Beyond the liver and deeper in the liver: Sub-organ level analysis of in vivo sequestration mechanism of silica nanocapsules.

Long-Lived Tissue-Resident Memory T and NK Cells in CLAD Explant Lung Parenchyma - Consequences for Tolerance and Rejection

The invisible burden: A meta-analysis of methodological evolutions and the reassessment of microplastic concentrations in human tissues.

Internode morphology critically determines bamboo wood quality. Phyllostachys edulis 'Heterocycla' (GJ) is a natural dwarf mutant of P. edulis (WT), yet regulatory mechanisms remain unclear. Here, we show that impaired parenchyma cell elongation is the primary cellular basis for dwarfism in GJ. This phenotype correlates with a marked reduction in gibberellin (GA) and an increase in abscisic acid (ABA) levels. Through weighted gene co-expression network analysis, we identified PeGA20ox1 as a critical regulator of internode elongation. In GJ, PeGA20ox1 undergoes alternative splicing (AS), producing a transcript, PeGA20ox1-GJ, with a premature termination codon. Moreover, we characterized that a bifunctional transcription factor PeRAP2-1 mediates GA/ABA homeostasis, which is an activator of PeABA3 and an inhibitor of PeGA20ox1 by binding to different cis-acting elements. Intriguingly, this AS event results in the loss of function of PeGA20ox1-GJ, which disrupts the PeRAP2-1-mediated hormonal balance, ultimately inhibiting internode elongation. This study suggests that the posttranscriptional regulation induced by AS may adapt to the rapid growth of bamboo plants and provides important genetic resources for the molecular breeding of bamboo with improved culm traits.

Effects of loss of CO2-Responsive CCT Protein interaction with 14-3-3 proteins and its ectopic expression on starch synthesis in rice.

Administration of ciliary neurotrophic factor (CNTF) reduces food intake and body weight in both humans and experimental animals, where it also ameliorates hyperglycemia, hyperinsulinemia, and dyslipidemia. To exert its anti-obesogenic and anti-diabetogenic effects, CNTF targets brain feeding centers as well as multiple peripheral organs inducing the phosphorylation of the transcription factor signal transducer and activator of transcription 3 (p-STAT3). However, data showing which peripheral cytotypes are specifically targeted by exogenous CNTF invivo in metabolically relevant organs are currently lacking. Here, we first evaluated the gene expression levels of the subunits of the tripartite CNTF receptor (Cntfr) complex, that is, the Cntfrα, the leukemia inhibitory factor receptor β (Lifrβ) and the glycoprotein 130 (gp130), by quantitative real-time PCR in metabolically relevant organs of adult male mice: gastrointestinal (GI) tract, pancreas, liver, visceral and subcutaneous white (WAT) and interscapular brown adipose tissue (iBAT), skeletal muscle and the sciatic nerve. We then quantified p-STAT3 by Western blotting in these organs after intraperitoneal administration of CNTF (0.3 mg/kg) or saline. Finally, we mapped CNTF-responsive cells by immunohistochemistry, followed by morphometric quantification and confocal microscopy in both CNTF- and saline-treated mice. Lifrβ and gp130 were ubiquitously detected across all the investigated organs; the Cntfrα showed the highest expression levels in the skeletal muscle, sciatic nerve, and iBAT, whereas it was found to be expressed to a lesser extent in the other sites. Administration of CNTF led to a significant increase of p-STAT3/STAT3 protein ratio in all organs examined, except the duodenum, and induced a distinctive pattern of cell nuclear p-STAT3 immunoreactivity. Notably, along the analyzed GI tract, CNTF induced nuclear STAT3 phosphorylation in neurons of the submucosal and myenteric plexuses of the enteric nervous system and in contractile cells of the muscularis externa, where the response peaked in the mesenteric gut and colon. In the pancreas, CNTF triggered a higher activation within the endocrine component compared to the exocrine parenchyma. In the liver, CNTF induced STAT3 phosphorylation not only in parenchymal cells but also in sinusoids and resident macrophages. The cytokine activated p-STAT3 in subcutaneous and visceral white adipocytes, but also in brown adipocytes, with a prominent response observed in the beige subcutaneous adipocytes; adipose-resident macrophages and endothelial cells of numerous blood vessels were also CNTF-responsive. Lastly, in skeletal muscle, a major site for glucose/lipid utilization, CNTF induced widespread nuclear p-STAT3 immunoreactivity in muscle fibers and in connective and Schwann cells of the peripheral nerves, including the sciatic nerve, supplying the gastrocnemius. In conclusion, our data indicate that CNTF acts across diverse cytotypes within metabolically relevant organs and tissues, likely fostering its peripheral metabolic effects through this cellular heterogeneity.

