131I]iodocarvedilol was nearly stable for a time up to one day. Biodistribution of [131I]iodocarvedilol was investigated in experimental animals. [131/123I]iodocarvedilol was located in the heart with a concentration of 19.6 ± 0.41% of the injected dose at 60 min post injection. It has a high heart uptake and heart to liver ratio, both of which are beneficial for high-quality SPECT (single-photon emission computerized tomography) myocardial imaging. [131/123I]iodocarvedilol solve most the drawbacks of the FDA (Food and Drug Administration) approved 99mTc-sestamibi. [ABSTRACT FROM AUTHOR]

Serial focussed ion beam scanning electron microscopy (FIB/SEM) enables imaging and assessment of subcellular structures on the mesoscale (10 nm to 10 µm). When applied to vitrified samples, serial FIB/SEM is also a means to target specific structures in cells and tissues while maintaining constituents' hydration shells for in situ structural biology downstream. However, the application of serial FIB/SEM imaging of non-stained cryogenic biological samples is limited due to low contrast, curtaining, and charging artefacts. We address these challenges using a cryogenic plasma FIB/SEM. We evaluated the choice of plasma ion source and imaging regimes to produce high-quality SEM images of a range of different biological samples. Using an automated workflow we produced three-dimensional volumes of bacteria, human cells, and tissue, and calculated estimates for their resolution, typically achieving 20-50 nm. Additionally, a tag-free localisation tool for regions of interest is needed to drive the application of in situ structural biology towards tissue. The combination of serial FIB/SEM with plasma-based ion sources promises a framework for targeting specific features in bulk-frozen samples (>100 µm) to produce lamellae for cryogenic electron tomography. [ABSTRACT FROM AUTHOR]

Autonomous experimentation is an emerging area of research, primarily related to autonomous vehicles, scientific combinatorial discovery approaches in materials science and drug discovery, and iterative research loops of planning, experimentation, and analysis. However, autonomous approaches developed in these contexts are difficult to apply to high-dimensional mapping technologies, such as scanning hyperspectral imaging of biological systems, due to sample complexity and heterogeneity. We briefly cover the history of adaptive sampling algorithms and surrogate modeling in order to define autonomous adaptive data acquisition as an objective-based, flexible building block for future biological imaging experimentation driven by intelligent infrastructure. We subsequently summarize the recent implementations of autonomous adaptive data acquisition (AADA) for scanning hyperspectral imaging, assess how these address the difficulties of autonomous approaches in hyperspectral imaging, and highlight the AADA design variation from a goal-oriented perspective. Finally, we present a modular AADA architecture that embeds AADA-driven flexible building blocks to address the challenge of time resolution for high-dimensional scanning hyperspectral imaging of nonequilibrium dynamical systems. In our example research-driven experimental design case, we propose an AADA infrastructure for time-resolved, noninvasive, and label-free scanning hyperspectral imaging of living biological systems. This AADA infrastructure can accurately target the correct state of the system for experimental workflows that utilize subsequent expensive, high-information-content analytical techniques. [ABSTRACT FROM AUTHOR]

Accurate monitoring of intracellular pH in living cells is critical for developing a better understanding of cellular activities. In the current study, label-free carbon dots (p-CDs), which were fabricated using a straightforward one-pot solvothermal treatment of p-phenylenediamine and urea, were employed to create a new ratiometric pH nanosensor. Under single-wavelength excitation (λex = 500 nm), the p-CDs gave dual emission bands at 525 and 623 nm. The fluorescent intensity ratio (I525/I623) was linearly related to pH over the range 4.0 to 8.8 in buffer solutions, indicating that the ratiometric fluorescence nanoprobe may be useful for pH sensing. In pH measurements, the p-CDs also demonstrated outstanding selectivity, reversibility, and photostability. Owing to the advantages outlined above, the nanoprobe was used to monitor the pH of HeLa cells effectively. The label-free CD-based ratiometric nanoprobe features comparatively easy manufacturing and longer excitation and emission wavelengths than the majority of previously reported CD-based ratiometric pH sensors, which is ultimately beneficial for applications in biological imaging. [ABSTRACT FROM AUTHOR]

