Mechanism of structural remodeling of cell-wall polysaccharides in peach during osmotic dehydration: a comparative investigation of sucrose, maltitol and their synergistic mixture.

Rank-aware agglomeration of foundation models for immunohistochemistry image cell counting.

Astrocytes are the most abundant glial cells within the central nervous system. They are highly specialized mechanosensors able to detect the kinetics and magnitude of external biomechanical stimuli (i.e., matrix substrate stiffness and shear/compressive/tensile biomechanical strains). They integrate these biomechanical cues through a complex interplay of integrins, focal adhesions, junctional proteins and mechanosensitive channels. In development, this crosstalk secures astrocyte fate maturation and heterogeneity. However, in mechanically induced neural injuries, these mechanosensing elements can drive aberrant signaling. The range of astrocyte mechanoresponses in pathology include cytoskeletal remodeling that impacts cellular morphology and stiffness, disrupted calcium signaling, altered metabolism, pro-inflammatory signaling, and disruptive matrix remodeling. Ultimately, these mechanoresponses can form positive feedback loops that aggravate reactive astrogliosis, damage neural tissue, and altogether inhibit neural regeneration in these diseases. This review synthesizes the current knowledge of astrocyte mechanobiology during development and disease, and highlights the importance of continued investigation into the therapeutic potential of mitigating astrocyte mechanodysfunction in disease.

Engineering complex phenotypes in Halomonas bluephagenesis TD01 via large-fragment manipulation and multiplex base editing.

Mechanistic investigation of modified Ma-Xing-Shi-Gan Decoction in the treatment of Klebsiella pneumoniae pneumonia via the V-ATPase/ATG16L1 pathway.

Targeting microglial dysfunction: The antioxidant potential of and the regulation of microglial responsiveness by dermatan sulfate from a marine invertebrate.

The progressive accumulation of senescent neuronal cells promotes sustained inflammatory responses and has been recognised as the key contributor to age-associated neurodegenerative changes. However, the molecular mechanisms underlying neuronal senescence and its impact on neural homeostasis remain poorly defined. Therefore, robust in vitro models recapitulating neuronal senescence are required for better mechanistic insight. Chemical inducers like Hydrogen peroxide (H₂O₂) and D-galactose (D-gal) are widely used to induce senescence in cell culture systems. In the present study, we systematically compared H₂O₂ and D-galactose induced senescence in human pre-neuroblastoma SH-SY5Y cells, to establish a reliable cellular model for neuronal aging and understand its implications in brain degeneration. Cellular senescence was assessed based on morphological alterations as well as senescence-associated markers like SA-β-gal activity, expression of senescence-associated genes and proteins such as p16, p21, p53, and γH2AX. SH-SY5Y cells exhibited altered cellular morphology and changes in senescence-associated molecular markers following treatment. H₂O₂ treatment induced senescence-associated changes within five days, whereas senescence was observed within 24h following D-galactose exposure. Both treatments resulted in increased cellular granularity and flattened or fibroblast-like morphology, although the kinetics of senescence induction differed between the two models. The results demonstrated that D-galactose- induced a more rapid and pronounced senescence phenotype in SH-SY5Y cells compared with hydrogen peroxide based on the molecular hallmarks of aging. Based on the study, it can be concluded that the D-galactose- induced model provides a more reliable and physiologically relevant in vitro model for investigating neuronal senescence .

Energy-equivalent cyclic pulsed electric fields enable reversible membrane permeabilization and sustainable protein release from microalgae.

Fabrication and biocompatibility evaluation of 3D printed tablets using Digital Light Processing (DLP) printing for the controlled release of ketoprofen.