In this review, we summarize the data on the cellular pigment lipofuscin that accumulates in liver tissue over time, due to aging and cellular stress. Despite the presence of these typical subcellular inclusions under various conditions, relatively little is known about their origins, roles, and effects on liver cell and tissue health. Pathologists use the presence of lipofuscin, in combination with other markers, to achieve differential diagnosis across various diseases. Routine histological stains reveal characteristic irregular shaped intracellular inclusions of lipofuscin that cannot be missed. Moreover, lipofuscin is autofluorescent and in transmission electron microscopy it appears in the cytoplasm as irregularly shaped structures containing fat and floccular material with varying electron density. Herein, we discuss the current state of knowledge concerning the origin and function of this pigment in the liver. Lipofuscin can distinctively be found in liver, although it has also been reported in cells in the heart, brain, and eye. Its biochemical composition is heterogeneous and varies depending on the tissue and the age of the organism. The liver parenchymal cells have efficient cellular waste disposal mechanisms, but they are still susceptible to aging. Lipofuscin accumulation in the liver may result from ongoing oxidative damage and impaired hepatic detoxification leading to cellular stress.

Intraorgan and Targeted Nanodelivery in Organ Transplantation of Non-Human Primates.

The current management of end-stage liver diseases relies on organ transplantation but is constrained by donor scarcity and immune rejection. Induced pluripotent stem cell (iPSC)-derived cells represent a promising alternative, as they can be expanded indefinitely, genome-edited to reduce immunogenicity and employed to develop more mature three-dimensional cultures. Here, we combined tissue engineering with hypoimmunogenic iPSC technology to generate off-the-shelf hepatic organoids that integrate parenchymal and non-parenchymal cells: hepatocytes, cholangiocytes, hepatic stellate cells, Kupffer cells and both vascular and sinusoidal endothelial cells. Lineage identity was validated by gene and protein expression analysis, electron microscopy and single-cell RNA sequencing. The complex microenvironment obtained recapitulates native liver composition and promotes hepatic maturation, as evidenced by the secretion of liver-specific proteins, including albumin and apolipoproteins. These properties, together with the presence of a functional endothelial network, represent a step towards universal cell therapies for end-stage liver diseases and hepatic metabolic disorders.

Neutrophil extracellular traps (NETs) are chromatin-based extracellular structures consisting of DNA, histones and multiple antimicrobial proteins, which exert dual biological effects in host defense and inflammation-triggered tissue injury. This review focuses on NET-associated signaling pathways and their regulatory crosstalk with diverse forms of regulated cell death (RCD) in inflammatory disorders. First, we summarize the structural characteristics and biogenesis pathways of NETs closely linked to inflammatory amplification, including lytic and non-lytic NETosis as well as reactive oxygen species (ROS)-dependent and ROS-independent mechanisms, and elaborate the functions of NADPH oxidase, myeloperoxidase, neutrophil elastase, peptidylarginine deiminase 4 and gasdermin D during these processes. Second, we discuss how NET-derived damage-associated molecular patterns, such as DNA, histones, granular proteases, ROS and mitochondrial DNA, interact with apoptosis, necroptosis, pyroptosis, ferroptosis, autophagy and cuproptosis. Under pathological conditions, excessive NET formation or impaired NET clearance leads to autoantigen exposure, accelerated thrombosis, enhanced inflammasome activation, parenchymal cell damage and modulated tumor progression. Finally, we outline therapeutic interventions targeting NET biogenesis, NET clearance and downstream NETs-driven signaling, with an emphasis on the translational potential and safety concerns of these strategies across distinct diseases. Future investigations are required to decipher context-dependent NETs-RCD regulatory circuits, standardize NETs detection protocols, and develop precision-targeted therapeutics that restrain pathological inflammation while preserving host antimicrobial defense.