Background: RNA interference (RNAi) provides a powerful way to investigate the role of genes in disease pathogenesis and modulate gene expression to treat disease. In 2018, the FDA approved patisiran, the first RNAi‐based drug, hence paving the way for a novel class of RNAi therapeutics. Harnessing RNAi to inhibit vaginal HIV transmission requires effective gene silencing in immune cells, which remains difficult. Knockdown in accessible mucosal tissues may be easier than systemic gene silencing. Vaginally applied cholesterol‐conjugated small interfering RNAs (chol‐siRNAs) blocked herpes simplex virus transmission in mice without tissue damage or immunostimulation. Objectives and methods: To investigate using flow cytometry, confocal microscopy, and quantitative imaging if chol‐siRNAs silence gene expression in vaginal immune cells in mice. Results: Although chol‐siRNAs and lipoplexed‐siRNAs silence gene expression in dendritic cells (DCs) in vitro, most internalized siRNAs concentrate within multivesicular bodies, where they are inaccessible to the cellular RNAi machinery. When applied intravaginally in vivo, chol‐siRNAs penetrate the vaginal mucosa, including the lamina propria, and are efficiently internalized by intraepithelial (IE) and lamina propria (LP) DCs, and CD11b+CD45+ cells, but not by T cells. Chol‐siRNAs induce partial gene silencing in IE and LP DCs throughout the genital mucosa in vivo but are inactive in F4/80+CD11b+ macrophages and T cells. Conclusion: As mucosal DCs play an essential role for mucosal viral entry and dissemination, chol‐siRNAs could be harnessed to target various host factors that are critical for viral uptake, DC migration and trans‐infection of virions to T cells, hence allowing the development of a preventive vaginal HIV microbicide. Furthermore, chol‐siRNAs could help elucidate the pathways of HIV transmission and understand the immunologic function of DCs in the genital tract. [ABSTRACT FROM AUTHOR]

Aggregation-induced emission (AIE)-active micelles are a type of fluorescent functional materials that exhibit enhanced emissions in the aggregated surfactant state. They have received significant interest due to their excellent fluorescence efficiency in the aggregated state, remarkable processability, and solubility. AIE-active micelles can be designed through the self-assembly of amphipathic AIE luminogens (AIEgens) and the encapsulation of non-emissive amphipathic molecules in AIEgens. Currently, a wide range of AIE-active micelles have been constructed, with a significant increase in research interest in this area. A series of advanced techniques has been used to characterize AIE-active micelles, such as cryogenic-electron microscopy (Cryo-EM) and confocal laser scanning microscopy (CLSM). This review provides an overview of the synthesis, characterization, and applications of AIE-active micelles, especially their applications in cell and in vivo imaging, biological and organic compound sensors, anticancer drugs, gene delivery, chemotherapy, photodynamic therapy, and photocatalytic reactions, with a focus on the most recent developments. Based on the synergistic effect of micelles and AIE, it is anticipated that this review will guide the development of innovative and fascinating AIE-active micelle materials with exciting architectures and functions in the future. [ABSTRACT FROM AUTHOR]

High Content Screening (HCS) combines high throughput techniques with the ability to generate cellular images of biological systems. The objective of this work is to evaluate the performance of predictive models using CNN to identify the number of cells present in digital contrast microscopy images obtained by HCS. One way to evaluate the algorithm was through the Mean Squared Error metric. The MSE was 4,335.99 in the A549 cell line, 25,295.23 in the Huh7 and 36,897.03 in the 3T3. After obtaining these values, different parameters of the models were changed to verify how they behave. By reducing the number of images, the MSE increased considerably, with the A549 cell line changing to 49,973.52, Huh7 to 79,473.88 and 3T3 to 52,977.05. Correlation analyzes were performed for the different models. In lineage A549, the best model showed a positive correlation with R = 0.953. In Huh7, the best correlation of the model was R = 0.821, it was also a positive correlation. In 3T3, the models showed no correlation, with the best model having R = 0.100. The models performed well in quantifying the number of cells, and the number and quality of the images interfered with this predictive ability. [ABSTRACT FROM AUTHOR]

Fluorescent hypochlorite probes with ratiometric imaging ability are highly desirable for imaging hypochlorite in biological systems. However, it is still challenging to develop new scaffolds for these probes. In this study, we demonstrate that phenothiazine-fused boron complexes are promising scaffolds for the design of ratiometric fluorescent hypochlorite probes. The synthesized complexes based on the scaffold show ultrafast and ratiometric absorption/fluorescence changes for hypochlorite. We also developed an endoplasmic reticulum-targeting probe and demonstrated that it has excellent real-time imaging ability for both endogenous and exogenous hypochlorite in the endoplasmic reticulum of living cells. [ABSTRACT FROM AUTHOR]