Primary aldosteronism is the most common form of secondary hypertension, and aldosterone-producing adenoma (APA) is the most prevalent surgically curable subtype of primary aldosteronism. Over the past decade, the identification of recurrent somatic mutations, most notably in KCNJ5, has transformed the molecular understanding of APA and has revealed substantial heterogeneity across tumors. In parallel, advances in transcriptomic, epigenomic, and metabolomic profiling have further expanded insights into genotype-dependent differences in steroidogenic activity, cellular morphology, and clinical presentation. More recently, the application of single-cell, single-nucleus, and spatial transcriptomics technologies has revealed an additional layer of complexity, demonstrating marked heterogeneity within individual APA tissues. These studies show that APAs are composed of transcriptionally and functionally diverse cell populations with variable steroidogenic capacities, extending beyond what can be explained by the somatic mutation status alone. This review summarizes the current evidence for both intertumoral and intratumoral heterogeneity in APA, integrating findings from genomic, epigenomic, metabolomic, and single-cell-based studies. By highlighting how mutation-driven differences intersect with cell-state diversity and differentiation processes, this review aimed to provide a comprehensive framework for understanding the pathophysiology of APA and discuss the potential implications for disease classification, biomarker development, and future therapeutic strategies.

The extracellular matrix (ECM) is a major regulator of cellular behavior, fate, and various mechanisms underlying homeostasis, development, and disease. Biophysical and biochemical properties of the ECM are known to affect three-dimensional (3D) cellular behavior and phenotype that regulate a wide range of pathological conditions. Tunable biomimetic hydrogels are extensively employed to investigate cell-matrix interactions in defined 3D microenvironments. However, the dynamics of these interactions in complex coupled multiparametric microenvironments has been relatively less studied. This study aims to provide a framework for correlating cellular and nuclear behavior as a function of specific matrix properties (adhesivity, degradability, porosity, and stiffness) in a biosynthetic hydrogel system with varying crosslinking mechanisms. Poly(ethylene glycol diacrylate)-fibrinogen (PF)-based hydrogels were fabricated with varying crosslinking profiles, resulting in a series of hydrogels with varying matrix properties. NIH3T3 mouse fibroblasts were cultured in 3D and their matrix-associated morphological responses were studied over time. Matrix adhesivity and degradability (collectively termed 'matrix permissiveness') were found to be the most influential parameters regulating cell and nuclear behavior. Cells in permissive matrices displayed high viability, high cell density, increased spreading and protrusivity, and large elongated nuclei. Cells in restrictive matrices displayed reduced viability and lower cell density, rounded morphology, lower spreading, and smaller rounded nuclei. Cell confinement and nuclear confinement determined from various morphological features were correlated to matrix permissiveness. Overall, this study provides insights into regulation of cellular and nuclear behavior through modulation of matrix properties which could be used for future applications in various disease contexts. STATEMENT OF SIGNIFICANCE: Biomimetic engineered hydrogels are commonly used to support three-dimensional (3D) tissue-level behavior to recapitulate various developmental processes and disease states. Specific biophysical and biochemical cues in the engineered microenvironment can be used to control cellular behavior, morphology, and function, thereby providing mechanistic insights into cell-matrix interactions. This study assesses the variations in the cellular and nuclear features of fibroblasts encapsulated in 3D hydrogel matrices with varying crosslinking mechanisms. Our results reveal the combinatorial role of matrix adhesivity and degradability in regulating cellular and nuclear confinement of fibroblasts in 3D restrictive microenvironments. Overall, these matrix guiding principles can be implemented in the future to design tunable biomimetic matrices to modulate cell state, behavior, and function.