The clearance of apoptotic cells by phagocytes is crucial for restoring tissue balance after injury. In autoimmune liver diseases like primary sclerosing cholangitis, cell death is thought to result from accumulation of toxic bile acids within parenchymal cells. Whether, in this context, bile acid-loaded dying cells impact the efficiency of phagocytic macrophages in restoring tissue balance remains unknown. Here, we demonstrate that in a murine model of cholangitis, bile acids accumulate in a subpopulation of efferocytic macrophages with pro-inflammatory features. Our in vitro results indicate that, upon their engulfment, apoptotic hepatocytes laden with bile acids can serve as Trojan horses, delivering bile acids into efferocytic macrophages and thereby shaping macrophage function. This contrasts with the characteristics of macrophages that engulf apoptotic parenchymal cells lacking bile acids. Together, our findings delineate a system in which the content of the phagocytosed dying cells, specifically bile acid-laden hepatocytes, drives a pro-inflammatory program in the corresponding efferocytic macrophages, potentially contributing to chronic hepatic inflammation.

Cancer stem cells (CSCs) drive recurrence and drug resistance in hepatocellular carcinoma (HCC), but their origin remains controversial: are they tumour-initiating cells or late-stage dedifferentiation products? Direct human single-cell evidence linking bipotent progenitors (BPs) and CSCs has been lacking. We integrated single-cell RNA sequencing (scRNA-seq) data from 109 samples (44 patients; 410,608 cells) across five public cohorts and generated EpCAM-enriched scRNA-seq from two additional HCC patients. Single-cell somatic mutations were inferred from the transcriptomic data, yielding 384,867 high-confidence variants across 31,908 cells from 20 patients. Clonal evolution was reconstructed through copy number variation (CNV) phylogenies and transcription-coupled-repair-based cell-of-origin inference. CSC and non-CSC subpopulations from Huh7 cells were flow-sorted before and after two weeks of culture and profiled by targeted bisulfite sequencing. A core-imprint risk score was evaluated in multiple cohorts and validated on a 97-case tissue microarray by multiplex immunofluorescence. Unexpectedly, BPs harboured higher mutation burdens than other non-malignant parenchymal cells, and CSCs harboured higher mutation burdens than most other tumour cells, challenging their role as genomically quiescent ancestors. CNV phylogenies and evolutionary distances placed CSCs at the most distal branches of the tumour tree, while cell-of-origin analysis identified BPs as a pre-malignant precursor arising from hepatocyte dedifferentiation. RNA velocity, pseudotime and SNP-integrated lineage reconstruction converged on this directionality, with CSCs arising at the terminus of tumour evolution, reproduced at single-patient resolution in the EpCAM-enriched samples. Mechanistically, CSCs upregulated DNA methyltransferases (DNMTs), and ~78% of CSC-specific methylation changes were stably retained after CSC differentiation but not reproduced during de novo stemness acquisition, indicating locked-in epigenetic memory. A 16-gene core-imprint risk score specifically predicted early recurrence (≤2 years), and CD13+ CD133+ CSCs showed elevated 5-hydroxymethylcytosine correlating with poor prognosis. We propose a framework in which hepatocytes dedifferentiate into BPs as a pre-malignant state, undergo malignant transformation, and a subset acquires stemness through DNMT-mediated reprogramming stabilized by epigenetic memory. These findings challenge the classical stem cell origin hypothesis, showing that CSCs in established HCC are late-stage dedifferentiation products, provide a rationale for targeting CSC epigenetic stability, and offer a biomarker for early recurrence.