Clinical medicine has experienced a rapid development in recent decades, during which therapies targeting specific cellular signaling pathways, or specific cell surface receptors, have been increasingly adopted. While these developments in clinical medicine call for improved precision in diagnosis and treatment monitoring, modern medical imaging methods are restricted mainly to anatomical imaging, lagging behind the requirements of precision medicine. Although positron emission tomography and single photon emission computed tomography have been used clinically for studies of metabolism, their applications have been limited by the exposure risk to ionizing radiation, the subsequent limitation in repeated and longitudinal studies, and the incapability in assessing downstream metabolism. Magnetic resonance spectroscopy (MRS) or spectroscopic imaging (MRSI) are, in theory, capable of assessing molecular activities in vivo, although they are often limited by sensitivity. Here, we review some recent developments in MRS and MRSI of multiple nuclei that have potential as molecular imaging tools in the clinic. [ABSTRACT FROM AUTHOR]

Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) hold great potential in the cardiovascular field for human disease modeling, drug development, and regenerative medicine. However, multiple hurdles still exist for the effective utilization of hiPSC-CMs as a human-based experimental platform that can be an alternative to the current animal models. To further expand their potential as a research tool and bridge the translational gap, we have generated a cardiac-specific hiPSC reporter line that differentiates into fluorescent CMs using CRISPR-Cas9 genome editing technology. The CMs illuminated with the mScarlet fluorescence enable their non-invasive continuous tracking and functional cellular phenotyping, offering a real-time 2D/3D imaging platform. Utilizing the reporter CMs, we developed an imaging-based cardiotoxicity screening system that can monitor distinct drug-induced structural toxicity and CM viability in real time. The reporter fluorescence enabled visualization of sarcomeric disarray and displayed a drug dose–dependent decrease in its fluorescence. The study also has demonstrated the reporter CMs as a biomaterial cytocompatibility analysis tool that can monitor dynamic cell behavior and maturity of hiPSC-CMs cultured in various biomaterial scaffolds. This versatile cardiac imaging tool that enables real time tracking and high-resolution imaging of CMs has significant potential in disease modeling, drug screening, and toxicology testing. [ABSTRACT FROM AUTHOR]

Understanding the physiological dynamics of gametes and embryos in the fallopian tube (oviduct) has significant implications for managing reproductive disorders and improving assisted reproductive technologies. Recent advancements in imaging of the mouse oviduct in vivo uncovered fascinating dynamics of gametes and embryos in their native states. These new imaging approaches and observations are bringing exciting momentum to uncover the otherwise-hidden processes orchestrating fertilization and pregnancy. For mechanistic investigations, in vivo imaging in genetic mouse models enables dynamic phenotyping of gene functions in the reproductive process. Here, we review these imaging methods, discuss insights recently revealed by in vivo imaging, and comment on emerging directions, aiming to stimulate new in vivo studies of reproductive dynamics. [ABSTRACT FROM AUTHOR]

Background: Schwann cells (SCs) are specialized glial cells of the peripheral nervous system that produce myelin and promote fast action potential propagation. In order to myelinate, SCs engage in a series of events that include migration and division along axons, followed by extensive cytoskeletal rearrangements that ensure axonal ensheathment and myelination. SCs are polarized and extend their processes along an abaxonal‐adaxonal axis. Here, we investigate the role of the apical polarity proteins, Pals1a, and aPKCλ, in SC behavior during zebrafish development. Results: We analyzed zebrafish nok and has mutants deficient for pals1a and aPKCλ function respectively. Using live imaging, transmission electron microscopy and whole mount immunostaining, we show that SCs can migrate and divide appropriately, exhibit normal radial sorting, express myelin markers and ensheath axons on time in has and nok mutants. Conclusions: Pals1a and aPKCλ are not essential for SC migration, division or myelination in zebrafish. Key Findings: Schwann cells are specialized myelinating cells of the peripheral nervous system.In this paper, we use live imaging, genetic, transmission electron microscopy and whole mount immunostaining to conduct a thorough analysis of the role of the two canonical polarity proteins Pals1 and aPKC in the development of Schwann cells in zebrafish.This study provides evidence that the canonical polarity proteins tested so far are not required for Schwann cell development in zebrafish underscoring strong differences between mice and zebrafish peripheral glial polarity.This strongly suggests, for the first time, that Schwann cell polarity is established differently in zebrafish and that the abaxonal‐adaxonal axis is independent of canonical polarity proteins.This highlights a novel example of how polarity might differ between cells (epithelial vs glial) and species (zebrafish vs mice). [ABSTRACT FROM AUTHOR]