Silver nanoparticles (AgNPs) have attracted considerable attention for their potent antiproliferative properties. The search for efficient anti-lung cancer therapeutics led to the biosynthesis of AgNPs from a cyanobacterium, Leptolyngbya valderiana (Gomont) Anagnostidis & Komárek. Within 72 h of exposure of algal biomass to 0.01 M AgNO3, a notable color change of extracellular medium from transparent to dark brown was demonstrated, indicating successful synthesis of Leptolyngbya valderiana-AgNPs (LV-AgNPs). Furthermore, LV-AgNPs were characterized revealing their crystalline nature, oval shape, with sizes averaging 50 ± 21.4 × 39 ± 14 nm and a specific Ag absorption peak at 3 keV. Biosynthesized nanosilver showed remarkable antiproliferative efficacy against lung cancer cells A549 with an IC50 value of 19.78 ± 1.296 μg/mL and had a negligible effect on BEAS-2B and HEK-293 normal cells. Prominent alterations in cellular and nuclear morphology revealed an increased number of disintegrated A549 cells with condensed and fragmented nuclei. Microscopic evaluation of cell death was followed by flow cytometric cell cycle analysis, where the expansion of the sub-G0 population reflected that LV-AgNPs targets G0/G1 phase. Cell death was also associated with the formation of endogenous ROS and disruption of mitochondrial membrane permeability. The above findings suggest that silver nanomaterials produced by L. valderiana can serve as an alternative nano-drug development for lung adenocarcinoma.

The global prevalence of multidrug-resistant bacteria has been rising at an alarming rate, posing a serious threat to both human and animal health. However, the mechanisms by which bacteria acquire antibiotic tolerance and subsequently develop resistance remain incompletely understood. In this study, Pasteurella multocida, a common pathogen in the animal husbandry industry, was exposed to enrofloxacin, and genome resequencing, transcriptomic, and metabolomic analyses were performed to elucidate the adaptive mechanisms of P. multocida under fluoroquinolone-induced stress. Compared with the wild-type strain, the enrofloxacin-tolerant strain exhibited an extended lag phase, a prolonged logarithmic phase, reduced sensitivity to polymyxin B, reduced biofilm formation, and an elongated cellular morphology. Multi-omics analysis revealed a deletion in the dusB gene of the tolerant strain, resulting in a truncated non-functional protein. The deletion of dusB enhanced tolerance by prolonging the lag phase and reducing the growth rate. Moreover, the expression of genes in the CAMP pathway was up-regulated, and deletion of cpxR further promoted tolerance by modulating ribosome-associated genes. Integrated transcriptomic and metabolomic analyses indicated activation of the tricarboxylic acid (TCA) cycle during tolerance development. This study identified dusB and cpxR as key genes mediating enrofloxacin tolerance in P. multocida, elucidated the association between the antibiotic tolerance, growth, and gene expression, and may provide potential targets for future strategies aimed at limiting tolerance-associated resistance development.

Extracellular and intracellular oxidative mechanisms in the combined UV and peracetic acid processes for inactivating intracellular phages in bacterial cells.

ObjectiveTo explore the effects and mechanisms of metandienone abuse on hepatotoxicity.MethodsHepG2 cells were treated with 1 μg/mL metandienone for 24 h. Cell viability and apoptosis were detected via Cell Counting Kit-8 and terminal deoxynucleotidyl transferase dUTP nick end labeling assays, respectively. Cellular morphology was assessed via transmission electron microscopy. Genetic changes were analyzed using transcriptome sequencing. Meanwhile, metandienone (0.1 mg·g−1·d−1) was administered via gavage to mice for 14 days. Liver pathology was examined via hematoxylin and eosin, periodic acid–Schiff, and transmission electron microscopy. Serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, albumin, total cholesterol, triglyceride, lactate dehydrogenase, and creatine kinase were measured.ResultsCell Counting Kit-8 assay confirmed that cell viability decreased to 88.01% after 24 h of metandienone treatment. Terminal deoxynucleotidyl transferase dUTP nick end labeling assay showed that the apoptosis rate increased to 25.08%, compared with 4.94% in the NC group. RNA transcriptome data showed that the expression levels of 221 genes in the metandienone group altered significantly; these genes were closely related to liver metabolism, oxidative stress, inflammatory response, and immune regulation. Meanwhile, exposure to metandienone-induced liver injury was characterized by hepatocyte steatosis and inflammation, accompanied by significant increases in serum alanine aminotransferase and aspartate aminotransferase.ConclusionThe abuse of anabolic hormones leads to liver function damage, and the mechanism may be related to inflammatory responses and metabolism.