Cardiac sarcoidosis: new insights beyond the granuloma using spatial proteomics.

Lignification is a crucial process for strengthening plant tissues, facilitating water transport, and providing defense against pathogens. In the Poaceae family, p-hydroxycinnamic acids are commonly incorporated into lignin, with acylation by p-coumarate (pCA) occurring during lignification. In this study, we performed DFRC and 2D HSQC-NMR analyses to investigate changes in lignin substructures and the degree of lignin pCA-acylation throughout bamboo stem development. Furthermore, immunohistochemical analysis was conducted to elucidate the spatial distribution of lignin substructures within different cell types. Our results revealed that, in young tissues, β-O-4-linked lignin units are predominantly derived from monolignol-pCA conjugates, specifically coniferyl- and sinapyl-pCA. Both lignin structure and the pattern of pCA acylation varied depending on the stage of cell wall formation and the cell type, particularly between vascular fiber cells and parenchyma cells. Based on our results, moso bamboo culms exhibit a distinctive feature during incipient lignification, in which monolignols are predominantly acylated with pCA. This feature has not been reported in other grasses, suggesting that extensive p-coumaroylation of monolignols plays an important role in the rapid elongation of bamboo culms.

Rapid revascularization is critical to tissue graft survival, as delayed reperfusion drives tissue ischemia and compromises cell viability and graft function. Although bulk hydrogels have been explored for promoting vessel formation, vascularization remains too slow to prevent ischemic injury to grafted tissues, highlighting the need for biomaterial platforms that accelerate graft revascularization and reperfusion. In this study, we present granular hydrogel composites (GHCs), where interstitial space is filled with fibrin and collagen to provide a vasculogenic matrix environment. GHCs supported the assembly of embedded endothelial cells into interconnected, lumenized networks in vitro which anastomosed with host vasculature and were systemically perfused 7 days after implantation. Careful optimization studies revealed that GHCs formed from covalently interlinked, RGD-functionalized microgels of 115 µm diameter best supported vascular network formation in vitro and intravascular blood perfusion in vivo . To test the utility of GHCs for the vascular integration of a demanding and therapeutically relevant parenchymal tissue, GHC-based ovarian tissue grafts were implanted in a murine xenograft model and successfully connected to host vasculature, restoring blood flow to embedded human ovarian tissues within 10 days post-implantation. Notably, endothelial cells seeded within GHCs formed viable vasculature without pre-culture. This work establishes GHCs as a biomaterial platform to rapidly connect parenchymal tissues to host vasculature, with broad translational potential across engineered tissue grafting applications.

A PSC-tailored deep learning model for liver segmentation on fat-saturated T2-weighted MR: Robustness to hepatic dysmorphia and multicentre generalisability.

Immunopharmacology of GPR35: Context-driven signaling and functional plasticity in host defense.

Cassava (Manihot esculenta Crantz) exhibits high photosynthetic efficiency and remarkable starch accumulation in its storage roots. The effective loading of photosynthates into the phloem from mesophyll cells in leaves is a critical determinant of yield; however, this process remains poorly understood. In this study, we propose a theoretical model of apoplastic sucrose phloem loading in cassava based on a multitechnique approach. The concentration of primary photoassimilates in leaf veins, analyzed using a [14C]CO2 tracer, and the existence of few plasmodesmata between bundle sheath cells/phloem parenchyma cells and sieve element-companion cell (SE-CC) complexes in minor veins suggest characteristic apoplastic phloem loading in cassava. We identified five sucrose transporters (MeSUTs) in the cassava genome, among which MeSUT1a exhibited the highest expression and the strongest sucrose intake activity. Subcellular localization analyses showed that MeSUT1a specifically localizes in the plasma membrane of the SE-CC complexes and that sugar transporter (MeSWEET2a) localizes on parenchyma cells, suggesting potential functional synergy. Interference with MeSUT1a expression led to a reduction in sucrose loading efficiency by more than 50%, resulting in abnormal sucrose and transient starch accumulation. This interference subsequently impaired chloroplast development and leaf photosynthesis, ultimately reducing storage root yield and starch content. RNA-seq analysis of MeSUT1a transgenic lines further revealed remarkable transcriptional changes in genes associated with sugar transport, carbohydrate metabolism, and photosynthesis. These results establish that MeSUT1a is essential for driving sucrose phloem loading and plays a key role in the distribution of photosynthetic assimilates and the coordination of source-sink dynamics in cassava.