Cell-in-water: complementary optical photothermal infrared and Raman microscopy for submicron chemical imaging of hydrated cells.

In situ spatial (ISS) sequencing can uncover co-variation between cellular morphology and gene expression in vivo. However, a principled and interpretable mathematical representation of morphology has not yet been applied in this context. In particular, current deep learning-based representations of cell images confound a cell’s shape with its size. We present an interpretable representation of cellular boundary contours, based on tangent principal component analysis (TPCA) in a Kendall shape manifold, that captures size-independent contour shape features. This approach successfully recovers shape-perturbing genes in an RNAi screen than a previous metric geometry-based approach. We build on TPCA to develop STITCH (Shape–TranscriptomIc Correlation and Harmonization), an approach to reveal covariation between cell morphology with gene expression in ISS datasets. In a Xenium dataset, STITCH outperforms a deep learning-based approach in both recovering the layered organization of keratinocytes and a spatial gradient in nuclear eccentricity. Across samples in a melanoma CosMx dataset, STITCH reproducibly associates elongated and triangular fibroblasts with proximity to malignant cells and myofibroblast-like transcriptional program. Finally, STITCH independently recovers a known link between mesenchymal-like malignant cell states and increased cell area in two melanoma cohorts. STITCH can thus yield interpretable morphology–transcriptome relationships across cell types, patients, and spatial transcriptomics platforms.

As the primary peripheral relay station for vibrissal tactile information, the trigeminal ganglion (TG) features heterogeneous three-dimensional (3D) cytoarchitecture that eludes full characterization using conventional two-dimensional methodologies. A high-resolution 3D imaging and reconstruction pipeline is thus required to unveil TG structural organization and define the spatial framework of target-related sensory neurons. Herein, we established a fluorescence micro-optical sectioning tomography (fMOST)-based workflow for 3D cytoarchitectural mapping of TG anatomy and validated its utility for profiling the distributions of TG neurons innervating vibrissae via single-axon tracing. fMOST imaging coupled with propidium iodide (PI) staining was applied to acquire whole-head anatomical data encompassing the vibrissae and the TG at cellular resolution. Based on clearly resolved cellular morphology and the spatial distribution of neuronal somata, we delineated the soma distribution of TG neurons and revealed a spatially heterogeneous 3D organization pattern, from which we operationally defined two anatomically distinct subdomains: the neuronal soma-rich region (NSRR) and the fiber-rich region (FRR). Furthermore, with retrograde viral/genetic labeling combined with neuronal tracing, TG neurons innervating the C2, D3, and δ vibrissae were observed in both NSRR and FRR, showing partially overlapping yet spatially biased distributions consistent with previous population-level observations of vibrissa-row-dependent topography. Notably, TG neurons innervating the δ vibrissa occupied a comparatively broader spatial extent along the anteroposterior plane in our dataset. Overall, this study facilitates an in-depth mechanistic and anatomical understanding of TG cytoarchitectural organization and underlying functional mechanisms.

FGD3, a member of the FGD family, regulates GTPase activity and cellular morphology, thereby influencing tumor proliferation and migration. However, its role across diverse cancer types remains insufficiently understood. Using TCGA and GTEx databases, FGD3 expression was compared between tumor and normal tissues to assess its relevance to cancer. Single-cell transcriptomic data from TISCH were used to characterize FGD3 expression across immune cell subsets. Univariate Cox regression was performed to evaluate associations between FGD3 expression and patient survival. TIMER2.0 Spearman correlation analysis examined its relationship with immune cell infiltration. Gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA) revealed FGD3's association with immune and metabolic pathways. Bioinformatics and molecular docking predicted interactions with anticancer drugs. Experimental validation demonstrated that FGD3 knockdown suppresses cancer cell migration and proliferation, indicating an anti-tumor role in breast cancer. FGD3 expression varies across tumors, with differing prognostic implications. In most malignancies, expression inversely correlates with copy number variation (CNV) and methylation and associates with immunotherapy biomarkers and responses. ESTIMATE and immune infiltration analyses suggest a role in immunosuppression and tumor immunity. Experimental downregulation of FGD3 significantly reduces cell proliferation and migration, highlighting its therapeutic potential. FGD3 plays an important role in tumorigenesis and progression and may serve as a promising biomarker for cancer prognosis. Its expression is inversely associated with tumor metastasis. Experimental findings further suggest that FGD3 may function as a migration-suppressive factor in breast cancer. Collectively, these results provide new insights into the biological role of FGD3 and suggest its potential as a therapeutic target.

Objective: To investigate the clinicopathological features of tuberous sclerosis complex (TSC)-associated multifocal micronodular pneumocyte hyperplasia (MMPH) and the mutation status of TSC1/TSC2 genes. Methods: A retrospective analysis was conducted on 8 MMPH cases diagnosed at Shanghai Pulmonary Hospital Affiliated to Tongji University from September 2020 to August 2024. Clinical information and imaging findings of these cases were collected. Histopathological analysis and next-generation sequencing (NGS) were performed. Results: Among the 8 patients, there were 4 males and 4 females, aged 44 (36, 50) years old. None of them had a definite family history of TSC. Only 1 patient presented with respiratory symptoms at the time of consultation. Four patients had TSC-related cutaneous lesions. Two had shagreen patches, 1 had hypomelanotic macules, and 1 had hypomelanotic macules combined with fibrous plaques on the scalp and angiofibromas. Three patients themselves or their first-degree relatives had a history of epilepsy. Five patients themselves or their first-degree relatives had liver/kidney cysts or a surgical history of renal angiomyolipoma. Computerized tomography scans showed multiple ground-glass nodules in both lungs, with an average diameter of 10.5 (8.5, 11.0) mm. Six cases were diagnosed via surgical resection, and 2 via transbronchial cryobiopsy. Intraoperative frozen sections of the 6 surgically resected cases were all misdiagnosed as early-stage lung adenocarcinoma (including adenocarcinoma in situ, minimally invasive adenocarcinoma, and invasive non-mucinous adenocarcinoma), but the diagnosis was corrected to MMPH postoperatively. One of these 6 cases was concurrent with a benign perivascular epithelioid cell tumor (PEComa). Microscopically, MMPH was characterized by multifocal proliferation of type Ⅱ alveolar epithelial cells, which showed bland cellular morphology without atypia or invasive growth. For the case concurrent with PEComa, clear epithelioid cells were observed growing in nests or sheets. Immunohistochemically, the proliferative epithelial cells were positive for TTF-1 and showed low expression of Ki-67. PEComa cells were positive for Melan A and MiTF. NGS showed that TSC1 mutations were detected in 6 cases, TSC2 mutation in 1 case (which also had a concurrent class Ⅲ missense mutation of BRAF G466E), and no mutation in 1 case, with an overall mutation detection ratio is 7/8. No other lung cancer-related driver-gene variations were found in any case. Follow-up data were available for 8 patients. During the follow-up, 1 patient developed scattered thin-walled lucencies in both lungs, but no pathological examination was performed. All patients had a favorable overall survival status, follow-up time was 14.0(9.5,47.0) months. Conclusions: MMPH is a rare, benign TSC-associated pulmonary lesion, often presenting as multiple ground-glass nodules in both lungs. It can be easily misdiagnosed as early-stage lung adenocarcinoma. The diagnosis requires a comprehensive judgment based on clinical data, imaging findings, histopathology, and TSC1/TSC2 gene mutation results.