Cancer stem cells (CSCs), defined as tumor cell populations with self-renewal and tumor-propagating capacity, contribute to tumor initiation and participate in progression, therapeutic resistance and relapse through pronounced metabolic plasticity. Although CSC metabolism has traditionally been regarded as a cell-intrinsic feature, accumulating evidence highlights the tumor microenvironment as a critical determinant of CSC metabolic states. Diverse stromal and tissue-specific parenchymal cell populations actively shape metabolic niches through context-dependent interactions, thereby reinforcing CSC stemness and adaptive potential. This Review synthesizes current insights into how widespread and organ-specific tumor microenvironment cell populations reprogram CSC metabolism via bidirectional crosstalk. Such a framework provides a mechanistic basis for intratumoral and organ-context-dependent heterogeneity, as well as differential therapeutic responses. Finally, we discuss the emerging potential of targeting CSC-supportive metabolic niches through drug repurposing, highlighting context-aware metabolic interventions as a pragmatic strategy to overcome CSC-driven treatment resistance.

ABSTRACTSmall signaling peptides (SSPs) are important regulators of plant growth, development, and responses to biotic and abiotic stress, yet their role in the C4 grass Sorghum bicolor is largely uncharacterized. To help fill this knowledge gap, 219 sorghum genes that encode SSPs were identified based on SSP sequences previously identified in Arabidopsis thaliana, Zea mays, Oryza sativa, Triticum aestivum, and Brachypodium distachyon. The 219 sorghum SSP‐encoding genes were assigned to 19 gene families, analyzed for the presence of motifs, and aligned with genes that encode SSPs in other plants using phylogenetic analysis. Sorghum genes in 12 of the 19 SSP gene families had not been previously characterized. Expression of the 219 SSP‐encoding genes in sorghum organs, during stem development, and in stem tissues and cell types revealed distinct spatial, temporal, and developmental patterns of expression. Genes associated with the SbCEP and SbRGF families were preferentially expressed in roots, whereas SbEPF genes were expressed in stem epidermal and pith parenchyma cells and panicles. The expression of genes during bioenergy sorghum stem growth and development was investigated because stems account for ~80% of harvested biomass and serve as conduits for water and nutrient transport between leaves and roots. During stem development, 28 SSP genes in several families (CLE, EPF, CEP, GASS, PSY, ES, PSK, CAPE, POE) were expressed at higher levels in zones of cell proliferation. For example, the TDIF homologs SbCLE41 and SbCLE42 were expressed at high levels in nascent stem nodes where they may regulate vascular bundle cambial activity and cell differentiation. A different set of 15 genes in the CIF, POE, CAPE, PSY, CEP, RALF, and CLE families were expressed at higher levels in zones of stem tissue differentiation highlighted by elevated expression of five SbRALFRs in the stem nodal plexus. Cell type–specific expression of many sorghum genes that encode SSPs was observed in fully elongated internodes indicating gene expression is regulated with high spatial resolution. Overall, the results provide a foundation of information for analysis of SSP function in sorghum that can be integrated with knowledge of sorghum gene regulatory networks to modulate traits important for production of sorghum crops.